The GenMark ePlex® Blood Culture Identification Gram-Negative (BCID-GN) Panel is a qualitative nucleic acid multiplex in vitro diagnostic test intended for use on GenMark's ePlex Instrument for simultaneous qualitative detection and identification of multiple potentially pathogenic gram-negative bacterial organisms and select determinants associated with antimicrobial resistance in positive blood culture. In addition, the ePlex BCID-GN Panel is capable of detecting several gram-positive bacteria (Pan Gram-Positive assay) and several Candida species (Pan Candida assay). The ePlex BCID-GN Panel is performed directly on blood culture samples identified as positive by a continuous monitoring blood culture system and which contain gram-negative organism.

The following bacterial organisms and genes associated with antibiotic resistance are identified using the ePlex BCID-GN Panel: Acinetobacter baumannii, Bacteroides fragilis, Citrobacter, Cronobacter sakazakii. Enterobacter cloacae complex, Enterobacter (non-cloacae complex), Escherichia coli, Fusobacterium necrophorum, Fusobacterium nucleatum, Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae group, Morganella morganii, Neisseria meningitidis, Proteus, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella, Serratia, Serratia marcescens, Stenotrophomas maltophilia, СТХ-М (blactх-м), IMP (blамм) , КРС (blakec) , NDM (bland), OXA (blaoxa) (OXA-23 and OXA-48 groups only), and VIM (blaviм).

The ePlex BCID-GN Panel contains assays for the detection of genetic determinants associated with resistance to antimicrobial agents including CTX-M(blactx.M), which is associated with resistance to extended spectrum beta-lactamase (ESBL)-mediated resistance to penicillins, cephalosporins and monobactams, as well as OXA (blaoxA) (OXA-23 and OXA-48 groups only), KPC (blakpc), and metallo-beta-lactamases IMP (blavM), VIM (blavM), and NDM (blaNDM), which is associated with carbapenemase-mediated resistance. The antimicrobial resistance gene detected may or may not be associated with the agent responsible for disease. Negative results for these select antimicrobial resistance assays do not indicate susceptibility, as there are multiple mechanisms of resistance in gram-negative bacteria.

The ePlex BCID-GN Panel also contains targets designed to detect a broad range of organisms with a potentially misleading Gram stain result or organisms that may be missed by Gram staining altogether, for example in the case of co-infections. These include a broad Pan Gram-Positive assay (which is designed to detect Bacillus cereus group, Bacillus subtilis group, Enterococcus, Staphylococcus, and Streptococcus), as well as a Pan Candida assay, which is designed to detect four Candida species: Candida albicans, Candida glabrata, Candida krusei, and Candida parapsilosis.

The detection and identification of specific bacterial and fungal nucleic acids from individuals exhibiting signs and/or symptoms of bloodstream infection aids in the diagnosis of bloodstream infection when used in conjunction with other clinical information. The results from the ePlex BCID-GN Panel are intended to be interpreted in conjunction with Gram stain results and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.

Negative results in the setting of a suspected bloodstream infection may be due to infection with pathogens that are not detected by this test. Positive results do not rule out co-infection with other organisms; the organism(s) detected by the ePlex BCID-GN Panel may not be the definite cause of disease. Additional laboratory testing (e.g. sub-culturing of positive blood cultures for identification of organisms not detected by ePlex BCID-GN Panel and for susceptibility testing, differentiation of mixed growth, and association of antimicrobial resistance marker genes to a specific organism) and clinical presentation must be taken into consideration in the final diagnosis of bloodstream infection.

Device Description

The ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel is based on the principles of competitive nucleic acid hybridization using a sandwich assay format, wherein a single-stranded target binds concurrently to a sequence-specific solution-phase signal probe and a solid-phase electrode-bound capture probe. The test employs nucleic acid extraction, target amplification via polymerase chain reaction (PCR) or reverse transcription PCR (RT-PCR) and hybridization of target DNA. In the process, the double-stranded PCR amplicons are digested with exonuclease to generate single-stranded DNA suitable for hybridization.

Nucleic acid extraction from biological samples occurs within the cartridge via cell lysis, nucleic acid capture onto magnetic beads, and release for amplification. The nucleic acid extraction is processed through microfluidic liquid handling. Once the nucleic acid targets are captured and inhibitors are washed away, the magnetic particles are delivered to the electrowetting environment on the printed circuit board (PCB) and the targets are eluted from the particles and amplified.

During hybridization, the single-stranded target DNA binds to a complementary, single-stranded capture probe immobilized on the working gold electrode surface. Single-stranded signal probes (labeled with electrochemically active ferrocenes) bind to specific target sequence / region adjacent to the capture probe. Simultaneous hybridization of target to signal probes and capture probe is detected by alternating current voltammetry (ACV). Each working electrode on the array contains specific capture probes, and sequential analysis of each electrode allows detection of multiple analyte targets.

AI/ML Overview

Here's a summary of the acceptance criteria and study information for the ePlex Blood Culture Identification Gram Negative (BCID-GN) Panel, extracted from the provided text:

Acceptance Criteria and Device Performance for ePlex Blood Culture Identification Gram Negative (BCID-GN) Panel

The ePlex BCID-GN Panel is a qualitative nucleic acid multiplex in vitro diagnostic test for the simultaneous qualitative detection and identification of multiple potentially pathogenic gram-negative bacterial organisms and select determinants associated with antimicrobial resistance in positive blood cultures. Performance characteristics were evaluated through clinical and analytical studies.

1. Table of Acceptance Criteria and Reported Device Performance

The acceptance criteria are generally implied by the achieved performance metrics, which show high Positive Percent Agreement (PPA) for sensitivity and Negative Percent Agreement (NPA) for specificity. While explicit numerical acceptance thresholds are not directly stated as "acceptance criteria" in a dedicated section, the provided tables demonstrate the device's performance against its intended use. For brevity, the "Overall" performance (combining prospective, retrospective, and contrived samples where applicable) is used here to represent the device's reported performance against implied high accuracy criteria.

Target (Organism/Resistance Gene)	Implied Acceptance Criteria (High Accuracy)	Reported Device Performance (Overall PPA % (95% CI))	Reported Device Performance (Overall NPA % (95% CI))
Acinetobacter baumannii	High Sensitivity & Specificity	100 (95.1-100)	99.9 (99.7-100)
Bacteroides fragilis	High Sensitivity & Specificity	95.6 (87.8-98.5)	99.9 (99.6-100)
Citrobacter	High Sensitivity & Specificity	98.6 (92.2-99.7)	99.8 (99.4-99.9)
Cronobacter sakazakii	High Sensitivity & Specificity	100 (92.3-100)	100 (99.8-100)
Enterobacter cloacae complex	High Sensitivity & Specificity	95.3 (89.4-98.0)	99.6 (99.2-99.8)
Enterobacter (non-cloacae complex)	High Sensitivity & Specificity	96.6 (88.3-99.0)	99.9 (99.7-100)
Escherichia coli	High Sensitivity & Specificity	96.9 (94.4-98.3)	99.8 (99.4-99.9)
Fusobacterium necrophorum	High Sensitivity & Specificity	98.0 (89.3-99.6)	100 (99.8-100)
Fusobacterium nucleatum	High Sensitivity & Specificity	100 (93.1-100)	99.9 (99.7-100)
Haemophilus influenzae	High Sensitivity & Specificity	100 (93.5-100)	100 (99.8-100)
Klebsiella oxytoca	High Sensitivity & Specificity	89.6 (80.0-94.8)	99.8 (99.5-99.9)
Klebsiella pneumoniae group	High Sensitivity & Specificity	97.9 (95.2-99.1)	99.7 (99.3-99.9)
Morganella morganii	High Sensitivity & Specificity	100 (94.2-100)	99.9 (99.7-100)
Neisseria meningitidis	High Sensitivity & Specificity	100 (92.0-100)	99.9 (99.7-100)
Proteus	High Sensitivity & Specificity	97.7 (92.0-99.4)	100 (99.8-100)
Proteus mirabilis	High Sensitivity & Specificity	97.6 (91.6-99.3)	100 (99.8-100)
Pseudomonas aeruginosa	High Sensitivity & Specificity	95.8 (90.6-98.2)	99.7 (99.4-99.9)
Salmonella	High Sensitivity & Specificity	96.4 (87.9-99.0)	100 (99.8-100)
Serratia	High Sensitivity & Specificity	100 (95.4-100)	99.9 (99.7-100)
Serratia marcescens	High Sensitivity & Specificity	100 (94.2-100)	99.9 (99.7-100)
Stenotrophomonas maltophilia	High Sensitivity & Specificity	94.0 (83.8-97.9)	99.9 (99.7-100)
Pan Candida	High Sensitivity & Specificity	62.5 (30.6-86.3)	99.7 (98.4-99.9) (Prospective All)
Pan Gram-Positive	High Sensitivity & Specificity	78.2 (67.8-85.9)	97.9 (95.6-99.0) (Prospective All)
CTX-M (Overall)	High Sensitivity & Specificity	93.1 (88.1-96.1)	100 (99.7-100)
IMP (Overall)	High Sensitivity & Specificity	100 (91.2-100)	100 (99.7-100)
KPC (Overall)	High Sensitivity & Specificity	98.1 (89.9-99.7)	99.9 (99.6-100)
NDM (Overall)	High Sensitivity & Specificity	100 (93.4-100)	100 (99.7-100)
OXA (Overall)	High Sensitivity & Specificity	94.0 (83.8-97.9)	99.8 (99.4-100)
VIM (Overall)	High Sensitivity & Specificity	100 (91.6-100)	100 (99.7-100)

2. Sample Sizes Used for the Test Set and Data Provenance

The test set comprised a combination of clinical and contrived samples:

Prospective Clinical Samples: 349 evaluable samples (167 fresh, 182 frozen). Patients were of all ages and genders. Collected at 7 clinical sites.
Retrospective Clinical Samples: 577 evaluable samples. Collected at 10 clinical sites.
Contrived Samples: 777 samples.

Data Provenance: The clinical samples (prospective and retrospective) were collected at multiple clinical sites, suggesting data from different countries/regions, but specific countries of origin are not detailed. Both prospective and retrospective collection methods were used.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts

The document does not explicitly state the number of experts or their specific qualifications (e.g., radiologist with 10 years of experience) used to establish the ground truth. It refers to "standard laboratory procedures for identification of blood culture isolates, including traditional and automated identification methods, MALDI-TOF IVD, and microbiological and biochemical techniques" as the comparator method. For Acinetobacter baumannii and Candida parapsilosis, as well as antibiotic resistance genes, PCR assays followed by bi-directional sequencing were used for confirmation. This implies a reliance on established laboratory protocols and potentially expert interpretation within those processes, but specific expert count or detailed qualifications are not provided.

4. Adjudication Method for the Test Set

The document does not describe an explicit adjudication method (e.g., 2+1, 3+1). The "Comparator Method" served as the reference standard and was used to determine true positive (TP), false negative (FN), true negative (TN), and false positive (FP) results. Discrepancies were investigated using PCR/sequencing (as noted in footnotes for various tables), suggesting a re-evaluation process for conflicting results but not a formal multi-reader adjudication scheme for the initial ground truth establishment.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done

No MRMC comparative effectiveness study involving human readers and AI assistance was performed or described in the provided text. The study focuses on the standalone performance of the ePlex BCID-GN Panel against laboratory reference methods.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done

Yes, the study described is a standalone performance study of the ePlex BCID-GN Panel. The performance metrics (PPA and NPA) are calculated by comparing the device's output directly against the "Comparator Method" without human interpretation influencing the device's result.

7. The Type of Ground Truth Used

The ground truth was established using expert consensus methods based on standard laboratory procedures for organism identification and analytically validated molecular assays (qPCR followed by bi-directional sequencing) for confirming specific organisms and resistance genes. This includes:

Traditional and automated identification methods
MALDI-TOF IVD
Microbiological and biochemical techniques
PCR/sequencing for specific organisms and resistance markers.

8. The Sample Size for the Training Set

The document does not mention a distinct "training set" in the context of the clinical performance evaluation. The clinical studies (prospective, retrospective, and contrived samples) are described as evaluation/test sets. However, the "Analytical Reactivity (Inclusivity)" and "Limit of Detection (LoD)" studies (Tables 56 and 57) describe testing of various strains and isolates to establish the device's analytical performance and broad reactivity. These analytical studies are crucial for the development and "training" (in a broader sense of assay design and validation) of such molecular diagnostic panels.

Specifically:

Limit of Detection (LoD): At least 20 replicates per target were tested for each condition using quantified reference strains (Table 56 lists 38 organisms/targets with specific strains).
Analytical Reactivity (Inclusivity): A panel of 336 strains/isolates was evaluated (Table 57 provides a partial list of these validated strains).
Predicted (in silico) Reactivity: Bioinformatic analysis covered many additional variants for genus/group assays and resistance markers (Tables 58-76).

9. How the Ground Truth for the Training Set Was Established

For the analytical "training" aspects:

LoD: The ground truth for LoD was established by using quantified reference strains with known concentrations in simulated blood culture sample matrix. The lowest concentration detected in ≥95% of tested replicates was defined as the LoD.
Analytical Reactivity (Inclusivity): The ground truth was established by testing well-characterized strains/isolates with known identities at specified concentrations, confirming their detection by the panel.
Predicted (in silico) Reactivity: This involved bioinformatic analysis of existing sequence data for various organisms and resistance gene variants to predict the panel's ability to detect them. This is a computational method for establishing theoretical ground truth based on genetic sequences.

Summary

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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo includes the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.

April 12, 2019

GenMark Diagnostics, Incorporated Alan Maderazo VP, Ouality, Regulatory & Clinical Affairs 5964 La Place Court Carlsbad, California 92008

Re: K182619

Trade/Device Name: ePlex Blood Culture Identification Gram Negative (BCID-GN) Panel Regulation Number: 21 CFR 866.3365 Regulation Name: Multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures Regulatory Class: Class II Product Code: PEN PAM PEO Dated: September 21, 2018 Received: September 24, 2018

Dear Alan Maderazo:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's

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requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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5.0 510(k) Summary

Summary of Safety and Effectiveness

Submitter Information

GenMark Diagnostics, Incorporated Submitter: 5964 La Place Court Carlsbad, CA 92008
Manufacturer: GenMark Diagnostics, Incorporated 5964 La Place Court Carlsbad, CA 92008

Establishment Registration Number: 3008632402

Contact:	Alan Maderazo, Ph.D., RACVice President, Quality, Regulatory and Clinical Affairs
Phone:	760-448-4308
Fax:	760-683-6961
E-mail:	Al.Maderazo@genmarkdx.com
Alternate Contact:	Beth StofkaSr. Regulatory Affairs Specialist
Phone:	760-579-4778
Fax:	760-683-6961
E-mail:	Beth.Stofka@genmarkdx.com
Date Prepared:	September 21, 2018

Name of Device and Classification

ePlex® Blood Culture Identification Gram-Negative (BCID-GN) Panel Product Name: Device Classification: 866.3980, Multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures, Class II PEN, PAM, PEO Product Code(s):

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Predicate Device

Predicate:

FilmArray Blood Culture Identification Panel: BioFire Diagnostics; K130914

Device Description

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Intended Use/Indications for Use

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Summary of Technological Characteristics of the Device Compared to the Predicate Device

The GenMark ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel ("Subject Device") and the legally marketed device, FilmArray Blood Culture Identification Panel, K130914, ("Predicate Device") are described below:

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Characteristic	Subject Device	Predicate Device
Product Name	ePlex BCID-GN Panel	FilmArray BCID Panel
Manufacturer	GenMark Diagnostics, Inc.	BioFire Diagnostics, Inc.
OrganismsDetected	Acinetobacter baumannii Bacteroides fragilis Citrobacter Cronobacter sakazakii Enterobacter cloacae complex Enterobacter (non-cloacae complex) Escherichia coli Fusobacterium necrophorum Fusobacterium nucleatum Haemophilus influenzae Klebsiella oxytoca Klebsiella pneumoniae Morganella morganii Neisseria meningitidis Proteus Proteus mirabilis Pseudomonas aeruginosa Salmonella Serratia Serratia marcescens Stenotrophomonas Saltophilia pyogenes (GAS)	Enterococci, Listeria monocytogenes,commonly encountered Staphylococci(including specific differentiation ofStaphylococcus aureus), commonlyencountered Streptococci (withspecific differentiation ofStreptococcus agalactiae,Streptococcus pneumoniae, andStreptococcus pyogenes),Acinetobacter baumannii, commonlyencountered Enterobacteriaceae(including specific differentiation ofthe Enterobacter cloacae complex,Escherichia coli, Klebsiella oxytoca,Klebsiella pneumoniae, Proteus, andSerratia marcescens), Haemophilusinfluenzae, Neisseria meningitidis(encapsulated), Pseudomonasaeruginosa, Candida albicans,Candida glabrata, Candida krusei,Candida parapsilosis, and Candidatropicalis.
Resistance GenesDetected	CTX-M, IMP, KPC, NDM, OXA, andVIM	Genetic determinants of resistance tomethicillin (mecA), vancomycin(vanA and vanB), and carbapenems(blaKPC)
Indication forUse	The ePlex BCID-GN Panel is indicatedas an aid in the diagnosis of specificagents of bacteremia. The use ofadditional laboratory testing (e.g. sub-culturing of positive blood cultures foridentification of organisms not detectedby the ePlex BCID-GN Panel and forsusceptibility testing, differentiation ofmixed growth, and association ofantimicrobial resistance marker genes toa specific organism) and clinicalpresentation must be taken intoconsideration in the final diagnosis ofblood stream infection.	FilmArray BCID is indicated as an aidin the diagnosis of specific agents ofbacteremia and fungemia and resultsshould be used in conjunction withother clinical and laboratory findings.
Characteristic	Subject Device	Predicate Device
Specimen Type	Blood culture samples identified aspositive by a continuous monitoringblood culture system thatdemonstrates the presence oforganisms as confirmed by Gramstain.	Blood culture samples identified aspositive by a continuous monitoringblood culture system thatdemonstrates the presence oforganisms as confirmed by Gramstain.
Chemistry	Reagents on cartridge include: samplelysis and nucleic acid extraction, PCRamplification and hybridization-basedelectrochemical detection reagents.	The FilmArray BCID pouch containsfreeze-dried reagents to performnucleic acid purification and nested,multiplex PCR with DNA meltanalysis.
Hardware	GenMark ePlex Instrument & SingleUse Cartridge	FilmArray Instrument and assay pouch
SoftwareInterfaceResult Reporting	GenMark ePlex System Software GenMark ePlex BCID-GN Panel Software	The FilmArray Software automaticallyinterprets the results of each DNAmelt curve analysis and combines thedata with the results of the internalpouch controls to provide a test resultfor each organism and antimicrobialresistance gene on the panel.

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Analysis of the similarities and differences indicate that the devices are substantially equivalent in their intended uses/indications for use, and are generally the same regarding user process, ease of use and general operator protocol. Comparison of technological similarities and differences between the proposed device and the predicate do not raise new or different questions of safety and effectiveness, and therefore render the proposed device as substantially equivalent to the predicate device.

Summarv of Performance Data

Expected Values

A prospective, multicenter clinical study was conducted to evaluate the clinical performance of the ePlex BCID-GN Panel in positive blood culture samples. A total of 349 samples were prospectively collected at 7 clinical sites in 2 phases from patients of all ages and genders. In the first phase from June 2014 through July 2016, 182 samples were prospectively collected and frozen; from June through July 2018, 167 samples were prospectively collected and tested fresh (never frozen). The expected values of individual analytes based on the ePlex BCID-GN Panel results in prospective samples are summarized by age group and by site in Tables 1 and 2 below.

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Table 1: Expected Value by Age Group (Prospective Samples)

Target	All Ages(N=349)n (%)	Age <1(N=7)n (%)	Age 1-17(N=10)n (%)	Age 18-44(N=50)n (%)	Age 45-64(N=124)n (%)	Age 65-84(N=125)n (%)	Age 85+(N=33)n (%)
Acinetobacter baumannii	4 (1.1)	0 (0.0)	0 (0.0)	1 (2.0)	2 (1.6)	1 (0.8)	0 (0.0)
Bacteroides fragilis	11 (3.2)	0 (0.0)	0 (0.0)	2 (4.0)	4 (3.2)	2 (1.6)	3 (9.1)
Citrobacter	8 (2.3)	0 (0.0)	0 (0.0)	2 (4.0)	1 (0.8)	2 (1.6)	3 (9.1)
Cronobacter sakazakii	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
Enterobacter (non-cloacaecomplex)	9 (2.6)	0 (0.0)	1 (10.0)	2 (4.0)	5 (4.0)	1 (0.8)	0 (0.0)
Enterobacter cloacae complex	23 (6.6)	3 (42.9)	1 (10.0)	6 (12.0)	5 (4.0)	8 (6.4)	0 (0.0)
Escherichia coli	132 (37.8)	2 (28.6)	2 (20.0)	16 (32.0)	41 (33.1)	55 (44.0)	16 (48.5)
Fusobacterium necrophorum	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
Fusobacterium nucleatum	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
Haemophilus influenzae	7 (2.0)	0 (0.0)	0 (0.0)	3 (6.0)	1 (0.8)	1 (0.8)	2 (6.1)
Klebsiella oxytoca	12 (3.4)	0 (0.0)	0 (0.0)	3 (6.0)	8 (6.5)	1 (0.8)	0 (0.0)
Klebsiella pneumoniae group	59 (16.9)	1 (14.3)	1 (10.0)	10 (20.0)	26 (21.0)	17 (13.6)	4 (12.1)
Morganella morganii	3 (0.9)	0 (0.0)	0 (0.0)	0 (0.0)	2 (1.6)	1 (0.8)	0 (0.0)
Neisseria meningitidis	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
Proteus	22 (6.3)	0 (0.0)	0 (0.0)	0 (0.0)	6 (4.8)	13 (10.4)	3 (9.1)
Proteus mirabilis	22 (6.3)	0 (0.0)	0 (0.0)	0 (0.0)	6 (4.8)	13 (10.4)	3 (9.1)
Pseudomonas aeruginosa	28 (8.0)	0 (0.0)	2 (20.0)	3 (6.0)	12 (9.7)	10 (8.0)	1 (3.0)
Salmonella	2 (0.6)	0 (0.0)	0 (0.0)	0 (0.0)	1 (0.8)	1 (0.8)	0 (0.0)
Serratia	10 (2.9)	0 (0.0)	1 (10.0)	0 (0.0)	5 (4.0)	4 (3.2)	0 (0.0)
Serratia marcescens	9 (2.6)	0 (0.0)	1 (10.0)	0 (0.0)	4 (3.2)	4 (3.2)	0 (0.0)
Stenotrophomonas maltophilia	3 (0.9)	0 (0.0)	0 (0.0)	3 (6.0)	0 (0.0)	0 (0.0)	0 (0.0)
Pan Candida	2 (0.6)	1 (14.3)	0 (0.0)	0 (0.0)	1 (0.8)	0 (0.0)	0 (0.0)
Pan Gram-Positive	24 (6.9)	1 (14.3)	2 (20.0)	5 (10.0)	7 (5.6)	7 (5.6)	2 (6.1)
CTX-M	24 (6.9)	0 (0.0)	0 (0.0)	2 (4.0)	7 (5.6)	12 (9.6)	3 (9.1)
IMP	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
КРС	3 (0.9)	0 (0.0)	0 (0.0)	1 (2.0)	1 (0.8)	1 (0.8)	0 (0.0)
NDM	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
OXA	1 (0.3)	0 (0.0)	0 (0.0)	0 (0.0)	1 (0.8)	0 (0.0)	0 (0.0)
VIM	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)

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Table 2: Expected Value by Collection Site (Prospective Samples)
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Target	All Sites(N=349)n (%)	Site 1(N=88)n (%)	Site 2(N=23)n (%)	Site 3(N=98)n (%)	Site 4(N=58)n (%)	Site 5(N=46)n (%)	Site 6(N=28)n (%)	Site 7(N=8)n (%)		Resistance Gene
Enterobacter(non-cloacae complex)	9 (2.6)	2 (2.3)	0 (0.0)	4 (4.1)	0 (0.0)	1 (2.2)	2 (7.1)	0 (0.0)	Organism	CTX-M	IMP	KPC	NDM	OXA	VIM
	Acinetobacter baumannii	X	X	X	X	X	X
Enterobacter cloacae complex	23 (6.6)	3 (3.4)	1 (4.3)	10 (10.2)	1 (1.7)	6 (13.0)	2 (7.1)	0 (0.0)	Bacteroides fragilis
Escherichia coli	132 (37.8)	30 (34.1)	8 (34.8)	37 (37.8)	25 (43.1)	17 (37.0)	12 (42.9)	3 (37.5)	Citrobacter	X	X	X	X	X	X
Fusobacterium necrophorum	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	Cronobacter sakazakii			X
Fusobacterium nucleatum	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	Enterobacter cloacae complex	X	X	X	X	X	X
Haemophilus influenzae	7 (2.0)	1 (1.1)	0 (0.0)	2 (2.0)	2 (3.4)	1 (2.2)	1 (3.6)	0 (0.0)	Enterobacter (non-cloacae complex)	X	X	X	X	X	X
Klebsiella oxytoca	12 (3.4)	5 (5.7)	0 (0.0)	3 (3.1)	1 (1.7)	2 (4.3)	1 (3.6)	0 (0.0)	Escherichia coli	X	X	X	X	X	X
Klebsiella pneumoniae group	59 (16.9)	17 (19.3)	3 (13.0)	20 (20.4)	5 (8.6)	7 (15.2)	4 (14.3)	3 (37.5)	Fusobacterium necrophorum
Morganella morganii	3 (0.9)	0 (0.0)	1 (4.3)	2 (2.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	Fusobacterium nucleatum
Neisseria meningitidis	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	Haemophilus influenzae
Proteus	22 (6.3)	9 (10.2)	0 (0.0)	5 (5.1)	5 (8.6)	2 (4.3)	1 (3.6)	0 (0.0)	Klebsiella oxytoca	X	X	X	X	X	X
Proteus mirabilis	22 (6.3)	9 (10.2)	0 (0.0)	5 (5.1)	5 (8.6)	2 (4.3)	1 (3.6)	0 (0.0)	Klebsiella pneumoniae	X	X	X	X	X	X
Pseudomonas aeruginosa	28 (8.0)	5 (5.7)	2 (8.7)	10 (10.2)	8 (13.8)	2 (4.3)	1 (3.6)	0 (0.0)	Morganella morganii	X	X	X	X	X	X
Salmonella	2 (0.6)	1 (1.1)	0 (0.0)	0 (0.0)	0 (0.0)	1 (2.2)	0 (0.0)	0 (0.0)	Neisseria meningitidis
Serratia	10 (2.9)	1 (1.1)	2 (8.7)	1 (1.0)	3 (5.2)	3 (6.5)	0 (0.0)	0 (0.0)	Proteus	X	X	X	X	X	X
Serratia marcescens	9 (2.6)	1 (1.1)	2 (8.7)	1 (1.0)	3 (5.2)	2 (4.3)	0 (0.0)	0 (0.0)	Proteus mirabilis	X	X	X	X	X	X
Stenotrophomonas maltophilia	3 (0.9)	1 (1.1)	0 (0.0)	0 (0.0)	1 (1.7)	0 (0.0)	1 (3.6)	0 (0.0)	Pseudomonas aeruginosa	X	X	X	X	X	X
Pan Candida	2 (0.6)	0 (0.0)	0 (0.0)	1 (1.0)	0 (0.0)	0 (0.0)	1 (3.6)	0 (0.0)	Salmonella	X	X	X	X	X	X
Pan Gram-Positive	24 (6.9)	15 (17.0)	1 (4.3)	5 (5.1)	1 (1.7)	0 (0.0)	2 (7.1)	0 (0.0)	Serratia	X	X	X	X	X	X
CTX-M	24 (6.9)	9 (10.2)	1 (4.3)	3 (3.1)	4 (6.9)	5 (10.9)	2 (7.1)	0 (0.0)	Serratia marcescens	X	X	X	X	X	X
IMP	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	Stenotrophomonas maltophilia	X
КРС	3 (0.9)	1 (1.1)	0 (0.0)	1 (1.0)	1 (1.7)	0 (0.0)	0 (0.0)	0 (0.0)
NDM	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
OXA	1 (0.3)	1 (1.1)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)
VIM	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)

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PERFORMANCE CHARACTERISTICS

Clinical Performance

Samples with final, valid ePlex BCID-GN Panel test results and a valid comparator result were evaluable and included in summaries and analyses of demographics, expected values (positivity rate), and performance characteristics. Evaluable samples included 167 prospective fresh and 182 prospective frozen samples as well as 577 retrospective samples and 777 contrived samples.

Comparator Method

The performance of the ePlex BCID-GN Panel was compared to standard laboratory procedures for identification of blood culture isolates, including traditional and automated identification methods, MALDI-TOF IVD, and microbiological and biochemical techniques. Identification for samples with Acinetobacter baumannii or Candida parapsilosis identified by standard laboratory procedures was confirmed using analytically validated PCR assays followed by bi-directional sequencing. For antibiotic resistance genes, the ePlex BCID-GN Panel was compared to analytically validated qPCR amplification assays followed by bi-directional sequencing in samples with an associated organism identified by culture (See Table 3 for organism associations).

Table 3: Resistance Marker Organism Associations

VOL 005 Page 9

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The comparator method(s) results were used to determine the Detected / Not Detected status for each target organism on the ePlex BCID-GN Panel. The comparator methods for each target are summarized in Table 4.

Target	Comparator Method
Acinetobacter baumannii	Standard laboratory procedures for organism ID.PCR/sequencing to confirm Acinetobacter baumannii or differentAcinetobacter species not included in this panel.
Bacteroides fragilis
Citrobacter
Cronobacter sakazakii
Enterobacter cloacae complex
Enterobacter non-cloacae complex
Escherichia coli
Fusobacterium necrophorum
Fusobacterium nucleatum
Haemophilus influenzae
Klebsiella oxytoca
Klebsiella pneumoniae	Standard laboratory procedures for organism identification.
Morganella morganii
Neisseria meningitidis
Proteus
Proteus mirabilis
Pseudomonas aeruginosa
Salmonella
Serratia
Serratia marcescens
Stenotrophomonas maltophilia
Pan Gram-Negative
Pan Candida	Standard laboratory procedures for organism ID.PCR/sequencing to confirm C. parapsilosis or identifyC. metapsilosis, C. orthopsilosis.
CTX-M, IMP, KPC, NDM, OXA, VIM	qPCR/sequencing in samples with associated organism detectedby comparator method. See Table 3 for organism associations.

Table 4: Comparator Method(s) by ePlex BCID-GN Panel Target

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Demographics of Clinical Samples

Clinical performance was evaluated in positive blood culture samples prospectively and retrospectively collected. Prospective samples were collected at 7 clinical sites in 2 phases. From June 2014 through July 2016, 183 samples were prospectively collected and frozen; from June through July 2018, 171 samples were prospectively collected and tested fresh (never frozen) for a total of 354 samples across the 2 phases. One of these samples was withdrawn due to organism identification from unacceptable methods. Of the 353 prospectively-collected samples eligible for testing, 349 were evaluable. Samples with final, valid ePlex BCID-GN Panel results and a valid comparator result were evaluable. Four samples were not evaluable because they did not have final, valid ePlex BCID-GN Panel results and were excluded from performance evaluations. Demographic information for prospectively-collected samples is described in Table 5. Subjects enrolled in this study were from a diverse demographic distribution and represent the intended patient population.

To supplement the number of positives for low prevalence targets in the prospective collection, 578 samples were collected retrospectively, 577 were evaluable. One sample was not evaluable because it did not have a final, valid ePlex BCID-GN Panel result and was excluded from performance evaluations. Demographic information for retrospectively-collected samples is described in Table 6.

	All SitesN = 349n (%)	Site 1N = 88n (%)	Site 2N = 23n (%)	Site 3N = 98n (%)	Site 4N = 58n (%)	Site 5N = 46n (%)	Site 6N = 28n (%)	Site 7N=8n (%)
Sex
Male	168 (48.1)	37 (42.0)	12 (52.2)	52 (53.1)	28 (48.3)	21 (45.7)	13 (46.4)	5 (62.5)
Female	181 (51.9)	51 (58.0)	11 (47.8)	46 (46.9)	30 (51.7)	25 (54.3)	15 (53.6)	3 (37.5)
Age
<1 yr	7 (2.0)	2 (2.3)	0 (0.0)	4 (4.1)	0 (0.0)	1 (2.2)	0 (0.0)	0 (0.0)
1-17 yrs	10 (2.9)	4 (4.5)	1 (4.3)	3 (3.1)	1 (1.7)	1 (2.2)	0 (0.0)	0 (0.0)
18-44 yrs	50 (14.3)	10 (11.4)	3 (13.0)	20 (20.4)	3 (5.2)	8 (17.4)	6 (21.4)	0 (0.0)
45-64 yrs	124 (35.5)	35 (39.8)	9 (39.1)	28 (28.6)	21 (36.2)	14 (30.4)	13 (46.4)	4 (50.0)
65-84 yrs	125 (35.8)	29 (33.0)	8 (34.8)	35 (35.7)	25 (43.1)	17 (37.0)	7 (25.0)	4 (50.0)
85+ yrs	33 (9.5)	8 (9.1)	2 (8.7)	8 (8.2)	8 (13.8)	5 (10.9)	2 (7.1)	0 (0.0)

Table 5: Demographic Data for Clinical Samples by Collection Site (Prospective Collection)

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	All SitesN = 577n (%)	Site 1N = 78n (%)	Site 2N = 73n (%)	Site 3N = 31n (%)	Site 4N = 93n (%)	Site 5N=1n (%)	Site 6N = 80n (%)	Site 7N = 67n (%)	Site 8N = 48n (%)	Site 9N = 29n (%)	Site 10N = 77n (%)
Sex
Male	307 (53.2)	36 (46.2)	41 (56.2)	15 (48.4)	49 (52.7)	0 (0.0)	47 (58.8)	38 (56.7)	29 (60.4)	19 (65.5)	33 (42.9)
Female	270 (46.8)	42 (53.8)	32 (43.8)	16 (51.6)	44 (47.3)	1 (100)	33 (41.3)	29 (43.3)	19 (39.6)	10 (34.5)	44 (57.1)
Age
<1 yr	9 (1.6)	1 (1.3)	0 (0.0)	0 (0.0)	3 (3.2)	0 (0.0)	2 (2.5)	0 (0.0)	1 (2.1)	0 (0.0)	2 (2.6)
1-17 yrs	20 (3.5)	1 (1.3)	0 (0.0)	1 (3.2)	8 (8.6)	0 (0.0)	6 (7.5)	0 (0.0)	0 (0.0)	1 (3.4)	3 (3.9)
18-44 yrs	78 (13.5)	13 (16.7)	7 (9.6)	2 (6.5)	10 (10.8)	1 (100)	15 (18.8)	8 (11.9)	8 (16.7)	6(20.7)	8 (10.4)
45-64 yrs	193 (33.4)	27 (34.6)	18 (24.7)	13 (41.9)	27 (29.0)	0 (0.0)	32 (40.0)	27 (40.3)	16 (33.3)	9 (31.0)	24 (31.2)
65-84 yrs	226 (39.2)	29 (37.2)	40 (54.8)	11 (35.5)	40 (43.0)	0 (0.0)	20 (25.0)	24 (35.8)	21 (43.8)	11 (37.9)	30 (39.0)
85+ yrs	49 (8.5)	7 (9.0)	8 (11.0)	4 (12.9)	5 (5.4)	0 (0.0)	5 (6.3)	6 (9.0)	2 (4.2)	2 (6.9)	10 (13.0)
Unknown	2 (0.3)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	0 (0.0)	2 (3.0)	0 (0.0)	0 (0.0)	0 (0.0)

Clinical Performance

Sensitivity or positive percent agreement (PPA) was calculated by dividing the number of true positive (TP) results by the sum of TP and false negative (FN) results, while specificity or negative percent agreement (NPA) was calculated by dividing the number of true negative (TN) results by the sum of TN and false positive (FP) result being defined as a sample where the detected ePlex BCID-GN Panel result matched the detected comparator method result, while a TN result was one where a negative ePlex BCID-GN Panel result matched a negative comparator method result. The two-sided 95% confidence interval was also calculated.

A total of 349 prospectively-collected samples (167 tested fresh and 182 tested after previously frozen) and 577 retrospectively collected samples from blood culture bottles flagged positive in a continuously monitoring blood culture system and removed from the system within 8 hours of positivity were evaluated for the ePlex BCID-GN Panel targets. Specimens evaluated were determined to contain gram-negative or gram-variable organisms based on Gram stain. A total of 777 contrived samples were prepared by spiking an isolate into a blood culture bottle with human whole blood and growing until flagged positive by a continuously monitoring blood culture system. Contrived samples were removed from the system within 8 hours of positivity and stored frozen until the time of testing. PPA and NPA results are summarized by target in Tables 7-34 below, and the strains used to contrive samples are summarized in Table 35.

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Target	Sample Type	TP/TP+FN	Sensitivity/PPA% (95% CI)	TN/TN+FP	Specificity/NPA% (95% CI)
	Prospective (Fresh)	0/0	---	167/167	100 (97.8-100)
Acinetobacterbaumannii	Prospective (Frozen)	4/4	100 (51.0-100)	178/178	100 (97.9-100)
	Prospective (All)	4/4	100 (51.0-100)	345/345	100 (98.9-100)
	Retrospective	15/15	100 (79.6-100)	560/561	99.8 (99.0-100)
	Prospective / Retrospective	19/19	100 (83.2-100)	905/906A	99.9 (99.4-100)
	Contrived	55/55	100 (93.5-100)	722/722	100 (99.5-100)
	Overall	74/74	100 (95.1-100)	1627/1628	99.9 (99.7-100)

CI= Confidence Interval

A. Acinetobacter baumanii was detected in the 1/1 false positive sample using PCR/sequencing.

Table 8: Clinical Performance for Bacteroides fragilis

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Bacteroides fragilis	Prospective (Fresh)	6/6	100 (61.0-100)	161/161	100 (97.7-100)
	Prospective (Frozen)	5/5	100 (56.6-100)	177/177	100 (97.9-100)
	Prospective (All)	11/11	100 (74.1-100)	338/338	100 (98.9-100)
	Retrospective	14/17	82.4 (59.0-93.8)	558/560	99.6 (98.7-99.9)
	Prospective / Retrospective	25/28A	89.3 (72.8-96.3)	896/898B	99.8 (99.2-99.9)
	Contrived	40/40	100 (91.2-100)	737/737	100 (99.5-100)
	Overall	65/68	95.6 (87.8-98.5)	1633/1635	99.9 (99.6-100)

A. B. fragilis was not detected in 2 false negative samples, but PCR/sequencing instead detected B. caccae and B. thetaiotaomicron, which were not identified by standard laboratory procedures.

B. B. fragilis was detected in 2/2 false positive samples using PCR/sequencing.

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Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Citrobacter	Prospective (Fresh)	3/3	100 (43.9-100)	163/164	99.4 (96.6-99.9)
	Prospective (Frozen)	2/2	100 (34.2-100)	178/180	98.9 (96.0-99.7)
	Prospective (All)	5/5	100 (56.6-100)	341/344	99.1 (97.5-99.7)
	Retrospective	20/21	95.2 (77.3-99.2)	555/556	99.8 (99.0-100)
	Prospective / Retrospective	25/26	96.2 (81.1-99.3)	896/9004	99.6 (98.9-99.8)
	Contrived	43/43	100 (91.8-100)	734/734	100 (99.5-100)
Citrobacter	Overall	68/69	98.6 (92.2-99.7)	1630/1634	99.8 (99.4-99.9)

Table 9: Clinical Performance for Citrobacter

A. Citrobacter braakii (2) and Citrobacter freundii (2) were detected in 44 false positive samples using PCR/sequencing.

Table 10: Clinical Performance for Cronobacter sakazakii

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Cronobacter sakazakii	Prospective (Fresh)	0/0	---	167/167	100 (97.8-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	0/0	---	349/349	100 (98.9-100)
	Retrospective	1/1	100 (20.7-100)	576/576	100 (99.3-100)
	Prospective / Retrospective	1/1	100 (20.7-100)	925/925	100 (99.6-100)
	Contrived	45/45	100 (92.1-100)	732/732	100 (99.5-100)
	Overall	46/46	100 (92.3-100)	1657/1657	100 (99.8-100)

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Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Enterobacter cloacaecomplex	Prospective (Fresh)	12/12	100 (75.8-100)	153/155	98.7 (95.4-99.6)
	Prospective (Frozen)	7/7	100 (64.6-100)	173/175	98.9 (95.9-99.7)
	Prospective (All)	19/19	100 (83.2-100)	326/330	98.8 (96.9-99.5)
	Retrospective	47/50	94.0 (83.8-97.9)	526/527	99.8 (98.9-100)
	Prospective / Retrospective	66/69A	95.7 (88.0-98.5)	852/857B	99.4 (98.6-99.8)
	Contrived	35/37C	94.6 (82.3-98.5)	739/740	99.9 (99.2-100)
	Overall	101/106	95.3 (89.4-98.0)	1591/1597	99.6 (99.2-99.8)

Table 11: Clinical Performance for Enterobacter cloacae complex

A. A species of the Enterobacter cloacae complex was not detected in 1 false negative sample, but PCR/sequencing and MALDI-TOF instead detected E. coli. Standard laboratory procedures identified E. cloacae only.

B. E. cloacae was detected in 2/5 false positive samples using PCR/sequencing.

C. E. cloacae complex was not detected in 2 samples containing Enterobacter asburiae.

Table 12: Clinical Performance for Enterobacter (non-cloacae complex)

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Enterobacter -non-cloacae complex	Prospective (Fresh)	3/3	100 (43.9-100)	163/164	99.4 (96.6-99.9)
	Prospective (Frozen)	5/7	71.4 (35.9-91.8)	175/175	100 (97.9-100)
	Prospective (All)	8/10	80.0 (49.0-94.3)	338/339	99.7 (98.3-99.9)
	Retrospective	12/12	100 (75.8-100)	565/565	100 (99.3-100)
	Prospective / Retrospective	20/22A	90.9 (72.2-97.5)	903/904B	99.9 (99.4-100)
	Contrived	36/36	100 (90.4-100)	741/741	100 (99.5-100)
	Overall	56/58	96.6 (88.3-99.0)	1644/1645	99.9 (99.7-100)

A. A species of the Enterobacter non-cloacae complex was not detected in 2 false negative samples. Standard laboratory procedures identified E. aerogenes and PCR/sequencing detected E. cloacae.,.

B. A species of the Enterobacter non-cloacae complex was not detected in the false positive sample using PCR/sequencing.

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Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Escherichia coli	Prospective (Fresh)	59/60	98.3 (91.1-99.7)	106/107	99.1 (94.9-99.8)
	Prospective (Frozen)	72/73	98.6 (92.6-99.8)	109/109	100 (96.6-100)
	Prospective (All)	131/133	98.5 (94.7-99.6)	215/216	99.5 (97.4-99.9)
	Retrospective	132/140	94.3 (89.1-97.1)	435/437	99.5 (98.3-99.9)
	Prospective / Retrospective	263/273	96.3 (93.4-98.0)	650/653A	99.5 (98.7-99.8)
	Contrived	52/52	100 (93.1-100)	725/725	100 (99.5-100)
	Overall	315/325	96.9 (94.4-98.3)	1375/1378	99.8 (99.4-99.9)

Table 13: Clinical Performance for Escherichia coli

A. E. coli was detected in 3/3 false positive samples using PCR/sequencing.

Table 14: Clinical Performance for Fusobacterium necrophorum

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Fusobacteriumnecrophorum	Prospective (Fresh)	0/0	---	167/167	100 (97.8-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	0/0	---	349/349	100 (98.9-100)
	Retrospective	1/1	100 (20.7-100)	576/576	100 (99.3-100)
	Prospective / Retrospective	1/1	100 (20.7-100)	925/925	100 (99.6-100)
	Contrived	47/48	97.9 (89.1-99.6)	729/729	100 (99.5-100)
	Overall	48/49	98.0 (89.3-99.6)	1654/1654	100 (99.8-100)

{18}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Fusobacteriumnucleatum	Prospective (Fresh)	0/0	---	167/167	100 (97.8-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	0/0	---	349/349	100 (98.9-100)
	Retrospective	5/5	100 (56.6-100)	571/572	99.8 (99.0-100)
	Prospective / Retrospective	5/5	100 (56.6-100)	920/921A	99.9 (99.4-100)
	Contrived	47/47	100 (92.4-100)	730/730	100 (99.5-100)
	Overall	52/52	100 (93.1-100)	1650/1651	99.9 (99.7-100)

Table 15: Clinical Performance for Fusobacterium nucleatum
------------------------------------------------------------

A. F. nucleatum was detected in the 1/1 false positive sample using PCR/sequencing.

Table 16: Clinical Performance for Haemophilus influenzae

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Haemophilusinfluenzae	Prospective (Fresh)	3/3	100 (43.9-100)	164/164	100 (97.7-100)
	Prospective (Frozen)	4/4	100 (51.0-100)	178/178	100 (97.9-100)
	Prospective (All)	7/7	100 (64.6-100)	342/342	100 (98.9-100)
	Retrospective	7/7	100 (64.6-100)	570/570	100 (99.3-100)
	Prospective / Retrospective	14/14	100 (78.5-100)	912/912	100 (99.6-100)
	Contrived	41/41	100 (91.4-100)	736/736	100 (99.5-100)
	Overall	55/55	100 (93.5-100)	1648/1648	100 (99.8-100)

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Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Klebsiella oxytoca	Prospective (Fresh)	4/6	66.7 (30.0-90.3)	160/161	99.4 (96.6-99.9)
	Prospective (Frozen)	7/7	100 (64.6-100)	175/175	100 (97.9-100)
	Prospective (All)	11/13	84.6 (57.8-95.7)	335/336	99.7 (98.3-99.9)
	Retrospective	29/34	85.3 (69.9-93.6)	541/543	99.6 (98.7-99.9)
	Prospective / Retrospective	40/47A	85.1 (72.3-92.6)	876/879B	99.7 (99.0-99.9)
	Contrived	20/20	100 (83.9-100)	757/757	100 (99.5-100)
	Overall	60/67	89.6 (80.0-94.8)	1633/1636	99.8 (99.5-99.9)

Table 17: Clinical Performance for Klebsiella oxytoca

A. K. oxytoca was not detected in 2 false negative samples using but 16S sequencing instead detected Raoultella ornithinolytica and Raoultella planticola, which were not identified by standard laboratory procedures.

B. Klebsiella oxytoca was detected in 3/3 false positive samples using PCR/sequencing.

Table 18: Clinical Performance for Klebsiella pneumoniae group

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Klebsiella pneumoniaegroup	Prospective (Fresh)	29/30	96.7 (83.3-99.4)	136/137	99.3 (96.0-99.9)
	Prospective (Frozen)	29/31	93.5 (79.3-98.2)	151/151	100 (97.5-100)
	Prospective (All)	58/61	95.1 (86.5-98.3)	287/288	99.7 (98.1-99.9)
	Retrospective	106/108	98.1 (93.5-99.5)	466/469	99.4 (98.1-99.8)
	Prospective / Retrospective	164/169A	97.0 (93.3-98.7)	753/757B	99.5 (98.6-99.8)
	Contrived	72/72	100 (94.9-100)	705/705	100 (99.5-100)
	Overall	236/241	97.9 (95.2-99.1)	1458/1462	99.7 (99.3-99.9)

A. K. pneumoniae was not detected in 1 false negative sample, but PCR/sequencing and MALDI-TOF instead detected K. oxyloca, which was not identified by standard laboratory procedures.

B. K. pneumoniae was detected in 4/4 false positive samples using PCR/sequencing.

{20}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Morganella morganii	Prospective (Fresh)	3/3	100 (43.9-100)	164/164	100 (97.7-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	3/3	100 (43.9-100)	346/346	100 (98.9-100)
	Retrospective	10/10	100 (72.2-100)	566/567	99.8 (99.0-100)
	Prospective / Retrospective	13/13	100 (77.2-100)	912/913A	99.9 (99.4-100)
	Contrived	49/49	100 (92.7-100)	728/728	100 (99.5-100)
	Overall	62/62	100 (94.2-100)	1640/1641	99.9 (99.7-100)

Table 19: Clinical Performance for Morganella morganii

A. M. morganii was detected in 1/1 false positive clinical samples using PCR/sequencing.

Table 20: Clinical Performance for Neisseria meningitidis

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Neisseria meningitidis	Prospective (Fresh)	0/0	---	167/167	100 (97.8-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	0/0	---	349/349	100 (98.9-100)
	Retrospective	0/0	---	576/577	99.8 (99.0-100)
	Prospective / Retrospective	0/0	---	925/926A	99.9 (99.4-100)
	Contrived	44/44	100 (92.0-100)	733/733	100 (99.5-100)
	Overall	44/44	100 (92.0-100)	1658/1659	99.9 (99.7-100)

A. N. meningitidis was not detected in the false positive sample using PCR/sequencing.

{21}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Proteus	Prospective (Fresh)	7/8	87.5 (52.9-97.8)	159/159	100 (97.6-100)
	Prospective (Frozen)	15/15	100 (79.6-100)	167/167	100 (97.8-100)
	Prospective (All)	22/23	95.7 (79.0-99.2)	326/326	100 (98.8-100)
	Retrospective	54/55	98.2 (90.4-99.7)	522/522	100 (99.3-100)
	Prospective / Retrospective	76/78	97.4 (91.1-99.3)	848/848	100 (99.5-100)
	Contrived	9/9	100 (70.1-100)	768/768	100 (99.5-100)
	Overall	85/87	97.7 (92.0-99.4)	1616/1616	100 (99.8-100)

Table 21: Clinical Performance for Proteus

Table 22: Clinical Performance for Proteus mirabilis

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Proteus mirabilis	Prospective (Fresh)	7/8	87.5 (52.9-97.8)	159/159	100 (97.6-100)
	Prospective (Frozen)	15/15	100 (79.6-100)	167/167	100 (97.8-100)
	Prospective (All)	22/23	95.7 (79.0-99.2)	326/326	100 (98.8-100)
	Retrospective	50/51	98.0 (89.7-99.7)	526/526	100 (99.3-100)
	Prospective / Retrospective	72/74	97.3 (90.7-99.3)	852/852	100 (99.6-100)
	Contrived	9/9	100 (70.1-100)	768/768	100 (99.5-100)
	Overall	81/83	97.6 (91.6-99.3)	1620/1620	100 (99.8-100)

{22}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Pseudomonasaeruginosa	Prospective (Fresh)	10/10	100 (72.2-100)	157/157	100 (97.6-100)
	Prospective (Frozen)	17/18	94.4 (74.2-99.0)	163/164	99.4 (96.6-99.9)
	Prospective (All)	27/28	96.4 (82.3-99.4)	320/321	99.7 (98.3-99.9)
	Retrospective	56/60	93.3 (84.1-97.4)	514/517	99.4 (98.3-99.8)
	Prospective / Retrospective	83/88	94.3 (87.4-97.5)	834/838A	99.5 (98.8-99.8)
	Contrived	32/32	100 (89.3-100)	745/745	100 (99.5-100)
	Overall	115/120	95.8 (90.6-98.2)	1579/1583	99.7 (99.4-99.9)

Table 23: Clinical Performance for Pseudomonas aeruginosa

A. P. aeruginosa was detected in 2/4 false positive samples using PCR/sequencing.

Table 24: Clinical Performance for Salmonella

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Salmonella	Prospective (Fresh)	2/2	100 (34.2-100)	165/165	100 (97.7-100)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	2/2	100 (34.2-100)	347/347	100 (98.9-100)
	Retrospective	18/19	94.7 (75.4-99.1)	558/558	100 (99.3-100)
	Prospective / Retrospective	20/21 A	95.2 (77.3-99.2)	905/905	100 (99.6-100)
	Contrived	34/35	97.1 (85.5-99.5)	742/742	100 (99.5-100)
	Overall	54/56	96.4 (87.9-99.0)	1647/1647	100 (99.8-100)

A. Salmonella was not detected in 1 false negative sample, but PCR sequencing instead detected E. coli, which was not identified by standard laboratory procedures.

{23}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Serratia	Prospective (Fresh)	6/6	100 (61.0-100)	161/161	100 (97.7-100)
	Prospective (Frozen)	4/4	100 (51.0-100)	178/178	100 (97.9-100)
	Prospective (All)	10/10	100 (72.2-100)	339/339	100 (98.9-100)
	Retrospective	34/34	100 (89.8-100)	542/543	99.8 (99.0-100)
	Prospective / Retrospective	44/44	100 (92.0-100)	881/8824	99.9 (99.4-100)
	Contrived	36/36	100 (90.4-100)	741/741	100 (99.5-100)
	Overall	80/80	100 (95.4-100)	1622/1623	99.9 (99.7-100)

Table 25: Clinical Performance for Serratia

A. S. marcescens was detected in the 1/1 false positive sample using PCR/sequencing.

Table 26: Clinical Performance for Serratia marcescens

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Serratia marcescens	Prospective (Fresh)	5/5	100 (56.6-100)	162/162	100 (97.7-100)
	Prospective (Frozen)	4/4	100 (51.0-100)	178/178	100 (97.9-100)
	Prospective (All)	9/9	100 (70.1-100)	340/340	100 (98.9-100)
	Retrospective	34/34	100 (89.8-100)	542/543	99.8 (99.0-100)
	Prospective / Retrospective	43/43	100 (91.8-100)	882/883A	99.9 (99.4-100)
	Contrived	19/19	100 (83.2-100)	758/758	100 (99.5-100)
	Overall	62/62	100 (94.2-100)	1640/1641	99.9 (99.7-100)

A. S. marcescens was detected in the 1/1 false positive sample using PCR/sequencing.

{24}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Stenotrophomonasmaltophilia	Prospective (Fresh)	2/3	66.7 (20.8-93.9)	164/164	100 (97.7-100)
	Prospective (Frozen)	1/1	100 (20.7-100)	181/181	100 (97.9-100)
	Prospective (All)	3/4	75.0 (30.1-95.4)	345/345	100 (98.9-100)
	Retrospective	8/10	80.0 (49.0-94.3)	566/567	99.8 (99.0-100)
	Prospective / Retrospective	11/14	78.6 (52.4-92.4)	911/912A	99.9 (99.4-100)
	Contrived	36/36	100 (90.4-100)	741/741	100 (99.5-100)
	Overall	47/50	94.0 (83.8-97.9)	1652/1653	99.9 (99.7-100)

Table 27: Clinical Performance for Stenotrophomonas maltophilia

A. S. maltophilia was detected in the 1/1 false positive sample using PCR/sequencing.

Table 28: Clinical Performance for CTX-M

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
CTX-M	Prospective (Fresh)	10/13	76.9 (49.7-91.8)	127/127	100 (97.1-100)
	Prospective (Frozen)	12/16	75.0 (50.5-89.8)	144/144	100 (97.4-100)
	Prospective (All)	22/29	75.9 (57.9-87.8)	271/271	100 (98.6-100)
	Retrospective	52/56	92.9 (83.0-97.2)	483/483	100 (99.2-100)
	Prospective / Retrospective	74/85A	87.1 (78.3-92.6)	754/754	100 (99.5-100)
	Contrived	75/75	100 (95.1-100)	437/437	100 (99.1-100)
	Overall	149/160	93.1 (88.1-96.1)	1191/1191	100 (99.7-100)

A. In 3/11 false negative samples, CTX-M signal was above the threshold for detection; however, an associated by the BCID-GN Panel and the CTX-M target was reported as 'N/A'. Further testing of the 8/11 remaining false negative samples indicated that 7 of the 8 samples may have been contaminated during the original comparator extraction process and misidentified as having CTX-M present. Specifically, results for 7 of the 8 samples were negative for CTX-M from the following additional testing of 2 repeat extractions from the original sample, 2) qPCR testing of an extraction from the isolate, and 3) testing of the original sample with an FDA-cleared multiplex assay. For these 7 samples, the originally extracted sample was re-tested with qPCR and CTX-M was again detected, suggesting contamination during the original extraction process. The remaining 8th sample was possitive for CTX-M from the repeat extractions, negative for CTX-M from the isolate, and negative for CTX-M when tested with an FDA-cleared multiplex assay. These inconsistent detection results suggest the 8th sample may be a true low-copy CTX-M positive sample.

Confidential

{25}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
IMP	Prospective (Fresh)	0/0	---	138/138	100 (97.3-100)
	Prospective (Frozen)	0/0	---	159/159	100 (97.6-100)
	Prospective (All)	0/0	---	297/297	100 (98.7-100)
	Retrospective	0/0	---	532/532	100 (99.3-100)
	Prospective / Retrospective	0/0	---	829/829	100 (99.5-100)
	Contrived	40/40	100 (91.2-100)	436/436	100 (99.1-100)
	Overall	40/40	100 (91.2-100)	1265/1265	100 (99.7-100)

Table 29: Clinical Performance for IMP

Table 30: Clinical Performance for KPC

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
KPC	Prospective (Fresh)	2/2	100 (34.2-100)	136/136	100 (97.3-100)
	Prospective (Frozen)	1/1	100 (20.7-100)	158/158	100 (97.6-100)
	Prospective (All)	3/3	100 (43.9-100)	294/294	100 (98.7-100)
	Retrospective	4/5	80.0 (37.6-96.4)	527/528	99.8 (98.9-100)
	Prospective / Retrospective	7/8	87.5 (52.9-97.8)	821/822	99.9 (99.3-100)
	Contrived	44/44	100 (92.0-100)	477/477	100 (99.2-100)
	Overall	51/52	98.1 (89.9-99.7)	1298/1299	99.9 (99.6-100)

{26}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
NDM	Prospective (Fresh)	0/0	---	138/138	100 (97.3-100)
	Prospective (Frozen)	0/0	---	159/159	100 (97.6-100)
	Prospective (All)	0/0	---	297/297	100 (98.7-100)
	Retrospective	0/0	---	532/532	100 (99.3-100)
	Prospective / Retrospective	0/0	---	829/829	100 (99.5-100)
NDM	Contrived	54/54	100 (93.4-100)	422/422	100 (99.1-100)
	Overall	54/54	100 (93.4-100)	1251/1251	100 (99.7-100)

Table 31: Clinical Performance for NDM

Table 32: Clinical Performance for OXA

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
OXA	Prospective (Fresh)	0/1	0.0 (0.0-79.3)	137/137	100 (97.3-100)
	Prospective (Frozen)	1/1	100 (20.7-100)	158/158	100 (97.6-100)
	Prospective (All)	1/2	50.0 (9.5-90.5)	295/295	100 (98.7-100)
	Retrospective	9/11	81.8 (52.3-94.9)	519/521	99.6 (98.6-99.9)
	Prospective / Retrospective	10/13A	76.9 (49.7-91.8)	814/816	99.8 (99.1-99.9)
	Contrived	37/37	100 (90.6-100)	439/439	100 (99.1-100)
	Overall	47/50	94.0 (83.8-97.9)	1253/1255	99.8 (99.4-100)

A. In 1/3 false negative samples, OXA signal was above the threshold for detection; however, an associated organism was not detected by the BCID-GN Panel and the OXA target was reported as 'N/A'. One additional false negative sample was tested with an FDA-cleared multiplex assay and OXA was not detected. The isolate from the remaining false negative for OXA-23 and OXA-48 by qPCR.

{27}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
VIM	Prospective (Fresh)	0/0	---	138/138	100 (97.3-100)
	Prospective (Frozen)	0/0	---	159/159	100 (97.6-100)
	Prospective (All)	0/0	---	297/297	100 (98.7-100)
	Retrospective	0/0	---	532/532	100 (99.3-100)
	Prospective / Retrospective	0/0	---	829/829	100 (99.5-100)
	Contrived	42/42	100 (91.6-100)	434/434	100 (99.1-100)
	Overall	42/42	100 (91.6-100)	1263/1263	100 (99.7-100)

Table 33: Clinical Performance for VIM

Pan Targets

In addition to the evaluable prospective and retrospective samples that contain gram-negative organisms, the clinical performance of the Pan Candida and Pan Gram-Positive targets was evaluated by testing an additional 741 non-intended use retrospective samples with grampositive or fungal organisms; these are denoted as Retrospective (Non-Intended Use) samples. Results for the Pan targets are summarized in Table 34 and results stratified by species are summarized in Table 36. A summary of the strains used to prepare contrived samples and the number of strains for each target is shown in Table 35.

{28}------------------------------------------------

Target	Sample Type	Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Pan Candida	Prospective (Fresh)	1/1	100 (20.7-100)	165/166	99.4 (96.7-99.9)
	Prospective (Frozen)	0/0	---	182/182	100 (97.9-100)
	Prospective (All)	1/1	100 (20.7-100)	347/348A	99.7 (98.4-99.9)
	Retrospective	4/7B	57.1 (25.0-84.2)	569/570C	99.8 (99.0-100)
	Retrospective (Non-Intended Use)	99/102D	97.1 (91.7-99.0)	638/639E	99.8 (99.1-100)
	Contrived	0/0	---	777/777	100 (99.5-100)
	Prospective (Fresh)	7/8	87.5 (52.9-97.8)	155/159	97.5 (93.7-99.0)
Pan Gram-Positive	Prospective (Frozen)	10/15	66.7 (41.7-84.8)	164/167	98.2 (94.9-99.4)
	Prospective (All)	17/23F	73.9 (53.5-87.5)	319/326G	97.9 (95.6-99.0)
	Retrospective	44/55H	80.0 (67.6-88.4)	512/522I	98.1 (96.5-99.0)
	Retrospective (Non-Intended Use)	567/571	99.3 (98.2-99.7)	165/170J	97.1 (93.3-98.7)
	Contrived	0/0	---	776/777	99.9 (99.3-100)

Table 34: Clinical Performance for Pan Targets

A. Candida glabrata was detected in the 1/1 false positive sample using PCR/sequencing.

B. 3 of 3 (100%) false negative results occurred in mixed infections with bacterial organisms where the BCID-GN Panel results were correct for the other infections in these samples.

C. Candida albicans was detected in the 1/1 false positive sample using PCR/sequencing.

D. 2 of 3 (67%) false negative results occurred in mixed infections with bacterial organisms where the BCID-GN Panel results were correct for the other infections in these samples.

E. Candida glabrata was detected in the 1/1 false positive sample using PCR/sequencing.
F. Bacillus (the gram-positive organism identified by standard laboratory procedures) was not detected in 2 false negative samples using PCR sequencing, but 16S sequencing instead detected Paenibacillus urinalis, which were not identified by standard laboratory procedures. 3 of the remaining 4 (75%) false negative results occurred in mixed infections with bacterial organisms where the BCID-GN Panel correctly detected the gram-negative organisms present.

G. Enterococcus (1), Staphylococus (3), or Streptococcus (2) were detected in 6/7 false positive samples using PCR/sequencing (testing was not performed for the remaining 1/7 false positive samples).

H. 11 of 11 (100%) false negative results occurred in mixed infections with bacterial organisms where the BCID-GN Panel correctly detected the gram-negative organisms present.

Enterococcus (2), Staphylococcus (1), or Streptococus (5) were detected in 8/10 false positive samples using PCR/sequencing (testing was I. not performed for the remaining 2/10 false positive samples).

Bacillus (1) or Streptococcus (1) were detected in 2/5 false positive samples using (testing was not performed for the J. remaining 3/5 false positive samples).

{29}------------------------------------------------

Table 35: Contrived Sample Summary

Target	Organism	Strain	Independent ContrivedSamples Tested
Acinetobacter baumannii	Acinetobacter baumannii	ATCC19606	2
		ATCCBAA-2093	4
		ATCCBAA-747	5
		NCIMB12457	2
		NCTC13302	3
		NCTC13303	4
	Acinetobacter baumannii	NCTC13423	17
	Acinetobacter baumannii, NDM	CDC#0033	5
		ATCCBAA-1605	5
	Acinetobacter baumannii, OXA-23	NCTC13304	5
		NCTC13305	3
	Acinetobacter baumannii Total		55
Bacteroides fragilis	Bacteroides fragilis	ATCC23745	8
		ATCC25285	8
		ATCC43860	8
		ATCC700786	8
		NCTC9343	8
	Bacteroides fragilis Total		40
Citrobacter	Citrobacter braakii	ATCC43162	4
		ATCC51113	4
		ATCC43864	4
	Citrobacter freundii	ATCC8090	4
		NCTC8581	4
		NCTC9750	5
	Citrobacter freundii, CTX-M	JMI2047	6
	Citrobacter freundii, KPC	CDC#0116	4
	Citrobacter koseri	ATCC27156	4
	Citrobacter youngae	ATCC29935	4
	Citrobacter Total		43
Cronobacter sakazakii	Cronobacter sakazakii	ATCC12868	2
Target	Organism	Strain	Independent ContrivedSamples Tested
		ATCC29004	2
		ATCC29544	4
		ATCCBAA-894	3
		FSLF6-0023	4
		FSLF6-0028	4
		FSLF6-0029	4
		FSLF6-0034	3
		FSLF6-0035	3
		FSLF6-0043	4
		FSLF6-0049	3
		FSLF6-0050	4
		FSLF6-0051	5
	Cronobacter sakazakii Total		45
		ATCC35953	2
		ATCC35955	1
	Enterobacter asburiae	ATCC35956	4
		ATCC35957	1
	Enterobacter cloacae, CTX-M	CDC#0038	4
Enterobacter cloacaecomplex		NCTC13464	3
	Enterobacter cloacae, CTX-M, KPC	CDC#0163	2
	Enterobacter cloacae, CTX-M, NDM	CDC#0038	1
		JMI53571	12
	Enterobacter cloacae, VIM	CDC#0154	6
	Enterobacter hormaechei, KPC	ATCCBAA-2082	1

	Enterobacter cloacae complex Total		37
		ATCC13048	3
Enterobacter (non-cloacae complex)	Enterobacter aerogenes	ATCC29010	3
		ATCC51697	3
	Enterobacter aerogenes, IMP	CDC#0161	5
	Enterobacter aerogenes, OXA-48	CDC#0074	12
	Enterobacter amnigenus	ATCC33072	3
Target	Organism	Strain	Independent ContrivedSamples Tested
		ATCC33731	3
		ATCC51816	4
	Enterobacter (non-cloacae complex) Total		36
	Escherichia coli, CTX-M	CDC#0086	3
		NCTC13452	3
		NCTC13461	6
		NCTC13463	2
	Escherichia coli, CTX-M, NDM	CDC#0137	6
		CDC#0151	2
Escherichia coli	Escherichia coli, IMP	NCTC13476	7
	Escherichia coli, KPC	ATCCBAA-2340	2
		CDC#0114	4
		CDC#0118	6
	Escherichia coli, NDM	CDC#0150	5
		CDC#0151	4
	Escherichia coli, VIM	JMI32465	2
	Escherichia coli Total		52
Fusobacteriumnecrophorum	Fusobacterium necrophorum subsp.funduliforme	ATCC51357	24
	Fusobacterium necrophorum subsp.necrophorum	ATCC27852	24
	Fusobacterium necrophorum Total		48
	Fusobacterium nucleatum	ATCC23726	8
Fusobacterium nucleatum		ATCC25586	20
		ATCC31647	19
	Fusobacterium nucleatum Total		47
	Haemophilus influenzae	ATCC10211	9
		ATCC43065	6
Haemophilus influenzae		ATCC49144	9
		NCTC12699	9
		NCTC8468	8
	Haemophilus influenzae Total		41
Target	Organism	Strain	Independent ContrivedSamples Tested
Klebsiella oxytoca	Klebsiella oxytoca	ATCC43086	4
		ATCC43165	4
		ATCC43863	2
		ATCC49131	4
		ATCC51817	3
		ATCC700324	3
	Klebsiella oxytoca Total		20
Klebsiella pneumoniaegroup	Klebsiella pneumoniae, CTX-M	NCTC13465	5
	Klebsiella pneumoniae, CTX-M, NDM	ATCCBAA-2146	3
	Klebsiella pneumoniae, CTX-M, OXA	CDC#0140	12
	Klebsiella pneumoniae, IMP	CDC#0034	8
		CDC#0080	6
		CDC#0112	1
		CDC#0113	1
		CDC#0115	4
	Klebsiella pneumoniae, KPC	CDC#0117	4
		CDC#0120	4
		CDC#0125	4
		CDC#0129	4
			LMC_DR00015
		Klebsiella pneumoniae, VIM	NCTC13439
		NCTC13440	5
	Klebsiella pneumoniae group Total		72
Morganella morganii	Morganella morganii	148-200	8
		148-204	8
		148-205	8
		148-206	9
		148-209	7
	Morganella morganii, CTX-M1, NDM	CDC#0057	5
	Morganella morganii, KPC	CDC#0133	4
	Morganella morganii Total		49
Target	Organism	Strain	Independent ContrivedSamples Tested
Neisseria meningitidis	Neisseria meningitidis	ATCC13077	9
		ATCC13090	8
		ATCC13102	8
		ATCC13113	3
		ATCC35561	10
		NCTC10026	6
	Neisseria meningitidis Total		44
Proteus mirabilis	Proteus mirabilis, KPC	CDC#0155	4
	Proteus mirabilis, NDM	CDC#0159	5
	Proteus mirabilis Total		9
Pseudomonas aeruginosa	Pseudomonas aeruginosa, IMP	CDC#0092	5
		CDC#0103	8
	Pseudomonas aeruginosa, KPC	CDC#0090	1
		CDC#0054	5
	Pseudomonas aeruginosa, VIM	CDC#0100	4
		CDC#0108	4
		NCTC13437	5
	Pseudomonas aeruginosa Total		32
Salmonella	Salmonella 4,5,12:I:-	FSLS5-0580	2
	Salmonella Heidelberg	ATCC8326	2
	Salmonella Infantis	ATCCBAA-1675	2
	Salmonella Javiana	ATCC10721	1
	Salmonella Montevideo	ATCC8387	8
	Salmonella Muenchen	ATCC8388	1
	Salmonella Newport	ATCC6962	6
	Salmonella Typhimurium	ATCC13311	7
	Salmonella enterica subspecies enterica Enteritidis (Group D1)	ATCCBAA-708	6
	Salmonella Total		35
Serratia	Serratia ficaria	ATCC33105	4
	Serratia grimesii	ATCC14460	3

{30}------------------------------------------------

{31}------------------------------------------------

{32}------------------------------------------------

{33}------------------------------------------------

{34}------------------------------------------------

Target	Organism	Strain	Independent ContrivedSamples Tested
	Serratia plymuthica	ATCC53858	3
	Serratia rubidaea	ATCC27593	4
		ATCC29025	3
	Serratia Total	17
		ATCC13880	2
		ATCC14041	3
	Serratia marcescens	ATCC14756	3
Serratia marcescens		ATCC43861	4
		ATCC43862	3
		ATCC8100	3
	Serratia marcescens, IMP	l
	Serratia marcescens Total	19
Stenotrophomonasmaltophilia		148-201	7
	Stenotrophomonas maltophilia	148-222	6
		148-223	7
		148-224	8
		148-225	8
	Stenotrophomonas maltophilia Total	36

{35}------------------------------------------------

Genus and Group Assay Species Stratification

The ePlex BCID-GN Panel reports genus or group level results for Citrobacter, Enterobacter cloacae complex, Enterobacter non-cloacae complex, Salmonella, Serratia, Pan Candida and Pan Gram-Positive targets. Sensitivity/PPA of these genus and group level targets for species as determined by comparator methods for all evaluable samples tested are summarized in Table 36 and for the Pan targets for non-intended use samples in Table 37.

{36}------------------------------------------------

	Prospective Samples		Retrospective Samples		Contrived Samples		Combined Samples
Target SpeciesDetected byComparator Method	Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA
	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)
Citrobacter	5/5	100 (56.6-100)	20/21	95.2 (77.3-99.2)	43/43	100 (91.8-100)	68/69	98.6 (92.2-99.7)
Citrobacter braakii			2/3	66.7 (20.8-93.9)	8/8	100 (67.6-100)	10/11	90.9 (62.3-98.4)
Citrobacter freundii	4/4	100 (51.0-100)	13/13	100 (77.2-100)	27/27	100 (87.5-100)	44/44	100 (92.0-100)
Citrobacter koseri	1/1	100 (20.7-100)	4/4	100 (51.0-100)	4/4	100 (51.0-100)	9/9	100 (70.1-100)
Citrobacter youngae			1/1	100 (20.7-100)	4/4	100 (51.0-100)	5/5	100 (56.6-100)
Enterobacter (non-cloacaecomplex)	8/10	80.0 (49.0-94.3)	12/12	100 (75.8-100)	36/36	100 (90.4-100)	56/58	96.6 (88.3-99.0)
Enterobacter aerogenes	7/9	77.8 (45.3-93.7)	12/12	100 (75.8-100)	26/26	100 (87.1-100)	45/47	95.7 (85.8-98.8)
Enterobacter amnigenus					10/10	100 (72.2-100)	10/10	100 (72.2-100)
Enterobacter gergoviae	1/1	100 (20.7-100)					1/1	100 (20.7-100)
Enterobacter cloacaecomplex	19/19	100 (83.2-100)	47/50	94.0 (83.8-97.9)	35/37	94.6 (82.3-98.5)	101/106	95.3 (89.4-98.0)
Enterobacter asburiae					6/8	75.0 (40.9-92.9)	6/8	75.0 (40.9-92.9)
Enterobacter cloacae	19/19	100 (83.2-100)	46/49	93.9 (83.5-97.9)	28/28	100 (87.9-100)	93/96	96.9 (91.2-98.9)
Enterobacter hormaechei			1/1	100 (20.7-100)	1/1	100 (20.7-100)	2/2	100 (34.2-100)
Proteus	22/23	95.7 (79.0-99.2)	54/55	98.2 (90.4-99.7)	9/9	100 (70.1-100)	85/87	97.7 (92.0-99.4)
Proteus mirabilis	22/23	95.7 (79.0-99.2)	50/51	98.0 (89.7-99.7)	9/9	100 (70.1-100)	81/83	97.6 (91.6-99.3)

Table 36: Species Detected in Genus and Group Assays by Comparator Methods

Enterobacter cloacae	19/19	100 (83.2-100)	46/49	93.9 (83.5-97.9)	28/28	100 (87.9-100)	93/96	96.9 (91.2-98.9)
Enterobacter hormaechei			1/1	100 (20.7-100)	1/1	100 (20.7-100)	2/2	100 (34.2-100)
Proteus	22/23	95.7 (79.0-99.2)	54/55	98.2 (90.4-99.7)	9/9	100 (70.1-100)	85/87	97.7 (92.0-99.4)
Proteus mirabilis	22/23	95.7 (79.0-99.2)	50/51	98.0 (89.7-99.7)	9/9	100 (70.1-100)	81/83	97.6 (91.6-99.3)
Proteus vulgaris			5/5	100 (56.6-100)			5/5	100 (56.6-100)
Salmonella	2/2	100 (34.2-100)	18/19	94.7 (75.4-99.1)	34/35	97.1 (85.5-99.5)	54/56	96.4 (87.9-99.0)
Salmonella	2/2	100 (34.2-100)	15/15	100 (79.6-100)			17/17	100 (81.6-100)
Salmonella 4,5,12:i:-					2/2	100 (34.2-100)	2/2	100 (34.2-100)
Salmonella Heidelberg					2/2	100 (34.2-100)	2/2	100 (34.2-100)
Salmonella Infantis					2/2	100 (34.2-100)	2/2	100 (34.2-100)
Salmonella Javiana					1/1	100 (20.7-100)	1/1	100 (20.7-100)
Salmonella Montevideo					7/8	87.5 (52.9-97.8)	7/8	87.5 (52.9-97.8)
Salmonella Muenchen					1/1	100 (20.7-100)	1/1	100 (20.7-100)
Salmonella Newport					6/6	100 (61.0-100)	6/6	100 (61.0-100)
Salmonella Typhimurium					7/7	100 (64.6-100)	7/7	100 (64.6-100)
Salmonella choleraesuissubsp. Arizonae			0/1	0.0 (0.0-79.3)			0/1	0.0 (0.0-79.3)
Salmonella enterica subsp.enterica Enteritidis (GroupD1)					6/6	100 (61.0-100)	6/6	100 (61.0-100)

	Prospective Samples		Retrospective Samples		Contrived Samples		Combined Samples
Target SpeciesDetected byComparator Method	Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA
	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)
Salmonella enterica subsp.enterica serovarTyphimurium			1/1	100 (20.7-100)			1/1	100 (20.7-100)
Salmonella Typhi			2/2	100 (34.2-100)			2/2	100 (34.2-100)
Serratia	10/10	100 (72.2-100)	34/34	100 (89.8-100)	36/36	100 (90.4-100)	80/80	100 (95.4-100)
Serratia ficaria					4/4	100 (51.0-100)	4/4	100 (51.0-100)
Serratia grimesii					3/3	100 (43.9-100)	3/3	100 (43.9-100)
Serratia liquefaciens	1/1	100 (20.7-100)					1/1	100 (20.7-100)
Serratia marcescens	9/9	100 (70.1-100)	34/34	100 (89.8-100)	19/19	100 (83.2-100)	62/62	100 (94.2-100)
Serratia plymuthica					3/3	100 (43.9-100)	3/3	100 (43.9-100)
Serratia rubidaea					7/7	100 (64.6-100)	7/7	100 (64.6-100)
Pan Candida	1/1	100 (20.7-100)	4/7	57.1 (25.0-84.2)	N/A	N/A	5/8	62.5 (30.6-86.3)
Candida albicans	1/1	100 (20.7-100)	2/4	50.0 (15.0-85.0)			3/5	60.0 (23.1-88.2)
Candida glabrata			1/2	50.0 (9.5-90.5)			1/2	50.0 (9.5-90.5)
Candida krusei			1/1	100 (20.7-100)			1/1	100 (20.7-100)
Candida parapsilosis
Pan Gram-Positive	17/23	73.9 (53.5-87.5)	44/55	80.0 (67.6-88.4)	N/A	N/A	61/78	78.2 (67.8-85.9)
Bacillus (unspeciated)	1/4	25.0 (4.6-69.9)					1/4	25.0 (4.6-69.9)
Enterococcus (unspeciated)			0/1	0.0 (0.0-79.3)			0/1	0.0 (0.0-79.3)
Enterococcus casseliflavus			0/1	0.0 (0.0-79.3)			0/1	0.0 (0.0-79.3)
Enterococcus faecalis	5/7	71.4 (35.9-91.8)	18/20	90.0 (69.9-97.2)			23/27	85.2 (67.5-94.1)
Enterococcus faecium	1/1	100 (20.7-100)	8/9	88.9 (56.5-98.0)			9/10	90.0 (59.6-98.2)
Staphylococcus(unspeciated)	3/3	100 (43.9-100)	4/6	66.7 (30.0-90.3)			7/9	77.8 (45.3-93.7)
Staphylococcus aureus	2/2	100 (34.2-100)	5/6	83.3 (43.6-97.0)			7/8	87.5 (52.9-97.8)
Staphylococcus cohnii	1/1	100 (20.7-100)					1/1	100 (20.7-100)
Staphylococcusepidermidis	1/1	100 (20.7-100)	2/3	66.7 (20.8-93.9)			3/4	75.0 (30.1-95.4)
Staphylococcushaemolyticus	2/2	100 (34.2-100)					2/2	100 (34.2-100)
Staphylococcus hominis	1/1	100 (20.7-100)	3/3	100 (43.9-100)			4/4	100 (51.0-100)
Streptococcus	0/1	0.0 (0.0-79.3)					0/1	0.0 (0.0-79.3)
Streptococcus - viridansgroup	1/1	100 (20.7-100)	0/1	0.0 (0.0-79.3)			1/2	50.0 (9.5-90.5)

{37}------------------------------------------------

{38}------------------------------------------------

Target Species	Prospective Samples		Retrospective Samples		Contrived Samples		Combined Samples
Detected byComparator Method	Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA		Sensitivity/PPA
	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)	TP/TP+FN	% (95% CI)
Streptococcus anginosusgroup	1/1	100 (20.7-100)	4/5	80.0 (37.6-96.4)			5/6	83.3 (43.6-97.0)
Streptococcus infantarius	1/1	100 (20.7-100)	1/1	100 (20.7-100)			1/1	100 (20.7-100)
Streptococcus mitis group	1/1	100 (20.7-100)	1/1	100 (20.7-100)			1/1	100 (20.7-100)
Streptococcus oralis	1/1	100 (20.7-100)	1/1	100 (20.7-100)			1/1	100 (20.7-100)
Streptococcus pneumoniae	1/1	100 (20.7-100)	1/1	100 (20.7-100)			1/1	100 (20.7-100)
Streptococcus salivarius	1/1	100 (20.7-100)	1/1	100 (20.7-100)			1/1	100 (20.7-100)

Table 37. Species Detected in Pan Assays by Comparator Methods for Samples in Retrospective (Non-Intended Use) Samples with Gram-Positive or Fungal Organisms

Target Species Detected by ComparatorMethod	Retrospective (Non-Intended Use) SamplesSensitivity/PPA
	TP/TP+FN	% (95% CI)
Pan Candida	99/102	97.1 (91.7-99.0)
Candida albicans	47/48	97.9 (89.1-99.6)
Candida glabrata	37/38	97.4 (86.5-99.5)
Candida krusei	3/3	100 (43.9-100)
Candida parapsilosis	15/16	93.8 (71.7-98.9)
Pan Gram-Positive	567/571	99.3 (98.2-99.7)
Bacillus (unspeciated)	4/4	100 (51.0-100)
Bacillus cereus	4/5	80.0 (37.6-96.4)
Bacillus cereus group - not anthracis	1/1	100 (20.7-100)
Enterococcus	1/1	100 (20.7-100)
Enterococcus faecalis	34/34	100 (89.8-100)
Enterococcus faecium	14/14	100 (78.5-100)
Coagulase-negative Staphylococci (CoNS)	14/14	100 (78.5-100)
Staphylococcus (unspeciated)	86/86	100 (95.7-100)
Staphylococcus aureus	173/174	99.4 (96.8-99.9)
Staphylococcus auricularis	3/3	100 (43.9-100)
Staphylococcus capitis	7/7	100 (64.6-100)
Staphylococcus cohnii	1/1	100 (20.7-100)
Target Species Detected by ComparatorMethod	Retrospective (Non-Intended Use) Samples
	Sensitivity/PPATP/TP+FN	% (95% CI)
Staphylococcus epidermidis	86/87	98.9 (93.8-99.8)
Staphylococcus haemolyticus	6/6	100 (61.0-100)
Staphylococcus hominis	19/19	100 (83.2-100)
Staphylococcus hominis ssp hominis	21/21	100 (84.5-100)
Staphylococcus intermedius	1/1	100 (20.7-100)
Staphylococcus lugdunensis	1/1	100 (20.7-100)
Staphylococcus saccharolyticus	1/1	100 (20.7-100)
Staphylococcus saprophyticus	1/1	100 (20.7-100)
Staphylococcus sciuri	1/1	100 (20.7-100)
Staphylococcus simulans	2/2	100 (34.2-100)
Staphylococcus warneri	4/4	100 (51.0-100)
Alpha Hemolytic Streptococcus	1/1	100 (20.7-100)
Beta Hemolytic Streptococci, Group G	1/1	100 (20.7-100)
Gamma Hemolytic Streptococcus	1/1	100 (20.7-100)
Streptococcus (unspeciated)	9/9	100 (70.1-100)
Streptococcus - viridans group	17/17	100 (81.6-100)
Streptococcus agalactiae	21/21	100 (84.5-100)
Streptococcus anginosus	2/2	100 (34.2-100)
Streptococcus bovis	2/2	100 (34.2-100)
Streptococcus constellatus ssp constellatus	1/1	100 (20.7-100)
Streptococcus dysgalactiae (Group G)	4/4	100 (51.0-100)
Streptococcus gordonii	1/1	100 (20.7-100)
Streptococcus intermedius	1/1	100 (20.7-100)
Streptococcus mitis	11/12	91.7 (64.6-98.5)
Streptococcus mitis group	2/2	100 (34.2-100)
Streptococcus pneumoniae	22/22	100 (85.1-100)
Streptococcus pyogenes	9/9	100 (70.1-100)
Streptococcus salivarius	2/2	100 (34.2-100)

{39}------------------------------------------------

{40}------------------------------------------------

Resistance Gene Assay Species Stratification

Test results for resistance genes are only reported when an associated organism assay is positive in the same sample. (See Table 3 for organisms specifically associated with the six resistance markers on the ePlex BCID-GN Panel).

CTX-M

The PPA and NPA of the BCID-GN Panel CTX-M target stratified by the organism identified by comparator methods for prospective, retrospective and contrived samples are shown in Table 38.

Species Detected by Comparator Method		Sensitivity/PPA		Specificity/NPA
	TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Acinetobacter baumannii	Prospective	0/0	---	4/4	100 (51.0-100)
	Retrospective	0/0	---	15/15	100 (79.6-100)
	Contrived	0/0	---	55/55	100 (93.5-100)
	Combined	0/0	---	74/74	100 (95.1-100)
Citrobacter	Prospective	0/0	---	5/5	100 (56.6-100)
	Retrospective	1/1	100 (20.7-100)	20/20	100 (83.9-100)
	Contrived	6/6	100 (61.0-100)	37/37	100 (90.6-100)
	Combined	7/7	100 (64.6-100)	62/62	100 (94.2-100)
Enterobacter (non-cloacae complex)	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	12/12	100 (75.8-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
	Combined	0/0	---	58/58	100 (93.8-100)
Enterobacter cloacae complex	Prospective	0/0	---	19/19	100 (83.2-100)
	Retrospective	0/0	---	50/50	100 (92.9-100)
	Contrived	22/22	100 (85.1-100)	15/15	100 (79.6-100)
	Combined	22/22	100 (85.1-100)	84/84	100 (95.6-100)
Escherichia coli	Prospective	16/18	88.9 (67.2-96.9)	115/115	100 (96.8-100)
	Retrospective	35/37	94.6 (82.3-98.5)	103/103	100 (96.4-100)
	Contrived	22/22	100 (85.1-100)	30/30	100 (88.6-100)
	Combined	73/77	94.8 (87.4-98.0)	248/248	100 (98.5-100)
Klebsiella oxytoca	Prospective	0/1	0.0 (0.0-79.3)	12/12	100 (75.8-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	20/20	100 (83.9-100)
	Combined	0/1	0.0 (0.0-79.3)	66/66	100 (94.5-100)
Klebsiella pneumoniae group	Prospective	5/5	100 (56.6-100)	56/56	100 (93.6-100)
	Retrospective	14/15	93.3 (70.2-98.8)	93/93	100 (96.0-100)
	Contrived	20/20	100 (83.9-100)	52/52	100 (93.1-100)
	Combined	39/40	97.5 (87.1-99.6)	201/201	100 (98.1-100)
Morganella morganii	Prospective	0/0	---	3/3	100 (43.9-100)
	Retrospective	0/0	---	10/10	100 (72.2-100)
	Contrived	5/5	100 (56.6-100)	44/44	100 (92.0-100)
	Combined	5/5	100 (56.6-100)	57/57	100 (93.7-100)
Proteus	Prospective	2/5	40.0 (11.8-76.9)	18/18	100 (82.4-100)
	Retrospective	2/3	66.7 (20.8-93.9)	52/52	100 (93.1-100)
	Contrived	0/0	---	9/9	100 (70.1-100)
	Combined	4/8	50.0 (21.5-78.5)	79/79	100 (95.4-100)

Table 38: Clinical Performance of CTX-M Target in Samples with Associated Organisms Detected by Comparator Methods

{41}------------------------------------------------

Species Detected by Comparator Method		Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Proteus mirabilis	Prospective	2/5	40.0 (11.8-76.9)	18/18	100 (82.4-100)
	Retrospective	2/3	66.7 (20.8-93.9)	48/48	100 (92.6-100)
	Contrived	0/0	---	9/9	100 (70.1-100)
	Combined	4/8	50.0 (21.5-78.5)	75/75	100 (95.1-100)
Pseudomonas aeruginosa	Prospective	0/1	0.0 (0.0-79.3)	27/27	100 (87.5-100)
	Retrospective	0/0	---	60/60	100 (94.0-100)
	Contrived	0/0	---	32/32	100 (89.3-100)
	Combined	0/1	0.0 (0.0-79.3)	119/119	100 (96.9-100)
Salmonella	Prospective	0/0	---	2/2	100 (34.2-100)
	Retrospective	1/1	100 (20.7-100)	18/18	100 (82.4-100)
	Contrived	0/0	---	35/35	100 (90.1-100)
	Combined	1/1	100 (20.7-100)	55/55	100 (93.5-100)
Serratia	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
	Combined	0/0	---	80/80	100 (95.4-100)
Serratia marcescens	Prospective	0/0	---	9/9	100 (70.1-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	19/19	100 (83.2-100)
	Combined	0/0	---	62/62	100 (94.2-100)
Stenotrophomonas maltophilia	Prospective	0/0	---	4/4	100 (51.0-100)
	Retrospective	0/0	---	10/10	100 (72.2-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
	Combined	0/0	---	50/50	100 (92.9-100)

A comparison of CTX-M identified by comparator methods versus the ePlex BCID-GN Panel

results are shown in Table 39 for prospective and retrospective samples.

	Comparator Method
BCID-GN	Org+/ARG+	Org+/ARG-	Org-	Total
Org+/ARG+	74	0	2	76
Org+/ARG-	8A	741	2	751
Org-	3B	13	83	99
Total	85	754	87	926

Table 39: Distribution of CTX-M in Clinical Samples

% Agreement (95% CI) for Org+/ARG+: 74/85=87.1% (78.3-92.6)

% Agreement (95% CI) for Org+/ARG-: 741/754=98.3% (97.1-99.0)

% Agreement (95% CI) for Org-: 83/87=95.4% (88.8-98.2)

A. Further testing of these 8 false negative samples indicated that 7 of the 8 samples may have been contaminated during the original extraction process and misidentified as having CTX-M present. Specifically, results for 7 of the 8 samples were negative for CTX-M from the following additional testing of 2 repeat extractions from the original sample, 2) qPCR testing of an extraction from the isolate, and 3) testing of the original sample with an FDA-cleared multiplex assay. For these 7 samples, the originally extracted sample was re-tested with qPCR and CTX-M was again detected, suggesting contamination during the original extraction process. The remaining 8th sample was positive for CTX-M from the repeat extractions, negative for CTX-M from the isolate, and negative for CTX-M when tested with an FDA-cleared multiplex assay. These inconsistent detection results suggest the 8th sample may be a true low-copy CTX-M positive sample.

B. For these 3 samples, CTX-M signal was above the threshold for detection; however, an associated organism was not detected by the ePlex BCID-GN Panel and the CTX-M target was reported as 'N/A'.

{42}------------------------------------------------

IMP

The PPA and NPA of the BCID-GN Panel IMP target stratified by the contrived organism are shown below in Table 40. No prospective or retrospective samples were found to contain IMP.

	Table 40: Clinical Performance of IMP Target in Contrived Samples

		Sensitivity/PPA		Specificity/NPA
Species Detected by Comparator Method	TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Acinetobacter baumannii	0/0	---	74/74	100 (95.1-100)
Citrobacter	0/0	---	69/69	100 (94.7-100)
Enterobacter (non-cloacae complex)	5/5	100 (56.6-100)	53/53	100 (93.2-100)
Enterobacter cloacae complex	0/0	---	106/106	100 (96.5-100)
Escherichia coli	7/7	100 (64.6-100)	318/318	100 (98.8-100)
Klebsiella oxytoca	0/0	---	67/67	100 (94.6-100)
Klebsiella pneumoniae group	14/14	100 (78.5-100)	227/227	100 (98.3-100)
Morganella morganii	0/0	---	62/62	100 (94.2-100)
Proteus	0/0	---	87/87	100 (95.8-100)
Proteus mirabilis	0/0	---	83/83	100 (95.6-100)
Pseudomonas aeruginosa	13/13	100 (77.2-100)	107/107	100 (96.5-100)
Salmonella	0/0	---	56/56	100 (93.6-100)
Serratia	1/1	100 (20.7-100)	79/79	100 (95.4-100)
Serratia marcescens	1/1	100 (20.7-100)	61/61	100 (94.1-100)

A comparison of IMP identified by comparator methods versus the ePlex BCID-GN Panel results are shown in Table 41 for prospective and retrospective samples.

Table 41: Distribution of IMP in Clinical Samples

	Comparator Method
BCID-GN	Org+/ARG+	Org+/ARG-	Org-	Total
Org+/ARG+	0	0	0	0
Org+/ARG-	0	812	4	816
Org-	0	17	93	110
Total	0	829	97	926

% Agreement (95% CI) for Org+/ARG+: 0/0= N/A

% Agreement (95% CI) for Org+/ARG-: 812/829=97.9% (96.7-98.7)

% Agreement (95% CI) for Org-: 93/97=95.9% (89.9-98.4)

{43}------------------------------------------------

KPC

The PPA and NPA of the BCID-GN Panel KPC target stratified by the organism identified by comparator methods for prospective, retrospective and contrived samples are shown in Table 42.

Table 42: Clinical Performance of KPC Target in Samples with Associated Organisms Detected by Comparator Methods

Species Detected by Comparator Method		Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Acinetobacter baumannii	Prospective	0/0	---	4/4	100 (51.0-100)
	Retrospective	0/0	---	15/15	100 (79.6-100)
	Contrived	0/0	---	55/55	100 (93.5-100)
	Combined	0/0	---	74/74	100 (95.1-100)
Citrobacter	Prospective	0/0	---	5/5	100 (56.6-100)
	Retrospective	0/0	---	21/21	100 (84.5-100)
	Contrived	4/4	100 (51.0-100)	39/39	100 (91.0-100)
	Combined	4/4	100 (51.0-100)	65/65	100 (94.4-100)
Cronobacter sakazakii	Prospective	---	---	---	---
	Retrospective	0/0	---	1/1	100 (20.7-100)
	Contrived	0/0	---	45/45	100 (92.1-100)
	Combined	0/0	---	46/46	100 (92.3-100)
Enterobacter (non-cloacae complex)	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	12/12	100 (75.8-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
	Combined	0/0	---	58/58	100 (93.8-100)
Enterobacter cloacae complex	Prospective	0/0	---	19/19	100 (83.2-100)
	Retrospective	0/1	0.0 (0.0-79.3)	49/49	100 (92.7-100)
	Contrived	3/3	100 (43.9-100)	34/34	100 (89.8-100)
	Combined	3/4	75.0 (30.1-95.4)	102/102	100 (96.4-100)
Escherichia coli	Prospective	1/1	100 (20.7-100)	132/132	100 (97.2-100)
	Retrospective	0/0	---	140/140	100 (97.3-100)
	Contrived	6/6	100 (61.0-100)	46/46	100 (92.3-100)
	Combined	7/7	100 (64.6-100)	318/318	100 (98.8-100)
Klebsiella oxytoca	Prospective	0/0	---	13/13	100 (77.2-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	20/20	100 (83.9-100)
	Combined	0/0	---	67/67	100 (94.6-100)
Klebsiella pneumoniae group	Prospective	2/2	100 (34.2-100)	59/59	100 (93.9-100)
	Retrospective	4/4	100 (51.0-100)	103/104	99.0 (94.8-99.8)
	Contrived	22/22	100 (85.1-100)	50/50	100 (92.9-100)
	Combined	28/28	100 (87.9-100)	212/213	99.5 (97.4-99.9)
Morganella morganii	Prospective	0/0	---	3/3	100 (43.9-100)
	Retrospective	0/0	---	10/10	100 (72.2-100)
	Contrived	4/4	100 (51.0-100)	45/45	100 (92.1-100)
	Combined	4/4	100 (51.0-100)	58/58	100 (93.8-100)
Proteus	Prospective	0/0	---	23/23	100 (85.7-100)
	Retrospective	0/0	---	55/55	100 (93.5-100)
	Contrived	4/4	100 (51.0-100)	5/5	100 (56.6-100)
	Combined	4/4	100 (51.0-100)	83/83	100 (95.6-100)
Proteus mirabilis	Prospective	0/0	---	23/23	100 (85.7-100)
	Retrospective	0/0	---	51/51	100 (93.0-100)
	Contrived	4/4	100 (51.0-100)	5/5	100 (56.6-100)
	Combined	4/4	100 (51.0-100)	79/79	100 (95.4-100)
Pseudomonas aeruginosa	Prospective	0/0	---	28/28	100 (87.9-100)
	Retrospective	0/0	---	60/60	100 (94.0-100)
	Contrived	1/1	100 (20.7-100)	31/31	100 (89.0-100)
	Combined	1/1	100 (20.7-100)	90/90	100 (96.9-100)
Species Detected by Comparator Method		Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Salmonella	Prospective	0/0	---	2/2	100 (34.2-100)
	Retrospective	0/0	---	19/19	100 (83.2-100)
	Contrived	0/0	---	35/35	100 (90.1-100)
	Combined	0/0	---	56/56	100 (93.6-100)
Serratia	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
	Combined	0/0	---	80/80	100 (95.4-100)
Serratia marcescens	Prospective	0/0	---	9/9	100 (70.1-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	19/19	100 (83.2-100)
	Combined	0/0	---	62/62	100 (94.2-100)

{44}------------------------------------------------

A comparison of KPC identified by comparator methods versus the ePlex BCID-GN Panel results are shown in Table 43 for prospective and retrospective samples.

Table 43: Distribution of KPC in Clinical Samples

	Comparator Method
BCID-GN	Org+/ARG+	Org+/ARG-	Org-	Total
Org+/ARG+	7	1	0	8
Org+/ARG-	1	804	4	809
Org-	0	17	92	109
Total	8	822	96	926

% Agreement (95% CI) for Org+/ARG+: 7/8=87.5% (52.9-97.8)

% Agreement (95% CI) for Org+/ARG-: 804/822=97.8% (96.6-98.6)

% Agreement (95% CI) for Org-: 92/96=95.8% (89.8-98.4)

{45}------------------------------------------------

NDM

The PPA and NPA of the BCID-GN Panel NDM target stratified by the contrived organism are shown below in Table 44. No prospective or retrospective samples were found to contain NDM.

Table 44: Clinical Performance of NDM Target in Contrived Samples by Organism

		Sensitivity/PPA		Specificity/NPA
Species Detected by Comparator Method	TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Acinetobacter baumannii	5/5	100 (56.6-100)	69/69	100 (94.7-100)
Citrobacter	0/0	---	69/69	100 (94.7-100)
Enterobacter (non-cloacae complex)	0/0	---	58/58	100 (93.8-100)
Enterobacter cloacae complex	13/13	100 (77.2-100)	93/93	100 (96.0-100)
Escherichia coli	23/23	100 (85.7-100)	302/302	100 (98.7-100)
Klebsiella oxytoca	0/0	---	67/67	100 (94.6-100)
Klebsiella pneumoniae group	3/3	100 (43.9-100)	238/238	100 (98.4-100)
Morganella morganii	5/5	100 (56.6-100)	57/57	100 (93.7-100)
Proteus	5/5	100 (56.6-100)	82/82	100 (95.5-100)
Proteus mirabilis	5/5	100 (56.6-100)	78/78	100 (95.3-100)
Pseudomonas aeruginosa	0/0	---	120/120	100 (96.9-100)
Salmonella	0/0	---	56/56	100 (93.6-100)
Serratia	0/0	---	80/80	100 (95.4-100)
Serratia marcescens	0/0	---	62/62	100 (94.2-100)

A comparison of NDM identified by comparator methods versus the ePlex BCID-GN Panel results are shown in Table 45 for prospective and retrospective samples.

Table 45: Distribution of NDM in Clinical Samples

	Comparator Method
BCID-GN	Org+/ARG+	Org+/ARG-	Org-	Total
Org+/ARG+	0	0	0	0
Org+/ARG-	0	812	4	816
Org-	0	17	93	110
Total	0	829	97	926

% Agreement (95% Cl) for Org+/ARG+: 0/0= N/A

% Agreement (95% Cl) for Org+/ARG-: 812/829=97.9% (96.7-98.7)

% Agreement (95% CI) for Org-: 93/97=95.9% (89.9-98.4)

{46}------------------------------------------------

OXA

The PPA and NPA of the BCID-GN Panel OXA target stratified by target identified by comparator methods for prospective, retrospective, and contrived samples are shown in Table 46.

			Sensitivity/PPA		Specificity/NPA
Species Detected by Comparator Method		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
	Prospective	1/1	100 (20.7-100)	3/3	100 (43.9-100)
Acinetobacter baumannii	Retrospective	7/7	100 (64.6-100)	8/8	100 (67.6-100)
	Contrived	13/13	100 (77.2-100)	42/42	100 (91.6-100)
	Combined	21/21	100 (84.5-100)	53/53	100 (93.2-100)
	Prospective	0/0	---	5/5	100 (56.6-100)
	Retrospective	0/0	---	21/21	100 (84.5-100)
Citrobacter	Contrived	0/0	---	43/43	100 (91.8-100)
	Combined	0/0	---	69/69	100 (94.7-100)
	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	12/12	100 (75.8-100)
Enterobacter (non-cloacae complex)	Contrived	12/12	100 (75.8-100)	24/24	100 (86.2-100)
	Combined	12/12	100 (75.8-100)	46/46	100 (92.3-100)
	Prospective	0/0	---	19/19	100 (83.2-100)
	Retrospective	0/0	---	50/50	100 (92.9-100)
Enterobacter cloacae complex	Contrived	0/0	---	37/37	100 (90.6-100)
	Combined	0/0	---	106/106	100 (96.5-100)
	Prospective	0/1	0.0 (0.0-79.3)	132/132	100 (97.2-100)
	Retrospective	1/2	50.0 (9.5-90.5)	138/138	100 (97.3-100)
Escherichia coli	Contrived	0/0	---	52/52	100 (93.1-100)
	Combined	1/3	33.3 (6.1-79.2)	322/322	100 (98.8-100)
	Prospective	0/0	---	13/13	100 (77.2-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
Klebsiella oxytoca	Contrived	0/0	---	20/20	100 (83.9-100)
	Combined	0/0	---	67/67	100 (94.6-100)
	Prospective	0/0	---	61/61	100 (94.1-100)
	Retrospective	0/1	0.0 (0.0-79.3)	107/107	100 (96.5-100)
Klebsiella pneumoniae group	Contrived	12/12	100 (75.8-100)	60/60	100 (94.0-100)
	Combined	12/13	92.3 (66.7-98.6)	228/228	100 (98.3-100)
	Prospective	0/0	---	3/3	100 (43.9-100)
	Retrospective	0/0	---	10/10	100 (72.2-100)
Morganella morganii	Contrived	0/0	---	49/49	100 (92.7-100)
	Combined	0/0	---	62/62	100 (94.2-100)
	Prospective	0/0	---	23/23	100 (85.7-100)
	Retrospective	1/1	100 (20.7-100)	53/54	98.1 (90.2-99.7)
Proteus	Contrived	0/0	---	9/9	100 (70.1-100)
	Combined	1/1	100 (20.7-100)	85/86	98.8 (93.7-99.8)
Proteus mirabilis	Prospective	0/0	---	23/23	100 (85.7-100)
	Retrospective	0/0	---	50/51	98.0 (89.7-99.7)
	Contrived	0/0	---	9/9	100 (70.1-100)
	Combined	0/0	---	82/83	98.8 (93.5-99.8)
	Prospective	0/0	---	28/28	100 (87.9-100)
	Retrospective	0/0	---	59/60	98.3 (91.1-99.7)
Pseudomonas aeruginosa	Contrived	0/0	---	32/32	100 (89.3-100)
	Combined	0/0	---	119/120	99.2 (95.4-99.9)
	Prospective	0/0	---	2/2	100 (34.2-100)
Salmonella	Retrospective	0/0	---	19/19	100 (83.2-100)
	Contrived	0/0	---	35/35	100 (90.1-100)
Species Detected by Comparator Method		Sensitivity/PPA		Specificity/NPA
		TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Serratia	Combined	0/0	---	56/56	100 (93.6-100)
	Prospective	0/0	---	10/10	100 (72.2-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	36/36	100 (90.4-100)
Serratia marcescens	Combined	0/0	---	80/80	100 (95.4-100)
	Prospective	0/0	---	9/9	100 (70.1-100)
	Retrospective	0/0	---	34/34	100 (89.8-100)
	Contrived	0/0	---	19/19	100 (83.2-100)
	Combined	0/0	---	62/62	100 (94.2-100)

Table 46: Clinical Performance of OXA Target in Samples with Associated Organisms Detected by Comparator Methods

{47}------------------------------------------------

A comparison of OXA identified by comparator methods versus the ePlex BCID-GN Panel

results are shown in Table 47 for prospective and retrospective samples.

Table 47: Distribution of OXA in Clinical Samples
---------------------------------------------------	--	--	--	--

BCID-GN	Comparator Method
	Org+/ARG+	Org+/ARG-	Org-	Total
Org+/ARG+	10	2	0	12
Org+/ARG-	2A	798	4	804
Org-	1B	16	93	110
Total	13	816	97	926

% Agreement (95% CI) for Org+/ARG+: 10/13=76.9% (49.7-91.8)

% Agreement (95% CI) for Org+/ARG-: 798/816=97.8% (96.5-98.6)

% Agreement (95% CI) for Org-: 93/97=95.9% (89.9-98.4)

A. One false negative sample was tested with an FDA-cleared multiplex assay and OXA was not detected. The isolate from the remaining false negative sample tested negative for OXA-23 and OXA-48 by qPCR.

B. For this sample, OXA signal was above the threshold for detection; however, an associated organism was not detected by the ePlex BCID-GN Panel and the OXA target was reported as 'N/A'.

{48}------------------------------------------------

VIM

The PPA and NPA of the BCID-GN Panel VIM target stratified by the contrived organism are shown below in Table 48. No prospective or retrospective samples were found to contain VIM.

Table 48: Clinical Performance of VIM Target in Contrived Samples by Organism

		Sensitivity/PPA	Specificity/NPA
Species Detected by Comparator Method	TP/TP+FN	% (95% CI)	TN/TN+FP	% (95% CI)
Acinetobacter baumannii	0/0	---	74/74	100 (95.1-100)
Citrobacter	0/0	---	69/69	100 (94.7-100)
Enterobacter (non-cloacae complex)	0/0	---	58/58	100 (93.8-100)
Enterobacter cloacae complex	6/6	100 (61.0-100)	100/100	100 (96.3-100)
Escherichia coli	2/2	100 (34.2-100)	323/323	100 (98.8-100)
Klebsiella oxytoca	0/0	---	67/67	100 (94.6-100)
Klebsiella pneumoniae group	16/16	100 (80.6-100)	225/225	100 (98.3-100)
Morganella morganii	0/0	---	62/62	100 (94.2-100)
Proteus	0/0	---	87/87	100 (95.8-100)
Proteus mirabilis	0/0	---	83/83	100 (95.6-100)
Pseudomonas aeruginosa	18/18	100 (82.4-100)	102/102	100 (96.4-100)
Salmonella	0/0	---	56/56	100 (93.6-100)
Serratia	0/0	---	80/80	100 (95.4-100)
Serratia marcescens	0/0	---	62/62	100 (94.2-100)

A comparison of VIM identified by comparator methods versus the ePlex BCID-GN Panel results are shown in Table 49 for prospective and retrospective samples.

Table 49: Distribution of VIM in Clinical Samples

BCID-GN	Org+/ARG-	Org-	Total
Org+/ARG+	0	0	0
Org+/ARG-	812	4	816
Org-	17	93	110
Total	829	97	926

% Agreement (95% CI) for Org+/ARG+: 0/0= N/A

% Agreement (95% CI) for Org+/ARG-: 812/829=97.9% (96.7-98.7)

% Agreement (95% CI) for Org-: 93/97=95.9% (89.9-98.4)

{49}------------------------------------------------

Resistance Markers and Antimicrobial Resistance Susceptibility

A supplemental comparison of the CTX-M BCID-GN Panel target versus phenotypic antimicrobial susceptibility testing (AST) for extended spectrum beta-lactamase (ESBL) activity, Ceftazidime, Ceftriaxone, and Aztreonam, and a combination of the 4 results (ESBL/Combo) is provided in Table 57 for clinical isolates with available AST results. In total, 162 isolates had ESBL confirmatory testing and 770 isolates had ESBL confirmatory testing and/or AST results for 1 or more of Ceftazidime, Ceftriaxone, or Aztreonam. A true positive (TP) result was defined where CTX-M was detected by the BCID-GN Panel and the isolate was positive for the ESBL confirmatory test or resistant (R) or intermediate (I) to the specific antimicrobial. A false negative (FN) result was defined similarly when CTX-M was not detected by the BCID-GN Panel. PPA was calculated as 100 x (TP/(TP+FN)). A true negative (TN) result was defined where CTX-M was not detected by the BCID-GN Panel and the isolate was negative for the ESBL confirmatory test or susceptible (S) to the specific antimicrobial. A false positive (FP) result was defined similarly when CTX-M was detected by the BCID-GN Panel. NPA was calculated as 100 x (TN/(TN+FP)). For the ESBL/Combo analysis, the phenotypic AST result was positive or negative based on the ESBL confirmatory test, if available. If the ESBL confirmatory test was not available, then the phenotypic AST result was positive if any of the 3 antimicrobials were resistant or intermediate, otherwise, the result was negative if any of the 3 antimicrobials were susceptible. Note: ESBL resistance can be due to mechanisms other than acquisition of the CTX-M resistance gene.

Table 50: Clinical Performance of the BCID-GN Panel target CTX-M to Phenotypic Antimicrobial Susceptibility Testing for ESBL, Ceftazidime, Ceftriaxone, and Aztreonam

AssociatedOrganism	ESBL confirmatory		Ceftazidime		Ceftriaxone		Aztreonam		ESBL/Combo
	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)
A. baumannii	---	---	0/10(0.0%)	3/3(100%)	0/11(0.0%)	1/1(100%)	0/3(0.0%)	0/0(%)	0/15(0.0%)	2/2(100%)
Citrobacter	---	---	0/2(0.0%)	14/15(93.3%)	0/3(0.0%)	21/22(95.5%)	0/1(0.0%)	16/17(94.1%)	0/3(0.0%)	21/22(95.5%)
Enterobacter	---	---	0/2(0.0%)	12/12(100%)	0/2(0.0%)	18/18(100%)	0/1(0.0%)	9/9(100%)	0/3(0.0%)	17/17 (100%)
E. cloacaecomplex	---	---	0/10(0.0%)	30/30(100%)	0/17(0.0%)	39/39(100%)	0/5(0.0%)	25/25(100%)	0/18(0.0%)	39/39 (100%)
E. coli	28/30(93.3%)	82/82(100%)	29/38(76.3%)	159/167(95.2%)	47/59(79.7%)	180/181(99.4%)	23/27(85.2%)	118/120(98.3%)	47/53(88.7%)	196/197(99.5%)
K. oxytoca	0/0	10/10(100%)	0/1(0.0%)	29/29(100%)	0/2(0.0%)	40/40(100%)	0/2(0.0%)	20/20(100%)	0/2(0.0%)	41/41 (100%)
K. pneumoniaegroup	7/11 (63.6%)	25/25(100%)	13/23(56.5%)	83/83(100%)	17/27(63.0%)	115/115(100%)	10/12(83.3%)	72/72(100%)	17/26(65.4%)	119/119(100%)

{50}------------------------------------------------

AssociatedOrganism	ESBL confirmatory		Ceftazidime		Ceftriaxone		Aztreonam		ESBL/Combo
	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)
M. morganii			0/1(0.0%)	6/6(100%)	0/2(0.0%)	9/9(100%)	0/0	4/4(100%)	0/2(0.0%)	9/9(100%)
Proteus	3/3(100%)	0/0	1/2(50.0%)	46/48(95.8%)	4/7(57.1%)	59/59(100%)	2/3(66.7%)	35/36(97.2%)	4/8(50.0%)	59/59 (100%)
P. mirabilis	3/3(100%)	0/0	1/2(50.0%)	43/45(95.6%)	4/5(80.0%)	56/56(100%)	2/3(66.7%)	32/33(97.0%)	4/6(66.7%)	56/56 (100%)
P. aeruginosa			0/7(0.0%)	46/46(100%)	0/20(0.0%)	0/0	0/9(0.0%)	25/25(100%)	0/32(0.0%)	41/41 (100%)
Salmonella	1/1(100%)	0/0	0/0	6/6(100%)	1/2(50.0%)	12/12(100%)	1/1(100%)	8/8(100%)	1/2(50.0%)	15/15 (100%)
Serratia			0/1(0.0%)	25/25(100%)	0/4(0.0%)	32/32(100%)	0/1(0.0%)	20/20(100%)	0/4(0.0%)	34/34 (100%)
S. marcescens			0/1(0.0%)	24/24(100%)	0/4(0.0%)	31/31(100%)	0/1(0.0%)	19/19(100%)	0/4(0.0%)	33/33 (100%)
S. maltophilia			0/2(0.0%)	2/2(100%)	0/3(0.0%)	0/0	0/5(0.0%)	0/0	0/5(0.0%)	2/2(100%)
Any Organism	42/48(87.5%)	117/117(100%)	44/102(43.1%)	528/541(97.6%)	73/168(43.5%)	613/615(99.7%)	38/74(51.4%)	403/408(98.8%)	73/183(39.9%)	684/686(99.7%)
CI	(75.3-94.1)	(96.8-100)	(33.9-52.8)	(95.9-98.6)	(36.2-51.0)	(98.8-99.9)	(40.2-62.4)	(97.2-99.5)	(33.1-47.1)	(98.9-99.9)

A supplemental comparison of the 5 BCID-GN Panel carbapenemase resistance gene (OXA, KPC, IMP, NDM, VIM) targets versus phenotypic antimicrobial susceptibility testing (AST) for Ertapenem, Imipenem, and Meropenem is provided in Table 51 for clinical isolates with available AST results. In total, 731 isolates had AST results for 1 or more of Ertapenem, Imipenem, or Meropenem. A true positive (TP) result was defined where OXA, KPC, IMP, NDM and/or VIM was detected by the BCID-GN Panel and the isolate was resistant (R) or intermediate (I) to Ertapenem, Imipenem, or Meropenem. A false negative (FN) result was defined similarly when OXA, KPC, IMP, NDM and/or VIM was not detected by the BCID-GN Panel. PPA was calculated as 100 x (TP/(TP+FN)). A true negative (TN) result was defined where OXA, KPC, IMP, NDM and/or VIM was not detected by the BCID-GN Panel and the isolate was susceptible (S) to Ertapenem, Imipenem, or Meropenem. A false positive (FP) result was defined similarly when OXA. KPC, IMP, NDM and/or VIM was detected by the BCID-GN Panel. NPA was calculated as 100 x (TN/(TN+FP)). Note: Carbapenemase resistance can be due to mechanisms other than acquisition of the OXA, KPC, IMP, NDM and/or VIM resistance genes.

{51}------------------------------------------------

Table 51: Clinical Performance of the BCID-GN Panel Resistance Gene Targets Compared to Phenotypic Antimicrobial Susceptibility Testing for Ertapenem, Imipenem, and Meropenem

AssociatedOrganism	OXA		KPC		IMP		NDM		VIM		Any ResistanceMarker
	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)	PPATP/TP+FN(%)	NPATN/TN+FP(%)
A. baumannii	8/9 (88.9%)	9/9(100%)	0/9(0.0%)	9/9 (100%)	0/9(0.0%)	9/9(100%)	0/9(0.0%)	9/9(100%)	0/9(0.0%)	9/9(100%)	8/9(88.9%)	9/9(100%)
Citrobacter	0/1(0.0%)	22/22(100%)	0/1(0.0%)	22/22(100%)	0/1(0.0%)	22/22(100%)	0/1(0.0%)	22/22(100%)	0/1(0.0%)	22/22(100%)	0/1(0.0%)	22/22 (100%)
C. sakazakii	---	---	---	1/1 (100%)	---	---	---	---	---	---	---	---
Enterobacter	0/0	15/15(100%)	0/0	15/15(100%)	0/0	15/15(100%)	0/0	15/15(100%)	0/0	15/15(100%)	0/0	15/15 (100%)
E. cloacaecomplex	0/1(0.0%)	51/51(100%)	0/1(0.0%)	51/51(100%)	0/1(0.0%)	51/51(100%)	0/1(0.0%)	51/51(100%)	0/1(0.0%)	51/51(100%)	0/1(0.0%)	51/51 (100%)
E. coli	0/2(0.0%)	247/248(99.6%)	1/2(50.0%)	248/248(100%)	0/2(0.0%)	248/248(100%)	0/2(0.0%)	248/248(100%)	0/2(0.0%)	248/248(100%)	1/2 (50.0%)	247/248(99.6%)
K. oxytoca	0/0	40/40(100%)	0/0	40/40(100%)	0/0	40/40(100%)	0/0	40/40(100%)	0/0	40/40(100%)	0/0	40/40 (100%)
K. pneumoniaegroup	0/8(0.0%)	136/136(100%)	5/8(62.5%)	135/136(99.3%)	0/8(0.0%)	136/136(100%)	0/8(0.0%)	136/136(100%)	0/8(0.0%)	136/136(100%)	5/8 (62.5%)	135/136(99.3%)
M. morganii	0/0	10/10(100%)	0/0	10/10(100%)	0/0	10/10(100%)	0/0	10/10(100%)	0/0	10/10(100%)	0/0	10/10 (100%)
Proteus	0/0	61/62(98.4%)	0/0	62/62(100%)	0/0	62/62(100%)	0/0	62/62(100%)	0/0	62/62(100%)	0/0	61/62(98.4%)
P. mirabilis	0/0	57/57(100%)	0/0	57/57(100%)	0/0	57/57(100%)	0/0	57/57(100%)	0/0	57/57(100%)	0/0	57/57 (100%)
P. aeruginosa	0/5(0.0%)	65/66(98.5%)	0/5(0.0%)	66/66(100%)	0/5(0.0%)	66/66(100%)	0/5(0.0%)	66/66(100%)	0/5(0.0%)	66/66(100%)	0/5(0.0%)	65/66(98.5%)
Salmonella	0/0	8/8 (100%)	0/0	8/8 (100%)	0/0	8/8 (100%)	0/0	8/8 (100%)	0/0	8/8 (100%)	0/0	8/8 (100%)
Serratia	0/0	38/38(100%)	0/0	38/38(100%)	0/0	38/38(100%)	0/0	38/38(100%)	0/0	38/38(100%)	0/0	38/38 (100%)
S. marcescens	0/0	37/37(100%)	0/0	37/37(100%)	0/0	37/37(100%)	0/0	37/37(100%)	0/0	37/37(100%)	0/0	37/37 (100%)
Any Organism	8/26(30.8%)	796/799(99.6%)	6/26(23.1%)	799/800(99.9%)	0/26(0.0%)	799/799(100%)	0/26(0.0%)	799/799(100%)	0/26(0.0%)	799/799(100%)	14/26(53.8%)	796/800(99.5%)
CI	(16.5-50.0)	(98.9-99.9)	(11.0-42.1)	(99.3-100)	(0.0-12.9)	(99.5-100)	(0.0-12.9)	(99.5-100)	(0.0-12.9)	(99.5-100)	(35.5-71.2)	(98.7-99.8)

CI= confidence interval

{52}------------------------------------------------

Co-detections in Clinical Samples

The ePlex BCID-GN Panel identified a total of 103 bacterial co-detections in prospective and retrospective samples. Of the 349 prospective samples, 320 (91.7%) had single detections, 22 (6.2%) had double detections, and 7 (2.0%) had triple detections. Of the 577 retrospective samples, 503 (87.2%) had single detections, 62 (10.7%) had double detections, and 12 (2.1%) had triple detections. Tables 52-55 below summarize co-detections detected in prospective and retrospective samples.

Distinct Co-Detection Combinations Detected by the ePlexBCID-GN Panel in Prospective Clinical Samples
Target 1	Target 2	Target 3	ResistanceMarker	Number ofSamples(NumberDiscrepant)	DiscrepantOrganism(s) / ResistanceMarker(s)A,B
A. baumannii	Pan Gram-Positive			2 (0)
Citrobacter	E. cloacae complex	K. oxytoca		2 (2)	Citrobacter (2), E. cloacaecomplex (2)
Citrobacter	K. oxytoca	K. pneumoniae group		1 (1)	Citrobacter (1)
Citrobacter	P. mirabilis	Pan Gram-Positive		1 (1)	Pan Gram-Positive (1)
E. cloacae complex	E. coli	K. pneumoniae group		1 (0)
E. cloacae complex	Pan Candida	Pan Gram-Positive		1 (0)
E. cloacae complex	Pan Gram-Positive			2 (0)
E. coli	K. oxytoca			2 (1)	K. oxytoca (1)
E. coli	K. pneumoniae group		CTX-M	1 (1)	E. coli (1)
E. coli	Pan Gram-Positive			2 (1)	Pan Gram-Positive (1)
Enterobacter	K. pneumoniae group			1 (1)	Enterobacter (1)
K. oxytoca	Pan Gram-Positive			1 (0)
K. oxytoca	S. marcescens			1 (0)
K. pneumoniae group	P. mirabilis			1 (0)
K. pneumoniae group	Pan Gram-Positive			2 (0)
K. pneumoniae group	Pan Gram-Positive		CTX-M, KPC	1 (1)	Pan Gram-Positive (1)
M. morganii	P. mirabilis			1 (0)

Table 52: Co-Detections Identified by the ePlex BCID-GN Panel (Prospective Samples)

{53}------------------------------------------------

Distinct Co-Detection Combinations Detected by the ePlexBCID-GN Panel in Prospective Clinical Samples				Number ofSamples(Number Discrepant)	DiscrepantOrganism(s) / ResistanceMarker(s)A,B
Target 1	Target 2	Target 3	ResistanceMarker
P. aeruginosa	P. mirabilis	Pan Gram-Positive		1 (0)
P. aeruginosa		Pan Gram-Positive		1 (0)
P. mirabilis		Pan Gram-Positive		3 (2)	Pan Gram-Positive (2)
P. mirabilis		Pan Gram-Positive	CTX-M	1 (0)

A. A discrepant organism or resistance marker is defined as one that was detected by the comparator method(s).

B. 12/13 false positive organisms were investigated using; the discrepant organism was detected in 1/13 and not detected in 1.

One false positive Pan Gram-Positive sample was not tested.

i. In 3/3 false positive Citrobacter samples, Citrobacter was detected.
ii. In 2/2 false positive E. cloacae complex samples, E. cloacae complex was detected.
1. An Enterobacter species was not detected in the 1 false positive Enterobacter (non-cloacae complex) sample.
iv. In 1/1 false positive E. coli sample, E. coli was detected.
In 1/1 false positive K. oxytoca sample, K. oxytoca was detected. v.
vi. In 4/4 false positive Pan Gram-Positive samples, a Pan Gram-Positive organism was detected.

Table 53: Co-Detections Identified by the ePlex BCID-GN Panel (Retrospective Samples)

	Distinct Co-Detection Combinations Detected by the ePlexBCID-GN Panel in Retrospective Clinical Samples			Number Samples(Number Discrepant)	Discrepant Organism(s) /Resistance Marker(s)A,B
Target 1	Target 2	Target 3	Resistance Marker
A. baumannii	K. pneumoniae group	Pan Gram-Positive	CTX-M, OXA	1 (1)	A. baumannii (1), K. pneumoniae group (1), Pan Gram-Positive (1)
A. baumannii	Pan Gram-Positive			2 (0)
A. baumannii	Pan Gram-Positive		OXA	4 (1)	Pan Gram-Positive (1)
B. fragilis	E. cloacae complex	Pan Gram-Positive		1 (1)	B. fragilis (1)
B. fragilis	E. coli			2 (1)	B. fragilis (1)
B. fragilis	Pan Gram-Positive			1 (0)
Citrobacter	E. cloacae complex			1 (1)	E. cloacae complex (1)
Citrobacter	E. coli			1 (0)
Citrobacter	K. oxytoca			1 (1)	Citrobacter (1)
Citrobacter	K. oxytoca	K. pneumoniae group		1 (1)	K. oxytoca (1)
Citrobacter	K. pneumoniae group			1 (0)
Distinct Co-Detection Combinations Detected by the ePlexBCID-GN Panel in Retrospective Clinical Samples				NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /Resistance Marker(s)A,B
Target 1	Target 2	Target 3	ResistanceMarker
Citrobacter	K. pneumoniae group	Pan Gram-Positive	CTX-M	1 (0)
Citrobacter	M. morganii	Pan Gram-Positive		1 (1)	M. morganii (1)
Citrobacter	Pan Gram-Positive			3 (2)	Pan Gram-Positive (2)
E. cloacaecomplex	K. pneumoniae group			1 (0)
E. cloacaecomplex	P. aeruginosa	Pan Gram-Positive		1 (1)	P. aeruginosa (1)
E. cloacaecomplex	Pan Candida			1 (1)	Pan Candida (1)
E. cloacaecomplex	Pan Gram-Positive			2 (1)	Pan Gram-Positive (1)
E. coli	K. oxytoca			1 (0)
E. coli	K. oxytoca	Pan Gram-Positive		1 (0)
E. coli	K. pneumoniae group			2 (0)
E. coli	M. morganii			1 (0)
E. coli	P. mirabilis			3 (0)
E. coli	P. mirabilis	Pan Gram-Positive		1 (0)
E. coli	Pan Gram-Positive			8 (2)	Pan Gram-Positive (2)
E. coli	Pan Gram-Positive		CTX-M	1 (0)
Enterobacter	Pan Candida			1 (0)
Enterobacter	Pan Gram-Positive			1 (0)
H. influenzae	N. meningitidis	P. aeruginosa		1 (1)	N. meningitidis (1), P.aeruginosa (1)
K. oxytoca	K. pneumoniae group			2 (1)	K. pneumoniae group (1)
K. oxytoca	Pan Gram-Positive			3 (2)	Pan Gram-Positive (2)
K. oxytoca	S. marcescens			1 (1)	S. marcescens (1)
K. pneumoniaegroup	Pan Gram-Positive			4 (1)	Pan Gram-Positive (1)
K. pneumoniaegroup	Pan Gram-Positive	S. marcescens		1 (1)	K. pneumoniae group (1)
Distinct Co-Detection Combinations Detected by the ePlexBCID-GN Panel in Retrospective Clinical Samples				NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /Resistance Marker(s)A,B
Target 1	Target 2	Target 3	ResistanceMarker
K. pneumoniaegroup	S. maltophilia			1 (0)
M. morganii	P. aeruginosa	Pan Gram-Positive		1 (1)	P. aeruginosa (1)
M. morganii	P. mirabilis			1 (0)
M. morganii	Pan Gram-Positive	Proteus		1 (0)
P. aeruginosa	Pan Gram-Positive			1 (0)
P. mirabilis	Pan Gram-Positive			5 (0)
Pan Candida	Pan Gram-Positive			2 (0)
Pan Gram-Positive	S. maltophilia			1 (0)
Pan Gram-Positive	S. marcescens			3 (0)

{54}------------------------------------------------

{55}------------------------------------------------

A. A discrepant organism or resistance marker is defined as one that was detected by the BCID-GN Panel but not by the comparator method(s).

B. 24/26 false positive organisms were investigated using the discrepant organism was detected in 21/24, not detected in 2, and was indeterminate for one organism.

i. In 1/1 false positive A.baumannii sample, A. baumannii was detected.

1. In 2/2 false positive B. fragilis samples, B. fragilis was detected.
  iii. In 1/1 false positive Citrobacter sample, Citrobacter was detected.
In the one false positive E. cloacae complex sample, PCR/sequencing was indeterminate. iv.
v. In 1/1 false positive K. oxytoca sample, K. oxytoca was detected.
In 3/3 false positive K. pneumoniae group samples, K. pneumoniae group was detected. V1.
vii. In 1/1 false positive sample, M. morganii was detected.
V111. N. meningitidis was not detected in the 1 N. meningitidis false positive sample.

In 2/3 false positive P. aeruginosa samples, P. aeruginosa was detected. P. aeruginosa was not detected in the remaining sample. ıx..

In 1/1 false positive pan Candida sample, Pan Candida was detected. X.

In 8/8 false positive Pan Gram-Positive samples, a Pan Gram-Positive organism was detected. xi.
In 1/1 false positive S. marcescens sample, S. marcescens was detected. xii.

Table 54: Additional Co-Detections Identified by the Comparator Method(s) (Prospective

Samples)

Distinct Co-Detection Combinations Detected by the ComparatorMethod(s) in Prospective Clinical Samples					NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /ResistanceMarker(s)A
Target 1	Target 2	Target 3	Target 4	ResistanceMarker
A. baumannii	E. faecium	Staphylococcus			1 (0)
A. baumannii	Staphylococcus				1 (0)
Achromobacter xylosoxidans*	E. cloacae				1 (0)
Distinct Co-Detection Combinations Detected by the ComparatorMethod(s) in Prospective Clinical Samples					NumberSamples	DiscrepantOrganism(s) /
Target 1	Target 2	Target 3	Target 4	ResistanceMarker	(NumberDiscrepant)	ResistanceMarker(s)A
Acinetobacterlwoffii*	Staphylococcushominis				1 (0)
Acinetobacterpittii*	S. aureus				1 (0)
Aerococcusviridans*	K. oxytoca	S. epidermidis	Staphylococcuscohnii		1 (0)
Aerococcusviridans*	Staphylococcushominis				1 (0)
B. fragilis	Clostridiumspecies*				1 (0)
Bacillus	E. cloacae				1 (0)
C. acnes*	E. coli				1 (0)
C. albicans	E. cloacae	E. faecalis			1 (0)
C. freundii	P. mirabilis	Providenciastuartii*			1 (0)
Candida lusitaniae*	S. liquefaciens				1 (0)
Citrobacteramalonaticus*	E. coli				1 (0)
E. aerogenes	K. oxytoca	Leclerciaadecarboxylata*			2 (2)	E. aerogenes (2)
E. aerogenes	P. aeruginosa				1 (1)	P. aeruginosa (1)
E. cloacae	E. coli	K. pneumoniae			1 (0)
E. cloacae	S. aureus				1 (0)
E. coli	E. faecalis				1 (0)
E. coli	K. pneumoniae				1 (1)	E. coli (1)
E. coli	P. mirabilis	Providenciastuartii*	S. anginosusgroup	CTX-M	1 (1)	E. coli (1)
E. faecalis	K. pneumoniae				3 (1)	E. faecalis (1)
E. faecalis	M. morganii	P. mirabilis			1 (1)	E. faecalis (1)
E. faecalis	Providenciastuartii*				1 (0)
Enterobacteriaceae*	K. pneumoniae				1 (0)
Distinct Co-Detection Combinations Detected by the Comparator					NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /ResistanceMarker(s)A
Method(s) in Prospective Clinical Samples
Target 1	Target 2	Target 3	Target 4	ResistanceMarker
K. pneumoniae	Staphylococcushaemolyticus	non-fermentingGram-Negativebacilli*			1 (1)	K. pneumoniae (1)
Lactococcus lactis*	P. mirabilis				1 (0)
Micrococcusluteus*	Sphingomonaspaucimobilis*				1 (0)
P. aeruginosa	P. mirabilis	Streptococcus -viridans group			1 (0)
P. aeruginosa	S. maltophilia				1 (1)	S. maltophilia (1)
P. aeruginosa	Staphylococcushaemolyticus				1 (0)
P. mirabilis	Staphylococcus				1 (0)
S. maltophilia	Streptococcus				1 (1)	Streptococcus (1)

{56}------------------------------------------------

{57}------------------------------------------------

Indicates an off-panel organism not detected by the BCID-GN Panel.

A. A discrepant organism or resistance marker is defined as one that was detected by the comparator method(s) but not by the BCID-GN Panel (excludes organisms not detected by the BCID-GN panel).

Table 55: Additional Co-Detections Identified by the Comparator Method(s)

(Retrospective Samples)

Distinct Co-Detection Combinations Detected by the Comparator Method(s) in Retrospective Clinical Samples					Number Samples (Number Discrepant)	Discrepant Organism(s) / Resistance Marker(s)A
Target 1	Target 2	Target 3	Target 4	Resistance Marker
A. baumannii	E. faecalis			OXA	2 (0)
A. baumannii	E. faecalis	S. aureus			1 (0)
A. baumannii	E. faecium			OXA	1 (0)
A. baumannii	Staphylococcus				1 (0)
Acinetobacter radioresistens*	P. vulgaris			OXA	1 (0)
Aeromonas caviae*	E. coli	Enterococcus casseliflavus	K. oxytoca		1 (1)	E. casseliflavus (1)
Aeromonas veronii*	E. cloacae				1 (1)	E. cloacae (1)
B. fragilis	S. anginosus group				1 (0)
Distinct Co-Detection Combinations Detected by the Comparator Method(s) inRetrospective Clinical Samples					NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /ResistanceMarker(s)A
Target 1	Target 2	Target 3	Target 4	ResistanceMarker
C. albicans	E. faecalis				1 (0)
C. albicans	E. faecium	Staphylococcushominis			1 (1)	C. albicans (1)
C. albicans	P. aeruginosa				1 (1)	C. albicans (1)
C. albicans	S. epidermidis				1 (0)
C. braakii	E. cloacae	K. oxytoca			1 (1)	C. braakii (1), K.oxytoca (1)
C. braakii	E. coli				1 (0)
C. braakii	Streptococcus oralis				1 (0)
C. freundii	Enterococcus				1 (1)	Enterococcus (1)
C. freundii	K. pneumoniae				2 (0)
C. freundii	K. pneumoniae	Staphylococcushominis		CTX-M	1 (0)
C. glabrata	E. aerogenes	Staphylococcus			1 (1)	Staphylococcus (1)
C. glabrata	P. mirabilis				1 (1)	C. glabrata (1)
C. koseri	E. faecalis				1 (0)
C. krusei	S. epidermidis				1 (1)	S. epidermidis (1)
C. youngae	K. oxytoca				1 (1)	K. oxytoca (1)
Clostridiumperfringens*	E. coli				1 (0)
E. aerogenes	S. anginosus group				1 (0)
E. cloacae	E. coli				1 (1)	E. coli (1)
E. cloacae	E. faecalis				1 (1)	E. faecalis (1)
E. cloacae	E. faecium				1 (0)
E. cloacae	E. faecium	Staphylococcushominis			1 (0)
E. cloacae	K. pneumoniae				1 (0)
E. cloacae	M. morganii				1 (1)	E. cloacae (1)
E. cloacae	S. anginosus group				1 (0)
E. cloacae	S. maltophilia				1 (1)	S. maltophilia (1)
E. coli	E. faecalis				2 (0)
Distinct Co-Detection Combinations Detected by the Comparator Method(s) inRetrospective Clinical Samples					NumberSamples(NumberDiscrepant)	DiscrepantOrganism(s) /ResistanceMarker(s)A
Target 1	Target 2	Target 3	Target 4	ResistanceMarker
E. coli	E. faecalis			CTX-M	1 (0)
E. coli	E. faecalis	K. pneumoniae			1 (1)	E. coli (1)
E. coli	E. faecalis	P. mirabilis			1 (0)
E. coli	E. faecium				1 (0)
E. coli	E. faecium			CTX-M	1 (1)	E. faecium (1)
E. coli	K. oxytoca	Streptococcusinfantarius			1 (0)
E. coli	P. aeruginosa				1 (1)	P. aeruginosa (1)
E. coli	P. mirabilis				1 (1)	E. coli (1)
E. coli	P. mirabilis	P. vulgaris	Streptococcus -viridans group		1 (1)	S. viridans group(1)
E. coli	Propionibacteria*				1 (0)
E. coli	S. anginosus gp				1 (1)	S. anginosus group(1)
E. coli	S. aureus				1 (0)
E. coli	S. pneumoniae				1 (0)
E. coli	Staphylococcus				1 (0)
E. faecalis	K. pneumoniae				1 (1)	K. pneumoniae (1)
E. faecalis	M. morganii				1 (0)
E. faecalis	M. morganii	P. vulgaris			1 (0)
E. faecalis	P. aeruginosa	S. aureus			1 (1)	P. aeruginosa (1)
E. faecalis	P. mirabilis				3 (0)
E. faecalis	S. maltophilia				1 (1)	E. faecalis (1)
E. faecalis	S. marcescens				1 (0)
E. faecium	K. pneumoniae				1 (0)
E. faecium	P. aeruginosa				1 (0)
E. faecium	P. mirabilis				1 (0)
K. oxytoca	S. anginosus group				1 (0)
K. pneumoniae	P. aeruginosa				1 (1)	P. aeruginosa (1)
K. pneumoniae	S. aureus				2 (1)	S. aureus (1)
Distinct Co-Detection Combinations Detected by the Comparator Method(s) inRetrospective Clinical Samples				NumberSamples	DiscrepantOrganism(s) /ResistanceMarker(s)A
Target 1	Target 2	Target 3	Target 4	(NumberDiscrepant)
K. pneumoniae	Staphylococcus			1 (1)	Staphylococcus (1)
P. aeruginosa	S. maltophilia			1 (1)	S. maltophilia (1)
P. mirabilis	Peptostreptococcusanaerobius*			1 (0)
P. mirabilis	Providenciastuartii*			2 (1)	P. mirabilis (1)
P. mirabilis	Staphylococcus			1 (0)
Pseudomonasputida*	S. epidermidis	S. maltophilia		1 (0)
S. aureus	S. marcescens			1 (0)
S. marcescens	Staphylococcus			1 (0)
S. marcescens	Streptococcus mitisgroup	Streptococcussalivarius		1 (0)

{58}------------------------------------------------

{59}------------------------------------------------

{60}------------------------------------------------

Indicates an off-panel organism not detected by the BCID-GN Panel.

A. A discrepant organism or resistance marker is defined as one that was detected by the comparator method(s) but not by the BCID-GN Panel (excludes organisms not detected by the BCID-GN panel).

Clinical Study ePlex Instrument Performance

A total of 2460 samples (including prospective, retrospective, and contrived samples) were initially tested in the clinical evaluations. Of these, 23/2460 (0.9%) did not complete the run and the sample was retested. After repeat testing, all 2460 samples completed testing and 2334/2460 (94.9%, 95% CI: 93.9%-95.7%) generated valid results and 126/2460 (5.1%, 95% CI: 4.3%-6.1%) generated invalid results on the first completed attempt.

Upon repeat testing of the 126 samples with initially invalid results, 1/126 (0.8%) did not complete the run and the sample was retested. After repeat testing, all 126 samples completed testing and 114/126 (90.5%) generated valid results. Overall, after final testing, 12/2460 (0.5%, 95% CI: 0.3%-0.9%) had final, invalid results, resulting in a final validity rate of 2448/2460 (99.5%, 95% CI: 99.1%-99.7%).

{61}------------------------------------------------

ANALYTICAL PERFORMANCE CHARACTERISTICS Limit of Detection (LoD)

The limit of detection (LoD), or analytical sensitivity, was identified and verified for each target on the BCID-GN Panel using quantified reference strains in simulated blood culture sample matrix, which is defined as a whole blood with EDTA added to a blood culture bottle in the same ratio as the manufacturer recommends and incubated for 8 hours. At least 20 replicates per target were tested for each condition. The limit of detection was defined as the lowest concentration of each target that is detected in ≥95% of tested replicates. The confirmed LoD for each ePlex BCID-GN Panel organism is shown in Table 56.

Target	Organism	Strain	LoDConcentration(CFU/mL)
Acinetobacter baumannii	Acinetobacter baumannii	NCTC 13421	1 x 106
Acinetobacter baumannii	Acinetobacter baumannii	NCTC 13304	1 x 106
Bacteroides fragilis	Bacteroides fragilis	ATCC 25285	1 x 105
Bacteroides fragilis	Bacteroides fragilis	ATCC 43860	1 x 104
Citrobacter	Citrobacter freundii	NCTC 9750	1 x 106
Citrobacter	Citrobacter koseri	ATCC 27156	1 x 106
Cronobacter sakazakii	Cronobacter sakazakii	ATCC 29544	1 x 105
Cronobacter sakazakii	Cronobacter sakazakii	ATCC 29004	1 x 106
Enterobacter (non-cloacae complex)	Enterobacter aerogenes	CDC#0074	1 x 106
Enterobacter (non-cloacae complex)	Enterobacter aerogenes	CDC#0161	1 x 105
Enterobacter cloacaeComplex	Enterobacter amnigenus	ATCC 33072	1 x 106
Enterobacter cloacaeComplex	Enterobacter cloacae	CDC#0154	1 x 106
Enterobacter cloacaeComplex	Enterobacter asburiae	ATCC 35957	1 x 106
Enterobacter cloacaeComplex	Enterobacter hormaechei	ATCC BAA-2082	1 x 106
Escherichia coli	Escherichia coli	CDC#0118	1 x 107
Escherichia coli	Escherichia coli	NCTC 13441	1 x 106
Target	Organism	Strain	LoD Concentration (CFU/mL)
Fusobacterium necrophorum	Fusobacterium necrophorum	ATCC 51357	1 x 108
Fusobacterium necrophorum	Fusobacterium necrophorum	ATCC 27852	1 x 107
	Fusobacterium nucleatum	Fusobacterium nucleatum	ATCC 25586
Fusobacterium nucleatum	Fusobacterium nucleatum	ATCC 23726	1 x 105
	Haemophilus influenzae	Haemophilus influenzae	ATCC 19418
Haemophilus influenzae	Haemophilus influenzae	ATCC 9006	1 x 107
	Klebsiella oxytoca	Klebsiella oxytoca	ATCC 43165
Klebsiella oxytoca	Klebsiella oxytoca	ATCC 8724	1 x 107
	Klebsiella pneumoniae group	Klebsiella pneumoniae	CDC#0160
Klebsiella pneumoniae group	Klebsiella pneumoniae	CDC#0107	1 x 106
	Morganella morganii	Morganella morganii	ATCC 25829
Morganella morganii	Morganella morganii	CDC#0133	1 x 107
	Neisseria meningitidis	Neisseria meningitidis	ATCC 13090
Neisseria meningitidis	Neisseria meningitidis	ATCC 13102	1 x 104
	Proteus	Proteus vulgaris	ATCC 6896
Proteus	Proteus vulgaris	ATCC 6380	1 x 107
	Proteus mirabilis	Proteus mirabilis	CDC#0159
Proteus mirabilis	Proteus mirabilis	ATCC 43071	1 x 106
	Pseudomonas aeruginosa	Pseudomonas aeruginosa	CDC#0103
Pseudomonas aeruginosa	Pseudomonas aeruginosa	NCTC 13437	1 x 106
	Salmonella	Salmonella bongori	ATCC 43975
Salmonella	Salmonella enterica	ATCC 6962	1 x 105
	Serratia	Serratia liquefaciens	ATCC 27592
Serratia	Serratia plymuthica	ATCC 53858	1 x 107
	Serratia marcescens	Serratia marcescens	ATCC 14041
Serratia marcescens	Serratia marcescens	ATCC 14756	1 x 105
	Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	ATCC 13637
Target	Organism	Strain	LoDConcentration(CFU/mL)
	Stenotrophomonas maltophilia	ATCC 17666	$1 x 10^7$
Pan Candida	Candia albicans	ATCC 10231	$1 x 10^6$
	Candia glabrata	ATCC 15126	$1 x 10^5$

Pan Gram-Positive	Enterococcus faecalis	ATCC 51575	$1 x 10^5$
	Enterococcus faecium	ATCC 31282	$1 x 10^7$
	Bacillus subtilis	ATCC 21008	$1 x 10^6$
	Staphylococcus aureus	ATCC BAA-2313	$1 x 10^5$
	Streptococcus agalactiae	ATCC 13813	$1 x 10^6$
	Streptococcus anginosus	ATCC 33397	$1 x 10^6$
CTX-M	Escherichia coli (CTX-M-15)	NCTC 13441	$1 x 10^4$
	Klebsiella pneumoniae (CTX-M-2)	CDC#0107	$1 x 10^5$
IMP	Enterobacter aerogenes (IMP-4)	CDC#0161	$1 x 10^6$
	Pseudomonas aeruginosa (IMP-1)	CDC#0103	$1 x 10^5$
KPC	Enterobacter hormaechei (KPCvariant not known)	ATCC BAA-2082	$1 x 10^6$
	Morganella morganii (KPC-2)	CDC#0133	$1 x 10^6$
NDM	Escherichia coli (NDM-1)	CDC#0118	$1 x 10^5$
	Proteus mirabilis (NDM-1)	CDC#0159	$1 x 10^5$
OXA	Acinetobacter baumannii (OXA-23)	NCTC 13421	$1 x 10^5$
	Acinetobacter baumannii (OXA-27)	NCTC 13304	$1 x 10^5$
	Enterobacter aerogenes (OXA-48)	CDC#0074	$1 x 10^6$
	Klebsiella pneumoniae (OXA-48)	CDC#0160	$1 x 10^6$
VIM	Enterobacter cloacae (VIM-1)	CDC#0154	$1 x 10^6$
	Pseudomonas aeruginosa (VIM-10)	NCTC 13437	$1 x 10^5$

Table 56: LoD Results Summary

{62}------------------------------------------------

{63}------------------------------------------------

{64}------------------------------------------------

Analytical Reactivity (Inclusivity)

A panel of 336 strains/isolates representing the genetic, temporal and geographic diversity of each target on the ePlex BCID-GN Panel was evaluated to demonstrate analytical reactivity. Bacteria were tested at 1 x 10° CFU/mL or less and fungal strains were tested at 1 x 10° CFU/mL. In the cases where the initial testing concentration did not result in a "Detected" result, the concentration was increased to the point where detection was observed (see footnotes for concentration of these strains). Organisms detected are shown in Table 57. Additional strains were detected as part of the Limit of Detection (Analytical Sensitivity) Study and can be found in Table 56. Citrobacter strains that were tested but not detected include the following: C. amalonaticus, C. farmer, C. gillenii, C. murliniae, and C. sedlakii. Serratia odorifera and Staphylococcus simulans were not detected at concentrations of 1 x 108 CFU/mL and only one of three replicates were detected at concentrations of 1 x 109 CFU/mL.

Organism	Strain	Organism	Strain
Acinetobacter baumannii	CDC#0052	Cronobacter sakazakii	ATCC
	NCTC 13302	Cronobacter sakazakii	ATCC
	NCTC 13303		FSL
Acinetobacter baumannii	NCTC 13305	Enterobacter (non-cloacae complex)
	NCTC 13420		ATCC
	NCTC 13422	Enterobacter aerogenes	ATCC
	NCTC 13423		ATCC
Acinetobacter baumannii (NDM-1)	CDC#0033	Enterobacter amnigenus	ATCC
	ATCC BAA-1605		ATCC
	CDC#0045	Enterobacter gergoviae	ATCC
Acinetobacter baumannii (OXA-23)	CDC#0056	Enterobacter cloacae complex
	NCTC 13301		ATCC
	NCTC 13424	Enterobacter asburiae	ATCC
Acinetobacter spp. (IMP only)	JMI4084A		ATCC
Bacteroides fragilis		Enterobacter cloacae (CTX-15)	CDC
	ATCC 23745	Enterobacter cloacae (CTX-9)	NCTC
Bacteroides fragilis	ATCC 700786	Enterobacter cloacae (CTX-15, KPC-2)	CDC
	NCTC 9343	Enterobacter cloacae (CTX, NDM)	JMI
Citrobacter		Enterobacter cloacae subsp. cloacae	ATCC
Citrobacter braakii	ATCC 43162		ATCC
	ATCC 51113	Enterobacter cloacae subsp. dissolvens	ATCC
Citrobacter freundii	ATCC 6879	Enterobacter hormaechei	ATCC
	ATCC 8090	Enterobacter hormaechei subsp. hormaechei	ATCC
Citrobacter freundii (CTX)	JMI2047	Enterobacter hormaechei subsp. oharae	ATCC
Citrobacter freundii (KPC-2)	CDC#0116	Enterobacter hormaechei subsp. steigerwaltii	CIP
	ATCC 25409	Enterobacter ludwigii	DSM
Citrobacter koseri	ATCC 27028	Escherichia coli
	ATCC 29225		ATCC
	ATCC 29936		ATCC
Citrobacter species (CTX-15, NDM-1)	CDC #0157	Escherichia coli	ATCC
Citrobacter werkmanii	ATCC 51114		ATCC

	Table 57: Analytical Reactivity (Inclusivity)

Strain

ATCC 12868 ATCC BAA-894 FSL F6-0023

ATCC 13048 ATCC 29010 ATCC 51697 ATCC 33731 ATCC 51816ª ATCC 33028 ATCC 33426

ATCC 35954 ATCC 35955 ATCC 35956 CDC#0038 NCTC 13464 CDC#0163 JMI53571 ATCC 23355 ATCC 35030 ATCC 23373 ATCC 700323 ATCC 49162 ATCC 49163 CIP108489T DSM-16688

ATCC 14948 ATCC 25922 ATCC 33605 ATCC 33876 ATCC 35150 ATCC 4157

{65}------------------------------------------------

Organism	Strain
	ATCC 43888
	ATCC 51446
	ATCC 51755
	ATCC 53498
	ATCC 700728
	NCIMB 8545
	NCTC 8620
	ATCC 9637
	ATCC BAA-196
	ATCC BAA-197
	ATCC BAA-198
	ATCC BAA-199
	ATCC BAA-200
	ATCC BAA-201
	ATCC BAA-202
	ATCC BAA-203
	ATCC BAA-204
	LMC 243094647
	LMC 243098776
	LMC 243098947
	LMC 243108047
	LMC 243109799
	LMC 243112411
	LMC 244006281
	LMC 244006433
	LMC 244008038
	LMC 244012579
	NCTC 13351
	NCTC 10279
	ATCC 10536
	ATCC 10538
	ATCC 10799
	ATCC 11229
	ATCC 13762
	ATCC 14169
Escherichia coli (CTX-14)	CDC#0086
	ATCC BAA-2326
	NCTC 13353
Escherichia coli (CTX-15)	NCTC 13400
	NCTC 13450
	NCTC 13451
Escherichia coli (CTX-3)	NCTC 13452
Escherichia coli (CTX-1)	NCTC 13461
Escherichia coli (CTX-2)	NCTC 13462
Escherichia coli (CTX-8)	NCTC 13463
Escherichia coli (CTX-15, NDM-6)	CDC#0137
Escherichia coli (CTX-15, NDM-7)	CDC#0162
Escherichia coli (IMP)	NCTC 13476
Escherichia coli (KPC)	ATCC BAA-2340
Escherichia coli (NDM-5)	CDC#0150
Escherichia coli (OXA)	LMC DR00012
Escherichia coli (VIM)	JMI32465
Fusobacterium necrophorum
	ATCC 25286
Fusobacterium necrophorum subsp.	NCTC 10575
necrophorum	NCTC 10577
Fusobacterium nucleatum
Fusobacterium nucleatum subsp. nucleatum	ATCC 31647
Fusobacterium nucleatum subsp. fusiforme	ATCC 51190

Organism	Strain
Fusobacterium nucleatum subsp. vincentii	ATCC 49256
Haemophilus influenzae	ATCC 33930
	ATCC 43065
	ATCC 43163
Haemophilus influenzae	NCTC 11931
	NCTC 12699
	NCTC 8143
Haemophilus influenzae Type b	ATCC 10211
Haemophilus influenzae Type c	ATCC 9007
Haemophilus influenzae Type d	ATCC 9332
Haemophilus influenzae Type e	NCTC 8472
Haemophilus influenzae Type f	ATCC 9833
Klebsiella oxytoca
	ATCC 43086
	ATCC 43863
Klebsiella oxytoca	ATCC 49131
	ATCC 700324
	ATCC 51817
Klebsiella oxytoca (KPC-3)	CDC#0147
Klebsiella pneumoniae group
Klebsiella pneumoniae (CTX-15)	CDC#0109
Klebsiella pneumoniae (CTX-25)	NCTC 13465
Klebsiella pneumoniae (CTX, KPC)	IMH-C2261309
Klebsiella pneumoniae (CTX, NDM-1)	NCTC 13443
Klebsiella pneumoniae (CTX-15; NDM-1; OXA-232)	CDC#0153
Klebsiella pneumoniae (CTX-15, OXA-232)	CDC#0075
	CDC#0066
	CDC#0039
Klebsiella pneumoniae (CTX-15, OXA-181)	CDC#0140
	CDC#0141
	CDC#0142
Klebsiella pneumoniae (IMP-4)	CDC#0034
	CDC#0080
	CDC#0125
Klebsiella pneumoniae (KPC-3)	CDC#0112
	CDC#0113
	ATCC BAA-1705
	IMH-C2260742
Klebsiella pneumoniae (KPC)	IMH-C3151729
	IMH-C4151728
	IMH-C4171868
Klebsiella pneumoniae (OXA-48)	NCTC 13442
Klebsiella pneumoniae (CTX-15; VIM-27)	CDC#0040
	CDC#0135
Klebsiella pneumoniae (VIM-1)	NCTC 13439
	NCTC 13440
Klebsiella pneumoniae subsp. Ozaenae	ATCC 11296
	ATCC 13883
	ATCC 27736
Klebsiella pneumoniae subsp. pneumoniae	ATCC 51503
	ATCC 51504
Klebsiella quasipneumoniae	ATCC 700603
Klebsiella pneumoniae subsp. rhinoscleromatis	ATCC 9436
Klebsiella variicola	Morganella morganii	ATCC BAA-830
Morganella morganii	ATCC 25830
	GM148-209

{66}------------------------------------------------

Organism	Strain
Morganella morganii (CTX-15; NDM-1)	CDC#0057c
Neisseria meningitidis
Neisseria meningitidis Serotype A	ATCC 13077
Neisseria meningitidis Serotype B	NCTC 10026
Neisseria meningitidis Serotype W135	NCTC 11203
Neisseria meningitidis Serotype Y	ATCC 35561
Proteus
Proteus hauseri	ATCC 13315
	ATCC 33583
Proteus mirabilis	ATCC BAA-663
Proteus mirabilis (IMP)	JMI955389
Proteus mirabilis (KPC-6)	CDC#0155
Proteus penneri	ATCC 35197
	ATCC 33420
	ATCC 49132
Proteus vulgaris	ATCC 8427
	NCTC 4636
Pseudomonas aeruginosa
Pseudomonas aeruginosa (IMP-14)	CDC#0092
Pseudomonas aeruginosa (IMP-1)	CDC#0241
Pseudomonas aeruginosa (IMP)	CDC#0439
Pseudomonas aeruginosa (KPC-5)	CDC#0090
Pseudomonas aeruginosa (VIM-2)	CDC#0100
Pseudomonas aeruginosa (VIM-4)	CDC#0054
Salmonella
Salmonella enterica serovar 4,[5],12:i	FSL S5-0580
Salmonella enterica serovar Agona	ATCC 51957
Salmonella enterica serovar Bareilly	ATCC 9115
Salmonella enterica serovar Braenderup	ATCC 700136
Salmonella enterica serovar Enteritidis	ATCC BAA-708
Salmonella enterica serovar Hadar	ATCC 51956
Salmonella enterica serovar Heidelberg	ATCC 8326
Salmonella enterica serovar Infantis	ATCC BAA-1675
Salmonella enterica serovar Javiana	ATCC 10721
Salmonella enterica serovar Montevideo	ATCC 8387
Salmonella enterica serovar Muenchen	ATCC 8388
Salmonella enterica serovar Oranienburg	ATCC 9239
Salmonella enterica serovar Paratyphi B	FSL S5-0447
Salmonella enterica serovar Saintpaul	ATCC 9712
Salmonella enterica serovar Thompson	ATCC 8391
Salmonella enterica serovar Typhi	ATCC 19430
Salmonella enterica subsp. arizonae	ATCC 13314
Salmonella enterica subsp. diarizonae	ATCC 12325
Salmonella enterica subsp. enterica serovar Typhimurium	ATCC 14028
Salmonella enterica subsp. houtenae	ATCC 29834
Salmonella enterica subsp. indica	ATCC BAA-1578
Salmonella enterica subsp. salamae	ATCC 6959
Salmonella enterica subsp. enterica serovar Mississippi	FSL A4-0633
Salmonella enterica subsp. enterica serovar Schwarzengrund	FSL S5-0458
Serratia
Serratia ficaria	ATCC 33105
Serratia fonticola	ATCC 29844
Serratia grimesii	ATCC 14460
	ATCC 13880
Serratia marcescens	ATCC 43861
	ATCC 43862	ATCC 43862
Serratia marcescens (CTX)	JMI10244
Organism	Strain
Serratia rubidaea	ATCC 27593
	ATCC 29025
Stenotrophomonas maltophilia
Stenotrophomonas maltophilia	ATCC 13636
	GM148-207
	GM148-208
Pan-Gram Positive
Bacillus amyloliquefaciens	ATCC 23845
Bacillus atrophaeus	ATCC 49337
Bacillus cereus	ATCC 10876
Bacillus licheniformis	ATCC 21039
Bacillus thuringiensis	ATCC 35646
Enterococcus avium	ATCC 14025
Enterococcus casseliflavus	ATCC 700327
Enterococcus faecalis	JMI876745
Enterococcus gallinarum	ATCC 49573
Enterococcus hirae	ATCC 49479
Enterococcus raffinosus	ATCC 49464
Enterococcus saccharolyticus	ATCC 43076D
Staphylococcus capitis	NRS866
Staphylococcus chromogenes	ATCC 43764
Staphylococcus cohnii	ATCC 29974
Staphylococcus epidermidis	ATCC 35984
Staphylococcus gallinarum	ATCC 700401
Staphylococcus haemolyticus	ATCC 29970
Staphylococcus hominis	ATCC 27844
Staphylococcus hyicus	ATCC 11249
Staphylococcus lentus	ATCC 700403
Staphylococcus lugdunensis	ATCC 49576
Staphylococcus pasteuri	ATCC 51128
Staphylococcus vitulinus	ATCC 51699
Streptococcus constellatus	ATCC 27513
Streptococcus dysgalactiae	ATCC 35666
Streptococcus equi	ATCC 9528
Streptococcus gallolyticus	ATCC 9809
Streptococcus gordonii	ATCC 35557
Streptococcus infantis	ATCC 700779
Streptococcus intermedius	ATCC 27335
Streptococcus mitis	ATCC 49456
Streptococcus oralis	ATCC 35037
Streptococcus parasanguinis	ATCC 15909
Streptococcus pneumoniae	ATCC 8335
Streptococcus pyogenes	ATCC 12344
Streptococcus salivarius	ATCC 7073
Streptococcus thoraltensis	ATCC 700865E
Pan Candida
Candida albicans	ATCC 24433
	ATCC 90028
	ATCC 2001
Candida glabrata	ATCC 66032
	ATCC 14243
Candida krusei	ATCC 32196
	ATCC 34135F
	ATCC 22019
Candida parapsilosis	ATCC 58895
	ATCC 90018G

{67}------------------------------------------------

A. Non-target species used to evaluate resistance marker.
B. 5/6 replicates detected at 2.0 x 108 CFU/mL.
C. 5/6 replicates detected at 4.5 x 108 CFU/mL.

D. Strain may have reduced sensitivity and was not 100% detected at concentrations <1 x 10° CFU/mL.

E. Strain may have reduced sensitivity and was not 100% detected at concentrations <4 x 10° CFU/mL.

F. In initial testing, 1/6 replicates was detected at 1x 10° CFUmL; during additional testing, 3/3 replicates detected at bottle positivity.

G. In intial testing, 2/6 replicates were detected at 1 x 10° CFU/mL; during additional testing, 6/6 replicates detected at bottle positivity.

Predicted (in silico) Reactivity for Genus and Group Assays

In addition to species-specific assays, the ePlex BCID-GN Panel contains a number of broader genus or group-level assays including: Citrobacter cloacae complex, Enterobacter (non-cloacae complex), Klebsiella pneumoniae group, Proteus, Serratia, Pan Candida, and Pan Gram-Positive assays. Tables 58-65 highlight the-predicted (in silico) reactivity (inclusivity) for these assay targets.

Note: the performance of the ePlex BCID-GN Panel has not been established for all of the organisms listed in the tables below. See the Analytical Reactivity section for data on organisms for which performance characteristics have been established (indicated with an asterisk in Tables 58-65). Some species were not assessed in silico due to lack of sequence data, though they may appear in the analytical sensitivity or specificity studies.

		Table 58: Predicted (in silico) Reactivity (Inclusivity) Results for Citrobacter

Detection Predicted for ≥95% of target sequences
Citrobacter koseri*	Citrobacter intermedius
Detection Predicted for 85%-94% of target sequences
Citrobacter freundii*	Citrobacter braakii*
Detection Predicted for <85.0% of target sequences
Citrobacter werkmanii* (66.7%)	Citrobacter youngae* (50.0%)
Detection Not Predicted
Citrobacter europaeus	Citrobacter gillenii	Citrobacter amalonaticusA
Citrobacter farmeri	Citrobacter sedlakii	Citrobacter murliniae

A. Detection predicted in silico, however ATCCBAA-2563 was not detected in wet testing.

{68}------------------------------------------------

Table 59: Predicted (in silico) Reactivity (Inclusivity) Results for Enterobacter cloacae
complex

Detection Predicted for ≥95% of target sequences
Enterobacter cloacae*	Enterobacter asburiae*	Enterobacter hormaechei*
Enterobacter xiangfangensis
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85.0% of target sequences
Enterobacter ludwigii* (68.4%)	Enterobacter nimipressuralis(25.0%)
Detection Not Predicted
Enterobacter kobei	Enterobacter cancerogenus

Table 60: Predicted (in silico) Reactivity (Inclusivity) Results for Enterobacter (non-cloacae complex)

Detection Predicted for ≥95% of target sequences
Enterobacter aerogenes*	Enterobacter gergoviae*
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85.0% of target sequences
Enterobacter amnigenus* (62.5%)
Detection Not Predicted
None Identified

{69}------------------------------------------------

Detection Predicted for ≥95% of target sequences
Klebsiella pneumoniae*	Klebsiella quasipneumoniae*	Klebsiella variicola*
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85.0% of target sequences
None Identified
Detection Not Predicted
None Identified

Table 61: Predicted (in silico) Reactivity (Inclusivity) Results for Klebsiella pneumoniae Group

Table 62: Predicted (in silico) Reactivity (Inclusivity) Results for Proteus

Detection Predicted for ≥95% of target sequences
Proteus mirabilis*	Proteus penneri*	Proteus vulgaris*
Proteus hauseri*	Proteus cibarius
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85.0% of target sequences
None Identified
Detection Not Predicted
Proteus myxofaciens

{70}------------------------------------------------

Detection Predicted for ≥95% of target sequences
Serratia marcescens*	Serratia grimesii*	Serratia rubidaea*
Serratia ficaria*	Serratia liquefaciens*	Serratia proteamaculans
Serratia fonticola*	Serratia plymuthica*
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85.0% of target sequences
Serratia quinivorans (33.3%)
Detection Not Predicted
Serratia nematodiphila	Serratia odoriferaA*	Serratia ureilytica

Table 63: Predicted (in silico) Reactivity (Inclusivity) Results for Serratia

B. Not predicted in silico, however ATCC 33077 was intermittently detected in wet testing. See Analytical Reactivity (Inclusivity) Study.

Table 64: Predicted (in silico) Reactivity (Inclusivity) Results for Pan Candida

Detection Predicted for ≥95% of target sequences
Candida albicans*	Candida glabrata*	Candida krusei*
Candida parapsilosis*
Detection Predicted for 85%-94% of target sequences
None Identified
Detection Predicted for <85% of target sequences
None Identified
Detection Not Predicted
Detection was not predicted by bioinformatic analysis for other Candida species for which sequence data was available.

{71}------------------------------------------------

Table 65: Predicted (in silico) Reactivity (Inclusivity) Results for Pan Gram-Positive
Detection Predicted for ≥95% of target sequences
Bacillus
Bacillus amyloliquefaciens*	Bacillus paralicheniformis	Bacillus toyonensis
Bacillus atrophaeus*	Bacillus siamensis	Bacillus vallismortis
Bacillus bombysepticus	Bacillus subtilis*	Bacillus velezensis
Bacillus licheniformis*	Bacillus tequilensis	Bacillus weihenstephanensis
Bacillus methylotrophicus	Bacillus thuringiensis*
Enterococcus
Enterococcus avium*	Enterococcus faecium*	Enterococcus raffinosus*
Enterococcus dispar	Enterococcus flavescens	Enterococcus saccharolyticus*
Enterococcus durans	Enterococcus malodoratus	Enterococcus thailandicus
Enterococcus faecalis*	Enterococcus pseudoavium
Staphylococcus
Staphylococcus aureus*	Staphylococcus haemolyticus*	Staphylococcus pseudintermedius
Staphylococcus agnetis	Staphylococcus hominis*	Staphylococcus pseudolugdunensis
Staphylococcus argensis	Staphylococcus hominis subsp.novobiosepticus	Staphylococcus pulvereri
Staphylococcus argenteus	Staphylococcus hyicus*	Staphylococcus rostri
Staphylococcus auricularis	Staphylococcus jettensis	Staphylococcus saprophyticus
Staphylococcus capitis*	Staphylococcus kloosii	Staphylococcus schleiferi
Staphylococcus caprae	Staphylococcus lentus*	Staphylococcus schweitzeri
Staphylococcus carnosus	Staphylococcus lugdunensis*	Staphylococcus sciuri
Staphylococcus chromogenes*	Staphylococcus lutrae	Staphylococcus simiae
Staphylococcus cohnii*	Staphylococcus massiliensis	Staphylococcus simulans
Staphylococcus delphini	Staphylococcus microti	Staphylococcus stepanovicii
Staphylococcus devriesei	Staphylococcus muscae	Staphylococcus succinus
Staphylococcus epidermidis*	Staphylococcus nepalensis	Staphylococcus vitulinus*
Staphylococcus equorum	Staphylococcus pasteuri*	Staphylococcus warneri
Staphylococcus felis	Staphylococcus petrasii	Staphylococcus xylosus
Staphylococcus fleurettii	Staphylococcus pettenkoferi
Staphylococcus gallinarum*Staphylococcus piscifermentans
Streptococcus
Streptococcus agalactiae	Streptococcus infantarius	Streptococcus phocae
Streptococcus alactolyticus	Streptococcus infantis*	Streptococcus pneumoniae*
Streptococcus anginosus	Streptococcus intermedius*	Streptococcus porcinus
Streptococcus australis	Streptococcus intestinalis	Streptococcus porcorum
Streptococcus caballi	Streptococcus lactarius	Streptococcus pseudopneumoniae
Streptococcus constellatus	Streptococcus loxodontisalivarius	Streptococcus pseudoporcinus
Streptococcus criceti	Streptococcus luteciae	Strepotococcus pyogenes
Streptococcus cristatus	Streptococcus lutetiensis	Streptococcus rifensis
Streptococcus danieliae	Streptococcus macedonicus	Streptococcus rubneri
Streptococcus dentasini	Streptococcus marimammalium	Streptococcus salivarius*
Streptococcus dentisani	Streptococcus massiliensis	Streptococcus saliviloxodontae
Streptococcus didelphis	Streptococcus mitis*	Streptococcus sanguinis
Streptococcus difficilis	Streptococcus moroccensis	Streptococcus seminale
Streptococcus dysgalactiae subsp.dysgalactiae	Streptococcus oligofermentans	Streptococcus sinensis
Streptococcus dysgalactiae subsp.equisimilis	Streptococcus oralis*	Streptococcus suis
Streptococcus dysgalactiae*	Streptococcus oricebi	Streptococcus thermophilus
Streptococcus equi*	Streptococcus orisratti	Streptococcus thoraltensis*
Streptococcus equinus	Streptococcus panodentis	Streptococcus tigurinus
Streptococcus fryi	Streptococcus parasanguinis*	Streptococcus troglodytae
Streptococcus gallolyticus*	Streptococcus parasuis	Streptococcus troglodytidis
Streptococcus gordonii*	Streptococcus parauberis	Streptococcus urinalis
Streptococcus himalayensis	Streptococcus pasteuri	Streptococcus ursoris
Streptococcus hongkongensis	Streptococcus pasteurianus	Streptococcus vestibularis
Streptococcus hyointestinalis	Streptococcus peroris	Streptococcus waiu
Detection Predicted for 85%-94% of target sequences
Bacillus cereus*	Enterococcus hirae*	Staphylococcus saccharolyticus
Enterococcus casseliflavus*	Staphylococcus arlettae	Streptococcus bovis
Enterococcus cecorum	Staphylococcus condimenti	Streptococcus uberis
Enterococcus gallinarum	Staphylococcus intermedius
Detection Predicted for <85.0% of target sequences
Bacillus mojavensis (77.8%)	Bacillus sonorensis (83.3%)	Streptococcus halichoeri (66.7%)
Streptococcus ratti (75.0%)
Detection Not Predicted
Bacillus pseudomycoides	Enterococcus sulfureus	Streptococcus hyovaginalis
Enterococcus aquimarinus	Enterococcus termitis	Streptococcus ictaluri
Enterococcus asini	Enterococcus ureasiticus	Streptococcus iniae
Enterococcus caccae	Enterococcus ureilyticus	Streptococcus lactis
Enterococcus camelliae	Enterococcus villorum	Streptococcus macacae
Enterococcus canintestini	Staphylococcus caseolyticus	Streptococcus marmotae
Enterococcus canis	Streptococcus acidominimus	Streptococcus merionis
Enterococcus columbae	Streptococcus azizii	Streptococcus milleri
Enterococcus devriesei	Streptococcus cameli	Streptococcus minor
Enterococcus haemoperoxidus	Streptococcus canis	Streptococcus oriloxodontae
Enterococcus hawaiiensis	Streptococcus castoreus	Streptococcus orisasini
Enterococcus hermanniensis	Streptococcus cremoris	Streptococcus orisuis
Enterococcus italicus	Streptococcus criae	Streptococcus ovis
Enterococcus mundtii	Streptococcus cuniculi	Streptococcus pharyngis
Enterococcus pallens	Streptococcus dentapri	Streptococcus pluranimalium
Enterococcus pernyi	Streptococcus dentiloxodontae	Streptococcus plurextorum
Enterococcus phoeniculicola	Streptococcus dentirousetti	Streptococcus plutanimalium
Enterococcus plantarum	Streptococcus devriesei	Streptococcus porci
Enterococcus quebecensis	Streptococcus downei	Streptococcus rupicaprae
Enterococcus ratti	Streptococcus entericus	Streptococcus sobrinus
Enterococcus rivorum	Streptococcus ferus	Streptococcus tangierensis
Enterococcus rotai	Streptococcus gallinaceus
Enterococcus silesiacus	Streptococcus henryi

Table 65: Predicted (in silico) Reactivity (Inclusivity) Results for Pan Gram-Positive

{72}------------------------------------------------

{73}------------------------------------------------

{74}------------------------------------------------

Predicted (in silico) Reactivity for Resistance Markers

The ePlex BCID-GN Panel contains six resistance markers that were each assessed for predicted in silico reactivity. Tables 66-76 highlight the-predicted (in silico) reactivity for CTX-M, IMP, KPC, NDM, OXA, and VIM. Strains that were tested as part of the Analytical Reactivity (Inclusivity) Study are marked with an asterisk in Tables 66-76. It is noted that the performance of the ePlex BCID-GN Panel has not been established for all organisms listed in Tables 66-76. Table 77 includes all variants that are not predicted to be detected by the BCID-GN Panel by in silico analysis.

The CTX-M assay on the ePlex BCID-GN Panel is designed to detect the following CTX-M groups: CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, and CTX-M-25.

Target	Associated Organism	Variant Detected
Acinetobacter baumannii	Acinetobacter baumannii	CTX-M-15
Citrobacter	Citrobacter freundii	CTX-M-3CTX-M-15*CTX-M-30CTX-M-55
	Citrobacter koseri	CTX-M-3CTX-M-15
Enterobacter (non-cloacae Complex)	Enterobacter aerogenes	CTX-M-3
	Enterobacter gergoviae	CTX-M-15
	Enterobacter asburiae	CTX-M-15
Enterobacter cloacae Complex	Enterobacter cloacae	CTX-M-3CTX-M-15*CTX-M-22CTX-M-37CTX-M-89CTX-M-177
	Enterobacter hormaechei	CTX-M-15CTX-M-1CTX-M-3CTX-M-10CTX-M-12CTX-M-14*
Escherichia coli	Escherichia coli	CTX-M-15*CTX-M-22CTX-M-23CTX-M-28CTX-M-29

Table 66: Predicted (in silico) Reactivity (Inclusivity) Results for CTX-M-1

{75}------------------------------------------------

Target	Associated Organism	Variant Detected	Target	Associated Organism	Variant Detected
		CTX-M-174CTX-M-175CTX-M-179CTX-M-180CTX-M-181CTX-M-182CTX-M-184CTX-M-186			CTX-M-176CTX-M-183CTX-M-197CTX-M-204
Klebsiella oxytoca	Klebsiella oxytoca	CTX-M-3CTX-M-15CTX-M-28CTX-M-162	Morganella morganii	Morganella morganii	CTX-M-3CTX-M-15*CTX-M-55
Klebsiella pneumoniae group	Klebsiella pneumoniae	CTX-M-1CTX-M-3CTX-M-10CTX-M-11CTX-M-12CTX-M-15*CTX-M-22CTX-M-28CTX-M-32CTX-M-52CTX-M-54CTX-M-55CTX-M-57CTX-M-60CTX-M-62CTX-M-71CTX-M-72CTX-M-96CTX-M-155CTX-M-156CTX-M-157CTX-M-173	Proteus mirabilis	Proteus mirabilis	CTX-M-1CTX-M-3CTX-M-15CTX-M-32CTX-M-66CTX-M-116CTX-M-136CTX-M-164CTX-M-167
		Pseudomonas aeruginosa	Pseudomonas aeruginosa	CTX-M-15CTX-M-28CTX-M-32
			Salmonella	Salmonella enterica	CTX-M-15CTX-M-53CTX-M-55CTX-M-57CTX-M-61CTX-M-88


				Salmonella sp.	CTX-M-3
				Salmonella typhimurium	CTX-M-37CTX-M-61
			Serratiamarcescens/Serratia	Serratia liquefaciens	CTX-M-22
				Serratia marcescens	CTX-M-3CTX-M-15CTX-M-55

			Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	CTX-M-15

Table 67: Predicted (in silico) Reactivity (Inclusivity) Results for CTX-M-2

Target	Associated Organism	Variant Detected
Acinetobacterbaumannii	Acinetobacterbaumannii	CTX-M-2CTX-M-5CTX-M-43CTX-M-115
Enterobacter cloacaecomplex	Enterobacter cloacae	CTX-M-5
Escherichia coli	Escherichia coli	CTX-M-2*CTX-M-44CTX-M-56CTX-M-92CTX-M-97CTX-M-124
Klebsiellapneumoniae Group	Klebsiellapneumoniae	CTX-M-2CTX-M-35CTX-M-59CTX-M-141CTX-M-165
Target	AssociatedOrganism	VariantDetected
Morganella morganii	Morganella morganii	CTX-M-200
Proteusmirabilis/Proteus	Proteus mirabilis	CTX-M-2
		CTX-M-20
		CTX-M-171
Pseudomonasaeruginosa	Pseudomonasaeruginosa	CTX-1-2
		CTX-M-2
Salmonella	Salmonella enterica	CTX-M-2
		CTX-M-4
		CTX-M-5
		CTX-M-6
		CTX-M-7
	Salmonella typhimurium	CTX-M-2
		CTX-M-4
		CTX-M-5
		CTX-M-7

{76}------------------------------------------------

Table 68: Predicted (in silico) Reactivity (Inclusivity) Results for CTX-M-8

Target	AssociatedOrganism	VariantDetected
Citrobacter	Citrobacteramalonaticus	CTX-M-8
Enterobacter cloacaeComplex	Enterobacter cloacae	CTX-M-8
Escherichia coli	Escherichia coli	CTX-M-8*

Target	Associated Organism	Variant Detected
Klebsiellapneumoniae Group	Klebsiellapneumoniae	CTX-M-8
Morganella morganii	Morganella morganii	CTX-M-63
Salmonella	Salmonella enterica	CTX-M-8

Table 69: Predicted (in silico) Reactivity (Inclusivity) Results for CTX-M-9

Target	AssociatedOrganism	VariantDetected	Target	AssociatedOrganism	VariantDetected
Citrobacter	Citrobacter freundii	CTX-M-14CTX-M-65			CTX-M-168CTX-M-173
Enterobacter non-cloacae Complex	Enterobacter aerogenes	CTX-M-9			CTX-M-174CTX-M-176CTX-M-177
Enterobacter cloacaeComplex	Enterobacter cloacae	CTX-M-9*CTX-M-13CTX-M-14CTX-M-64CTX-M-125CTX-M-1/CTX-M-65CTX-M-9CTX-M-13CTX-M-14CTX-M-14/CTX-M-15CTX-M-15	Klebsiella pneumoniaeGroup	Klebsiella pneumoniae	CTX-M-191CTX-M-195CTX-M-196CTX-M-198CTX-M-199CTX-M-9CTX-M-13
		CTX-M-16			CTX-M-14
		CTX-M-19			CTX-M-17
		CTX-M-21			CTX-M-18
		CTX-M-24			CTX-M-19
		CTX-M-27			CTX-M-24
		CTX-M-38			CTX-M-38
		CTX-M-47			CTX-M-46CTX-M-48
		CTX-M-51			CTX-M-49
		CTX-M-64			CTX-M-50
		CTX-M-65			CTX-M-65
Escherichia coli	Escherichia coli	CTX-M-67	Proteusmirabilis/Proteus	Proteus mirabilis	CTX-M-81
		CTX-M-73			CTX-M-99
		CTX-M-82			CTX-M-104
		CTX-M-87			CTX-M-147
		CTX-M-93			CTX-M-159
		CTX-M-97			CTX-M-201
		CTX-M-98			CTX-M-13
		CTX-M-102			CTX-M-14
		CTX-M-104	Salmonella	Salmonella enterica	CTX-M-24
		CTX-M-105			CTX-M-65
		CTX-M-106			CTX-M-90
		CTX-M-121			CTX-M-9
		CTX-M-122			CTX-M-14
		CTX-M-126CTX-M-129			CTX-M-25
CTX-M-130	Serratia	Serratialiquefaciens	CTX-M-27
CTX-M-132			CTX-M-65
CTX-M-134				CTX-M-83
CTX-M-137CTX-M-148CTX-M-161				CTX-M-84CTX-M-85CTX-M-86CTX-M-143CTX-M-14

{77}------------------------------------------------

Target	AssociatedOrganism	VariantDetected
Escherichia coli	Escherichia coli	CTX-M-25
		CTX-M-39
		CTX-M-94
		CTX-M-100
Klebsiellapneumoniae Group	KlebsiellapneumoniaeA*	CTX-M-26

						Table 70: Predicted (in silico) Reactivity (Inclusivity) Results for CTX-M-25
--	--	--	--	--	--	-------------------------------------------------------------------------------

Target	AssociatedOrganism	VariantDetected
Proteusmirabilis/Proteus	Proteus mirabilis	CTX-M-41
		CTX-M-89
		CTX-M-91
		CTX-M-160
Salmonella	Salmonella enterica	CTX-M-25

A. CTX-M-25 detected in Analytical Reactivity (Inclusivity) study.

Table 71: Predicted (in silico) Reactivity (Inclusivity) Results for IMP

Target	Associated Organism	Variant Detected	Target	Associated Organism	Variant Detected
Acinetobacterbaumannii	Acinetobacterbaumannii	IMP-1IMP-2IMP-4IMP-5IMP-8IMP-10IMP-11IMP-14IMP-19IMP-55IMP-61	Proteusmirabilis/Proteus	Proteus mirabilis	IMP-19IMP-26IMP-32IMP-38IMP-1IMP-27IMP-64IMP-1*IMP-2IMP-4IMP-6
Citrobacter	Citrobacter freundii	IMP-1IMP-4IMP-8IMP-23IMP-38			IMP-7IMP-9IMP-10IMP-11IMP-13IMP-14*IMP-15
Enterobacter non-cloacae Complex	Enterobacter aerogenes	IMP-4			IMP-16IMP-17IMP-18IMP-19
Enterobacter cloacaeComplex	Enterobacter cloacae	IMP-1IMP-4IMP-8IMP-11IMP-26IMP-34IMP-60			IMP-20IMP-21IMP-22
Enterobacter cloacaeComplex	Enterobacterhormaechei	IMP-13IMP-14	Pseudomonasaeruginosa	Pseudomonasaeruginosa	IMP-25IMP-26IMP-29IMP-30IMP-33
Escherichia coli	Escherichia coli	IMP-1IMP-4IMP-6IMP-8IMP-14IMP-30IMP-52IMP-59IMP-66			IMP-34IMP-37IMP-40IMP-41IMP-43
Klebsiella oxytoca	Klebsiella oxytoca	IMP-1IMP-4IMP-8IMP-28IMP-34			IMP-44IMP-45IMP-48IMP-49IMP-51
Klebsiella pneumoniaeGroup	Klebsiella pneumoniae	IMP-1IMP-4*IMP-6IMP-8IMP-10IMP-13	Salmonella	Salmonella entericaSerratia marcescens	IMP-53IMP-54IMP-56IMP-62IMP-63IMP-4IMP-1

{78}------------------------------------------------

Target	AssociatedOrganism	VariantDetected
Serratiamarcescens/Serratia		IMP-2
		IMP-6
		IMP-8
Target	AssociatedOrganism	VariantDetected
		IMP-24
Stenotrophomonasmaltophilia	Stenotrophomonasmaltophilia	IMP-25

Table 72: Predicted (in silico) Reactivity (Inclusivity) Results for KPC

Target	Associated Organism	VariantDetected
Acinetobacter baumannii	Acinetobacter baumannii	KPC-2KPC-3KPC-10
Citrobacter	Citrobacter freundii	KPC-2*KPC-3KPC-18
Enterobacter non-cloacaeComplex	Enterobacter aerogenes	KPC-2KPC-3
Enterobacter cloacaeComplex	Enterobacter cloacae	KPC-2*KPC-3KPC-4KPC-13KPC-18
Escherichia coli	Escherichia coli	KPC-2KPC-3KPC-4KPC-5KPC-9KPC-18KPC-21KPC-28
Klebsiella oxytoca	Klebsiella oxytoca	KPC-2KPC-3*
Klebsiella pneumoniaeGroup	Klebsiella pneumoniae	KPC-1KPC-2KPC-3*KPC-4
Target	Associated Organism	VariantDetected
		KPC-5
		KPC-6
		KPC-7
		KPC-8
		KPC-11
		KPC-12
		KPC-14
		KPC-15
		KPC-16
		KPC-17
		KPC-19
		KPC-22
		KPC-24
		KPC-25
		KPC-26
		KPC-27
		KPC-30
Proteus mirabilis/Proteus	Proteus mirabilis	KPC-2
		KPC-6*
Pseudomonas aeruginosa	Pseudomonas aeruginosa	KPC-2
		KPC-5*
Salmonella	Salmonella enterica	KPC-2
Serratiamarcescens/Serratia	Serratia marcescens	KPC-2
		KPC-3

{79}------------------------------------------------

Target	Associated Organism	Variant Detected	Target	Associated Organism	Variant Detected
Acinetobacter baumannii	Acinetobacter baumannii	VIM-1VIM-2VIM-6VIM-11	Morganella morganii	Morganella morganii	VIM-39VIM-42VIM-51VIM-52
Citrobacter	Citrobacter freundii	VIM-1VIM-2VIM-4VIM-23	Proteus mirabilis/Proteus	Proteus mirabilis	VIM-4VIM-1VIM-1
Enterobacter non-cloacae Complex	Enterobacter aerogenes	VIM-1	Pseudomonas aeruginosa	Pseudomonas aeruginosa	VIM-2VIM-3VIM-4VIM-5VIM-6VIM-8VIM-9VIM-10VIM-11VIM-14VIM-15VIM-16VIM-17VIM-18VIM-20VIM-28VIM-30VIM-36VIM-37VIM-41VIM-43VIM-44VIM-45VIM-46VIM-48VIM-50
Enterobacter cloacae Complex	Enterobacter cloacae	VIM-1VIM-2VIM-4VIM-23VIM-31VIM-40
	Enterobacter hormaechei	VIM-1VIM-4VIM-23
	Enterobacter xiangfangensis	VIM-1
Escherichia coli	Escherichia coli^	VIM-1VIM-2VIM-19VIM-29
Klebsiella oxytoca	Klebsiella oxytoca	VIM-1VIM-2VIM-4VIM-32VIM-35
Klebsiella pneumoniae Group	Klebsiella pneumoniae	VIM-1VIM-2VIM-4VIM-12VIM-19VIM-24VIM-26VIM-27VIM-33VIM-34
			Salmonella	Salmonella enterica	VIM-1VIM-2
			Serratia marcescens/Serratia	Serratia marcescens	VIM-4VIM-54
	Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	VIM-2

A. Unspecified VIM variant detected in Analytical Reactivity (Inclusivity) study.

{80}------------------------------------------------

Target	AssociatedOrganism	VariantDetected
Acinetobacterbaumannii	Acinetobacterbaumannii	OXA-23*
		OXA-49
		OXA-23/OXA-104
		OXA-23/OXA-64
		OXA-23/OXA-66
		OXA-23/OXA-69
		OXA-27
		OXA-65/OXA-239
		OXA-68
		OXA-146
		OXA-165
		OXA-166
		OXA-167
		OXA-168
		OXA-169
		OXA-170
		OXA-171
		OXA-183

Target	Associated Organism	Variant Detected
		OXA-225
		OXA-366
		OXA-398
		OXA-422
		OXA-423
		OXA-435
		OXA-440
		OXA-469
		OXA-481
		OXA-482
		OXA-483
		OXA-565
Escherichia coli	Escherichia coli	OXA-23
Klebsiellapneumoniae Group	Klebsiellapneumoniae	OXA-73
Proteus mirabilis	Proteusmirabilis	OXA-23

Table 75: Predicted (in silico) Reactivity (Inclusivity) Results for OXA-48

Target	AssociatedOrganism	VariantDetected
Acinetobacter baumannii	Acinetobacterbaumannii	OXA-48
Citrobacter	Citrobacterfreundii	OXA-181
Enterobacter non-cloacae Complex	Enterobacteraerogenes	OXA-244
Enterobacter cloacaeComplex	Enterobactercloacae	OXA-48OXA-163OXA-181
	Enterobacterhormaechei	OXA-370
	Enterobacterludwigii	OXA-48
Escherichia coli	Escherichia coli^	OXA-48OXA-163OXA-181OXA-204OXA-232OXA-244OXA-438OXA-439OXA-566OXA-1/OXA-48
Target	AssociatedOrganism	VariantDetected
Klebsiella pneumoniaeGroup	Klebsiellapneumoniae	OXA-48OXA-10OXA-162OXA-181OXA-204OXA-232*OXA-244OXA-245OXA-247OXA-484OXA-505OXA-517OXA-519
	Klebsiellavariicola	OXA-181
Morganella morganii	Morganellamorganii	OXA-181
Proteus mirabilis/Proteus	Proteus mirabilis	OXA-48
Proteus mirabilis/Proteus	Proteus mirabilis	OXA-244
Serratia	Serratia	OXA-48
marcescens/Serratia	marcescens	OXA-405

A. Unspecified OXA variant detected in the Analytical Reactivity (Inclusivity) study.

{81}------------------------------------------------

Target	AssociatedOrganism	VariantDetected
Acinetobacterbaumannii	Acinetobacterbaumannii	NDM-1*
Citrobacter	Citrobacter braakii	NDM-2
Citrobacter	Citrobacter freundii	NDM-4
		NDM-1A
		NDM-4
		NDM-6
		NDM-7
Enterobacter non-cloacae Complex	Enterobacteraerogenes	NDM-1
		NDM-4
		NDM-5
		NDM-7
Enterobacter cloacaeComplex	EnterobactercloacaeB	NDM-1
		NDM-4
		NDM-7
	Enterobacterhormaechei	NDM-1
	Enterobacterludwigii	NDM-1
Escherichia coli	Escherichia coli	NDM-1
		NDM-3
		NDM-4
		NDM-5*
		NDM-6*
		NDM-7*
		NDM-8
		NDM-11
		NDM-12
		NDM-13
Target	AssociatedOrganism	VariantDetected
		NDM-15
		NDM-16
		NDM-17
		NDM-18
		NDM-19
Klebsiella oxytoca	Klebsiella oxytoca	NDM-1
		NDM-4
		NDM-1*
		NDM-4
Klebsiellapneumoniae Group	Klebsiellapneumoniae	NDM-5
		NDM-6
		NDM-7
		NDM-10
		NDM-16
		Morganella morganii
Proteusmirabilis/Proteus	Proteus mirabilis	NDM-1
Pseudomonasaeruginosa	Pseudomonasaeruginosa	NDM-1
		NDM-5
Salmonella	Salmonella enterica	NDM-1
		NDM-5
Serratiamarcescens/Serratia	Serratia marcescens	NDM-1
Stenotrophomonasmaltophilia	Stenotrophomonasmaltophilia	NDM-1

Table 76: Predicted (in silico) Reactivity (Inclusivity) Results for NDM

A. Detected in a Citrobacter species in the Analytical Reactivity (Inclusivity) study.

B. Unspecified NDM variant detected in the Analytical Reactivity (Inclusivity) study.

C. NDM-1 was detected in Morganii in the Analytical Reactivity (Inclusivity) study but no sequences were available for in silico analysis.

{82}------------------------------------------------

Table 77: Predicted (in silico) Reactivity (Inclusivity) Results for Variants Not Detected

ResistanceMarker	Variant NotDetected	AssociatedOrganism	No. ofSequences
CTX-M-1	CTX-M-80	Klebsiellapneumoniae	3
	CTX-M-15
	Not Specified
IMP	IMP-31	Pseudomonasaeruginosa	2
	IMP-35
	IMP-7		1
NDM	NDM-1	Escherichia coli	6
		Klebsiellavariicola	3
		Salmonellaenterica	1
	NDM-3	Acinetobacterbaumannii	1
	NDM-4	Escherichia coli	1
		Cronobactersakazakii	1
	NDM-9	Escherichia coli	1
		Klebsiellapneumoniae	2
	Not specified	Escherichia coli	2
		Klebsiellapneumoniae	1
		Klebsiella sp	1
		Pseudomonasaeruginosa	1
VIM	VIM-1	Pseudomonasaeruginosa	3
		Providenciavermicola	1
	VIM-2	Klebsiellapneumoniae	1
	VIM-5	Enterobactercloacae	2
		Klebsiellapneumoniae	3
	VIM-7	Pseudomonasaeruginosa	4
	VIM-13	Pseudomonasaeruginosa	3
	VIM-25	Acinetobacterbaumannii	1
		Proteus mirabilis	2
ResistanceMarker	Variant NotDetected	AssociatedOrganism	No. ofSequences
	VIM-38		2
	VIM-47	Pseudomonasaeruginosa	2
	VIM-49		2
	Not specified		1
OXA-48	OXA-232	Escherichia coli	1

{83}------------------------------------------------

Analytical Specificity (Cross-Reactivity and Exclusivity)

Cross-reactivity of on-panel and off-panel analytes was evaluated with the BCID-GN Panel. Bacterial targets were tested in triplicate at a concentration of ~1x10° CFU/mL while fungi were tested in triplicate at a concentration of ~1x107 CFU/mL. If the target concentration could not be reached, the organism was diluted 2-fold from stock for use (indicated with an asterisk in Tables 78-81).

No cross reactivity was observed for any of the on-panel organisms. The following off-panel organisms showed cross reactivity: Acinetobacter anitratus (at a concentration of >1x104 CFU/mL) cross-reacts with the Acinetobacter baumannii assay, Enterobacter cowanii (at a concentration of >1x108 CFU/mL) cross-reacts with the Enterobacter cloacae complex assay, Escherichia hermanii cross-reacts with the Enterobacter (non-cloacae complex) assay (at a concentration of >1x106 CFU/mL) and with the Serratia assay (at a concentration of >1x107 CFU/mL), Fusobacterium periodonticum (at a concentration of 5x10° CFU/mL) and Fusobacterium simiae (at a concentration of 2.9x10° CFU/mL) cross-react with the Fusobacterium nucleatum assay, and Shigella (at a concentration of 1x109 CFU/mL) cross-reacts with the Escherichia coli assay (off-panel organisms showing cross-reactivity are bolded in the tables below). See Table 56 for a summary of the on-panel strains tested and Tables 78-81 for a summary of off-panel strains tested.

Additional in silico analysis was performed to identify any off-panel gram-negative and grampositive organisms that may cross-react with the BCID-GN Panel (Tables 82-83).

Note: the performance of the ePlex BCID-GN Panel has not been established for organisms evaluated by in silico analysis alone.

{84}------------------------------------------------

Off-Panel Exclusivity

Table 78: Off-Panel Gram-Negative Organisms Assessed for Cross-reactivity with the ePlex BCID-GN Panel (Exclusivity)

Gram Negative Organism	Strain ID
Acinetobacter haemolyticus	ATCC 19002
Acinetobacter lwoffii	ATCC 15309
Acinetobacter junii	ATCC 17908
Acinetobacter anitratusA	ATCC 49139
Aeromonas hydrophila	JMI 938982
Aeromonas salmonicida	ATCC 33658
Aeromonas sobria	ATCC 35993
Bacteroides distasonis (Parabacteroides)	ATCC 8503
Bacteroides merdae	ATCC 43184
Bacteroides thetaiotaomicron	ATCC 29741
Bacteroides vulgatus*	ATCC 8482
Bacteroides caccae	ATCC 700189
Bacteroides eggerthii	ATCC 27754
Bacteroides ovatus*	ATCC BAA-1296
Bacteroides ureolyticus*	ATCC 33387
Bordetella pertussis	ATCC 9797
Burkholderia cepacia	ATCC 25416
Citrobacter amalonaticus	ATCC BAA-2563
Citrobacter gillenii	ATCC 51640
Citrobacter sedlakii	ATCC 51493
Citrobacter farmer	ATCC 51112
Citrobacter murliniae	ATCC 51642
Edwardsiella tarda	ATCC 15947
Enterobacter kobei	ATCC BAA-260
Enterobacter cancerogenus	ATCC 35315
Enterobacter cowaniiB	DSM-18146
Escherichia albertii	DSM-17582
Escherichia fergusonii	ATCC 35469
Escherichia hermaniiC	ATCC 33650
Ewingella americana*	ATCC 33853
Eikenella corrodens	ATCC BAA-1152
Fusobacterium naviforme*	ATCC 25832
Fusobacterium gonidiaformans	ATCC 25563
Fusobacterium necrogenes*	ATCC 25556
Fusobacterium periodonticum*D	ATCC 33693
Fusobacterium simiae*D	ATCC 33568
Fusobacterium varium	ATCC 27725
Fusobacterium russii*	ATCC 25533
Fusobacterium ulcerans	ATCC 49186
Haemophilus haemolyticus	ATCC 33390
Haemophilus parahaemolyticus	ATCC 10014
Hafnia alvei	ATCC 51815
Kingella kingae*	ATCC 23331
Gram Negative Organism	Strain ID
Kluyvera cochleae	ATCC 51609
Legionella pneumoniae	ATCC 33823
Leclercia adecarboxylata	ATCC 700325
Methylobacterium mesophilicum*	ATCC 29983
Neisseria gonorrhoeae	ATCC 19424
Neisseria mucosa	ATCC 19695
Neisseria sicca	ATCC 29193
Neisseria flavecens	ATCC 13115
Neisseria lactamica	ATCC 23970
Neisseria perflava	ATCC 14799
Ochrobactrum anthropi	ATCC BAA-749
Pantoea agglomerans	ATCC 14537
Pantoea ananatis	NRRL B-41502
Pasteurella aerogenes	ATCC 27883
Pasteurella multicida subsp multicide	ATCC 12945
Prevotella intermedia	ATCC 15032
Prevotella corporis*	ATCC 33547
Prevotella oralis*	ATCC 33269
Prevotella nigrescens*	ATCC 33563
Providencia rettgeri	ATCC 9250
Providencia stuartii	ATCC 33672
Providencia alcalifaciens	ATCC 9886
Pseudomonas fluorescens	ATCC 13525
Pseudomonas putida	ATCC 49128
Pseudomonas alcaligenes	ATCC 14909
Ralstonia insidiosa	ATCC 49129
Ralstonia pickettii	ATCC 27511
Raoultella planticola (Klebsiella planticola)	ATCC 31900
Raoultella ornithinolytica	CDC# 0134
Raoultella terrigena (Klebsiella terrigena)	ATCC 55553
Shigella boydiiE	ATCC 9207
Shigella sonneiE	ATCC 25931
Shigella flexneriE	ATCC 9199
Vibrio furnissii	NCTC11218
Vibrio alginolyticus	ATCC 17749
Vibrio parahaemolyticus	ATCC 17802
Yersinia enterocolitica subsp enterocolitica	ATCC 9610
Yersinia ruckeri	ATCC 29473
Yersinia kristensenii	ATCC 33639

A. Cross-reactivity seen with Acinetobacter baumanii at a concentration > 1x104 CFU/mL.

B. Cross-reactivity seen with Enterobacter cloacae complex at a concentration > 1x10° CFU/mL.

C. Cross-reactivity seen with Enterobacter (non-cloacae complex) at a concentration > 1x106 CFU/mL and Serration of > 1x10' CFU/mL.

D. Cross-reactivity seen with the Fusobacterium nucleatum assay.

E. Cross-reactivity seen with the Escherichia coli assay.

{85}------------------------------------------------

Table 79: Off-Panel Gram-Positive Organisms Assessed for Cross-reactivity with the ePlex BCID-GN Panel (Exclusivity)

Gram Positive Organisms	Strain ID
Actinomyces odontolyticus	ATCC 17929
Clostridium perfringens	ATCC 13124
Corynebacterium jeikeium	ATCC BAA-949
Corynebacterium renale	ATCC 19412
Corynebacterium ulcerans	ATCC 51799
Corynebacterium xerosis*	ATCC 373
Corynebacterium durum	ATCC 33449
Corynebacterium diphtheriae*	ATCC 13812
Corynebacterium pseudodiphtheriticum	ATCC 10700
Corynebacterium striatum*	ATCC 43735
Corynebacterium urealyticum	ATCC 43044
Lactobacillus casei	ATCC 39392
Lactobacillus paracasei*	ATCC 25598
Lactobacillus acidophilus*	ATCC 314
Lactobacillus crispatus	ATCC 33197
Lactobacillus rhamnosus	ATCC 39595
Lactococcus lactis	ATCC 49032
Listeria innocua	ATCC 33090
Listeria monocytogenes	ATCC 7644
Micrococcus luteus	ATCC 10240
Peptostreptococcus anaerobius	ATCC 27337
Propionibacterium acnes	ATCC 11827
Rothia mucilaginosa	ATCC 25296

{86}------------------------------------------------

Fungal Pathogens	Strain ID	Concentration Tested
Aspergillus fumigatus*	ATCC 204305	2.50 x 106 CFU/mL
Candida orthopsilosis	ATCC 96139	1 x 107 CFU/mL
Candida metapsilosis	ATCC 96144	1 x 107 CFU/mL
Candida tropicalis	ATCC 1369	1 x 107 CFU/mL
Cryptococcus grubii	ATCC 208821	1 x 107 CFU/mL
Cryptococcus gattii	ATCC 76108	1 x 107 CFU/mL
Cryptococcus neoformans	ATCC 14116	1 x 107 CFU/mL
Geotrichum capitatum	ATCC 10663	1 x 107 CFU/mL
Histoplasma capsulatum	In silico	N/A
Penicillium marneffei	ATCC 200050	1 x 107 CFU/mL
Rhodotorula glutinis	ATCC 32765	1 x 107 CFU/mL
Rhodotorula mucilaginosa	ATCC 9449	1 x 107 CFU/mL
Rhodotorula minuta	ATCC 36236	1 x 107 CFU/mL
Saccharomyces cervisiae*	ATCC 18824	5.55 x 106 CFU/mL
Trichosporon dermatis	ATCC MYA-4294	1 x 107 CFU/mL
Trichosporon mucoides	ATCC 90046	1 x 107 CFU/mL

Table 80: Off-Panel Fungal Organisms Assessed for Cross-reactivity with the ePlex BCID-GN Panel (Exclusivity)

Table 81: Off-Panel Resistance Genes Assessed for Cross-reactivity with the ePlex BCID-GN Panel (Exclusivity)

Antimicrobial Resistance Genes	Strain ID	Concentration Tested
FOX (Carried by Klebsiella oxytoca)*A	JMI 954306	8 x 108 CFU/mL
MOX (Carried by Aeromonas hydrophila)	JMI 938982	1 x 109 CFU/mL
SME (Carried by Serratia marcescens)A	CDC #0091	1 x 109 CFU/mL
SHV (Carried by Klebsiella pneumoniae)A	CDC #0087	1 x 109 CFU/mL
TEM (Carried by Escherichia coli)A	NCTC 13351	1 x 109 CFU/mL

A. The on-panel organism associated with the resistance gene was detected by the BCID-GN panel as expected

{87}------------------------------------------------

Table 82: Off-Panel Gram-Negative Organisms Assessed for Cross-Reactivity with the ePlex BCID-GN Panel based on In Silico Analysis

Cross-reactive Organism	ePlex BCID-GN Target	No. ofSequences	Predicted Cross-Reactive Sequences*n (%)
Fusobacterium hwasookii	F. nucleatum	10	5 (50%)
Haemophilus aegyptius	H. influenzae	3	3 (100%)
Klebsiella michiganensis	Klebsiella oxytoca	40	40 (100%)
Pseudomonas denitrificans	Pseudomonas aeruginosa	17	16 (94.1%)

Table 83: Off-Panel Gram-Positive Organisms Assessed for Cross-Reactivity with the Pan Gram-Positive Assay based on In Silico Analysis

Organism	Number ofSequences	Predicted Cross-ReactiveSequencesn (%)
Brevibacterium halotolerans	3	3 (100%)
Domibacillus indicus	1	1 (100%)
Domibacillus robiginosus	1	1 (100%)
Salinibacillus aidingensis	2	1 (50%)
Terribacillus aidingensis	1	1 (100%)
Terribacillus halophilus	2	1 (50%)
Terribacillus saccharophilus	1	1 (100%)
Planomicrobium okeanokoites	1	1 (100%)
Lactococcus chungangensis	4	4 (100%)
Lactococcus laudensis	1	1 (100%)
Lactococcus piscium	18	18 (100%)
Lactococcus plantarum	6	5 (83.8%)
Lactococcus raffinolactis	49	46 (93.9%)
Okadaella gastrococcus	4	4 (100%)

{88}------------------------------------------------

Bottle Positivity

Several representative bacterial and fungal organisms were spiked into blood culture bottles along with the manufacturer's recommended volume of human whole blood and grown to positivity in a commercially-available continuously monitoring blood culture system. Bottles were removed from the incubator within two hours of being identified as positive as well as eight hours after bottle positivity. At least two independent positive blood culture replicates and three blood replicates were quantified for each organism on culture plates. Organisms tested and approximate bottle positivity concentrations are summarized in Table 84. Concentrations shown below represent approximate levels that may be observed in a clinical setting. All estimated bottle positivity concentrations are equivalent or greater than the established Limit of Detection (LOD) for each of the assays of the ePlex BCID-GP Panel. The following bottle types were used for the Bottle Positivity Study: BD BACTEC Plus Aerobic/F blood culture bottle (E. faecium, S. aureus, S.anginosus, A. baumannii, E. cloacae, E. coli, H. influenzae, K. oxytoca, N. meningitidis, P. auerginosa, and S. marcescens), BD BACTEC Lytic/10 Anaerobic/F (B. fragilis and F. nucleatum), and BD BACTEC Myco F/Lytic blood culture bottle (C. albicans).

Organism	Strain ID	Mean Bottle PositivityConcentration	Mean Bottle Positivity+8 hours Concentration
Gram-positive Organisms
Enterococcus faecium	ATCC BAA-2317	$4.9 x 10^7 CFU/mL$	$3.6 x 10^7 CFU/mL$
Staphylococcus aureus	NRS 483	$2.8 x 10^7 CFU/mL$	$2.1 x 10^7 CFU/mL$
Streptococcus anginosus	ATCC 33397	$4.1 x 10^7 CFU/mL$	$4.0 x 10^8 CFU/mL$
Gram-negative Organisms
Acinetobacter baumannii	NCTC 13301	$4.4 x 10^8 CFU/mL$	$3.8 x 10^8 CFU/mL$
Bacteroides fragilis	ATCC 700786	$4.7 x 10^8 CFU/mL$	$6.7 x 10^9 CFU/mL$
Enterobacter cloacae	NCTC 13464	$2.8 x 10^8 CFU/mL$	$7.7 x 10^8 CFU/mL$
Escherichia coli	NCTC 13476	$2.3 x 10^8 CFU/mL$	$1.5 x 10^9 CFU/mL$
Fusobacterium nucleatum	ATCC 31647	$6.5 x 10^7 CFU/mL$	$4.9 x 10^8 CFU/mL$
Haemophilus influenzae	ATCC 19418	$6.9 x 10^8 CFU/mL$	$1.2 x 10^9 CFU/mL$
Klebsiella oxytoca	CDC #0147	$9.3 x 10^8 CFU/mL$	$1.5 x 10^9 CFU/mL$
Neisseria meningitidis	ATCC 13102	$3.2 x 10^7 CFU/mL$	$2.1 x 10^8 CFU/mL$
Pseudomonas aeruginosa	NCTC 13476	$1.6 x 10^8 CFU/mL$	$8.4 x 10^8 CFU/mL$
Serratia marcescens	ATCC 14041	$1.2 x 10^9 CFU/mL$	$2.2 x 10^9 CFU/mL$
Fungal Organisms
Candida albicans	ATCC 90082	$1.6 x 10^6 CFU/mL$	$1.4 x 10^6 CFU/mL$

			Table 84: Bottle Positivity Concentrations
--	--	--	--------------------------------------------

{89}------------------------------------------------

Reproducibility

Three positive mixes including 11 on-panel organisms and 5 antibiotic resistance genes representing 17 targets at two concentrations and one negative mix including an off-panel organism were tested. Concentrations in the positive mixes reflected those observed at time of bottle positivity plus 8 hours and time of bottle positivity, and the negative mix contained Cutibacterium granulosum grown in BD BACTEC Lytic/10 Anaerobic/F blood culture bottles to bottle positivity and bottle positivity plus eight hours, which is expected to yield a negative result. Bottle concentrations used in this study are summarized in Table 85. Each of the three positive mixes at two concentrations and the one negative mix were tested a minimum of 108 times. Testing occurred at three sites, with two operators testing the mixes over six days using three cartridge lots. For the negative mix, agreement with the expected negative result was 100% for all targets in the ePlex BCID-GN Panel.

Organism	Bottle PositivityConcentration	Bottle Positivity +8Hours Concentration
Acinetobacter baumannii (OXA)	1 x 108 CFU/mL	1 x 109 CFU/mL
Enterobacter cloacae (CTX-M, KPC)	1 x 108 CFU/mL	1 x 109 CFU/mL
Escherichia coli (IMP)	1 x 108 CFU/mL	1 x 109 CFU/mL
Fusobacterium nucleatum	1 x 107 CFU/mL	1 x 108 CFU/mL
Haemophilus influenzae	1 x 108 CFU/mL	1 x 109 CFU/mL
Klebsiella oxytoca	1 x 108 CFU/mL	1 x 109 CFU/mL
Neisseria meningitidis	3 x 107 CFU/mL	3 x 108 CFU/mL
Pseudomonas aeruginosa (VIM)	1 x 108 CFU/mL	1 x 109 CFU/mL
Serratia marcescens	1 x 108 CFU/mL	1 x 109 CFU/mL
Candida albicans (Pan Candida target)	1 x 106 CFU/mL	1 x 107 CFU/mL
Staphylococcus aureus (Pan Gram-Positive target)	1 x 107 CFU/mL	1 x 108 CFU/mL

Table 85: Bottle Positivity Concentrations

The percent agreement of each target with the expected result is summarized in Tables 86-102. The ePlex BCID-GN assay demonstrates a high level of agreement (≥98%) with the expected results.

{90}------------------------------------------------

Concentration of Acinetobacterbaumannii	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	179/179	100	(97.9-100)
	2	178/179	99.4	(96.9-99.9)
	3	180/180	100	(97.9-100)
	All	537/538	99.8	(99.0-100)

Table 86: Percent Agreement for Acinetobacter baumannii

CI=Confidence Interval

Table 87: Percent Agreement for Enterobacter cloacae complex

Concentration ofEnterobacter cloacae	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	35/35	100	(90.1-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Negative	1	180/180	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

{91}------------------------------------------------

Concentration ofEscherichia coli	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/35	100	(90.1-100)
	3	36/36	100	(90.4-100)
	All	107/107*	100	(96.5-100)
Negative	1	179/179	100	(97.9-100)
	2	180/180	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

Table 88: Percent Agreement for Escherichia coli

Two samples had a false positive Bacteroides fragilis result.

Table 89: Percent Agreement for Fusobacterium nucleatum

Concentration of Fusobacteriumnucleatum	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/35	100	(90.1-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(1x107 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108*	100	(96.6-100)
Negative	1	179/179	100	(97.9-100)
	2	180/180	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

One sample had a false positive Fusobacterium necrophorum result.

{92}------------------------------------------------

Concentration ofHaemophilus influenzae	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	179/179	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	538/538	100	(99.3-100)

Table 90: Percent Agreement for Haemophilus influenzae

Table 91: Percent Agreement for Klebsiella oxytoca

Concentration ofKlebsiella oxytoca	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	179/179	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	538/538	100	(99.3-100)

{93}------------------------------------------------

Concentration ofNeisseria meningitidis	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(3x108 CFU/mL)	1	35/35	100	(90.1-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(3x107 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	180/180	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

Table 92: Percent Agreement for Neisseria meningitidis

Table 93: Percent Agreement for Pseudomonas aeruginosa

Concentration of Pseudomonasaeruginosa	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/35	100	(90.1-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	179/179	100	(97.9-100)
	2	180/180	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

{94}------------------------------------------------

	Table 94: Percent Agreement for Serratia

Concentration ofSerratia marcescens	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	35/35	100	(90.1-100)
Bottle Positive+ 8 Hours(1x109 CFU/mL)	2	36/36	100	(90.4-100)
Bottle Positive+ 8 Hours(1x109 CFU/mL)	3	36/36	100	(90.4-100)
Bottle Positive+ 8 Hours(1x109 CFU/mL)	All	107/107	100	(96.5-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
Bottle Positive(1x108 CFU/mL)	2	33/33	100	(89.6-100)
Bottle Positive(1x108 CFU/mL)	3	33/33	100	(89.6-100)
Bottle Positive(1x108 CFU/mL)	All	102/102	100	(96.4-100)
Negative	1	180/180	100	(97.9-100)
Negative	2	179/179	100	(97.9-100)
Negative	3	180/180	100	(97.9-100)
Negative	All	539/539	100	(99.3-100)

Table 95: Percent Agreement for Serratia marcescens

Concentration ofSerratia marcescens	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	35/35	100	(90.1-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
		All	107/107	100
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
		All	108/108	100
Negative	1	180/180	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
		All	539/539	100

{95}------------------------------------------------

Concentration ofCandida albicans	Site	Agreement with Expected Results
	Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x10 $^{7}$ CFU/mL)	1	35/35	100	(90.1-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(1x10 $^{6}$ CFU/mL)	1	35/36	97.2	(85.8-99.5)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/108	99.1	(94.9-99.8)
Negative	1	180/180	100	(97.9-100)
	2	179/179	100	(97.9-100)
	3	180/180	100	(97.9-100)
	All	539/539	100	(99.3-100)

Table 96: Percent Agreement for Pan Candida

Table 97: Percent Agreement for Pan Gram-Positive

Concentration of	Site	Agreement with Expected Results
Staphylococcus aureus		Agreed / N	%	95% CI
	1	36/36	100	(90.4-100)
Bottle Positive+ 8 Hours	2	36/36	100	(90.4-100)
(1x108 CFU/mL)	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
	l	34/36	94.4	(81.9-98.5)
Bottle Positive	2	35/35	100	(90.1-100)
(1x107 CFU/mL)	3	36/36	100	(90.4-100)
	All	105/107	98.1	(93.4-99.5)
	l	179/179	100	(97.9-100)
Negative	2	179/180	99.4	(96.9-99.9)
	3	180/180	100	(97.9-100)
	All	538/539	99.8	(99.0-100)

{96}------------------------------------------------

Concentration of	Site	Agreement with Expected Results
Enterobacter cloacae(CTX-M+, KPC+)		Agreed / N	%	95% CI
	1	36/36	100	(90.4-100)
Bottle Positive+ 8 Hours	2	36/36	100	(90.4-100)
(1x109 CFU/mL)	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
	1	35/35	100	(90.1-100)
Bottle Positive	2	36/36	100	(90.4-100)
(1x108 CFU/mL)	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
	1	144/144	100	(97.4-100)
	2	143/143	100	(97.4-100)
Negative	3	144/144	100	(97.4-100)
	All	431/431	100	(99.1-100)

Table 98: Percent Agreement for CTX-M

Table 99: Percent Agreement for IMP

Concentration ofEscherichia coli (IMP+)	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/36	97.2	(85.8-99.5)
	3	36/36	100	(90.4-100)
	All	107/108	99.1	(94.9-99.8)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/35	100	(90.1-100)
	3	35/36	97.2	(85.8-99.5)
	All	106/107	99.1	(94.9-99.8)
Negative	1	143/143	100	(97.4-100)
	2	144/144	100	(97.4-100)
	3	144/144	100	(97.4-100)
	All	431/431	100	(99.1-100)

{97}------------------------------------------------

Concentration ofEnterobacter cloacae(CTX-M+, KPC+)	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(1x108 CFU/mL)	1	35/35	100	(90.16-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.4-100)
Negative	1	144/144	100	(97.4-100)
	2	143/143	100	(97.4-100)
	3	144/144	100	(97.4-100)
	All	431/431	100	(99.1-100)

Table 100: Percent Agreement for KPC

Table 101: Percent Agreement for OXA

Concentration of	Site	Agreement with Expected Results
Acinetobacter baumannii(OXA-23+)		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	143/143	100	(97.4-100)
	2	143/143	100	(97.4-100)
	3	144/144	100	(97.4-100)
	All	430/430	100	(99.1-100)

{98}------------------------------------------------

Table 102: Percent Agreement for VIM

Concentration of	Site	Agreement with Expected Results
Pseudomonas aeruginosa(VIM+)		Agreed / N	%	95% CI
Bottle Positive+ 8 Hours(1x109 CFU/mL)	1	36/36	100	(90.4-100)
	2	35/35	100	(90.1-100)
	3	36/36	100	(90.4-100)
	All	107/107	100	(96.5-100)
Bottle Positive(1x108 CFU/mL)	1	36/36	100	(90.4-100)
	2	36/36	100	(90.4-100)
	3	36/36	100	(90.4-100)
	All	108/108	100	(96.6-100)
Negative	1	143/1437	100	(97.4-100)
	2	144/144	100	(97.4-100)
	3	144/144	100	(97.4-100)
	All	431/431	100	(99.1-100)

{99}------------------------------------------------

Interfering Substances and Sample Matrix Equivalency (Bottle Evaluation)

Three organism mixes consisting of 12 on-panel organisms representing 16 targets and negative blood matrix were used to assess potentially interfering substances and bottle types for interference. The concentration of each organism tested is summarized in Table 103.

Table 103: Organism Concentrations for Interfering Substance and Bottle Equivalencv Evaluations

Organism	Concentration
Acinetobacter baumanii	4 x 108 CFU/mL
Bacteroides fragilis	4 x 108 CFU/mL
Enterobacter aerogenes	2 x 108 CFU/mL
Enterobacter cloacae (CTX-M)	2 x 108 CFU/mL
Escherichia coli (OXA)	2 x 108 CFU/mL
Haemophilus influenzae	6 x 108 CFU/mL
Klebsiella oxytoca	9 x 108 CFU/mL
Neisseria meningitidis	3 x 107 CFU/mL
Pseudomonas aeruginosa (IMP)	1 x 108 CFU/mL
Serratia marcescens	1 x 109 CFU/mL
Staphylococcus aureus (Pan Gram-Positive target)	2 x 107 CFU/mL
Candida albicans (Pan Candida target)	1 x 106 CFU/mL

Interfering Substances

Eighteen substances were used to assess the ePlex BCID-GN Panel for potential interference. The organisms in Table 103 were spiked into negative blood matrix and tested in triplicate with and without each potentially interfering substance. Negative blood matrix was tested to control for potential positive interference. Potentially interfering substances are summarized in Table 104. None of the eighteen substances commonly found in blood culture specimens or as medications commonly used to treat skin or bloodstream infections were found to inhibit the ePlex BCID-GN Panel at clinically relevant concentrations. The effect of interfering substances has only been evaluated for the substances listed in Table 104. Interference due to substances other than those described in this section can lead to erroneous results.

{100}------------------------------------------------

Endogenous Substances	Testing Concentration
Bilirubin	60 µg/mL
Hemoglobin	0.6 g/L
Human Genomic DNA	6 x 105 copies/mL
Triglycerides	1000 mg/dL
γ-globulin	1.7 g/dL
Exogenous Substances	Testing Concentration
Amoxicillin/Clavulanate	3.5 µg/mL
Amphotericin B	2 µg/mL
Caspofungin	5 µg/mL
Ceftriaxone	0.23 mg/mL
Ciprofloxacin	3 mg/L
Fluconazole	25 mg/L
Flucytosine	90 µg/mL
Gentamicin sulfate	3 µg/mL
Heparin	0.9 U/mL
Imipenem	83 µg/mL
Sodium Polyanethol Sulfonate	0.25% w/v
Tetracycline	5 mg/L
Vancomycin	30 mg/L

Table 104: Potentially Interfering Substances: Substance List

Sample Matrix Equivalency (Bottle Evaluation)

Thirteen bottle types were tested for interference with each of the organisms listed in Table103.

Five replicates of each organism were tested in each of two bottle lots. Negative blood matrix was run as a negative control. Twelve bottle types tested showed no interference for any of the targets tested. One of three lots of the BACTEC™ Lytic Anaerobic bottles tested showed reduced sensitivity for some targets. A summary of the bottle types assessed and the study outcomes is found in Table 105.

{101}------------------------------------------------

Manufacturer	Bottle Brand	Bottle Type	Study Outcome
BD	BACTECTM	Plus Aerobic	No interference observed
BD	BACTEC	Plus Anaerobic	No interference observed
BD	BACTEC	Standard Aerobic	No interference observed
BD	BACTEC	Standard Anaerobic	No interference observed
BD	BACTEC	Peds Plus™	No interference observed
BD	BACTEC	Lytic Anaerobic*	False negative results were observed for PanCandida, Enterobacter cloacae, Escherichiacoli, CTX-M and OXA in one of three lots.*
bioMérieux	BACT/ALERT®	SA Standard Aerobic	No interference observed
bioMérieux	BACT/ALERT	SN Standard Anaerobic	No interference observed
bioMérieux	BACT/ALERT	FA Plus	No interference observed
bioMérieux	BACT/ALERT	FN Plus	No interference observed
bioMérieux	BACT/ALERT	PF Plus	No interference observed
Thermo Scientific™	VersaTREK™	REDOX™ 1 EZ Draw Aerobic	No interference observed
Thermo Scientific	VersaTREK	REDOX 2 EZ Draw Anaerobic	No interference observed

Table 105: Sample Matrix Equivalency (Bottle Evaluation) Bottle Types

2/15 replicates were false negative for Pan Canades was false negative for Enterobacter cloacag; 1/15 replicates was false negative for Escherichia coli (OXA-48); 2/15 replicates were false negative for CTX-M

Carryover and Cross-Contamination

Carryover and cross-contamination were evaluated for the ePlex BCID-GN Panel within and between runs by alternating high positive and negative samples across multiple runs over 5 rounds of testing. A high-titer mix of OXA positive Escherichia coli, CTX-M and KPC positive Enterobacter cloacae, Salmonella enterica, and Enterococcus faecalis (a Pan Gram-Positive target organism) was prepared at 1 x 10° CFU/mL each as well as Candida krusei (a Pan Candida target organism) at 1x107 CFU/mL to simulate clinically relevant high positive samples for positive testing. Negative blood culture matrix was used to represent negative samples. Over 120 runs, all valid positive runs resulted in detection of Escherichia coli, Enterobacter cloacae complex, Salmonella, OXA, CTX-M, KPC, Pan Gram-Positive, and Pan Candida and no false positives were detected in the negative runs.

{102}------------------------------------------------

Competitive Inhibition Study

Competitive inhibition was evaluated for the ePlex BCID-GN Panel by pairing eight clinically relevant organisms (including a Pan Gram-Positive assay target and an off-panel gram-positive organism) in four simulated dual infection sample mixes. Each dual infection mix was tested in combination with each of the three other mixes, such that all organisms were tested at low titer (concentrations expected at bottle positivity) while in the presence of other organisms at higher titer (concentrations expected at 8 hours beyond bottle positivity, or ~ one log higher than that expected at bottle positivity). No competitive inhibition was observed in any replicates of the twelve testing conditions. A summary of the organisms assessed and testing concentrations is found in Table 106.

Organism	HighConcentration	LowConcentration
Klebsiella pneumoniae	1 x 109 CFU/mL	9 x 108 CFU/mL
Escherichia coli (CTX-M+)	1 x 109 CFU/mL	2 x 108 CFU/mL
Enterobacter cloacae (VIM+)	7 x 108 CFU/mL	2 x 108 CFU/mL
Klebsiella oxytoca (KPC+)	1 x 109 CFU/mL	9 x 108 CFU/mL
Pseudomonas aeruginosa (IMP+)	8 x 108 CFU/mL	1 x 108 CFU/mL
Serratia marcescens	2 x 109 CFU/mL	1 x 109 CFU/mL
Staphylococcus aureus	1 x 108 CFU/mL	2 x 107 CFU/mL
Corynebacterium striatumA	2 x 109 CFU/mL	4 x 106 CFU/mL

Table 106: Competitive Inhibition Organisms and Concentrations Tested

Off-panel organism A.

{103}------------------------------------------------

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Regulation Number and Section

§ 866.3365 Multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures.

(a)
Identification. A multiplex nucleic acid assay for identification of microorganisms and resistance markers from positive blood cultures is a qualitative in vitro device intended to simultaneously detect and identify microorganism nucleic acids from blood cultures that test positive by Gram stain or other microbiological stains. The device detects specific nucleic acid sequences for microorganism identification as well as for antimicrobial resistance. This device aids in the diagnosis of bloodstream infections when used in conjunction with other clinical and laboratory findings. However, the device does not replace traditional methods for culture and susceptibility testing.(b)
Classification. Class II (special controls). The special control for this device is FDA's guideline document entitled “Class II Special Controls Guideline: Multiplex Nucleic Acid Assay for Identification of Microorganisms and Resistance Markers from Positive Blood Cultures.” For availability of the guideline document, see § 866.1(e).