Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The description focuses on the biological and chemical processes of nucleic acid detection and identification, with no mention of AI or ML algorithms for data analysis or interpretation.

Therapeutic

No.
This device is an in vitro diagnostic test used for the qualitative detection and identification of microbial organisms and antimicrobial resistance genes from positive blood cultures. It aids in diagnosis and is not directly used for treatment or therapy.

Diagnostic

Yes

The text explicitly states: "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." and "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."

SaMD (Software as a Medical Device)

No

The device description clearly outlines a physical in vitro diagnostic test that involves nucleic acid extraction, PCR, hybridization, and electrochemical detection using a cartridge and an instrument (ePlex Instrument). This involves significant hardware components and processes beyond just software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Explicitly Stated: The "Intended Use / Indications for Use" section clearly states: "The GenMark ePlex® Blood Culture Identification Gram-Negative (BCID-GN) Panel is a qualitative nucleic acid multiplex in vitro diagnostic test..."
Purpose: The device is intended 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 culture." This is a diagnostic purpose performed outside of the body (in vitro) on a biological sample (blood culture).
Sample Type: It is performed directly on "blood culture samples," which are biological specimens.
Methodology: The "Device Description" details the use of "nucleic acid extraction, target amplification via polymerase chain reaction (PCR) or reverse transcription PCR (RT-PCR) and hybridization of target DNA," all of which are common techniques used in in vitro diagnostic tests.
Clinical Context: The results are intended to "aid in the diagnosis of bloodstream infection when used in conjunction with other clinical information." This further confirms its diagnostic role.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

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 oxvtoca, Klebsiella pneumoniae group, Morganella morganii, Neisseria meningitidis, Proteus, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella, Serratia, Serratia marcescens, Stenotrophomonas maltophilia, CTX-M (blacrx-M), IMP (blanм) , KPC (blakec) , NDM (blandM), 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(blacrx-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 (blakec), and metallo-beta-lactamases IMP (blanм), VIM (blaviм), 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 gramnegative 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.

Product codes (comma separated list FDA assigned to the subject device)

PEN, PAM, PEO

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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

The study included patients of all ages, as indicated by the "Expected Value by Age Group (Prospective Samples)" table, which categorizes data for ages

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).

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: on the left, there is a symbol representing the Department of Health & Human Services, and on the right, there is the text "FDA U.S. FOOD & DRUG ADMINISTRATION" in blue. The FDA logo is a recognizable symbol of the agency responsible for regulating food and drug products in the United States.

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

April 27, 2022

Re: K213236

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 29, 2021 Received: September 30, 2021

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

1

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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Noel Gerald, Ph.D. Chief Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K213236

Device Name

ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel

Indications for Use (Describe)

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 sakazakii, Enterobacter cloacae complex, Enterobacter (non-cloacae complex), Escherichia coli, Fusobacterium necrophorum, Fusobacterium nucleatum, Haemophilus influenzae, Klebsiella oxytoca, Klebsiella pneumoniae group, Morganii, Neisseria meningitidis, Proteus, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella, Serratia marcescens, Stenotrophomonas maltophilia, CTX-M (blaCTX-M), IMP (blaMP) , KPC (blaKPC) , NDM (blaNDM), OXA (blaOXA) (OXA-23 and OXA-48 groups only), and VIM (blaVIM).

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 betalactamase (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 (blaIMP), 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 gramnegative 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 coinfections. These include a broad Pan Gram-Positive assay (which is designed to detect Bacillus cereus group, Bacillus subtilis group, Enterococcus, Staphylococus, and Streptococcus), as well as a Pan Candida assay, which is designed to detect four Candida species: Candida albicans, 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 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.

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Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

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5.0 510(k) Summary

The 510(k) Summary was updated to align with the most current revision of the package insert, which incorporates changes in the Limit of Detection (LoD) section based on the results from studies conducted to support implementation of a design change (i.e., addition of oligonucleotides to improve robustness and inclusivity of the E.coli, Citrobacter, Enterococcus, and P. aeruginosa assays). The addition of oligonucleotides to specific PCR reactions introduces performance risks to the resident assays contained in those affected PCR reactions. As a result, the following studies were conducted to mitigate the identified risks to product performance.

1. Evaluation of overall BCID-GN Panel Performance: this study utilized a multianalyte test mix (containing a representative analyte from each of the eight multiplex PCR pools) to systematically assess overall performance of the BCID-GN Panel. This design was utilized to ensure that the proposed change does not introduce any unexpected issues that result in a systematic assay failure.
1. LoD Verification Study: this study verified that the LoDs of the targets affected by the change are not adversely impacted. These results demonstrate that the analytical sensitivity of the test remains equivalent.
1. Clinical Sample Evaluation: this study utilized characterized clinical samples to verify that the proposed change does not adversely impact clinical performance.

All studies met the predetermined acceptance criteria demonstrating no adverse impact to the BCID-GN Panel performance, which supports implementation of the proposed change.

The changes to the 510(k) Summary include identification of the following strains (in Table 56 of the 510(k) Summary) that were tested as part of the LoD verification study:

H. influenzae (ATCC33930) .
. N. meningitidis (NCTC10026)
E. coli (JHU01-D80401147) .
. P. aeruginosa (SDx071)

All other information in the 510(k) Summary are unchanged.

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510(k) Summary

Summary of Safety and Effectiveness

Submitter Information

| Submitter: | GenMark Diagnostics, Incorporated
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., RAC
Vice President, Quality, Regulatory and Clinical Affairs |
| Phone: | 760-448-4308 |
| Fax: | 760-683-6961 |
| E-mail: | Al.Maderazo@genmarkdx.com |
| Alternate Contact: | Beth Stofka
Sr. Regulatory Affairs Specialist |
| Phone: | 760-579-4778 |
| Fax: | 760-683-6961 |
| E-mail: | Beth.Stofka@genmarkdx.com |
| Date Prepared: | April 4, 2022 |

Name of Device and Classification

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

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Predicate Device

Predicate: The ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel; GenMark Diagnostics, Inc .; K182619

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.

7

Intended Use/Indications for Use

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 oxvtoca, Klebsiella pneumoniae group, Morganella morganii, Neisseria meningitidis, Proteus, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella, Serratia, Serratia marcescens, Stenotrophomonas maltophilia, СТХ-М (blacтх-м), IMP (blaмг) , КРС (blakec) , NDM (bland), OXA (blaoxa) (OXA-23 and OXA-48 groups only), and VIM (blavim).

The ePlex BCID-GN Panel contains assays for the detection of genetic determinants associated with resistance to antimicrobial agents including CTX-M(blacrx-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 (blakec), and metallo-beta-lactamases IMP (blanм), VIM (blaviм), 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.

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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.

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

The updated GenMark ePlex Blood Culture Identification Gram-Negative (BCID-GN) Panel (identified as the "SubjectDevice") and the legally marketed device, the original GenMark ePlex BCID-GN Panel (K182619) (identified as the "Predicate Device") are described below:

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Characteristic	Predicate Device (K182619)	Subject Device (Updated)
Product Name	ePlex BCID-GN Panel	Same
Manufacturer	GenMark Diagnostics, Inc.	Same
Organisms
Detected	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 Saltophiliapyogenes (GAS)	Same
Additional strains were tested as part of the Limit of Detection (LoD) study which include the following: H. influenzae (ATCC33930) N. meningitidis (NCTC10026) E. coli (JHU01-D80401147) P. aeruginosa (SDx071)
Resistance Genes
Detected	CTX-M, IMP, KPC, NDM, OXA, andVIM	Same
Indication for
Use	The ePlex BCID-GN Panel is indicated as an aid in the diagnosis of specific agents of bacteremia. The use of additional laboratory testing (e.g. sub- culturing of positive blood cultures for identification of organisms not detected by the ePlex BCID-GN Panel and for susceptibility testing, differentiation of mixed growth, and association of antimicrobial resistance marker genes toa specific organism) and clinical presentation must be taken into consideration in the final diagnosis of blood stream infection.	Same
Specimen Type	Blood culture samples identified as positive by a continuous monitoringblood culture system that demonstrates the presence of organisms as confirmed by Gram stain.	Same
Characteristic	Predicate Device (K182619)	Subject Device (Updated)
Chemistry	Reagents on cartridge include: sample
lysis and nucleic acid extraction, PCR
amplification and hybridization-based
electrochemical detection reagents.	Same
Hardware	GenMark ePlex Instrument & SingleUse
Cartridge	Same
SoftwareInterface
Result Reporting	GenMark ePlex System Software GenMark ePlex BCID-GN Panel
Software	Same

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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.

Summary 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)

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. 12/13 false positive organisms were investigated using PCR/sequencing; the discrepant organism was 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.
iii. 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.
v. In 1/1 false positive K. oxytoca sample, K. oxytoca was detected.
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 ePlex
BCID-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 ePlex | Number
Samples | Discrepant | | |
| BCID-GN Panel in Retrospective Clinical Samples | | | | (Number
Discrepant) | Organism(s) /
Resistance Marker(s)A,B |
| Target 1 | Target 2 | Target 3 | Resistance
Marker | | |
| 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. cloacae
complex | K. pneumoniae group | | | 1 (0) | |
| E. cloacae
complex | P. aeruginosa | Pan Gram-Positive | | 1 (1) | P. aeruginosa (1) |
| E. cloacae
complex | Pan Candida | | | 1 (1) | Pan Candida (1) |
| E. cloacae
complex | 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. pneumoniae
group | Pan Gram-Positive | | | 4 (1) | Pan Gram-Positive (1) |
| K. pneumoniae
group | Pan Gram-Positive | S. marcescens | | 1 (1) | K. pneumoniae group (1) |
| Distinct Co-Detection Combinations Detected by the ePlex
BCID-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 | | |
| K. pneumoniae
group | 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) | |

57

58

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

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.

In 2/2 false positive B. fragilis samples, B. fragilis was detected. 11.

In 1/1 false positive Citrobacter sample, Citrobacter was detected. 111.

In the one false positive E. cloacae complex sample, PCR/sequencing was indeterminate. 1V.

In 1/1 false positive K. oxytoca sample, K. oxytoca was detected. v.

In 3/3 false positive K. pneumoniae group samples, K. pneumoniae group was detected. vi.

V11. In 1/1 false positive sample, M. morganii was detected.

N. meningitidis was not detected in the 1 N. meningitidis false positive sample. V111.

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

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. X1.

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 Comparator
Method(s) in Prospective 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. faecium | Staphylococcus | | | 1 (0) | |
| A. baumannii | Staphylococcus | | | | 1 (0) | |
| Achromobacter
xylosoxidans * | E. cloacae | | | | 1 (0) | |
| Distinct Co-Detection Combinations Detected by the Comparator
Method(s) in Prospective Clinical Samples | | | | | Number
Samples
(Number
Discrepant) | Discrepant
Organism(s) /
Resistance
Marker(s)A |
| Target 1 | Target 2 | Target 3 | Target 4 | Resistance
Marker | | |
| Acinetobacter
lwoffii* | Staphylococcus
hominis | | | | 1 (0) | |
| Acinetobacter
pittii* | S. aureus | | | | 1 (0) | |
| Aerococcus
viridans* | K. oxytoca | S. epidermidis | Staphylococcus
cohnii | | 1 (0) | |
| Aerococcus
viridans* | Staphylococcus
hominis | | | | 1 (0) | |
| B. fragilis | Clostridium
species* | | | | 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 | Providencia
stuartii* | | | 1 (0) | |
| Candida lusitaniae* | S. liquefaciens | | | | 1 (0) | |
| Citrobacter
amalonaticus* | E. coli | | | | 1 (0) | |
| E. aerogenes | K. oxytoca | Leclercia
adecarboxylata* | | | 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 | Providencia
stuartii* | S. anginosus
group | 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 | Providencia
stuartii* | | | | 1 (0) | |
| Enterobacteriaceae* | K. pneumoniae | | | | 1 (0) | |
| Distinct Co-Detection Combinations Detected by the Comparator | | | | | | Discrepant
Organism(s) /
Resistance
Marker(s)A |
| Method(s) in Prospective Clinical Samples | | | | | | |
| Target 1 | Target 2 | Target 3 | Target 4 | Resistance
Marker | Number
Samples
(Number
Discrepant) | |
| K. pneumoniae | Staphylococcus
haemolyticus | non-fermenting
Gram-Negative
bacilli * | | | 1 (1) | K. pneumoniae (1) |
| Lactococcus lactis * | P. mirabilis | | | | 1 (0) | |
| Micrococcus
luteus * | Sphingomonas
paucimobilis * | | | | 1 (0) | |
| P. aeruginosa | P. mirabilis | Streptococcus -
viridans group | | | 1 (0) | |
| P. aeruginosa | S. maltophilia | | | | 1 (1) | S. maltophilia (1) |
| P. aeruginosa | Staphylococcus
haemolyticus | | | | 1 (0) | |
| P. mirabilis | Staphylococcus | | | | 1 (0) | |
| S. maltophilia | Streptococcus | | | | 1 (1) | Streptococcus (1) |

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).

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 | | | Samples | Discrepant
Organism(s) /
Resistance
Marker(s)A |
|--------------------------------------------------------------------------------------------------------------|------------ | Target 1 | Target 2 Marker | (Number
Discrepant) | |
| A. baumannii | E. faecalis | A. baumannii | E. faecalis | A. baumannii | E. faecium | A. baumannii | Staphylococcus | Acinetobacter radioresistens* | P. vulgaris | Aeromonas caviae* | E. coli | Aeromonas veronii* | E. cloacae | B. fragilis | S. anginosus group | Distinct Co-Detection Combinations Detected by the Comparator Method(s) in
Retrospective Clinical Samples | | | Samples | Discrepant
Organism(s) / | |
| Target 1 | Target 2 Marker | (Number
Discrepant) | Resistance
Marker(s)A |
| C. albicans | E. faecalis | C. albicans | E. faecium hominis | | | 1 (1) | C. albicans | P. aeruginosa | C. albicans | S. epidermidis | C. braakii | E. cloacae oxytoca (1) |
| C. braakii | E. coli | C. braakii | Streptococcus oralis | C. freundii | Enterococcus | C. freundii | K. pneumoniae | C. freundii | K. pneumoniae hominis | | CTX-M | 1 (0) | C. glabrata | E. aerogenes | C. glabrata | P. mirabilis | C. koseri | E. faecalis | C. krusei | S. epidermidis | C. youngae | K. oxytoca | Clostridium
perfringens* | E. coli | E. aerogenes | S. anginosus group | E. cloacae | E. coli | E. cloacae | E. faecalis | E. cloacae | E. faecium | E. cloacae | E. faecium hominis | | | 1 (0) | E. cloacae | K. pneumoniae | E. cloacae | M. morganii | E. cloacae | S. anginosus group | E. cloacae | S. maltophilia | E. coli | E. faecalis | Distinct Co-Detection Combinations Detected by the Comparator Method(s) in
Retrospective Clinical Samples | | | Samples | Discrepant
Organism(s) / | |
| Target 1 | Target 2 Marker | (Number
Discrepant) | Resistance
Marker(s)A |
| E. coli | E. faecalis | E. coli | E. faecalis | E. coli | E. faecalis | E. coli | E. faecium | E. coli | E. faecium | E. coli | K. oxytoca infantarius | | | 1 (0) | E. coli | P. aeruginosa | E. coli | P. mirabilis | E. coli | P. mirabilis viridans group | | 1 (1) (1) |
| E. coli | Propionibacteria* | E. coli | S. anginosus gp (1) |
| E. coli | S. aureus | E. coli | S. pneumoniae | E. coli | Staphylococcus | E. faecalis | K. pneumoniae | E. faecalis | M. morganii | E. faecalis | M. morganii | E. faecalis | P. aeruginosa | E. faecalis | P. mirabilis | E. faecalis | S. maltophilia | E. faecalis | S. marcescens | E. faecium | K. pneumoniae | E. faecium | P. aeruginosa | E. faecium | P. mirabilis | K. oxytoca | S. anginosus group | K. pneumoniae | P. aeruginosa | K. pneumoniae | S. aureus | Distinct Co-Detection Combinations Detected by the Comparator Method(s) in
Retrospective Clinical Samples | | | Samples
(Number
Discrepant) | Discrepant
Organism(s) /
Resistance
Marker(s)^A | |
| Target 1 | Target 2 Marker | | | K. pneumoniae | Staphylococcus | P. aeruginosa | S. maltophilia | P. mirabilis | Peptostrept anaerobius* | | | | P. mirabilis | Providencia stuartii* | | | | P. mirabilis | Staphylococcus | Pseudomonas
putida * | S. epidermidis | S. maltophilia | S. aureus | S. marcescens | S. marcescens | Staphylococcus | S. marcescens | Streptococcus group | Streptococcus
salivarius | | | 1 (0) | | Number
-----------------------|-----------------------------------|-----------------------------------|---------------------------------------------|----------------------------------------------------------|---------------------------------------------------------|
| Target 3 | Target 4 | Resistance
| | | OXA | 2 (0) | |
| S. aureus | | | 1 (0) | |
| | | OXA | 1 (0) | |
| | | | 1 (0) | |
| | | OXA | 1 (0) | |
| Enterococcus casseliflavus | K. oxytoca | | 1 (1) | E. casseliflavus (1) |
| | | | 1 (1) | E. cloacae (1) |
| | | | 1 (0) | |
| Number
| Target 3 | Target 4 | Resistance
| | | | 1 (0) | |
| Staphylococcus
| C. albicans (1) |
| | | | 1 (1) | C. albicans (1) |
| | | | 1 (0) | |
| K. oxytoca | | | 1 (1) | C. braakii (1), K.
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (1) | Enterococcus (1) |
| | | | 2 (0) | |
| Staphylococcus
| |
| Staphylococcus | | | 1 (1) | Staphylococcus (1) |
| | | | 1 (1) | C. glabrata (1) |
| | | | 1 (0) | |
| | | | 1 (1) | S. epidermidis (1) |
| | | | 1 (1) | K. oxytoca (1) |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (1) | E. coli (1) |
| | | | 1 (1) | E. faecalis (1) |
| | | | 1 (0) | |
| Staphylococcus
| |
| | | | 1 (0) | |
| | | | 1 (1) | E. cloacae (1) |
| | | | 1 (0) | |
| | | | 1 (1) | S. maltophilia (1) |
| | | | 2 (0) | |
| Number
| Target 3 | Target 4 | Resistance
| | | CTX-M | 1 (0) | |
| K. pneumoniae | | | 1 (1) | E. coli (1) |
| P. mirabilis | | | 1 (0) | |
| | | | 1 (0) | |
| | | CTX-M | 1 (1) | E. faecium (1) |
| Streptococcus
| |
| | | | 1 (1) | P. aeruginosa (1) |
| | | | 1 (1) | E. coli (1) |
| P. vulgaris | Streptococcus -
| S. viridans group
| | | | 1 (0) | |
| | | | 1 (1) | S. anginosus group
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (1) | K. pneumoniae (1) |
| | | | 1 (0) | |
| P. vulgaris | | | 1 (0) | |
| S. aureus | | | 1 (1) | P. aeruginosa (1) |
| | | | 3 (0) | |
| | | | 1 (1) | E. faecalis (1) |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (1) | P. aeruginosa (1) |
| | | | 2 (1) | S. aureus (1) |
| Number
| Target 3 | Target 4 | Resistance
|
| | | | 1 (1) | Staphylococcus (1) |
| | | | 1 (1) | S. maltophilia (1) |
ococcus
| 1 (0) | |

| 2 (1) | P. mirabilis (1) |
| | | | 1 (0) | |
| | | 1 (0) | |
| | | | 1 (0) | |
| | | | 1 (0) | |
mitis
| |

61

62

63

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%).

64

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 | LoD
Concentration
(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 cloacae | Enterobacter amnigenus | ATCC 33072 | 1 x 106 |
| Enterobacter cloacae | Enterobacter cloacae | CDC#0154 | 1 x 106 |
| Enterobacter cloacae | Enterobacter cloacae | CDC#0154 | 1 x 106 |

Table 56: LoD Results Summary

Enterobacter (non-cloacae complex)	Enterobacter aerogenes	CDC#0161	$1 x 10^5$
	Enterobacter amnigenus	ATCC 33072	$1 x 10^6$
Enterobacter cloacae
Complex	Enterobacter cloacae	CDC#0154	$1 x 10^6$
	Enterobacter asburiae	ATCC 35957	$1 x 10^6$
	Enterobacter hormaechei	ATCC BAA-2082	$1 x 10^6$
Escherichia coli	Escherichia coli	CDC#0118	$1 x 10^7$
	Escherichia coli	NCTC 13441	$1 x 10^6$
	Escherichia coli	JHU01-D80401147	$1 x 10^7$

65

| Target | Organism | Strain | LoD
Concentration
(CFU/mL) |
|------------------------------|----------------------------------------------------|---------------|----------------------------------|
| Fusobacterium necrophorum | Fusobacterium necrophorum | ATCC 51357 | $1 x 10^8$ |
| Fusobacterium necrophorum | Fusobacterium necrophorum | ATCC 27852 | $1 x 10^7$ |
| Fusobacterium nucleatum | Fusobacterium nucleatum | ATCC 25586 | $1 x 10^7$ |
| Fusobacterium nucleatum | Fusobacterium nucleatum | ATCC 23726 | $1 x 10^5$ |
| Haemophilus influenzae | Haemophilus influenzae | ATCC 19418 | $1 x 10^5$ |
| Haemophilus influenzae | Haemophilus influenzae | ATCC 9006 | $1 x 10^7$ |
| Haemophilus influenzae | Haemophilus influenzae | ATCC 33930 | $1 x 10^4$ |
| Klebsiella oxytoca | Klebsiella oxytoca | ATCC 43165 | $1 x 10^7$ |
| Klebsiella oxytoca | Klebsiella oxytoca | ATCC 8724 | $1 x 10^7$ |
| Klebsiella pneumoniae group | Klebsiella pneumoniae | CDC#0160 | $1 x 10^6$ |
| Klebsiella pneumoniae group | Klebsiella pneumoniae | CDC#0107 | $1 x 10^6$ |
| Morganella morganii | Morganella morganii | ATCC 25829 | $1 x 10^7$ |
| Morganella morganii | Morganella morganii | CDC#0133 | $1 x 10^7$ |
| Neisseria meningitidis | Neisseria meningitidis | ATCC 13090 | $1 x 10^5$ |
| Neisseria meningitidis | Neisseria meningitidis | ATCC 13102 | $1 x 10^4$ |
| Neisseria meningitidis | Neisseria meningitidis | NCTC 10026 | $1 x 10^4$ |
| Proteus | Proteus vulgaris | ATCC 6896 | $1 x 10^7$ |
| Proteus | Proteus vulgaris | ATCC 6380 | $1 x 10^7$ |
| Proteus mirabilis | Proteus mirabilis | CDC#0159 | $1 x 10^6$ |
| Proteus mirabilis | Proteus mirabilis | ATCC 43071 | $1 x 10^6$ |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa | CDC#0103 | $1 x 10^6$ |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa | NCTC 13437 | $1 x 10^6$ |
| Pseudomonas aeruginosa | Pseudomonas aeruginosa | SDx071 | $1 x 10^5$ |
| Salmonella | Salmonella bongori | ATCC 43975 | $1 x 10^5$ |
| Salmonella | Salmonella enterica | ATCC 6962 | $1 x 10^5$ |
| Serratia | Serratia liquefaciens | ATCC 27592 | $1 x 10^6$ |
| Serratia | Serratia plymuthica | ATCC 53858 | $1 x 10^7$ |
| Serratia marcescens | Serratia marcescens | ATCC 14041 | $1 x 10^7$ |
| Serratia marcescens | Serratia marcescens | ATCC 14756 | $1 x 10^5$ |
| Stenotrophomonas maltophilia | Stenotrophomonas maltophilia | ATCC 13637 | $1 x 10^6$ |
| Stenotrophomonas maltophilia | Stenotrophomonas maltophilia | ATCC 17666 | $1 x 10^7$ |
| Target | Organism | Strain | LoD
Concentration
(CFU/mL) |
| Pan Candida | Candida albicans | ATCC 10231 | 1 x 106 |
| | Candida glabrata | ATCC 15126 | 1 x 105 |
| Pan Gram-Positive | Enterococcus faecalis | ATCC 51575 | 1 x 105 |
| | Enterococcus faecium | ATCC 31282 | 1 x 107 |
| | Bacillus subtilis | ATCC 21008 | 1 x 106 |
| | Staphylococcus aureus | ATCC BAA-2313 | 1 x 105 |
| | Streptococcus agalactiae | ATCC 13813 | 1 x 106 |
| | Streptococcus anginosus | ATCC 33397 | 1 x 106 |
| CTX-M | Escherichia coli (CTX-M-15) | NCTC 13441 | 1 x 104 |
| | Klebsiella pneumoniae (CTX-M-2) | CDC#0107 | 1 x 105 |
| IMP | Enterobacter aerogenes (IMP-4) | CDC#0161 | 1 x 106 |
| | Pseudomonas aeruginosa (IMP-1) | CDC#0103 | 1 x 105 |
| KPC | Enterobacter hormaechei (KPC
variant not known) | ATCC BAA-2082 | 1 x 106 |
| | Morganella morganii (KPC-2) | CDC#0133 | 1 x 106 |
| NDM | Escherichia coli (NDM-1) | CDC#0118 | 1 x 105 |
| | Proteus mirabilis (NDM-1) | CDC#0159 | 1 x 105 |
| OXA | Acinetobacter baumannii (OXA-23) | NCTC 13421 | 1 x 105 |
| | Acinetobacter baumannii (OXA-27) | NCTC 13304 | 1 x 105 |
| | Enterobacter aerogenes (OXA-48) | CDC#0074 | 1 x 106 |
| | Klebsiella pneumoniae (OXA-48) | CDC#0160 | 1 x 106 |
| VIM | Enterobacter cloacae (VIM-1) | CDC#0154 | 1 x 106 |
| | Pseudomonas aeruginosa (VIM-10) | NCTC 13437 | 1 x 105 |

66

67

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 100 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
Acinetobacter baumannii	CDC#0052
Acinetobacter baumannii	NCTC 13302
Acinetobacter baumannii	NCTC 13303
Acinetobacter baumannii	NCTC 13305
Acinetobacter baumannii	NCTC 13420
Acinetobacter baumannii	NCTC 13422
Acinetobacter baumannii	NCTC 13423
Acinetobacter baumannii (NDM-1)	CDC#0033
Acinetobacter baumannii (NDM-1)	ATCC BAA-1605
Acinetobacter baumannii (OXA-23)	CDC#0045
Acinetobacter baumannii (OXA-23)	CDC#0056
Acinetobacter baumannii (OXA-23)	NCTC 13301
Acinetobacter baumannii (OXA-23)	NCTC 13424
Acinetobacter spp. (IMP only)	JMI4084^
Bacteroides fragilis	ATCC 23745
Bacteroides fragilis	ATCC 700786
Bacteroides fragilis	NCTC 9343
Citrobacter braakii	ATCC 43162
Citrobacter braakii	ATCC 51113
Citrobacter freundii	ATCC 6879
Citrobacter freundii	ATCC 8090
Citrobacter freundii (CTX)	JMI2047
Citrobacter freundii (KPC-2)	CDC#0116
Citrobacter koseri	ATCC 25409
Citrobacter koseri	ATCC 27028
Citrobacter koseri	ATCC 29225
Citrobacter koseri	ATCC 29936
Citrobacter species (CTX-15, NDM-1)	CDC #0157
Citrobacter werkmanii	ATCC 51114
Citrobacter youngae	ATCC 29935

Table 57: Analytical Reactivity (Inclusivity)
-----------------------------------------------

Organism	Strain
Cronobacter sakazakii
Cronobacter sakazakii	ATCC 12868
Cronobacter sakazakii	ATCC BAA-894
	FSL F6-0023
Enterobacter (non-cloacae complex)
Enterobacter aerogenes	ATCC 13048
Enterobacter aerogenes	ATCC 29010
Enterobacter amnigenus	ATCC 51697
Enterobacter amnigenus	ATCC 33731
Enterobacter amnigenus	ATCC 51816B
Enterobacter gergoviae	ATCC 33028
Enterobacter gergoviae	ATCC 33426
Enterobacter cloacae complex
Enterobacter asburiae	ATCC 35954
Enterobacter asburiae	ATCC 35955
Enterobacter asburiae	ATCC 35956
Enterobacter cloacae (CTX-15)	CDC#0038
Enterobacter cloacae (CTX-9)	NCTC 13464
Enterobacter cloacae (CTX-15, KPC-2)	CDC#0163
Enterobacter cloacae (CTX, NDM)	JMI53571
Enterobacter cloacae subsp. cloacae	ATCC 23355
Enterobacter cloacae subsp. cloacae	ATCC 35030
Enterobacter cloacae subsp. dissolvens	ATCC 23373
Enterobacter hormaechei	ATCC 700323
Enterobacter hormaechei subsp. hormaechei	ATCC 49162
Enterobacter hormaechei subsp. oharae	ATCC 49163
Enterobacter hormaechei subsp. steigerwaltii	CIP108489T
Enterobacter ludwigii	DSM-16688
Escherichia coli
	ATCC 14948
	ATCC 25922
Escherichia coli	ATCC 33605
	ATCC 33876
	ATCC 35150
	ATCC 4157

68

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. necrophorum	NCTC 10575
	NCTC 10577
Fusobacterium nucleatum
Fusobacterium nucleatum subsp. nucleatum	ATCC 31647
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
	CDC#0075
Klebsiella pneumoniae (CTX-15, OXA-232)	CDC#0066
	CDC#0039
	CDC#0140
Klebsiella pneumoniae (CTX-15, OXA-181)	CDC#0141
	CDC#0142
	CDC#0034
Klebsiella pneumoniae (IMP-4)	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
Klebsiella pneumoniae subsp. pneumoniae	ATCC 27736
	ATCC 51503
	ATCC 51504
Klebsiella quasipneumoniae	ATCC 700603
Klebsiella pneumoniae subsp.
rhinoscleromatis	ATCC 9436
Klebsiella variicola	ATCC BAA-830
Morganella morganii
Morganella morganii	ATCC 25830
	GM148-209

69

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
Proteus hauseri	ATCC 33583
Proteus mirabilis	ATCC BAA-663
Proteus mirabilis (IMP)	JMI955389
Proteus mirabilis (KPC-6)	CDC#0155
Proteus penneri	ATCC 35197
Proteus penneri	ATCC 33420
Proteus penneri	ATCC 49132
Proteus vulgaris	ATCC 8427
Proteus vulgaris	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
Serratia grimesii	ATCC 13880
Serratia marcescens	ATCC 43861
Serratia marcescens	ATCC 43862
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
Candida glabrata	ATCC 2001
ATCC 66032
Candida krusei	ATCC 14243
ATCC 32196
ATCC 34135F
Candida parapsilosis	ATCC 22019
ATCC 58895

70

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.

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

| Target | Associated
Organism | Variant
Detected |
|------------------------------|------------------------|---------------------|
| Proteus
mirabilis/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 | | |
|---------------------------------|---------------------------------|---------------------------------|--------------------------------------------|------------------------|----------------------------|--------------------------------------|--------------------------------------|
| Acinetobacter
baumannii | Acinetobacter
baumannii | IMP-1 | Proteus
mirabilis/Proteus | Proteus mirabilis | IMP-19 | | |
| | | IMP-2 | | | IMP-26 | | |
| | | IMP-4 | | | IMP-32 | | |
| | | IMP-5 | | | IMP-38 | | |
| | | IMP-8 | | | IMP-1 | | |
| | | IMP-10 | | | IMP-27 | | |
| | | IMP-11 | Pseudomonas
aeruginosa | Pseudomonas aeruginosa | IMP-64 | | |
| | | IMP-14 | | | IMP-1* | | |
| | | IMP-19 | | | IMP-2 | | |
| IMP-55 | Citrobacter | Citrobacter freundii | | | IMP-4 | | |
| IMP-61 | | | | | IMP-6 | | |
| Citrobacter | | | | | Citrobacter freundii | | |
| | | | | | | IMP-38 | Enterobacter non-
cloacae Complex |
| | | | | | | Enterobacter non-
cloacae Complex | |
| | | | | | | | |
| | IMP-4 | Enterobacter cloacae
Complex | | | | | Enterobacter cloacae |
| | IMP-8 | | | | | | |
| IMP-11 | | | | | | | |
| IMP-26 | | | | | | | |
| IMP-34 | | | | | | | |
| IMP-60 | Enterobacter cloacae
Complex | | | | Enterobacter
hormaechei | IMP-21 | |
| Enterobacter cloacae
Complex | | | | | | Enterobacter
hormaechei | |
| | | IMP-14 | | | | | IMP-25 |
| Escherichia coli | Escherichia coli | IMP-1 | IMP-26 | | | | |
| | | IMP-4 | IMP-29 | | | | |
| | | IMP-6 | IMP-30 | | | | |
| | | IMP-8 | IMP-33 | | | | |
| | | IMP-14 | IMP-34 | | | | |
| | | IMP-30 | IMP-37 | | | | |
| | | IMP-52 | IMP-40 | | | | |
| | | IMP-59 | IMP-41 | | | | |
| | | IMP-66 | IMP-43 | | | | |
| Klebsiella oxytoca | Klebsiella oxytoca | IMP-1 | IMP-44 | | | | |
| | | IMP-4 | IMP-45 | | | | |
| | | IMP-8 | IMP-48 | | | | |
| | | IMP-28 | IMP-49 | | | | |
| | | IMP-34 | IMP-51 | | | | |
| Klebsiella pneumoniae
Group | Klebsiella pneumoniae | IMP-1 | IMP-53 | | | | |
| | | IMP-4* | IMP-54 | | | | |
| | | IMP-6 | IMP-56 | | | | |
| | | IMP-8 | IMP-62 | | | | |
| | | IMP-10 | IMP-63 | | | | |
| | | IMP-13 | IMP-4 | | | | |
| | | Salmonella | Salmonella enterica
Serratia marcescens | IMP-1 | | | |

81

| Target | Associated
Organism | Variant
Detected |
|-----------------------------------------|---------------------------------|---------------------|
| Serratia
marcescens/Serratia | | IMP-2 |
| | | IMP-6 |
| | | IMP-8 |
| Target | Associated
Organism | Variant
Detected |
| | | IMP-24 |
| Stenotrophomonas
maltophilia | Stenotrophomonas
maltophilia | IMP-25 |

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

| Target | Associated Organism | Variant
Detected |
|-------------------------------------|-------------------------|---------------------|
| Acinetobacter baumannii | Acinetobacter baumannii | KPC-2 |
| | | KPC-3 |
| | | KPC-10 |
| Citrobacter | Citrobacter freundii | KPC-2* |
| | | KPC-3 |
| | | KPC-18 |
| Enterobacter non-cloacae
Complex | Enterobacter aerogenes | KPC-2 |
| | | KPC-3 |
| | | KPC-2* |
| Enterobacter cloacae
Complex | Enterobacter cloacae | KPC-3 |
| | | KPC-4 |
| | | KPC-13 |
| | | KPC-18 |
| Escherichia coli | Escherichia coli | KPC-2 |
| | | KPC-3 |
| | | KPC-4 |
| | | KPC-5 |
| | | KPC-9 |
| | | KPC-18 |
| | | KPC-21 |
| | | KPC-28 |
| Klebsiella oxytoca | Klebsiella oxytoca | KPC-2 |
| | | KPC-3* |
| Klebsiella pneumoniae
Group | Klebsiella pneumoniae | KPC-1 |
| | | KPC-2 |
| | | KPC-3* |
| | | KPC-4 |
| Target | Associated Organism | Variant
Detected |
| | | 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 |
| Serratia
marcescens/Serratia | Serratia marcescens | KPC-2 |
| | | KPC-3 |

82

Enterobacter cloacae

Escherichia coli

Klebsiella oxytoca

Klebsiella pneumoniae

Group

Complex

Associated

Organism

Variant

Detected

| Target | Associated
Organism | Variant
Detected | Target |
|--------------------------------------|----------------------------|-----------------------------------|-------------------------------------------------|
| Acinetobacter baumannii | Acinetobacter
baumannii | VIM-1
VIM-2
VIM-6
VIM-11 | |
| Citrobacter | Citrobacter freundii | VIM-1
VIM-2
VIM-4
VIM-23 | Morganella morganii
Proteus mirabilis/Protei |
| Enterobacter non-
cloacae Complex | Enterobacter
aerogenes | VIM-1
VIM-1 | |

Enterobacter cloacae

Enterobacter

xiangfangensis

Escherichia coli^

Klebsiella oxytoca

hormaechei

VIM-2 VIM-4

VIM-23 VIM-31

VIM-40

VIM-1

VIM-4

VIM-23

VIM-1

VIM-1 VIM-2

VIM-19 VIM-29 VIM-1 VIM-2 VIM-4

VIM-32 VIM-35 VIM-1* VIM-2 VIM-4 VIM-12 VIM-19

VIM-24

VIM-26 VIM-27* VIM-33 VIM-34

Table 73: Predicted (in silico) Reactivity (Inclusivity) Results for VIM

	Organism	Detected
		VIM-39
		VIM-42
		VIM-51
		VIM-52
Morganella morganii	Morganella morganii	VIM-4
Proteus mirabilis/Proteus	Proteus mirabilis	VIM-1
Pseudomonas aeruginosa	Pseudomonas aeruginosa	VIM-1
		VIM-2*
		VIM-3
		VIM-4*
		VIM-5
		VIM-6
		VIM-8
		VIM-9
		VIM-10
		VIM-11
		VIM-14
		VIM-15
		VIM-16
		VIM-17
		VIM-18
		VIM-20
		VIM-28
		VIM-30
		VIM-36
		VIM-37
		VIM-41
		VIM-43
		VIM-44
		VIM-45
		VIM-46
		VIM-48
		VIM-50
Salmonella	Salmonella enterica	VIM-1
		VIM-2
Serratia marcescens/Serratia	Serratia marcescens	VIM-4
		VIM-54
Stenotrophomonas maltophilia	Stenotrophomonas maltophilia	VIM-2

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

Klebsiella pneumoniae

83

| Target | Associated
Organism | Variant
Detected |
|----------------------------|----------------------------|---------------------|
| Acinetobacter
baumannii | Acinetobacter
baumannii | 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 |
| | Acinetobacter
baumannii | 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
Klebsiella pneumoniae Group	Klebsiella pneumoniae	OXA-73
Proteus mirabilis	Proteus mirabilis	OXA-23

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

| Target | Associated
Organism | Variant
Detected |
|--------------------------------------|----------------------------|----------------------------------------------------------------------------------------------------------------|
| Acinetobacter baumannii | Acinetobacter
baumannii | OXA-48 |
| Citrobacter | Citrobacter
freundii | OXA-48
OXA-181 |
| Enterobacter non-
cloacae Complex | Enterobacter
aerogenes | OXA-244 |
| Enterobacter cloacae
Complex | Enterobacter
cloacae | OXA-48
OXA-163
OXA-181 |
| | Enterobacter
hormaechei | OXA-370 |
| | Enterobacter
ludwigii | OXA-48 |
| Escherichia coli | Escherichia coliA | OXA-48
OXA-163
OXA-181
OXA-204
OXA-232
OXA-244
OXA-438
OXA-439
OXA-566
OXA-1/OXA-48 |
| Target | Associated
Organism | Variant
Detected |
| Klebsiella pneumoniae
Group | Klebsiella
pneumoniae | OXA-48* |
| | | OXA-10 |
| | | OXA-162 |
| | | OXA-181* |
| | | OXA-204 |
| | | OXA-232* |
| | | OXA-244 |
| | | OXA-245 |
| | | OXA-247 |
| | | OXA-484 |
| | | OXA-505 |
| | | OXA-517 |
| | | OXA-519 |
| | Klebsiella
variicola | OXA-181 |
| Morganella morganii | Morganella
morganii | OXA-181 |
| Proteus mirabilis/Proteus | Proteus mirabilis | OXA-48 |
| | Proteus mirabilis | OXA-244 |
| Serratia
marcescens/Serratia | Serratia
marcescens | OXA-48 |
| | Serratia
marcescens | OXA-405 |

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

84

| Target | Associated
Organism | Variant
Detected |
|--------------------------------------|---------------------------------|---------------------|
| Acinetobacter
baumannii | Acinetobacter
baumannii | NDM-1* |
| Citrobacter | Citrobacter braakii | NDM-2 |
| Citrobacter | Citrobacter braakii | NDM-4 |
| Citrobacter | Citrobacter freundii | NDM-1A |
| Citrobacter | | NDM-4 |
| Citrobacter | | NDM-6 |
| Citrobacter | | NDM-7 |
| Enterobacter non-
cloacae Complex | Enterobacter
aerogenes | NDM-1 |
| Enterobacter non-
cloacae Complex | | NDM-4 |
| Enterobacter non-
cloacae Complex | | NDM-5 |
| Enterobacter non-
cloacae Complex | | NDM-7 |
| Enterobacter cloacae
Complex | Enterobacter
cloacaeB | NDM-1 |
| | Enterobacter
cloacaeB | NDM-4 |
| | Enterobacter
cloacaeB | NDM-7 |
| Enterobacter cloacae
Complex | Enterobacter
hormaechei | NDM-1 |
| Enterobacter cloacae
Complex | Enterobacter
ludwigii | 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 | Associated
Organism | Variant
Detected |
| | | NDM-15 |
| | | NDM-16 |
| | | NDM-17 |
| | | NDM-18 |
| | | NDM-19 |
| Klebsiella oxytoca | Klebsiella oxytoca | NDM-1 |
| Klebsiella oxytoca | Klebsiella oxytoca | NDM-4 |
| Klebsiella
pneumoniae Group | Klebsiella
pneumoniae | NDM-1* |
| | | NDM-4 |
| | | NDM-5 |
| | | NDM-6 |
| | | NDM-7 |
| | | NDM-10 |
| | | NDM-16 |
| Morganella morganii | Morganella
morganii | NDM-1c |
| Proteus
mirabilis/Proteus | Proteus mirabilis | NDM-1 |
| Pseudomonas
aeruginosa | Pseudomonas
aeruginosa | NDM-1 |
| Pseudomonas
aeruginosa | Pseudomonas
aeruginosa | NDM-5 |
| Salmonella | Salmonella enterica | NDM-1 |
| Salmonella | Salmonella enterica | NDM-5 |
| Serratia
marcescens/Serratia | Serratia marcescens | NDM-1 |
| Stenotrophomonas
maltophilia | Stenotrophomonas
maltophilia | 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.

85

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

| Resistance
Marker | Variant Not
Detected | Associated
Organism | No. of
Sequences |
|----------------------|-------------------------|----------------------------|---------------------|
| CTX-M-1 | CTX-M-80 | Klebsiella
pneumoniae | 3 |
| | CTX-M-15 | | |
| | Not Specified | | |
| IMP | IMP-31 | Pseudomonas
aeruginosa | 2 |
| | IMP-35 | | 2 |
| | IMP-7 | | 1 |
| NDM | NDM-1 | Escherichia coli | 6 |
| | NDM-1 | Klebsiella
variicola | 3 |
| | NDM-1 | Salmonella
enterica | 1 |
| | NDM-3 | Acinetobacter
baumannii | 1 |
| | NDM-4 | Escherichia coli | 1 |
| | NDM-9 | Cronobacter
sakazakii | 1 |
| | NDM-9 | Escherichia coli | 1 |
| | NDM-9 | Klebsiella
pneumoniae | 2 |
| | Not specified | Escherichia coli | 2 |
| | | Klebsiella
pneumoniae | 1 |
| | | Klebsiella sp | 1 |
| | | Pseudomonas
aeruginosa | 1 |
| | | | |
| VIM | VIM-1 | Pseudomonas
aeruginosa | 3 |
| | VIM-1 | Providencia
vermicola | 1 |
| | VIM-2 | Klebsiella
pneumoniae | 1 |
| | VIM-5 | Enterobacter
cloacae | 2 |
| | VIM-5 | Klebsiella
pneumoniae | 3 |
| | VIM-7 | Pseudomonas
aeruginosa | 4 |
| | VIM-13 | | 3 |
| | VIM-25 | Acinetobacter
baumannii | 1 |
| | VIM-25 | Proteus mirabilis | 2 |
| Resistance
Marker | Variant Not
Detected | Associated
Organism | No. of
Sequences |
| | VIM-38 | | 2 |
| | VIM-47 | Pseudomonas
aeruginosa | 2 |
| | VIM-49 | | 2 |
| | Not specified | | 1 |
| OXA-48 | OXA-232 | Escherichia coli | 1 |

Confidential

86

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 >1x100 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.

87

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 boydii	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

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

B. Cross-reactivity seen with Enterobacter cloacae complex at a concentration > 1x108 CFU/mL.

C. Cross-reactivity seen with Enterobacter (non-cloacae complex) at a concentration > 1x106 CFU/mL and Serration of > 1x107 CFU/mL.

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

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

88

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

89

Fungal Pathogens	Strain ID	Concentration Tested
Aspergillus fumigatus*	ATCC 204305	$2.50 x 10^6$ CFU/mL
Candida orthopsilosis	ATCC 96139	$1 x 10^7$ CFU/mL
Candida metapsilosis	ATCC 96144	$1 x 10^7$ CFU/mL
Candida tropicalis	ATCC 1369	$1 x 10^7$ CFU/mL
Cryptococcus grubii	ATCC 208821	$1 x 10^7$ CFU/mL
Cryptococcus gattii	ATCC 76108	$1 x 10^7$ CFU/mL
Cryptococcus neoformans	ATCC 14116	$1 x 10^7$ CFU/mL
Geotrichum capitatum	ATCC 10663	$1 x 10^7$ CFU/mL
Histoplasma capsulatum	In silico	N/A
Penicillium marneffei	ATCC 200050	$1 x 10^7$ CFU/mL
Rhodotorula glutinis	ATCC 32765	$1 x 10^7$ CFU/mL
Rhodotorula mucilaginosa	ATCC 9449	$1 x 10^7$ CFU/mL
Rhodotorula minuta	ATCC 36236	$1 x 10^7$ CFU/mL
Saccharomyces cervisiae*	ATCC 18824	$5.55 x 10^6$ CFU/mL
Trichosporon dermatis	ATCC MYA-4294	$1 x 10^7$ CFU/mL
Trichosporon mucoides	ATCC 90046	$1 x 10^7$ 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	$8 x 10^8$ CFU/mL
MOX (Carried by Aeromonas hydrophila)	JMI 938982	$1 x 10^9$ CFU/mL
SME (Carried by Serratia marcescens)A	CDC #0091	$1 x 10^9$ CFU/mL
SHV (Carried by Klebsiella pneumoniae)A	CDC #0087	$1 x 10^9$ CFU/mL
TEM (Carried by Escherichia coli)A	NCTC 13351	$1 x 10^9$ CFU/mL

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

90

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. of
Sequences | 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 of
Sequences | Predicted Cross-Reactive
Sequences
n (%) |
|------------------------------|------------------------|------------------------------------------------|
| 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%) |

91

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. oxvtoca, 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 Positivity
Concentration | 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
--------------------------------------------	--	--	--	--

92

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 Positivity
Concentration | Bottle Positivity +8
Hours 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.

93

| Concentration of Acinetobacter

baumannii	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 of

Enterobacter 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) |

94

| Concentration of

Escherichia 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 Fusobacterium

nucleatum	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.

95

| Concentration of

Haemophilus influenzae		Agreement with Expected Results
	Site	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 of

Klebsiella 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) |

96

| Concentration of

Neisseria 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 Pseudomonas

aeruginosa	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) |

97

Table 94: Percent Agreement for Serratia
	Observer 1	Observer 2
Observer 1	-	96.7
Observer 2	96.7	-

| Concentration of

Serratia 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 | (96.5-100) |
| Bottle Positive
(1x108 CFU/mL) | 1 | 36/36 | 100 | (90.4-100) |
| | 2 | 33/33 | 100 | (89.6-100) |
| | 3 | 33/33 | 100 | (89.6-100) |
| | All | 102/102 | 100 | (96.4-100) |
| 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 95: Percent Agreement for Serratia marcescens

| Concentration of

Serratia 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 |

98

| Concentration of

Candida albicans	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive

8 Hours
(1x107 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
(1x106 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

Staphylococcus aureus	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive

8 Hours
(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 |
| Bottle Positive
(1x107 CFU/mL) | 1 | 34/36 | 94.4 | (81.9-98.5) |
| | 2 | 35/35 | 100 | (90.1-100) |
| | 3 | 36/36 | 100 | (90.4-100) |
| | All | | 105/107 | 98.1 |
| Negative | 1 | 179/179 | 100 | (97.9-100) |
| | 2 | 179/180 | 99.4 | (96.9-99.9) |
| | 3 | 180/180 | 100 | (97.9-100) |
| | All | | 538/539 | 99.8 |

99

| Concentration of
Enterobacter 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 | 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 | 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 98: Percent Agreement for CTX-M

Table 99: Percent Agreement for IMP

| Concentration of

Escherichia 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) |

100

| Concentration of
Enterobacter 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
Acinetobacter baumannii

(OXA-23+)	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 | 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) |

101

Table 102: Percent Agreement for VIM

| Concentration of
Pseudomonas aeruginosa

(VIM+)	Site	Agreement with Expected Results
		Agreed / N	%	95% CI
Bottle Positive

8 Hours
(1x109 CFU/mL) | 1 | 36/36 | 100 | (90.4-100) |
| Bottle Positive
8 Hours
(1x109 CFU/mL) | 2 | 35/35 | 100 | (90.1-100) |
| 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 | 36/36 | 100 | (90.4-100) |
| Bottle Positive
(1x108 CFU/mL) | 3 | 36/36 | 100 | (90.4-100) |
| Bottle Positive
(1x108 CFU/mL) | All | 108/108 | 100 | (96.6-100) |
| Negative | 1 | 143/143 | 100 | (97.4-100) |
| Negative | 2 | 144/144 | 100 | (97.4-100) |
| Negative | 3 | 144/144 | 100 | (97.4-100) |
| Negative | All | 431/431 | 100 | (99.1-100) |

102

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 Equivalency 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.

103

Endogenous Substances	Testing Concentration
Bilirubin	60 µg/mL
Hemoglobin	0.6 g/L
Human Genomic DNA	$6 x 10^5$ 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.

104

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 PlusTM	No interference observed
BD	BACTEC	Lytic Anaerobic*	False negative results were observed for Pan
Candida, Enterobacter cloacae, Escherichia
coli, 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 ScientificTM	VersaTREKTM	REDOXTM 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 Candida, 1/15 replicates was false negative for Enterobacter cloacae; 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 1x10' 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.

105

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 | High
Concentration | Low
Concentration |
|-------------------------------|-----------------------|----------------------|
| 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