The Great Basin Staph ID/R Blood Culture Panel is a qualitative, multiplex, nucleic acid-based in vitro diagnostic assay intended for the simultaneous identification of nucleic acid from Staphylococcus lugdumensis and various Staphylococcus species to the genus level and the detection of the mecA gene for methicillin resistance directly from patient positive blood culture specimens. The test utilizes automated hot-start enabled polymerase chain reaction (PCR) for the amplification of specific DNA targets detected by hybridization probes immobilized on a silicon chip surface. The assay is performed directly on positive blood culture specimens identified as positive by continuous monitoring blood culture system that demonstrates the presence of organisms as determined by Gram stain to contain gram-positive cocci in clusters (GPCC) or gram-positive cocci in singles (GPC). The test may be performed using blood culture bottles. The Staph ID/R Blood Culture Panel identifies Staphylococcus aureus (SA), and Staphylococcus lugdunensis, and detects other Staphylococcus species without identification to species level.

The Portrait Staph ID/R Blood Culture Panel is indicated for use in conjunction with other clinical or laboratory findings to aid in the diagnosis of bacterial bloodstream infections; however, it is not used to monitor these infections. Subculturing positive blood cultures is necessary to recover viable organisms for further identification, susceptibility testing, or epidemiological typing to identify organisms in the blood culture that are not detected by the Great Basin Staph ID/R Blood Culture Panel. If detected, mecA may or may not be associated with Staphylococcus spp. detected or the agent responsible for the disease. Negative results for mecA antimicrobial resistance gene assays do not always indicate susceptibility, as other mechanisms of resistance to methicillin exist.

Device Description

The Great Basin PA500 Portrait™ Analyzer System is a fully automated system that includes the Portrait Analyzer, single-use Staph ID/R Blood Culture Panel Test Cartridges, and the Portrait data analysis software. The PA500 Portrait Analyzer System is designed to perform automated sample preparation, PCR, and optical chip-based detection with integrated data analysis in approximately 110 minutes.

AI/ML Overview

Here's an analysis of its acceptance criteria and the study that proves the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

Feature/Metric	Acceptance Criteria (Stated or Implied)	Reported Device Performance
Analytical Sensitivity (LoD) - S. aureus	Not explicitly stated, but high accuracy expected at LoD.	LoDs for 6 S. aureus strains ± mecA range from 3.5-8.2 x 10⁵ CFU/mL, with 20/20 to 23/23 correct results for individual strains.
Analytical Sensitivity (LoD) - S. epidermidis	Not explicitly stated, but high accuracy expected at LoD.	LoDs for 6 S. epidermidis strains ± mecA range from 2.2-7.1 x 10⁵ CFU/mL, with 20/20 to 27/27 correct results for individual strains.
Analytical Sensitivity (LoD) - S. lugdunensis	Not explicitly stated, but high accuracy expected at LoD.	LoDs for 3 S. lugdunensis strains ± mecA range from 2.8-4.7 x 10⁵ CFU/mL, with 22/23 to 23/23 correct results for individual strains.
Analytical Reactivity (Inclusivity)	100% detection of target strains.	Correctly detected all 48 additional well-characterized S. aureus strains (USA100-1200 and SCCmecA I-XI types) and 104 untyped strains (S. aureus, S. epidermidis, S. lugdunensis, and other Staphylococcus species) tested in the challenge panel.
Analytical Specificity (Exclusivity)	No cross-reactivity with off-panel organisms.	The vast majority of 116 off-panel microflora strains tested 'Staphylococcus NEGATIVE'. 6 'Staphylococcus POSITIVE' calls (identified as single contamination events and resolved upon re-testing as 'Staphylococcus NEGATIVE'). 27 'invalid' calls (resolved upon re-testing as 'Staphylococcus NEGATIVE').
Microbial Interference (Off-Panel)	No interference with Staphylococcus detection.	For 'valid' runs, off-panel microorganisms did not interfere with detection of Staphylococcus strains (2/2 correct calls for most combinations). In cases of initial miscalls, re-testing at higher concentrations resolved the issue.
Microbial Interference (Staphylococcus)	No interference with Staphylococcus detection when co-cultured.	Staphylococcus interference observed in some cases at initial concentrations, but resolved upon re-testing at higher concentrations (within 2-3X LoD). The majority of tests showed no interference (2/2 correct calls).
Interfering Substances (Chemical Interference)	No interference with Staphylococcus detection.	All 16 tested substances did not interfere with the detection of Staphylococcus strains or the E. faecalis (Negative) control, resulting in expected Positive or Negative calls. A few 'Invalid' runs (max 1 invalid out of 2) were observed in specific cases, but those were still considered acceptable for the purpose of the study.
Carry-over/Cross-Contamination	No evidence of contamination.	All results were in concordance with expected calls; no evidence of contamination was found.
Reproducibility	High agreement with expected results across sites, operators, and lots.	- Overall valid results: 98.1% (630/642). - Staphylococcus positivity: 100% (540/540) for positive, 96.7% (87/90) for negative. - Specific Staphylococcus detection: 99.6% (538/540) for positive, 99.3% (1341/1350) for negative. - mecA detection (no organism): 99.7% (359/360) for present, 100% (270/270) for absent.
Evaluation of Blood Culture Bottle Types	Compatibility with various bottle types.	All 13 tested blood culture bottle types were compatible with the Staph ID/R Blood Culture Panel, with no false negative results. One false positive S. aureus and one false positive Staphylococcus species OTHER than S. aureus or S. lugdunensis were observed in a single test run each, attributed to contamination. Nine invalid calls for E. faecalis.
Prospective Clinical Performance - S. aureus	PPA ≥ 95%, NPA ≥ 95% (implied).	- PPA: 98.6% (211/214) (95% CI: 96.0 - 99.5%) - NPA: 99.5% (548/551) (95% CI: 98.4 - 99.8%)
Prospective Clinical Performance - S. lugdunensis	PPA ≥ 95%, NPA ≥ 95% (implied).	- PPA: 100.0% (3/3) (95% CI: 43.9 - 100%) - NPA: 99.9% (761/762) (95% CI: 99.3 - 99.9%)
*Prospective Clinical Performance - Other Staphylococcus* spp.**	PPA ≥ 95%, NPA ≥ 95% (implied).	- PPA: 98.9% (444/449) (95% CI: 97.4 - 99.5%) - NPA: 97.2% (307/316) (95% CI: 94.7 - 98.5%)
Prospective Clinical Performance - mecA with S. aureus	PPA ≥ 90%, NPA ≥ 98% (implied).	- PPA: 94.4% (68/72) (95% CI: 86.6 - 97.8%) - NPA: 98.8% (682/690) (95% CI: 97.7 - 99.4%)
*Prospective Clinical Performance - mecA* with Other Staphylococcus spp.**	PPA ≥ 90%, NPA ≥ 95% (implied).	- PPA: 92.7% (243/262) (95% CI: 88.1 - 97.1%) - NPA: 96.2% (481/500) (95% CI: 92.4 - 98.0%)
Invalid Rate - Clinical	Acceptable low rate, resolvable on retest.	Initial Invalid Rate: 1.39% (11/789). Final Invalid Rate: 0.00% after retest.
Abort Rate - Clinical	Acceptable low rate, resolvable on retest.	Initial Abort Rate: 3.30% (26/789). Final Abort Rate: 0.00% after retest.

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

Prospective Clinical Study:
- Sample Size: A total of 762 prospective specimens were tested. Additionally, 69 archived frozen specimens were tested after the prospective study, making the overall clinical sample set significantly larger. A 'Low Prevalence' panel of 102 contrived or 'simulated' blood culture specimens was also tested.
- Data Provenance: The prospective specimens were collected at three geographically diverse U.S. sites.
Analytical Studies (LoD, Inclusivity, Exclusivity, Interference, Bottle Types, Reproducibility):
- These studies used various numbers of cultured bacterial strains (ATCC, NCTC, CCUG, Clinical isolates where specified) and simulated blood culture specimens, often with multiple replicates per strain (ranging from 2 to 20+ replicates per strain for LoD, 2 replicates per strain for exclusivity, and 90 replicates per analyte for reproducibility). The specific number of unique strains and replicates varies per study as detailed in the tables (e.g., 22 strains for LoD, 116 off-panel strains for exclusivity, 7 simulated blood culture specimens for reproducibility).
- Data Provenance: Not explicitly stated for each analytical study, but context suggests laboratory-controlled experiments. Strains were from recognized culture collections (ATCC, NCTC, CCUG) or clinical sources.

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

The document does not specify the number of experts or their qualifications used to establish the ground truth for the clinical or analytical test sets.

For the clinical study, the ground truth (Staphylococcus species identification and mecA gene presence) was established using "Reference Method(s)". While not explicitly defined, these reference methods typically involve conventional microbiology techniques such as:

Sub-culturing of positive blood cultures.
Phenotypic identification (e.g., Gram stain, catalase, coagulase tests, biochemical panels).
Molecular methods (e.g., gene sequencing, PCR assays with a different target).
Antimicrobial susceptibility testing (e.g., oxacillin MIC) to confirm methicillin resistance (though the device detects the mecA gene, not phenotypic resistance itself).

For analytical studies, bacterial strains from recognized collections (ATCC, NCTC, CCUG) and presumably well-characterized clinical isolates were used, implying their identity and mecA status were already established by standard microbiological and genetic methods.

4. Adjudication Method for the Test Set

The document does not explicitly describe an adjudication method (like 2+1 or 3+1) for the clinical test set. The comparison is made directly against "Reference Method(s)". For discrepant results in the exclusivity and microbial interference studies, repeat testing was performed (e.g., a minimum of six (6) repeat tests for 'Staphylococcus POSITIVE' calls in exclusivity, sometimes at higher CFU/mL input for interference studies to resolve miscalls).

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

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not conducted. This device is an automated in vitro diagnostic assay (algorithm only), not an AI-assisted human reading system. Therefore, there is no human-in-the-loop performance to measure improvement with AI assistance.

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

Yes, the studies conducted demonstrate the standalone performance of the Great Basin Staph ID/R Blood Culture Panel. This device is an automated, qualitative, molecular diagnostic system, meaning its output is generated by the instrument itself (the Portrait Analyzer and its software) without direct human interpretation of complex images or data that would typically benefit from AI assistance. The performance metrics (PPA, NPA) directly reflect the device's accuracy against reference methods.

7. The Type of Ground Truth Used

The primary ground truth for both analytical and clinical performance studies was established through reference microbiology methods. This includes:

Culture-based identification: Sub-culturing, Gram staining, and phenotypic tests for species identification.
Molecular identification: Implied use of validated molecular techniques or genetic characterization (e.g., for mecA gene presence and confirmation of species for various strains).
Antimicrobial Susceptibility Testing (AST): Specifically, oxacillin MIC was used in the Analytical Reactivity (Inclusivity) Challenge Panel to determine phenotypic resistance, which indirectly supports the genotypic mecA detection.
For the analytical studies, well-characterized strains from culture collections (ATCC, NCTC, CCUG) served as a strong foundation for ground truth.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of device development or any specific AI/machine learning models. This is typical for traditional molecular diagnostic assays, where assay design (primer/probe sequences, reaction conditions) is based on known genetic targets and verified through extensive analytical validation rather than data-driven machine learning training. The analytical studies (LoD, inclusivity, exclusivity) serve to validate the analytical performance of the developed assay against a wide range of relevant organisms and conditions.

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

As no explicit "training set" is described for an AI/ML model, this question is not directly applicable. If one considers the development of the assay (design of specific DNA targets and hybridization probes) as analogous to "training," then the ground truth would have been established through extensive molecular biology research, genetic sequencing, and characterization of Staphylococcus species and the mecA gene. This would involve identifying conserved and variable regions of target genes across a broad collection of known Staphylococcus strains to ensure specificity and sensitivity.

Summary

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

April 26, 2016

GREAT BASIN SCIENTIFIC, INC. CHUCK OWEN DIRECTOR, REGULATORY AFFAIRS & QUALITY ASSURANCE 2441 S. 3850 WEST SALT LAKE CITY, UT 84120

Re: K152470 Trade/Device Name: Great Basin Staph ID/R Blood Culture 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: II Product Code: PAM, OOI Dated: February 22, 2016 Received: February 25, 2016

Dear Mr. Owen:

This letter corrects our substantially equivalent letter of March 25, 2016.

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 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. 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

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Page 2 - Mr. Owen

comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR 801 and 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

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

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely vours.

Kristian M. Roth -S

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

Enclosure

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Indications for Use

510(k) Number (if known) K152470

Device Name Great Basin Staph ID/R Blood Culture Panel

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
☒ Prescription Use (Part 21 CFR 801 Subpart D)
☐ Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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March 23, 2016

510(k) Summary: Great Basin Staph ID/R Blood Culture Panel

A. Submitted by:

Great Basin Corporation 2441 South 3850 West Salt Lake City, Utah 84120 Phone: 801-990-1055 Fax: 801-990-1051

Contact Information

Chuck Owen, Director of Requlatory Affairs Phone: 385-215-3313 801-990-1051 Fax: Email: cowen@gbscience.com

B. Name of Device

Proprietary Name:	Great Basin Staph ID/R Blood Culture Panel
-------------------	--------------------------------------------

Common or Usual Names:	Staph ID/R Blood Culture PanelStaph AssaySIDR
------------------------	-------------------------------------------------------

C. Regulatory Information:

a. Regulation Section:	21 CFR 866.3365 – Multiplex Nucleic Acid Assayfor Identification of Microorganisms and ResistanceMarkers from Positive Blood Cultures21 CFR 862.2570 – Instrumentation for clinicalmultiplex test systems
b. Classification:	Class II (Staph ID/R Blood Culture Panel; non-exempt)Class II (PA500 Portrait Analyzer System)
c. Classification panel:	Microbiology Devices, OIVD (83) Microbiology
d. Product Code:	PAM (Gram-positive bacteria and their resistance markers)OOI (Real-time nucleic acid amplification system)

D. Intended use(s)/Indications for Use:

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amplification of specific DNA targets detected by hybridization probes immobilized on a silicon chip surface. The assay is performed directly on positive blood culture specimens identified as positive by continuous monitoring blood culture system that demonstrates the presence of organisms as determined by Gram stain to contain gram-positive cocci in clusters (GPCC) or gram-positive cocci in singles (GPC). The test may be performed using blood culture bottles. The Staph ID/R Blood Culture Panel identifies Staphylococcus aureus (SA), and Staphylococcus lugdunensis, and detects other Staphylococcus species without identification to species level.

The Portrait Staph ID/R Blood Culture Panel is indicated for use in conjunction with other clinical or laboratory findings to aid in the diagnosis of bacterial bloodstream infections; however, it is not used to monitor these infections. Sub-culturing positive blood cultures is necessary to recover viable organisms for further identification, susceptibility testing, or epidemiological typing to identify organisms in the blood culture that are not detected by the Great Basin Staph ID/R Blood Culture Panel. If detected, mecA may or may not be associated with Staphylococcus spp. detected or the agent responsible for the disease. Negative results for mecA antimicrobial resistance gene assays do not always indicate susceptibility, as other mechanisms of resistance to methicillin exist.

E. Device Description:

Test Principle:

The Great Basin Staph ID/R Blood Panel on the PA500 Portrait™ Analyzer System utilizes automated hot-start enabled polymerase chain reaction (PCR) amplification technology to amplify specific nucleic acid sequences that are detected using species specific Staphylococcal DNA hybridization probes immobilized on a modified silicon chip surface.

Target genomic DNA is extracted from microbial cells and diluted to reduce potential inhibitors of the PCR reaction. During the PCR process, double-stranded DNA is separated and target nucleic acid sequences are amplified by thermal cycling. Biotin-labeled primers direct amplification of specific nucleic acid sequences within a variable region of the tuf gene for identification of coagulase-neqative Staphylococcus species, within conserved region of the thermonuclease (nuc) gene for specific identification of Staphylococcus aureus, and the mecA gene for detecting oxacillin/methicillin resistance. Following the PCR process, biotin-labeled, amplified target DNA sequences are hybridized to an array of probes immobilized on the silicon chip surface, then incubated with antibody conjugated to the horseradish peroxidase enzyme (HRP). The unbound conjugate is removed by washing and tetramethylbenzidine (TMB) is added to produce a colored precipitate at the location of the probe/target sequence complex. The resulting signal is detected by the automated Portrait Optical Reader within the Portrait System.

Test Device:

The Great Basin PA500 Portrait Analyzer System is a fully automated system that includes the Portrait Analyzer, single-use Staph ID/R Blood Culture Panel Test Cartridges, and the Portrait data analysis software. The PA500 Portrait Analyzer System is designed to perform automated sample preparation, PCR, and optical chip-based detection with integrated data analysis in approximately 110 minutes.

F. Substantial Equivalence Information:

a. Predicate Device: Verigene Gram Positive Blood Culture Nucleic Acid Test (BC-GP)
b. Predicate 510(k) number: K113450

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c. Comparison with Predicate

Item	Staph ID/R Blood Culture Panel	Predicate (K113450)
Manufacturer	Great Basin Scientific, Inc.	Nanosphere, Inc.
Trade Name	Portrait™ Staph ID/R Blood Culture Panel	Verigene Gram Positive Blood Culture Nucleic Acid Test (BC-GP)
510(k) Number	K152470	K113450
	Similarities
Classification	Class II	same
IntendedUse/Indicationsfor Use	The Great Basin Staph ID/R Blood Culture Panel is a qualitative, multiplex,nucleic acid-based in vitro diagnostic assay intended for the simultaneousidentification of nucleic acid from Staphylococcus aureus, Staphylococcuslugdunensis and various Staphylococcus species to the genus level and thedetection of the mecA gene for methicillin resistance directly from patientpositive blood culture specimens. The test utilizes automated hot-startenabled polymerase chain reaction (PCR) for the amplification of specificDNA targets detected by hybridization probes immobilized on a silicon chipsurface. The assay is performed directly on positive blood culture specimensidentified as positive by continuous monitoring blood culture system thatdemonstrates the presence of organisms as determined by Gram stain tocontain gram-positive cocci in clusters (GPCC) or gram-positive cocci insingles (GPC). The test may be performed using blood culture bottles. TheStaph ID/R Blood Culture Panel identifies Staphylococcus aureus (SA), andStaphylococcus lugdunensis, and detects other Staphylococcus specieswithout identification to species level. The Staph ID/R Blood Culture Panel isindicated for use in conjunction with other clinical or laboratory findings toaid in the diagnosis of bacterial bloodstream infections; however, it is notused to monitor these infections. Sub-culturing positive blood cultures isnecessary to recover viable organisms for further identification, susceptibilitytesting, or epidemiological typing to identify organisms in the blood culturethat are not detected by the Great Basin Staph ID/R Blood Culture Panel. Ifdetected, mecA may or may not be associated with Staphylococcus spp.detected or the agent responsible for the disease. Negative results for mecAantimicrobial resistance gene assays do not always indicate susceptibility, asother mechanisms of resistance to methicillin exist.	The Verigene® Gram Positive Blood Culture Nucleic Acid Test (BC-GP)performed using the sample-to-result Verigene System is a qualitative,multiplexed in vitro diagnostic test for the simultaneous detection andidentification of potentially pathogenic gram-positive bacteria which maycause bloodstream infection (BSI). BC-GP is indicated for use in conjunctionwith other clinical and laboratory findings, such as culture, to aid in thediagnosis of bacterial bloodstream infections; however, it is not used tomonitor bloodstream infections. BC-Gp detects the following bacterialgenera and species: Staphylococcus spp., Staphylococcus aureus ,Staphylococcus epidermidis , and Staphylococcus lugdunensis , Streptococcusspp., Streptococcus pneumonia , Streptococcus pyogenes , Streptococcusagalactiae , Streptococcus anginosus group, Entercoccus faecalis ,Enterococcus faecium , Listeria spp. In addition, BC-GP detects the mecAresistance marker, and the vanA and vanB resistance markers, inferringvanA/vanB mediated vancomycin resistance to either E. faecalis or E.faecium , or the mecA -mediated methicillin resistance to either S. aureus orS. epidermidis . BC-GP is indicated for use in conjunction with other clinicalor laboratory findings to aid in the diagnosis of bacterial bloodstreaminfections; however, it is not used to monitor these infections. Sub-culturingof positive blood cultures is necessary to recover organisms for susceptibilitytesting, identification of organisms not detected by BC-GP, differentiation ofmixed growth, association of antimicrobial marker genes to a specificorganism, or for epidemiological testing.
Qualitative/Quantitative	Qualitative	same
Single-Use TestCartridge	Disposable, single-use, self-contained fluidic test cartridge	same
Automated	Yes	same
Test Principle	Multiplex nucleic acid array-based detection	same
Sample Types	Direct blood culture positive by Gram stain for GPCC or GPC	same
Organism andResistanceMarker Detection	S. aureus , S. lugdunensis , Staphylococcus spp., mecA	S. aureus , S. lugdunensis , S. epidermidis , Staphylococcus spp., mecA
Controls	One Internal Processing Control (whole organism complete assaycontrol)	Two Internal Processing Controls (whole organism complete assaycontrol and single-stranded DNA Hybridization control)
Calibration	Not required	same
	Differences
IntendedUse/Indicationsfor Use	Identification of S. aureus , S. lugdunensis , detection of otherStaphylococcus spp. Detection of the mecA gene for methicillinresistance in all Staphylococcus organisms.	Tests for same Staphylococcus targets. Detection of mecA gene formethicillin reistance in S. aureus and S. epidermidis only. Tests foradditional gram-positive bacteria including Streptococcus spp.,Streptococcus pneumonia , Streptococcus pyogenes , Streptococcusagalactiae , Streptococcus anginosus group, Entercoccus faecalis ,Enterococcus faecium , and Listeria spp. Tests for additional resistancemarkers including the vanA and vanB resistance markers, inferringvanA/vanB mediated vancomycin resistance to either E. faecalis or E.faecium .
Test Principle/Technology	Fully automated multiplex PCR and detection by target-specificcapture oligonucleotides immobilized in a macroarray format onto achip surface for probe-based end-point.	Fully automated multiplex DNA detection of specific nucleic acidsequences in a microarray format using target-specific capture andmediator oligonucleotides for probe-based end-point detection.
Instrument	PA500 Portrait Analyzer	Verigene Reader and Processor SP

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G. Performance Data - Analytical Studies

a. Analytical Sensitivity

The limit of detection (LoD) of the Staph ID/R Blood Culture Panel to Staphylococcus with or without mecA was assessed and confirmed with twenty-two (22) different strains. For these studies, overnight incubations of each bacterial stock in Tryptic Soy Broth (TSB) were measured by optical density to estimate the inoculum, serially diluted and spiked into a BACTEC Plus Aerobic/F blood bottle containing neqative blood. Spiked bottles were incubated until alarm positivity in a BACTEC Blood Culture System. Alarm positive blood cultures were gram stained, diluted, plated on agar plates and colonies were counted the following day. The enumerated blood culture bottles were diluted to a 1x108-10° targeted sample input CFU/mL range in BACTEC Plus Aerobic/F media containing neqative blood, tested on device and plated on agar plates to enumerate the actual sample input CFU/mL. The estimations of the sample input CFU/mL were revised to reflect the correct CFU/mL by this colony counting method.

The LoDs for six (6) S. aureus strains ± mecA are reported as 3.5-8.2x10° CFU/mL; the LoD for each S. aureus strain is listed in Table 1.

Table 1. Performance of the Staph ID/R Blood Culture Panel on a minimum of 20 replicates of six (6) Staphylococcus aureus strains ± mecA for establishing LoD.

Species	ATCC #	Sample Input(CFU/mL)	Correct Staph ID/R BloodCulture Panel Results
S. aureus, mecA +	BAA-1680	3.5E+05	23/23
	BAA-1682	4.0E+05	20/20
	BAA-1684	8.2E+05	22/22
S. aureus, mecA-	25923	5.1E+05	21/21
	6538	3.9E+05	22/22
	11632	6.2E+05	20/20*

*This set of test runs also contained 1 "Invalid" run

The LoDs for six (6) S. epidermidis strains ± mecA are reported as 2.2-7.1x108 CFU/mL; the LoD for each S. epidermidis strain is listed in Table 2.

Table 2. Performance of the Staph ID/R Blood Culture Panel on a minimum of 20 replicates of six (6) Staphylococcus epidermidis strains ± mecA for establishing LoD.

Species	ATCC #	Sample Input(CFU/mL)	Correct Staph ID/R BloodCulture Panel Results
S. epidermidis, mecA +	35984	3.6E+05	20/20
S. epidermidis, mecA +	51625	4.0E+05	22/22
S. epidermidis, mecA +	700562	4.3E+05	27/27
S. epidermidis, mecA +	700566	5.8E+05	23/23
S. epidermidis, mecA-	700583	2.2E+05	23/23
S. epidermidis, mecA-	12228	7.1E+05	22/23

Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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The LoDs for three (3) S. lugdunensis strains without mecA are reported as 2.8-4.7x108 CFU/mL; the LoD for each S. lugdunensis strain is listed in Table 3.

Table 3. Performance of the Staph ID/R Blood Culture Panel on a minimum of 20 replicates of three (3) Staphylococcus lugdunensis strains ± mecA for establishing LoD.

Species	ATCC #	Sample Input(CFU/mL)	Correct Staph ID/R BloodCulture Panel Results
S. lugdunensis, mecA-	43809	2.8E+05	22/23
	49576	4.5E+05	23/23
	7990	4.7E+05	23/23

LoDs for seven (7) Staphylococcus species ± mecA, excluding S. aureus, S. epidermidis, and S. lugdunensis, are reported as 1.5-5.3x10* CFU/mL. The species were selected for inclusion in the LoD studies based on tuf sequence variation from in silico analysis of the target sequence. The LoD for each Staphylococcus strain is listed in Table 4.

Table 4. Performance of the Staph ID/R Blood Culture Panel on a minimum of 20 replicates of seven (7) Staphylococcus strains ± mecA for establishing LoD.

Species	ATCC #	Sample Input(CFU/mL)	Correct Staph ID/R BloodCulture Panel Results
S. capitis, mecA -	35661	1.5E+05	20/20*
S. haemolyticus, mecA+	BAA-1693	3.1E+05	19/20
S. hominis, mecA -	27844	5.3E+05	23/23*
S. pasteuri, mecA -	51129	5.3E+05	21/22
S. sciuri, mecA -	29060	4.3E+05	21/21**
S. simulans, mecA -	27848	2.0E+05	23/23
S. warneri, mecA -	27830	4.0E+05	22/22
Staphylococcus species;ATCC, CCUG, NRS, Clinical #	SCCmec, PFGE type /source	Sample Input(CFU/mL)	Correct Staph ID/RBlood CulturePanel Result
S. aureus BAA-38	I, Denmark	1.0x106	2/2
S. aureus BAA-44	I, Iberian	9.2x105	2/2
S. aureus 700698	II, Japan	1.1x106	2/2
S. aureus BAA-41	II, USA100	5.9x106	2/2
S. aureus BAA-1681	II, USA100	1.3x106	2/2
S. aureus BAA-1682	II, USA100	2.7x106	2/2
S. aureus BAA-1761	II, USA100	1.2x106	2/2
S. aureus NRS660	II, USA100	4.1x106	2/2
S. aureus BAA-1720	II, USA200	2.3x106	2/2
S. aureus BAA-1750	II, USA200	9.5x105	2/2
S. aureus BAA-1760	II, USA200	2.4x106	2/2
S. aureus NRS651	II, USA200	3.1x106	2/2
S. aureus BAA-39	III, Hungary	6.4x106	2/2
S. aureus 35592	III, ST239	1.0x106	2/2
S. aureus BAA-1680	IV, USA300	2.4x106	2/2
S. aureus BAA-1717	IV, USA300	3.5x106	2/2
S. aureus NRS643	IV, USA300	3.4x106	2/2
S. aureus NRS662	IV, USA300	5.8x106	2/2
S. aureus NRS688	IV, USA300	4.4x106	2/2
S. aureus NRS716	IV, USA300	4.4x106	2/2
S. aureus BAA-1683	IV, USA400	1.2x106	2/2
S. aureus BAA-1696	IV, USA400	4.1x106	2/2
S. aureus BAA-1707	IV, USA400	4.1x106	2/2
S. aureus BAA-1752	IV, USA400	4.5x106	2/2
S. aureus BAA-1684	IV, USA500	1.8x106	2/2
S. aureus BAA-1689	IV, USA500	2.5x106	2/2
S. aureus BAA-1763	IV, USA500	6.4x106	2/2
S. aureus NRS685	IV, USA500	3.5x106	2/2
S. aureus BAA-1754	IV, USA600	6.7x106	2/2
S. aureus BAA-1755	IV, USA700	6.9x106	2/2
S. aureus BAA-1758	IV, USA800	8.8x105	2/2
S. aureus BAA-1768	IV, USA800	8.8x105	2/2
S. aureus NRS692	IV, USA800	2.0x106	2/2
S. aureus NRS675	IV, USA800	3.1x106	2/2
S. aureus BAA-1747	IV, USA1000	1.5x106	2/2
S. aureus NRS483	IV, USA1000	6.2x106	2/2
S. aureus NRS730	IV, USA1000	2.6x106	2/2

S. aureus BAA-1764	IV, USA1100	9.8x105	2/2
S. aureus NRS484	IV, USA1100	4.4x106	2/2
S. aureus BAA-1766	V, USA700	4.4x106	2/2
S. aureus BAA-2094	V, WA-MRSA	1.0x106	2/2
S. aureus BAA-42	VI, USA800	4.2x106	2/2
S. aureus (mecC) BAA-2313	XI, CC130	4.0x106	2/2
S. aureus BAA-1751	Untyped, USA600	4.0x106	2/2
S. aureus BAA-1771	Untyped, USA800	4.7x106	2/2
S. aureus BAA-1718	NA, USA300	4.6x106	2/2
S. aureus BAA-1749	NA, USA900	2.5x106	2/2
S. aureus BAA-1765	NA, USA1200	1.1x106	2/2
S. aureus BAA-40	Untyped, Lisbon	6.5x106	2/2
S. aureus BAA-1685	Untyped, ATCC	4.8x106	2/2
S. aureus BAA-1708	Untyped, ATCC	2.4x106	2/2
S. aureus BAA-1721	Untyped, UK	9.0x105	2/2
S. aureus 6538	Untyped, ATCC	3.1x106	2/2
S. aureus 11632	Untyped, ATCC	1.4x106	2/2
S. aureus 12600	Untyped, ATCC	2.1x106	2/2
S. aureus 13150	Untyped, ATCC	9.0x105	2/2
S. aureus 14775	Untyped, ATCC	3.1x106	2/2
S. aureus 14776	Untyped, ATCC	4.1x106	2/2
S. aureus 14993	Untyped, ATCC	2.4x106	2/2
S. aureus 25923	Untyped, ATCC	4.5x106	2/2
S. aureus 29213	Untyped, ATCC	4.0x106	2/2
S. aureus 29247	Untyped, ATCC	3.0x106	2/2
S. aureus 43300	Untyped, ATCC	3.2x106	2/2
S. aureus 700699	Untyped, ATCC	3.6x106	2/2
S. aureus BORSA MCW1	Untyped, Wisconsin,Ledeboer Lab	1.9x106	2/2
S. aureus BORSA MCW2	Untyped, Wisconsin,Ledeboer Lab	3.6x106	2/2
S. aureus BORSA 23737	Untyped, New Jersey,Kreiswirth Lab	2.9x106	2/2
S. aureus BORSA 23739	Untyped, New Jersey,Kreiswirth Lab	3.4x106	2/2
S. aureus Empty Mec Cassette 45	Untyped, Iowa,Diekema lab	6.3x105	2/2
S. aureus Empty Mec Cassette 46	Untyped, Iowa,Diekema lab	2.7x106	2/2
S. aureus Empty Mec Cassette 50	Untyped, Iowa,Diekema lab	3.7x106	2/2
S. aureus Empty Mec Cassette 51	Untyped, Iowa,Diekema lab	2.2x106	2/2

S. auricularis 33751	ATCC	3.2x105	2/2
S. auricularis 33753	ATCC	3.2x105	2/2
S. capitis subsp. capitis 35661	ATCC	7.2x105	2/2
S. capitis subsp. ureolyticus 49326	ATCC	2.2x106	2/2
S. caprae 35538	ATCC	9.2x105	2/2
S. caprae 51548	ATCC	1.3x106	2/2
S. chromogenes 43764	ATCC	4.8x106	2/2
S. chohnii subsp. cohnii 29972	ATCC	4.3x106	2/2
S. chohnii subsp. urealyticus 49328	ATCC	9.3x105	2/2
S. condimenti 4753	ATCC	1.2x106	2/2
S. carnosus 51365	ATCC	4.9x106	2/2
S. delphini 49171	ATCC	1.7x105	2/2
S. epidermidis 12228	ATCC	1.2x106	2/2
S. epidermidis 35983	ATCC	1.8x106	2/2
S. epidermidis 35984	ATCC	4.6x106	2/2
S. epidermidis 51625	ATCC	1.2x106	2/2
S. epidermidis 700562	ATCC	2.0x106	2/2
S. epidermidis 700563	ATCC	2.7x106	2/2
S. epidermidis 700565	ATCC	1.1x106	2/2
S. epidermidis 700566	ATCC	6.6x105	2/2
S. epidermidis 700567	ATCC	3.9x106	2/2
S. epidermidis 700568	ATCC	9.4x105	2/2
S. epidermidis 700576	ATCC	8.5x105	2/2
S. epidermidis 700583	ATCC	3.5x105	2/2
S. equorum 43958	ATCC	4.2x106	2/2
S. felis 49163	ATCC	1.1x106	2/2
S. fleuretti BAA-274	ATCC	1.3x106	2/2
S. gallinarum 33539	ATCC	2.2x106	2/2
S. gallinarum 49148	ATCC	2.5x106	2/2
S. haemolyticus BAA-1693	ATCC	3.5x106	2/2
S. haemolyticus 43253	ATCC	1.2x106	2/2
S. haemolyticus 29968	ATCC	4.8x106	2/2
S. haemolyticus 29970	ATCC	3.9x106	2/2
S. haemolyticus 700564	ATCC	3.5x106	2/2
S. hominis 25615	ATCC	3.2x106	2/2
S. hominis 27844	ATCC	1.6x106	2/2
S. hominis 51624	ATCC	3.0x106	2/2
S. hominis 700586	ATCC	2.4x106	2/2
S. hominis subsp. novobiosepticus 700237	ATCC	2.2x106	2/2
S. intermedius 29663	ATCC	1.1x106	2/2
S. intermedius 49052	ATCC	1.0x106	2/2
S. intermedius 51874
	ATCC	2.5x10°	2/2
S. kloosii 43959	ATCC	2.4x10°	2/2
S. lentus 29070	ATCC	6.8x105	2/2
S. lugdunensis 4436	CCM, Czech	2.9x10°	2/2
S. lugdunensis 7990	ATCC, NCTC	3.0x10°	2/2
S. lugdunensis 43809	ATCC	2.0x10°	2/2
S. lugdunensis 48413	CCUG, Sweden	4.3×10°	2/2
S. lugdunensis 49576	ATCC	8.2x109	2/2
S. lugdunensis 700328	ATCC	3.0x10°	2/2
S. lutrae 700373	ATCC	8.4×10*	2/2
S. massiliensis 7895	ATCC	1.8×10°	2/2
S. muscae 49912	ATCC	1.4x10°	2/2
S. nepalensis 48992	ATCC	3.3x10°	2/2
S. pasteuri 51129	ATCC	5.1x10°	2/2
S. pettenkoferi 36	Indianapolis, Denys Lab	2.0x10°	2/2
S. piscifermentans 51183	ATCC	2.4x10°	2/2
S. pulvereri 33938	ATCC	1.2×10°	2/2
S. pseudintermidus 49444	ATCC	1.7×10°	2/2
S. saccharolyticus 14953	ATCC	7.3x103	2/2
S. saprophyticus BAA-750	ATCC	2.1x10°	2/2
S. saprophyticus 15305	ATCC	1.7×10°	2/2
S. schleiferi subsp. coagulans 49545	ATCC	1.7×10°	2/2
S. schleiferi subsp. schleiferi 43808	ATCC	1.3×10°	2/2
S. sciuri 29061	ATCC	5.5x10-	2/2
S. sciuri 29060	ATCC	1.1x10°	2/2
S. sciuri 700013	ATCC	2.9x10°	2/2
S. sciuri subsp. carnaticus 700058	ATCC	2.1x10°	2/2
S. sciuri subsp. rodentium 700063	ATCC	1.8x106	2/2
S. simulans 27841	ATCC	5.3x105	2/2
S. simulans 27848	ATCC	5.7x108	2/2
S. simiae 7213	ATCC	1.2×10°	2/2
S. succinus subsp. succinus 700337	ATCC	1.6x105	2/2
S. vitulinus 51162	ATCC	1.1x106	2/2
S. warneri 10209	ATCC	2.4x106	2/2
	ATCC	1.1x106	2/2
S. warneri 25614
S. warneri 27836	ATCC	4.2x106	2/2
S. warneri 49454	ATCC	1.4×10°	2/2
S. xylosus 35633	ATCC	1.7×10€	2/2

*This set of test runs also contained 1 "Invalid" run

**This set of test runs also contained 2 "Invalid" runs

b. Analytical Reactivity (Inclusivity)

The analytical reactivity of the Staph ID/R Blood Culture Panel was tested against an additional 48 well characterized S. aureus strains from ATCC representing USA100-1200 and SCCmecA I-VI, XI types representative of temporal and geographical diversity. In addition, 104 untyped strains, representing S. aureus, S. epidermidis, S. lugdunensis, and other various Staphylococcus species were tested in the Staph ID/R Blood Culture Panel.

The Staph ID/R Blood Culture Panel correctly detected all of the additional Staphylococcus strains, mecA present or absent (Table 5).

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Table 5. Analytical Reactivity (Inclusivity) Panel. Staphylococcus strains and results for inclusivity by the Staph ID/R Blood Culture Panel.

Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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In addition to these studies, a subset of eight (8) S. aureus strains representing SCCmecA subtypes I-V, one (1) mecc strain, four (4) Borderline Oxacillin Resistant S. aureus (BORSA), four (4) Empty Cassette S. aureus variants, and multiple S. epidermidis and S. lugdunensis strains were selected to be part of a "Challenge Panel". The challenge panel was tested for oxacillin MIC using BD Phoenix ID. The results from the MIC determination and card results are listed in Table 6.

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Table 6. Analytical Reactivity (Inclusivity) Challenge Panel. Staphylococcus strains tested for oxacillin MIC and for inclusivity by the Staph ID/R Blood Culture Panel.

Staphylococcusspecies	Strain	SCCmec Type	PFGE/Type Strain	Significance	OxacillinMIC (µg/mL)	Staph ID/R Blood Culture Panel Result
S. aureus	BAA-38	I	Unknown	MRSA	>2	S. aureus, mecA Present
S. aureus	700699	II	Genome sequenced	MRSA	>2	S. aureus, mecA Present
S. aureus	BAA-1682	II	USA100	MRSA	>2	S. aureus, mecA Present
S. aureus	BAA-1681	II	USA100	MRSA	>2	S. aureus, mecA Present
S. aureus	33592	III	ST239	MRSA	>2	S. aureus, mecA Present
S. aureus	BAA-1680	IV	USA300	MRSA	>2	S. aureus, mecA Present
S. aureus	BAA-1684	IV	USA500	MRSA	>2	S. aureus, mecA Present
S. aureus	BAA-2094	V	WA-MRSA	MRSA	>1	S. aureus, mecA Present
S. aureus	BAA-2313	XI (mecC)	CC130	mecC MRSA	2	S. aureus, mecA Absent
S. aureus	20723.046	NA	Unknown	Empty Cassette	0.5	S. aureus, mecA Absent
S. aureus	20723.051	NA	Unknown	Empty Cassette	0.5	S. aureus, mecA Absent
S. aureus	20723.045	NA	Unknown	Empty Cassette	≤0.25	S. aureus, mecA Absent
S. aureus	20723.050	NA	Unknown	Empty Cassette	≤0.25	S. aureus, mecA Absent
S. aureus	23736	NA	Unknown	BORSA	>2	S. aureus, mecA Absent
S. aureus	23739	NA	Unknown	BORSA	>2	S. aureus, mecA Absent
S. aureus	23737	NA	Unknown	BORSA	1	S. aureus, mecA Absent
S. aureus	MCW1	NA	Unknown	BORSA	0.5	S. aureus, mecA Absent
S. aureus	MCW2	NA	Unknown	BORSA	≤0.25	S. aureus, mecA Absent
S. aureus	12600	NA	serotype 3	MSSA	0.5	S. aureus, mecA Absent
S. aureus	14993	NA	Unknown	MSSA	0.5	S. aureus, mecA Absent
S. aureus	11632	NA	Unknown	MSSA	≤0.25	S. aureus, mecA Absent
S. aureus	6538	NA	Unknown	MSSA	≤0.25	S. aureus, mecA Absent
S. aureus	25923	NA	Unknown	MSSA	≤0.25	S. aureus, mecA Absent
S. epidermidis	35984	NA	Genome sequenced	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	35983	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	700562	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	700565	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	700567	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	700566	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	700576	NA	Unknown	MRSE	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	51625	NA	Unknown	MRSE	1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. epidermidis	12228	NA	Unknown	MSSE	≤0.25	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Absent
S. epidermidis	700583	NA	Unknown	MSSE	≤0.25	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Absent
S. hominis	700586	NA	Unknown	MR Staph ssp	>1	Staphylococcus species OTHER thanS. aureus or S. lugdunensis, mecA Present
S. lugdunensis	49576	NA	Unknown	MS - Lug	≤0.25	S. lugdunensis, mecA Absent
S. lugdunensis	700328	NA	Unknown	MS - Lug	≤0.25	S. lugdunensis, mecA Absent
S. lugdunensis	NCTC 7990	NA	Unknown	MS - Lug	≤0.25	S. lugdunensis, mecA Absent
S. lugdunensis	43809	NA	Unknown	MS - Lug	≤0.25	S. lugdunensis, mecA Absent

Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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All strains showed expected oxacillin MIC results, including the BORSA strains, which showed a range of oxacillin resistance (0.25-2 µg/mL) as expected for strains resistant by alternative mechanisms other than mecA. In addition, all empty cassette strains, which lack mecA, were sensitive to oxacillin (0.5-0.25 ug/mL). Samples were correctly detected as detected as mecA Present or Absent in the Staph ID/R Blood Culture Panel. The Staph ID/R did not detect mecA for mecC strain BAA-2313, empty cassette and BORSA strains as expected.

c. Analytical Specificity (Exclusivity)

Studies were performed to assess the potential cross-reactivity of the Staph ID/R Blood Culture Panel with 116 off-panel microflora (bacterial, yeast, and mycoplasma strains). BACTEC Plus Aerobic/F or Anaerobic/F media (for anaerobic strains) containing negative blood were inoculated with isolates and incubated in a BACTEC Blood Culture System until alarm positivity. Alarm positive samples were incubated for additional time in the Blood Culture System consistent with specimen stability studies to obtain a target microorganism bottle load ≥10° CFU/mL. The positive alarm bottles were Gram stained, diluted, plated and counted to confirm all organisms were tested at 1x10° CFU/mL or higher. For two (2) organisms, alarm positive bottles samples were substituted with genomic DNA at a final concentration of ≥10° copies/mL. Genomic DNA was spiked into a matrix of negative blood and BACTEC Plus Aerobic/F media.

A minimum of two (2) replicates was tested in the Staph ID/R Blood Culture Panel for each of the bacterial, fungal and mycoplasma strains evaluated and these data are summarized in Table 7.

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Table 7. Analytical Specificity (Exclusivity) Panel. Non-Staphylococcus microorganisms or DNA from micro-organisms tested for exclusivity by the Staph ID/R Blood Culture Panel.

Exclusivity Species	Strain (ATCC,CCUG, Clinical)	Sample Input inCFU/mL orcopies/mL (gDNA)	Correct Staph ID/RBlood Panel Result
Gram Positive Bacteria
Actinomyces odontolyticus	17929	1.7x109	2/2 (1)
Abiotrophia defectiva	49176	1.4x108	2/2
Aerococcus urinae	51268	6.1x108	2/2
Arcanobacterium haemolyticum	BAA-1784	6.0x108	10/11 (2)
Bacillus cereus	14579	2.4x109	2/2
Corynebacterium diphtheriae	11051	1.5x109	2/2
Corynebacterium jeikeium	43734	6.0x108	2/2
Enterococcus avium	14025	1.4x109	2/2
Enterococcus casseliflavus	700327	1.7x109	2/2
Enterococcus durans	6056	7.8x108	2/2
Enterococcus faecalis	29212	1.7x109	2/2
Enterococcus faecalis	19433	1.4x109	2/2
Enterococcus faecalis, van A	1MC	3.1x108	2/2
Enterococcus faecalis, van B	51575	1.5x109	2/2
Enterococcus faecium	19434	4.0x108	2/2
Enterococcus faecium	6057	8.2x108	2/2
Enterococcus faecium, van A	700221	2.6x108	2/2
Enterococcus gallinarum	700425	4.1x108	2/2
Enterococcus gallinarum	49573	6.4x108	2/2
Enterococcus hirae	8043	6.9x108	2/2
Enterococcus raffinosus	49464	9.9x108	2/2
Gemella morbillorum	27824	1.0x109	2/2
Globicatella sanguinis	51174	6.2x108	2/2
Kocuria kristinae	BAA-752	1.7x109	2/2
Kocuria rosea	186	2.5x108	2/2
Kytococcus schroeteri (oxacillin resistant)	BAA-2410	1.8x108	2/2
Lactobacillus acidophilus	4356	4.8x108	2/2
Lactococcus lactis	11454	6.9x108	2/2
Lactococcus lactis	40932	2.1x109	2/2
Leuconostoc mesenteroides subsp. mesenteroides	8293	1.1x109	2/2
Leuconostoc mesenteroides subsp. mesenteroides	19254	9.7x108	2/2
Leuconostoc pseudomesenteroides	12291	1.7x109	2/2
Listeria grayi	19120	1.1x109	2/2
Listeria innocua	33090	1.3x108	2/2
Listeria ivanovii	19119	1.5x109	2/2
Listeria monocytogenes	15313	4.9x108	2/2
Listeria seeligeri	35967	1.1x109	2/2
Macrococcus caseolyticus	13548	8.3x108	12/13 (1,2)
Micrococcus luteus	10240	2.5x108	2/2
Micrococcus lylae	27567	5.8x108	2/2
Mycobacterium avium	700898	1.9x108 (gDNA)	2/2
Pediococcus damnosus	29358	1.2x109	2/2
Pediococcus pentosaceus	33316	2.9x108	2/2
Peptostreptococcus anaerobius	27337	2.3x108	2/2
Planococcus citreus	14404	9.2x108	2/2
Planococcus kocurii	43650	8.7x108	2/2
Propionibacterium acnes	11827	2.4x108	2/2
Rhodococcus equi	6939	6.8x108	2/2
Rothia dentocariosa	BAA-907	2.3x108	2/2
Rothia mucilaginosa	49040	9.6x108	2/2
Streptococcus agalactiae	BAA-611	8.7x108	4/4
Streptococcus agalactiae	13813	7.6x108	2/2
Streptococcus angunosis	NCTC 10713	7.0x108	2/2
Streptococcus constellatus	27823	1.1x109	2/2
Streptococcus dysagalactiae	43078	4.8x108	2/2
Streptococcus equi	9528	6.8x108	2/2
Streptococcus gallolyticus	9809	1.9x109	2/2
Streptococcus gallolyticus	49475	2.3x109	2/2
Streptococcus mitis	6249	1.0x108	2/2
Streptococcus mutans	25175	4.2x108	2/2
Streptococcus mutans	35668	5.1x108	12/13 (2,3)
Streptococcus parasanguinis	15909	2.2x108	2/2
Streptococcus pneumoniae	ARUP	1.6x108	2/2
Streptococcus pyogenes	49399	1.5x108	2/2
Streptococcus pyogenes	12344	1.0x108	2/2
Streptococcus pyogenes	4543	1.1x109	2/2
Streptococcus sanguinis	10556	9.0x108	2/2
Streptococcus thoraltensis	700865	1.1x109	2/2
Streptococcus uberis	9927	1.3x108	2/2
Gram Negative Bacteria
Acinetobacter baumannii	19606	9.0x108	2/2
Acinetobacter calcoaceticus	23055	6.5x108	2/2
Acinetobacter haemolyticus	19002	3.0x108	2/2
Acinetobacter Iwoffi	17925	1.4x109	2/2
Bacteriodes fragilis	23745	7.6x108	2/2
Bordetella pertussis	9797	1.2x109	2/2
Burkholderia cepacia	25416	4.4x108	2/2
Citrobacter amalonaticus	25405	3.3x108	2/2
Citrobacter freundii	8090	1.1x109	2/2
Citrobacter koseri	27156	1.0x109	2/2
Enterobacter aerogenes	15038	1.1x109	2/2
Enterobacter cloacae	13047	1.0x109	2/2
Escherichia coli	BAA-199	1.5x109	2/2 (1)
Escherichia coli	4157	1.1x109	2/2
Fusobacterium nucleatum	25586	6.7x108	2/2
Haemophilus haemolyticus	33390	1.4x109	2/2
Hafnia alvei	13337	1.6x109	2/2
Klebsiella oxytoca	13182	1.7x109	2/2
Klebsiella pneumoniae	700603	2.3x109	2/2
Klebsiella pneumoniae	BAA-1705	2.2x109	2/2
Kluyvera intermedia	33421	4.4x108	2/2 (1)
Moraxella catarrhalis	23246	2.1x109	2/2 (1)
Morganella morganii	25829	1.2x109	2/2
Neisseria gonorrhoeae	19424	2.3x109	2/2
Neisseria meningitidis	13077	1.8x109	2/2 (1)
Neisseria subflava	49275	3.5x107	2/2 (1)
Oligella urethralis	17960	3.1x108	2/2
Proteus mirabilis	25933	1.3x109	11/13 (1,4)
Proteus vulgaris	6896	2.8x108	2/2
Providencia rettgeri	9250	1.5x108	2/2
Providencia rustigianii	13159	7.3x108	2/2
Pseudomonas aeruginosa	10145	1.0x109	2/2
Pseudomonas putida	49128	1.2x109	2/2
Salmonella enterica	14028	2.1x109	2/2
Salmonella typhimurium	13311	1.3x109	2/2 (1)
Serratia liquefaciens	27592	6.0x108	14/15 (2,5)
Serratia marcescens	13880	4.4x108	2/2
Shigella sonnei	29930	7.6x108	2/2
Stenotrophomonas maltophilia	13637	1.2x109	2/2
Yersinia enterocolitica	9610	1.6x109	2/2
Yeast
Candida albicans	18804	$3.5x10^9$	2/2
Candida glabrata	66032	$2.2x10^9$	2/2
Candida krusei	24210	$9.3x10^8$	2/2
Candida parapsilosis	14054	$6.0x10^8$	2/2
Candida tropicalis	ARUP 2	$1.5x10^9$	2/2
Cryptococcus neoformans	90112	$1.1x10^9$	2/2
Mycoplasma
Mycoplasma pneumoniae	15531	$2.7x10^8$ (gDNA)	2/2 (6)

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Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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The vast majority of strains tested 'Staphylococcus NEGATIVE,' indicating no crossreactivity or interference with internal controls. The exceptions were twenty-seven (27) 'invalid' calls and six (6) 'Staphylococcus POSITIVE' calls, all noted in Table 7.

One (1) 'invalid' call out of two (2) tests was observed for a single strain of the following species: Actinomyces odontolyticus, Escherichia coli, Kluyvera intermedia, Moraxella catarrhalis, Neisseria meningitides, Neisseria subflava, Proteus mirabilis, Salmonella typhimurium, and Streptococcus uberis. Each invalid case resolved upon re-testing as ' Staphylococcus NEGATIVE'.

Two (2) 'invalid' calls out of two (2) tests were observed for Mycoplasma pneumoniae upon initial testing. Two (2) valid calls out of two (2) tests were obtained upon retesting as 'Staphylococcus NEGATIVE'.

One (1) 'Staphylococcus POSITIVE' call out of two (2) tests was observed for a single strain of the following species: Arcanobacterium haemolyticum, Macrococcus caseolyticus, Streptococcus mutans, Proteus mirabilis, Serratia liquefaciens. Each positive result resolved upon re-testing as 'Staphylococcus NEGATIVE' with a minimum of six (6) repeat tests, indicating the positive results previously obtained were likely a single contamination event in one card. One or more invalid calls were observed upon re-testing for the following species: Macrococcus caseolyticus, Streptococcus mutans and Serratia liquefaciens as noted in Table 7.

d. Microbial Interference

Off-Panel Microbial Interference: As a follow up to the previous exclusivity and inclusivity studies, the Staph ID/R Blood Culture Panel was further evaluated for the ability to detect low level Staphylococcus species in the presence of fourteen (14) "offpanel" microorqanism strains that should not be detected. The "off-panel" strains represent Gram Positive, Gram Negative, Yeast and likely skin contaminants. BACTEC Plus Aerobic/F or Anaerobic/F Bottles containing blood were inoculated with competing "off-panel" microorganisms. The "off-panel" strains were grown to high concentrations by incubating 8 hours past bottle ring 'On-board', consistent with incubation time and temperatures tested in the specimen stability study. The bottle contents were confirmed by Gram stain and serial dilutions plated on agar and counted the following day to

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confirm a concentration of >10° CFU/mL. Bottles were stored at 4°C and tested within 72 hours in combination with Staphylococcus TSB cultures at approximately 1-2.5x10° CFU/mL for each strain. The concentration of the Staphylococcus strains were verified by plating serial dilutions on agar and performing colony counts the following day. Results for the studies are shown in Table 8.

Table 8. Microbial Interference Panel (Off-panel): Non-Staphylococcus microbial strains tested for microbial interference in detecting five (5) Staphylococcus strains by the Staph ID/R Blood Culture Panel.

"Off-Panel" Microorganisms Species,Sample Input ≥ $10^8$ CFU/mL;ATCC/NCTC strain #	S. aureusBAA-16820.7-1.2x $10^6$	S. aureus116320.6-1.6x $10^6$	S. epidermidis7005620.8-0.9x $10^6$	S. epidermidis7005830.9-2.2x $10^6$	S. lugdunensis495761-1.2x $10^6$
Corynebacterium jeikeium 43734	2/2	2/2	2/2	2/2	2/2
Enterococcus faecalis 19433	2/2	2/2	2/2"	2/2	2/2
Enterococcus faecium 19434	2/2	2/2	2/2"	2/2	2/2
Listeria monocytogenes 19115	2/2	2/2	2/2"	2/2"	2/2
Micrococcus luteus 10240	2/2	2/2	2/2	2/2	2/2
Propionibacterium acnes 11827	2/2	2/2	2/2	2/2	2/2
Streptoccocus agalactiae 13813	2/2	2/2	2/2*	2/2	2/2
Streptococcus anginosus 10713	2/2	2/2	2/2	2/2	2/2"
Streptococcus pneumoniae 27336	2/2	2/2	2/2	2/2	2/2
Streptococcus pyogenes 49399	2/2	2/2	2/2"	2/2	2/2
Escherichia coli 4157	2/2	2/2	2/2	2/2"	2/2*
Klebsiella pneumoniae 700603	2/2	2/2	2/2	2/2	2/2
Pseudomonas aeruginosa 10145	2/2	2/2	2/2	2/2	2/2
Candida albicans 18804	2/2	2/2	2/2	2/2	2/2

*This set of test runs also contained 1 "Invalid" run

"This set of test runs initially miscalled, but called correctly with a higher CFU/mL input of the low level target

For the 'valid' runs tested, the potentially interfering 'off-panel' microorqanisms did not interfere with the detection of the Staphylococcus strains, resulting in 'POSITIVE' calls as expected. In some cases, a miscall was observed, and the low-target Staphylococcus strains were re-tested at a higher concentration and resulted in a positive result as expected. The re-tested concentrations are included in the table and were within the 2-3X LoD range for each species.

Staphylococcus Microbial Interference: Twelve (12) Staphylococcus species expected to be co-detected with the low level Staphylococcus species. BACTEC Plus Aerobic/F bottles containing blood were inoculated with Staphylococcus isolates. The bacteria were grown to high concentrations by incubating 8 hours past bottle ring 'Onboard', consistent with incubation time and temperatures tested in the specimen stability study. The bottle contents were confirmed by Gram stain and serial dilutions plated on agar and counted the following day to confirm a concentration of >10° CFU/mL for competing Staphylococcus strains. Bottles were stored at 4°C and tested within 72 hours in combination with Staphylococcus TSB cultures at approximately 1-2.5x100 CFU/mL for each strain. The concentration of the Staphylococcus strains were verified

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by plating serial dilutions on agar and performing colony counts the following day. Results for the studies are shown in Table 9.

Microbial Interference Panel (Staphylococcus): Staphylococcus microbial Table 9. strains tested for microbial interference in detecting five (5) different low level Staphylococcus strains by the Staph ID/R Blood Culture Panel.

Microbial InterferenceStaphylococcus species,Sample Input ≥ 108 CFU/mL;ATCC, Clinical #	Species; ATCC Strain #; Sample Input (CFU/mL)	S. aureusBAA-16820.7-1.2x106	S. aureus116320.6-1.6x106	S. epidermidis7005620.2-0.9x106	S. epidermidis7005830.9-2.2x106
S. aureus BAA-1682, mecA+	--	2/2	2/2"	2/2"	2/2
S. aureus 11632, mecA -	2/2	--	2/2"	2/2"	2/2
S. epidermidis 700562, mecA+	2/2	2/2	--	2/2"	2/2
S. epidermidis 700583, mecA-	2/2"	2/2"	2/2"	--	2/2"
S. lugdunensis 49576, mecA -	2/2	2/2	2/2"	2/2"	--
S. capitis 35661, mecA -	2/2	2/2	2/2"	2/2	2/2
S. caprae 35538, mecA -	2/2"	1/2*	2/2	2/2	2/2
S. hominis 27844, mecA -	2/2	2/2	2/2"	2/2	2/2"
S. haemolyticus BAA-1693, mecA +	2/2	2/2*	2/2"	2/2"	2/2
S. pettenkoferii Denys 38, mecA +	2/2	2/2	2/2	2/2	2/2
S. simulans 27848, mecA -	2/2	2/2	2/2"	2/2	2/2
S. warneri 27830, mecA -	2/2	2/2	2/2	2/2	2/2

*This set of test runs also contained 1 "Invalid" run

"This set of test runs initially miscalled, but called correctly with a higher CFU/mL input of the low level target

Staphylococcus interference was observed for S. aureus with S. epidermidis and S. caprae at initial concentrations tested (5.9E+05 CFU/mL), but the interference was resolved upon re-testing at higher concentrations (1.5E+06 CFU/mL, within 2-3X LoD). There were 13 cases of interference with S. epidermidis at initial concentrations (2.2-2.5E+05 CFU/mL) when tested against S. aureus, S. epidermidis, S. lugdunensis, S. capitis, S. hominis, S. haemolyticus, and S. simulans. Higher concentrations of S. epidermidis resolved the interference (8.5-9.4E+05 CFU/mL, within 2-3X LoD). Two (2) cases of interference were observed with S. lugdunensis: one (1) case with S. epidermidis, one (1) case with S. hominis. Both cases resolved at higher concentrations of S. Jugdunensis (1.2E+06 CFU/mL, within 2-3X LoD).

e. Interfering Substances (Chemical Interference)

The Staph ID/R Blood Culture Panel was evaluated for interference by a panel of sixteen (16) different substances. Substances were spiked into BACTEC Plus Aerobic/F (with resin) or Standard Aerobic/F (without resin) media containing neqative blood incubated 24 hours in a BACTEC Blood Culture Device. Target Staphylococcus cells were combined with the substances at low positive concentrations at approximately 2-3X LoD (1-2.5x10° CFU/mL). The CFU concentrations for each strain were estimated by optical density measurements and then confirmed by colony counting. The studies assessed the detection of the same ten (10) Staphylococcus ATCC strains used for analytical sensitivity and microbial interference: S. aureus, mecA+ strains BAA-1680 and BAA-1682, S. aureus, mecA+ strains 11632 and 6538, S. epidermidis, mecA+ strains 700562 and 51625, S. epidermidis, mecA- strains 700583 and S. lugdunensis, mecA-

Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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strains 49576 and 43809. ATCC strain, E. faecalis, mecA- 29212, an off-panel "Negative" in the Staph ID/R Blood Culture Panel, was also included in the study to assess chemical interference with the sample processing control and all downstream detection steps.

Similarly to previous studies, a minimum of two replicate assays was performed for each Staphylococcus strain using each substance in a background of BACTEC Plus Aerobic bottles (with resin) or Standard Aerobic bottles (without resin), see Tables 10 and 11.

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Table 10. Interfering Substances Panel (BACTEC Plus with Resin). Staph ID/R Blood Culture Panel performance evaluation for chemical interference in detecting ten (10) different Staphylococcus strains and one (1) off-panel E. faecalis strain.

	Species, ATCC strain, Sample Input (CFU/mL)											Substance Input	Species, ATCC strain, Sample Input (CFU/mL)
Substance InputConcentration intoPlus Aerobic Media(with resin)	S. aureus, mecA +		S. aureus, mecA-		S. lugdunensis, mecA -		S. epidermidis, mecA +		S. epidermidis, mecA-		E. faecalis (Neg)	Concentration intoStandard AerobicMedia (without resin)	S. aureus, mecA +		S. aureus, mecA-		S. lugdunensis, mecA -		S. epidermidis, mecA +		S. epidermidis, mecA-		E. faecalis (Neg)
	BAA-16800.9-1.8x106	BAA-16821.4-1.6x106	116321.2-2.2x106	65381.8-2.2x106	438091.1-1.7x106	495760.9-1.8x106	516250.3-1.5x106	7005620.3-0.8x106	122280.9-1.2x106	7005830.3-1.9x106	292122.4x107	resin)	BAA-16800.9-1.8x106	BAA-16821.4-1.6x106	116321.2-2.2x106	65381.8-2.2x106	438091.1-1.7x106	495760.9-1.8x106	516250.3-1.5x106	7005620.3-0.8x106	122280.9-1.2x106	7005830.3-1.9x106	292122.4x107
Whole Blood in ACD(≥35% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Whole Blood in ACD(≥35% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Whole Blood in EDTA(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Whole Blood in EDTA(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Whole Blood inHeparin(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Whole Blood inHeparin(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Whole Blood inSodium Citrate(≥35% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Whole Blood inSodium Citrate(≥35% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Human Plasma(≥40%, v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Human Plasma(≥40%, v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
SodiumPolyanetholsulfonate(≥0.20% w/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	SodiumPolyanetholsulfonate(≥0.20% w/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Hemoglobin(≥10 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Hemoglobin(≥10 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
y-Globulin(≥40 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	y-Globulin(≥40 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Triglycerides(≥10 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2*	Triglycerides(≥10 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
White Blood Cells(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	White Blood Cells(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Platelets(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Platelets(≥40% v/v)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
UnconjugatedBilirubin (≥0.075mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	UnconjugatedBilirubin (≥0.075mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Conjugated Bilirubin(≥0.075 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Conjugated Bilirubin(≥0.075 mg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2*
Vancomycin(≥50 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Vancomycin(≥50 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Ciprofloxacin(≥7.5 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Ciprofloxacin(≥7.5 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2
Cefoxitin(≥125 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	Cefoxitin(≥125 µg/mL)	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2	2/2

*This set of test runs also contained 1 "Invalid" run

In Table 10, the substances did not interfere with detection of strains at the concentrations listed, resulting in Positive or Negative calls as expected.

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Table 11. Interfering Substances Panel (BACTEC Standard without Resin). Staph ID/R Blood Culture Panel performance evaluation for chemical interference in detecting ten (10) different Staphylococcus strains and one (1) off-panel E. faecalis strain.

*This set of test runs also contained 1 "Invalid" run

As summarized in Table 11, the chemical substances tested did not interfere with detection of the majority of strains, resulting in Positive or Negative calls as expected.

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Carry-over/Cross-Contamination Study f.

A study was performed to assess the cross-contamination of the Staph ID/R Blood Culture Panel by alternatively testing high titer S. aureus, mecA+ ATCC strain BAA-1682 and off-target negative E. faecalis ATCC strain 29212. BACTEC Plus Aerobic/F blood bottle containing negative blood were inoculated with strain isolates and incubated until alarm positivity in a BACTEC Blood Culture System. Alarm positive samples were incubated past positivity consistent with timeframes used during the specimen stability studies to obtain a high titer. Alarm positive blood cultures were Gram stained, diluted, plated on aqar plates and colonies were counted the following day to confirm target concentrations >107 for S. aureus (3.0x107 CFU/mL) and >10° E. faecalis (5.8x10' CFU/mL). Aliquots from the high titer blood culture bottles were stored at -20°C until testing. Carry-over/cross-contamination was tested by running a series of alternating runs of high titer positive and negative samples on multiple Portrait Analyzers.

In conclusion, all of the 'calls' were in concordance with expected 'calls'. Therefore, there was no evidence of contamination in any of the tests.

g. Reproducibility

A multicenter, blinded, reproducibility study was performed to determine reproducibility of the Staph ID/R Blood Culture Panel. Testing occurred at three sites using a panel of seven simulated blood culture specimens, each spiked with a single organism. Specimens were prepared in a matrix of whole blood culture media. Half of the replicates for the three Staphylococcus positive samples were consistent with the level of organism present at the time of positivity (low) and half were at a concentration similar to that observed after 8 hours of positivity (high). For the one off-panel organism (E. faecalis; Staph ID/R neqative) the concentration was "high."

The study incorporated several variables including six different operators at three sites (two operators/site), five different cartridge lots, and 89 different Portrait Analyzers (15 at site 1, and 12 at site 3, 62 at site 4). Over the course of 10 weeks, samples were tested on 12 different days, for a total of 90 replicates per analyte per concentration.

Valid results were attained for 630 of 642 (98.1%) runs (see Table 12). For the detection of Staphylococcus positivity (Test result "Positive"), expected positive results were obtained for 540/540 runs (100%), and expected Staphylococcus neqative results (Test result "Negative") were obtained in 87/90 runs (96.7%). For the detection of specific Staphylococcus, expected positive results were obtained for 538/540 runs (99.6%) and expected negative results were obtained for 1341/1350 (99.3%) results (Table 12).

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Table 12. Summary of Reproducibility Study.

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Staph ID/RBlood CulturePanel Result	Species, Bacteria Load (Low or High),Sample Input (CFU/mL)	Test Site	Detected	Not Detected	%Agreement withExpected Result
S. aureus,mecA present	S. aureus (mecA +), Low;$4.0x10^6$	1	30/30	0/30	90/90; 100%
		3	30/30	0/30
		4	30/30	0/30
		Total	90/90	0/90
S. aureus,mecA present	S. aureus (mecA +) High;$4.0x10^7$	1	30/30	0/30	90/90; 100%
		3	30/30	0/30
		4	30/30	0/30
		Total	90/90	0/90
	Negative	1	1/150 (1)	149/150	448/450; 99.6%
		3	1/150 (2)	149/150
		4	0/150	150/150
		Total	2/450	448/450
Staphylococcusspecies OTHERthan S. aureus orS. lugdunensis,mecA present	S. epidermidis (mecA +), Low;$8.5x10^6$	1	30/30	0/30	90/90; 100%
		3	30/30	0/30
		4	30/30	0/30
		Total	90/90	0/90
Staphylococcusspecies OTHERthan S. aureus orS. lugdunensis,mecA present	S. epidermidis (mecA +), High;$7.0x10^7$	1	30/30	0/30	90/90; 100%
		3	30/30	0/30
		4	30/30	0/30
		Total	90/90	0/90
	Negative	1	2/150 (3)	148/150	446/450; 99.1%
		3	0/150	150/150
		4	2/150 (4)	148/150
		Total	4/450	446/450
S. lugdunensis,mecA absent	S. lugdunensis (mecA - ), Low;$6.0x10^7$	1	28/30 (3)	2/30	88/90; 97.8%
		3	30/30	0/30
		4	30/30	0/30
		Total	88/90	2/90
S. lugdunensis,mecA absent	S. lugdunensis (mecA - ), High;$5.1x10^8$	1	30/30	0/30	90/90; 100%
		3	30/30	0/30
		4	30/30	0/30
		Total	90/90	0/90
	Negative	1	2/150 (5,6)	148/150	447/450; 99.3%
		3	1/150 (5)	149/150
		4	0/150	150/150
		Total	3/450	447/450
StaphylococcusPositive	S. aureus , S. epidermidis , S. lugdunensis,Low and High	1	180/180	0/180	540/540; 100%
		3	180/180	0/180
		4	180/180	0/180
		Total	540/540	0/540
StaphylococcusNegative	E. faecalis, (mecA - ), High;$1.1x10^9$	1	1/30 (6)	29/30	87/90; 96.7%
		3	0/30	30/30
		4	2/30 (4)	28/30
		Total	3/90	87/90
Staph ID/RBlood CulturePanel Result	Species, Bacteria Load (Low orHigh), Sample Input (CFU/mL)	Test Site	Detected	Not Detected	%Agreement withExpected Result
S. aureus, mecApresent	S. aureus (mecA+) , Low;$4.0x10^6$	Site 1	30/30	0/30	90/90; 100%
		Site 3	30/30	0/30
		Site 4	30/30	0/30
		Total	90/90	0/90
S. aureus, mecApresent	S. aureus (mec A+) High;$4.0x10^7$	Site 1	30/30	0/30	90/90; 100%
		Site 3	30/30	0/30
		Site 4	30/30	0/30
		Total	90/90	0/90
	Negative	Site 1	0/150	150/150	450/450; 100%
		Site 3	0/150	150/150
		Site 4	0/150	150/150
		Total	0/450	450/450
Staphylococcusspecies OTHERthan S. aureus orS. lugdunensis ,mecA present	S. epidermidis (mec A+) , Low;$8.5x10^6$	Site 1	30/30	0/30	90/90; 100%
		Site 3	30/30	0/30
		Site 4	30/30	0/30
		Total	90/90	0/90
	S. epidermidis (mec A+) , High;$7.0x10^7$	Site 1	29/30 (1)	1/30	89/90; 98.9%
		Site 3	30/30	0/30
		Site 4	30/30	0/30
		Total	89/90	1/90
	Negative	Site 1	0/150	150/150	450/450; 100%
		Site 3	0/150	150/150
		Site 4	0/150	150/150
		Total	0/450	450/450
mecA Present;no organismassociated	S. aureus (mec A+) ,S. epidermidis (mecA +)Low and High	Site 1	119/120	1/120	359/360; 99.7%
		Site 3	120/120	0/120
		Site 4	120/120	0/120
		Total	359/360	1/360
mecA Absent; noorganismassociated	S. lugdunensis (mecA -) ,E. faecalis (mecA -)Low and High	Site 1	0/90	90/90	270/270; 100%
		Site 3	0/90	90/90
		Site 4	0/90	90/90
		Total	0/270	270/270

(1) Sample detected as "Staphylococcus aureus in mixed Staph infection (NOT S. lugdunensis)"

(2) One S. lugdunensis specimen additionally detected S. aureus

(3) Two S. lugdunensis specimens detected as Staphylococcus OTHER than S. aureus or S. lugdunensis

(4) Two E. faecalis specimens detected as Staphylococus OTHER than S. aureus or S. lugdunensis

(5) One specimen detected S. aureus correctly, but additionally detected S. lugdunensis

(6) One E. faecalis specimen detected as S. lugdunensis

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For detection of the mecA gene with no associated organism, positive results (mecA Present) were detected in 359/360 (99.7%) runs and negative results (mecA absent) were detected in 270/270 (100%) runs (see Table 13). For S. aureus (mec4+), positive mecA results were obtained for 180/180 (100%) runs and negative mecA results were obtained for 450/450 (100%) runs. For S. epidermidis (mecA+), positive mecA results were obtained for 179/180 (99.4%) runs and negative mecA results were obtained for 450/450 (100%) runs.

	Table 13. Summary of mecA results from Reproducibility Studies.

(1) One Staphylococcus species OTHER than S. aureus or S. lugdunensis, mecA present reported as mecA absent

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In the study, ten (10) discrepant results were obtained (see Table 14).

Correct Result	Staph ID/R Blood Culture Panel Result	DiscrepantResult	Site	Operator	TestDay
Staphylococcus lugdunensis, mecA absent	Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staph. species	1	1	3
Staphylococcus lugdunensis, mecA absent	Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staph. species	1	2	4
Negative, mecA N/A	Staphylococcus lugdunensis, mecA absent	S. lugdunensis	1	2	5
Staphylococcus aureus, mecA present	Staphylococcus aureus ; Staphylococcus lugdunensis,mecA present	S. lugdunensis	1	2	5
Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	mecA absent	1	2	5
Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staphylococcus aureus in mixed Staph infection(NOT S. lugdunensis ), mecA present	S. aureus	1	2	5
Staphylococcus aureus, mecA present	Staphylococcus aureus ; Staphylococcus lugdunensis,mecA present	S. lugdunensis	3	1	1
Staphylococcus lugdunensis, mecA absent	Staphylococcus aureus ; Staphylococcus lugdunensis,mecA absent	S. aureus	3	2	2
Negative, mecA N/A	Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staph. species	4	2	4
Negative, mecA N/A	Staphylococcus OTHER than S. aureus orS. lugdunensis, mecA absent	Staph. species	4	2	5

Table 14. Summary of discrepant results from Reproducibility Studies.

h. Evaluation of Blood Culture Bottle Types

To determine the effect of blood culture bottle type on Staph ID/R Blood Culture Panel, thirteen (13) unique bottle types were tested in the presence of target and non-target orqanisms. Staphylococcus isolates or E. faecalis (neqative) at bottle rinq load levels (2x10°-1x10° CFU/mL) consistent with the specimen stability studies. All bottles were tested with the highest volume of negative blood recommended by the manufacturer (e.g., if the recommended blood volume was 8-10 mL of blood was spiked into the bottle). Bottles with blood were pre-incubated 18 hours at 35-37℃ in a BACTEC Blood Culture System or in a shaking incubator prior to testing. Bacteria isolates were incubated >18 hrs in TSB, and the CFU concentrations for each strain were estimated by optical density measurements, confirmed by serial dilution and colony counting.

The studies assessed the detection of seven (7) Staphylococcus ATCC strains used for analytical sensitivity: S. aureus, mecA+ BAA-1680, S. aureus, mecA+ 1682, S. aureus, mecA- 11632, S. epidermidis, mecA+ 51625, S. epidermidis, mecA- 12228, S. lugdunensis, mecA- 49576, S. capitis, mecA- 35661. The studies also assessed performance of the Staph ID/R Blood Culture Panel in the presence of E. faecalis 29212 (Negative).

The following bottle types were tested in the study: BACTEC (Standard 10 Aerobic/F, Standard Anaerobic 10/F, Plus Aerobic/F, Plus Anaerobic/F, Lytic 10/F, and PEDS Plus/F), BacT/Alert (SA Standard Aerobic, SN Standard Anaerobic, FA FAN Aerobic, FN FAN Anaerobic, and PF Pediatric FAN), and Versa Trek (Redox 1 and Redox 2). Three bottles of each bottle type were used for each strain, and the samples tested three

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times in the Staph ID/R Blood Culture Panel for a total of 9 runs for each strain and bottle type combination.

For the 'valid' runs tested, all of the potential blood bottle types were compatible with the Staph ID/R Blood Culture Panel, with no false negative results (see Table 15).

Bottle Type	S. aureus (mecA +)BAA-1680$1.9x10^7$	S. aureus (mecA +)BAA-1682$1.2x10^7$	S. aureus (mecA -)11632$7.5x10^7$	S. lugdunensis (mecA -)49576$1.9x10^8$	S. epidermidis (mecA -)12228$2.9x10^7$	S. epidermidis (mecA +)51625$6.5x10^7$	S. capitis (mecA-)35661$2.3x10^6$	E. faecalis (mecA -)29212$2.1-4.5x10^7$
BACTEC Standard 10 Aerobic/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BACTEC Standard Anaerobic 10/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BACTEC Plus Aerobic/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BACTEC Plus Anaerobic/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BACTEC Lytic 10/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BACTEC PEDS Plus/F	9/9	9/9	9/9	9/9	9/9	9/9	9/9	14/14****
BacT/Alert SA Aerobic	9/9	9/9	9/9	9/9	9/9	9/9	9/9	16/16**
BacT/Alert SN Anaerobic	9/9	9/9	9/9	9/9	9/9	9/9	9/9	16/17*"
BacT/Alert FA FAN Aerobic	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BacT/Alert FN FAN Anaerobic	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9
BacT/Alert PF Pediatric FAN	9/9	9/9	9/9	9/9	9/9	9/9	9/9	17/17**
Versa Trek Redox 1	9/9	9/9	8/9'	9/9	9/9	9/9	9/9	9/9
Versa Trek Redox 2	9/9	9/9	9/9	9/9	9/9	9/9	9/9	9/9

*This set of test runs also contained 1 "Invalid" run

**This set of test runs also contained 2 "Invalid" runs

***This set of test runs also contained 4 "Invalid" runs

'This set of test runs contained 1 false positive result for mecA

"This set of test runs contained 1 false positive Staphylococcus, species undetermined result

Table 15. Evaluation of blood culture bottle types study results.

A false positive " S. aureus, mecA Present" result was observed in one test run from one Versa Trek Redox 1 bottle for S. aureus, mecA- 11632. The discrepant result is thought to be a contamination event, since all other samples (8/9) gave the correct "S. aureus present, mecA Absent" call, including 2/3 correct results from the same bottle with the discrepant call. All test results did result in a correct S. aureus result, suggesting that the bottle type did not interfere with the assay.

A false positive " Staphylococcus Positive, Staphylococcus species OTHER than S. aureus or S. lugdunensis, mecA absent" result was observed for E. faecalis 29212 in one test run from one BacT/Alert SN Anaerobic bottle. The discrepant result is also thought to be a random contamination event, because the sixteen (16) other "valid" test runs returned a correct "Staphylococcus Negative" result, including 2/3 correct initial results and 3/3 correct re-test results from the same bottle with the discrepant call.

The only other exceptions observed during this study were nine (9) 'Invalid' call runs that are noted in Table 15, all with E. faecalis 29212. When tested with E. faecalis 29212, one (1) invalid run was observed for BACTEC PEDS Plus/F, one (1) invalid run for BacT/Alert SA Aerobic, and one (1) invalid run for BacT/Alert PF Pediatric FAN bottle types. In all of these cases, extra test runs were performed to evaluate any possible

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interference with the assay in a Negative sample (evaluating SPC only). Re-test with nine (9) cards resulted in an additional three (3) invalid runs for BACTEC PEDS Plus/F, one (1) invalid run for BacT/Alert SA Aerobic, and one (1) invalid run for BacT/Alert SN Anaerobic (retested due to false positive contamination result). Overall, it appears that the bottle types with invalid results have an elevated invalid rate (5-28%) compared to the prospective study (2%).

H. Performance Data - Prospective Clinical Studies

Specimens for the clinical study were collected prospectively at three geographically diverse U.S. sites. Eligible study subjects included individuals receiving routine care requiring blood culture testing. Blood culture specimens were collected from the patients and incubated on the BACTEC continuous monitoring blood culture system. Bottles that were flagged positive by the instrument were Gram stained and then bottles confirmed to contain gram-positive cocci in clusters (GPCC) or gram-positive cocci in singles (GPC) were then tested with the Staph ID/R Blood Culture Panel. A total of 853 samples were collected for all three sites combined. Twenty-two (22) specimens were excluded from the Staph ID/R Blood Culture Panel clinical study dataset. The remaining 831 clinical specimens met the inclusion criteria and were used in the prospective study to evaluate the performance of the Staph ID/R Blood Culture Panel. A total of 762 prospective specimens were tested in the clinical trial, while the remaining 69 archived frozen specimens were tested after the prospective clinical trial.

In addition, 102 Staph ID/R Blood Culture Panel assays were performed on a 'Low Prevalence' panel of contrived or 'simulated' blood culture specimens, consisting of low prevalence Staphylococcus species and gram-positive negatives. These specimens were prepared by spiking blood culture bottles containing whole blood with bacterial suspensions of bacterial isolates. Prepared blood culture bottles were then grown to positivity on the BACTEC blood culture system until flagged positive. Gram stain was performed to verify the presence of gram-positive cocci in clusters (GPCC) or gram-positive cocci in singles (GPC) and then testing was performed with the Staph ID/R Blood Culture Panel. Results from the studies of all three clinical sites combined are summarized in Table 16.

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Table 16. Summary of Clinical Performance of Staph ID/R Blood Culture Panel versus Reference Method(s) – Prospective and Simulated/Supplemental Blood Cultures.

	% Agreement
All sites combined		TP/TP +FN	PPA95% CI	TN/TN + TP	NPA95% CI
Detection of Staphylococcus aureus	Prospective	211/214	98.6%96.0 - 99.5%	548/551	99.5%98.4 - 99.8%
	Simulated	0	N/A	102/102	100.0%96.4 - 100%
	Overall	211/214	98.6%96.0 - 99.5%	650/653	99.5%98.7 - 99.8%
Detection of Staphylococcus lugdunensis	Prospective	3/3	100.0%43.9 - 100%	761/762	99.9%99.3 - 99.9%
	Simulated	30/30	100.0%88.7 - 100%	72/72	100.0%94.9 - 100%
	Overall	33/33	100.0%89.6 - 100%	833/834	99.9%99.3 - 99.9%
Detection of Staphylococcus species OTHER than S. aureusor S. lugdunensis	Prospective	444/449	98.9%97.4 - 99.5%	307/316	97.2%94.7 - 98.5%

Detection of mecA with Staphylococcus aureus	Prospective	68/72	94.4%86.6 - 97.8%	682/690	98.8%97.7 - 99.4%
	Frozen	35/35	100.0%90.1 - 100.0%	34/34	100.0%89.9 - 100.0%
	Overall	103/107	96.3%90.8 - 98.5%	716/724	98.9%97.8 - 99.4%
Detection of mecA with Staphylococcus lugdunensis	Prospective	0/0	N/A	762/762	100.0%99.5 - 100%
Detection of mecA with Staphylococcus species OTHERthan S. aureus or S. lugdunensis	Prospective	243/262	92.7%88.1 - 97.1%	481/500	96.2%92.4 - 98.0%

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Table 17 lists all polymicrobial specimens from the prospective performance data determined by the Staph ID/R Blood Culture Panel and/or Reference Method(s).

Table 17. Polymicrobials - Mixed Specimen Combinations Detected by Staph ID/R Blood
Culture Panel and Reference Method(s).

Staph ID/R Result			Reference Result			Discrepant Result Description
Site	Sample ID	Species Identification	mecA Result	Organism 1	Cefoxitin Result	Organism 2	Cefoxitin Result	Organism 3	Species Identification	mecA Result
Polymicrobial for both Staph ID/R Blood Culture Panel AND Reference Results
Site 2 (Daly)	DALY125	S. aureus in mixed Staph infection (NOT S. lugdunensis)	Present	S. aureus	Sensitive	S. epidermidis	Resistant		Correct species call	Correct mecA call
Polymicrobial for Reference Results
Site 1, IU (Denys)	DNYS031	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. aureus	Sensitive	S. epidermidis	Sensitive	S. hominis	FN for S. aureus; Correct for Staph call	Correct mecA call
Site 1, IU (Denys)	DNYS033	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. capitis	Sensitive	S. epidermidis	No growth		Correct species call	Correct mecA call
Site 1, IU (Denys)	DNYS047	Staph. species OTHER than S. aureus or S. lugdunensis	Present	S. capitis	Sensitive	S. epidermidis	Resistant		Correct species call	Correct mecA call
Site 1, IU (Denys)	DNYS071	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. epidermidis	Sensitive	S. haemolyticus	Sensitive		Correct species call	Correct mecA call
Site 1, IU (Denys)	DNYS123	Staph. species OTHER than S. aureus or S. lugdunensis	Present	S. hominis	Resistant	S. epidermidis	Resistant		Correct species call	Correct mecA call
Site 1, IU (Denys)	DNYS191	Staph. species OTHER than S. aureus or S. lugdunensis	Present	S. hominis	Resistant	S. epidermidis	Resistant		Correct species call	Correct mecA call
Site 1, IU (Denys)	DNYS202	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. epidermidis	Sensitive	S. epidermidis	Resistant		Correct species call	FN mecA
Site 1, IU (Denys)	DNYS288	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. capitis	Sensitive	S. pettenkoferi	Resistant		Correct species call	FN mecA
Site 1, IU (Denys)	DNYS297	Staph. species OTHER than S. aureus or S. lugdunensis	Present	S. hominis	Resistant	S. capitis	Sensitive		Correct species call	Correct mecA call
Site 1, IU (Denys)	YNG109	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. epidermidis	Sensitive	S. capitis	Sensitive		Correct species call	Correct mecA call
Site 3, TriCore (Young)	YNG129	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. epidermidis	Sensitive	S. warneri	Sensitive		Correct species call	Correct mecA call
Site 3, TriCore (Young)	YNG172	Staph. species OTHER than S. aureus or S. lugdunensis	Present	S. epidermidis	Resistant	S. haemolyticus	Sensitive		Correct species call	Correct mecA call
Site 3, TriCore (Young)	YNG197	Staph. species OTHER than S. aureus or S. lugdunensis	Absent	S. hominis	Sensitive	S. capitis	Sensitive		Correct species call	Correct mecA call
Polymicrobial for Staph ID/R Blood Culture Panel - 'mixed Staph infections'
Site 1, IU (Denys)	DNYS026	S. aureus in mixed Staph infection (NOT S. lugdunensis)	Present	S. hominis	Resistant				FP for S. aureus; Correct for Staph call	Correct mecA call
Site 1, IU (Denys)	DNYS045	S. aureus in mixed Staph infection (NOT S. lugdunensis)	Present	S. aureus	Sensitive				Correct for S. aureus; FP for mixed	FP mecA
Site 3, TriCore (Young)	YNG207	S. aureus in mixed Staph infection (NOT S. lugdunensis)	Present	S. epidermidis	Resistant				FP for S. aureus; Correct for Staph call	Correct mecA call
Site 3, TriCore (Young)	YNG299	S. aureus in mixed Staph infection (NOT S. lugdunensis)	Absent	S. epidermidis	Sensitive				FP for S. aureus; Correct for Staph call	Correct mecA call

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Table 18 summarizes the prospective and 'Low Prevalence' simulated performance data for Staphylococcus genus-level analyte in the Staph ID/R Blood Culture Panel, i.e., the 'Staphylococcus OTHER than S. aureus or S. lugdunensis' analyte. The performance data are stratified by individual Staphylococcus species as determined by the Reference Method(s).

Table 18. Summary of Staphylococcus Genus-level Analyte for "Staphylococcus OTHER than S. aureus or S. lugdunensis versus Reference Method(s).

	% Agreement (95% CI)
Species	Prospective	Simulated
S. arlettae	-	3/3100%43.9-100%
S. auricularis	-	3/3100%43.9-100%
S. capitis	35/35100%90.1-100%	-
S. carnosus	1/1100%20.7-100%	-
S. cohnii	2/2100%34.2-100%	3/3100%43.9-100%
S. equorum	2/2100%34.2-100%	-
S. haemolyticus	15/15100%79.6-100%	3/3100%43.9-100%
S. hominis	96/9797.9%92.8-99.4%	-
S. intermedius	-	3/3100%43.9-100%
S. pettenkoferi	7/7100%64.6-100%	-
S. saprophyticus	8/8100%67.6-100%	-
S. schleiferi	2/2100.0%67.6-100%	3/3100%43.9-100%
S. sciuri	-	3/3100%43.9-100%
S. simulans	2/2100%34.2-100%	3/3100.0%43.9-100%
S. species	2/2100%34.2-100%	-
S. warneri	8/8100%67.6-100%	3/3100%43.9-100%
S. xylosus	-	3/3100%43.9-100%

Great Basin Scientific, Inc. 2441 S. 3850 W. Salt Lake City, UT 84120 www.gbscience.com phone: 801-990-1055 fax: 801-990-1051

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I. Invalid and Abort Rates

Invalid rates and run abort rates for the prospective clinical studies are found in Table 19. The overall initial Invalid rate in the prospective clinical studies was 1.39%. Valid results were achieved after a single retest for all of the Invalid runs, resulting in a final Invalid rate of 0%. For run aborts, the overall abort rate for the prospective clinical studies was 3.30%. All of these specimens were later run successfully resulting in a final abort rate of 0.00%. All run aborts are referred as "Test Incomplete" by the Portrait Staph ID/R Blood Culture Panel software.

Table 19. Invalids and Aborts with Prospective Clinical Specimens.

Clinical Site	# Runs	# ofInitialInvalids	InitialInvalid Rate	FinalInvalidRate	# ofInitialAborts	Initial AbortRate	FinalAbortRate
Site 1, IU (Denys)	332	4	1.20%	0.00%	10	3.01%	0.00%
Site 2, PCMC (Daly)	157	4	2.55%	0.00%	6	3.82%	0.00%
Site 3, TriCore (Young)	300	3	1.00%	0.00%	10	3.33%	0.00%
Overall	789	11	1.39%	0.00%	26	3.30%	0.00%

No Call rates (invalid + abort rates) for the prospective clinical studies are found in Table 20. The overall initial No Call Rate was 4.69%. All Invalid runs and run aborts were later run successfully, resulting in a final No Call rate of 0.00%.

Table 20. Prospective Clinical Specimens with "No Call" (Invalids + Aborts).

Clinical Site	# of Runs	Initial No Call(Invalids + Aborts)	Initial No Call Rate(Invalids + Aborts)	Final No Call Rate(Invalids + Aborts)
Site 1, IU (Denys)	332	14	4.22%	0.00%
Site 2, PCMC (Daly)	157	10	6.37%	0.00%
Site 3, TriCore (Young)	300	13	4.33%	0.00%
Overall	789	37	4.69%	0.00%

J. Conclusion

The submitted information in this product notification is complete and supports a substantial equivalence decision.

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).