Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description focuses on automated real-time PCR and embedded calculation algorithms for interpreting fluorescent signals, which are standard techniques in molecular diagnostics and do not indicate the use of AI/ML. There is no mention of AI, ML, or related concepts in the document.

Therapeutic

No.
The device is an in vitro diagnostic test for detecting specific gene sequences related to carbapenem-nonsusceptible bacteria. While it aids clinicians in determining therapeutic strategies, it does not directly treat or provide therapy to patients.

Diagnostic

Yes
The "Intended Use / Indications for Use" section explicitly states that the device is a "qualitative in vitro diagnostic test designed for the detection and differentiation of gene sequences". It is also intended "as an aid for infection control" and "as an aid to clinicians in determining appropriate therapeutic strategies for patients." These phrases clearly indicate its function as a diagnostic tool.

SaMD (Software as a Medical Device)

No

The device description clearly states that the GenePOC™ Carba assay is an in vitro diagnostic test that includes physical components such as single-use, disposable microfluidic cartridges (PIEs), Sample Buffer Tubes (SBT), and Disposable Transfer Tools (DTT), and is performed on the revogene™ instrument, which is a piece of hardware. While software is involved in interpreting results, the device is not solely software.

IVD (In Vitro Diagnostic)

Based on the provided text, the device is indeed an IVD (In Vitro Diagnostic).

Here's why:

Intended Use/Indications for Use: The text explicitly states it is a "qualitative in vitro diagnostic test designed for the detection and differentiation of the blakec. blaNDM, blackA-48-like, and blamp gene sequences associated with carbapenem-nonsusceptible pure colonies..." This directly aligns with the definition of an in vitro diagnostic device, which is used to examine specimens taken from the human body to provide information for diagnosis, monitoring, or screening.
Device Description: The description details a system that processes biological samples (bacterial colonies) to detect specific genetic material using PCR, a common in vitro diagnostic technique. It also mentions "in vitro diagnostic test" again in the description.
Performance Studies: The document describes clinical and analytical performance studies, which are standard requirements for demonstrating the safety and effectiveness of IVD devices.
Predicate Device: The mention of a "Predicate Device(s)" with a K number (K152614) and name (Xpert® Carba-R Assay) strongly indicates that this device is being compared to a previously cleared IVD device, a common process for regulatory submission of new IVDs.

Therefore, all the evidence points to the GenePOC™ Carba assay being an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The GenePOC™ Carba assay, performed on the revogene™ instrument, is a qualitative in vitro diagnostic test designed for the detection and differentiation of the blakec. blaNDM, blackA-48-like, and blamp gene sequences associated with carbapenem-nonsusceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa, when grown on blood agar or MacConkey agar. The test utilizes automated real-time Polymerase Chain Reaction (PCR).

The GenePOC™ Carba assay should be used in conjunction with other laboratory tests including phenotypic antimicrobial susceptibility testing. A negative GenePOC™ Carba assay result does not preclude the presence of other resistance mechanisms.

The GenePOC™ Carba assay is intended as an aid for infection control in the detection of carbapenem-non-susceptible bacteria that colonize patients in healthcare settings. The identification of a bland, blaym or blaviM metallo-B-lactamase gene (i.e., the genes that encode the IMP, NDM and VIM metallo-ß-lactamases, respectively) may be used as an aid to clinicians in determining appropriate therapeutic strategies for patients with known or suspected carbapenem non-susceptible infections.

Special conditions for use statement(s):
Prescription Use Only

Organisms should be identified and carbapenem non-susceptibility status should be determined prior to testing with the GenePOCTM Carba assay.

The GenePOC™ Carba assay detects blakec, blandM, blavisM, blaoxA-48-like, and blamp resistance genes from pure colonies and is not for bacterial identification.

The performance of the GenePOC™ Carba assay with bacteria other than Enterobacteriaceae, Acinetobacter baumannii or Pseudomonas aeruginosa, has not been evaluated.

The GenePOC™ Carba assay is not a sub-typing tool and does not report variants of the blakpc, blandm, blaym, blaoxa-48-like, and blandp

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

PMY, OOI

Device Description

The GenePOC™ Carba assay is an automated real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection and differentiation of the blaype, blayDM, blavin, blaoxA-48-ike, and blamp gene sequences in carbapenem-non-susceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa grown on blood agar or MacConkey agar. The GenePOC™ Carba assay is designed to be performed on the revogene™ instrument. The revogene™ is an instrument that automates sample homogenization, sample dilution, cells lysis, DNA amplification, and detection of the amplified PCR products.

The GenePOC™ Carba assay kit is comprised of single-use, disposable microfluidic cartridges (PIEs), Sample Buffer Tubes (SBT), and Disposable Transfer Tools (DTT). These components are used to dilute the sample, extract, amplify, and simultaneously detect blayDM, blavin, blaoxA-48-like, and blamp DNA. A Process Control (PrC) is also incorporated into each PIE to verify sample processing and amplification steps. The PrC allows for the verification of potential inhibitor substances as well as microfluidic, instrument or reagent failure.

Each GenePOC™ Carba assay kit contains 24 individual pouches. Each pouch has components for one (1) test including one (1) Carba PIE, one (1) SBT, and one (1) DTT. The single-use, multi-chambered fluidic cartridges are designed to complete sample preparation and real-time PCR for the detection of the blaker, blands, blaoxa-48-like, and blanyp gene sequences from isolates of pure cultures of carbapenem-non-susceptible gram-negative bacteria in approximately 70 minutes. User intervention is only required for preparing the standardized 0.5 McFarland bacterial suspension from characterized carbapenem-non-susceptible isolated colonies, inoculating the bacterial suspension into the Sample Buffer Tube (SBT), transferring the sample into the single-use disposable PIE, and loading/unloading the PIEs into the revogene™ carousel. Each PIE is a completely integrated closed device in which a sample is dispensed and processed through different microfluidic chambers and channels that allow for the sample processing and subsequent real-time PCR steps.

Upon completion of a run, the results are interpolated by the revogene™ from measured fluorescent signals and embedded calculation algorithms. The output results include positive, negative, indeterminate, and unresolved. Upon completion of a run, the user removes the used cartridges and disposes of them in normal biological waste. The results are automatically generated at the end of the process in a report that can be viewed and printed.

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

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

A total of 532 bacterial isolates (475 clinical stock isolates and 57 prospectively collected fresh isolates) were enrolled in the clinical study. Sixteen (16) isolates were excluded from the analysis of performance for the following reasons: eleven (11) did not meet the inclusion criteria for species identification (either they were not among the targeted organisms or organism identification was inconclusive), three (3) were unavailable for analysis due to laboratory error and two (2) were found to be susceptible to all four (4) carbapenems tested. Four (4) additional isolates were also excluded because they were associated with a Negative External Control failure on initial testing and produced Indeterminate results upon repeat. Thus, 512 compliant isolates remained in the final analysis.

A total of 512 fully compliant isolates were tested with both the Reference Method and the GenePOC™ Carba assay for the performance estimation study. The primary objective of the trial was to establish the performance characteristics of the GenePOC™ Carba assay for its use in determining the presence of blaype, blaym, blaoxA-48-like, and blann gene DNA sequences in carbapenem-non-susceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa, when grown on blood agar and MacConkey agar, and to establish the sensitivity and the specificity in comparison to the Reference Method.

For Reference Method testing, well-isolated colonies grown on blood agar plates were diluted to a standardized 0.5 McFarland (McF) suspension before testing. For GenePOC™ Carba assay testing, well-isolated colonies grown on each of the agar types were suspended to a standardized 0.5 McF suspension. If discordant results between the GenePOC™ Carba assay and Reference Method were observed, discrepant testing was performed using alternative PCR for each of the five (5) assay analytes followed by bi-directional sequencing on isolates from blood agar plates.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Study Type: Analytical Performance - Reproducibility/Precision
Sample Size: 10 positive bacterial carbapenem-non-susceptible strains (2 per resistance gene) and 1 negative bacterial carbapenem-non-susceptible strain.
Key Results:

Between-Laboratory Reproducibility:
- Negative samples: 98.8% agreement (237/240; 95% CI: [96.4-99.6%])
- Positive samples: 100.0% agreement (120/120; 95% CI: [96.9-100.0%]) for each panel member.
Between-Lot Reproducibility:
- Negative samples: 99.6% agreement (239/240; 95% CI: [97.7-99.9%])
- Positive samples: 100.0% agreement (120/120; 95% CI: [96.9-100.0%]) for each panel member.
Within-Laboratory Precision:
- Negative samples: 100.0% agreement (80/80; 95% CI: [95.4-100.0%])
- Positive samples: 100.0% agreement (40/40; 95% CI: [91.2-100.0%]) for each panel member.

Study Type: Analytical Performance - Analytical Reactivity (Inclusivity)
Sample Size: 58 carbapenem-non-susceptible isolates of Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii.
Key Results: The ability of the GenePOC™ Carba assay to detect multiple variants of the blaxec, blaNDM, blaoxA-48-like, and blaiMP carbapenemase genes in the 58 strains was demonstrated.

Study Type: Analytical Performance - Analytical Specificity
Sample Size: 50 bacterial strains of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa.
Key Results: Only one (1) strain (Klebsiella pneumoniae CCUG 59359 blaTEM-52) out of 50 showed cross-reactivity when tested at 1.13x10^7 CFU/mL of SB, but not at a 10-fold dilution (1.13x10^6 CFU/mL of SB). All other strains yielded negative results.

Study Type: Analytical Performance - Interfering Substances
Sample Size: 5 positive bacterial strains and 1 negative bacterial strain tested with 12 combinations of culture media and saline solutions.
Key Results: The study demonstrated the compatibility of the GenePOCTM Carba assay with culture media and colony diluent from different manufacturers.

Study Type: Analytical Performance - Carry-over and Cross-Contamination Studies
Sample Size: Negative and high positive samples (n=40 each) for both carry-over and cross-contamination studies.
Key Results: No unexpected positive results for the blaxpc target were detected in negative samples in either study.

Study Type: Clinical Study
Sample Size: 512 compliant isolates.
Key Results:

Non-reportable rates on initial run: 1.7% (9/516) from blood agar and 1.4% (7/516) from MacConkey agar. Final non-reportable rate was 0.8% (4/516) across all media and gene targets.
Performance on Isolated Colonies Grown on Blood Agar Relative to Reference Method:
- NDM: Sensitivity 98.9% (96.2 - 99.7%), Specificity 99.4% (97.8 - 99.8%)
- KPC: Sensitivity 99.1% (95.2 - 99.8%), Specificity 99.2% (97.8 - 99.7%)
- OXA-48-like: Sensitivity 100.0% (94.4 - 100.0%), Specificity 99.3% (98.0 - 99.8%)
- IMP: Sensitivity 100.0% (87.5 - 100.0%), Specificity 96.1% (94.0 - 97.5%)
- VIM: Sensitivity 100.0% (93.1 - 100.0%), Specificity 99.8% (98.8 - 100.0%)
Performance on Isolated Colonies Grown on MacConkey Agar Relative to Reference Method (Sensitivity/Specificity not explicitly provided, but data similar to Blood Agar for most targets and organisms):
- NDM: TP 186, FP 21, TN 322, FN 26
- KPC: TP 114, FP 32, TN 395, FN 0
- OXA-48-like: TP 65, FP 23, TN 445, FN 0
- IMP: TP 27, FP 214, TN 464, FN 0
- VIM: TP 52, FP 15, TN 459, FN 0

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Performance of GenePOC™ Carba Assay on Isolated Colonies Grown on Blood Agar Relative to Reference Method:

NDM: Sensitivity 98.9% [96.2 - 99.7%], Specificity 99.4% [97.8 - 99.8%]
KPC: Sensitivity 99.1% [95.2 - 99.8%], Specificity 99.2% [97.8 - 99.7%]
OXA-48-like: Sensitivity 100.0% [94.4 - 100.0%], Specificity 99.3% [98.0 - 99.8%]
IMP: Sensitivity 100.0% [87.5 - 100.0%], Specificity 96.1% [94.0 - 97.5%]
VIM: Sensitivity 100.0% [93.1 - 100.0%], Specificity 99.8% [98.8 - 100.0%]

Performance of GenePOC™ Carba Assay by Organism Category and by Target Gene, for Isolated Colonies Grown on Blood Agar Relative to Reference Method:

Enterobacteriaceae
- NDM: Sensitivity 98.8% [93.7 - 99.8%], Specificity 99.5% [97.5 - 99.9%]
- KPC: Sensitivity 99.1% [95.2 - 99.8%], Specificity 98.4% [95.5 - 99.5%]
- OXA-48-like: Sensitivity 100.0% [94.3 - 100.0%], Specificity 98.8% [96.4 - 99.6%]
- IMP: Sensitivity 100.0% [78.5 - 100.0%], Specificity 98.3% [96.1 - 99.3%]
- VIM: Sensitivity 100.0% [75.8 - 100.0%], Specificity 100.0% [98.7 - 100.0%]
Pseudomonas aeruginosa
- NDM: Sensitivity 100.0% [87.1 - 100.0%], Specificity 98.8% [93.3 - 99.8%]
- KPC: Sensitivity -, Specificity 100.0% [96.5 - 100.0%]
- OXA-48-like: Sensitivity 100.0% [20.7 - 100.0%], Specificity 100.0% [96.5 - 100.0%]
- IMP: Sensitivity 100.0% [56.6 - 100.0%], Specificity 86.3% [78.3 - 91.6%]
- VIM: Sensitivity 100.0% [91.0 - 100.0%], Specificity 100.0% [94.7 - 100.0%]
Acinetobacter baumannii
- NDM: Sensitivity 98.7% [92.9 - 99.8%], Specificity 100.0% [85.7 - 100.0%]
- KPC: Sensitivity 100.0% [20.7 - 100.0%], Specificity 100.0% [96.2 - 100.0%]
- OXA-48-like: Sensitivity -, Specificity 100.0% [96.3 - 100.0%]
- IMP: Sensitivity 100.0% [67.6 - 100.0%], Specificity 100.0% [96.0 - 100.0%]
- VIM: Sensitivity 100.0% [20.7 - 100.0%], Specificity 99.0% [94.4 - 99.8%]

Performance of GenePOC™ Carba Assay by Organism Category and by Target Gene, on Isolated Colonies Grown on MacConkey Agar Relative to Reference Method (values from the table for MacConkey repeat for some metrics):

Enterobacteriaceae
- NDM: Sensitivity 98.8% [93.7 - 99.8%], Specificity 99.5% [ 97.5 - 99.9%]
- KPC: Sensitivity 100.0% [96.7 - 100.0%], Specificity 98.4% [95.5 - 99.5%]
- OXA-48-like: Sensitivity 100.0% [94.3 - 100.0%], Specificity 99.2% [97.0 - 99.8%]
- IMP: Sensitivity 100.0% [78.5 - 100.0%], Specificity 98.3% [96.1 - 99.3%]
- VIM: Sensitivity 100.0% [75.8 - 100.0%], Specificity 99.7% [98.1 - 99.9%]
Pseudomonas aeruginosa
- NDM: Sensitivity 100.0% [87.1 - 100.0%], Specificity 98.8% [93.3 - 99.8%]
- KPC: Sensitivity -, Specificity 100.0% [96.5 - 100.0%]
- OXA-48-like: Sensitivity 100.0% [20.7 - 100.0%], Specificity 100.0% [96.5 - 100.0%]
- IMP: Sensitivity 100.0% [56.6 - 100.0%], Specificity 86.3% [78.3 - 91.6%]
- VIM: Sensitivity 100.0% [91.0 - 100.0%], Specificity 100.0% [94.7 - 100.0%]
Acinetobacter baumannii
- NDM: Sensitivity 98.7% [92.9 - 99.8%], Specificity 100.0% [85.7 - 100.0%]
- KPC: Sensitivity 100.0% [20.7 - 100.0%], Specificity 100.0% [96.2 - 100.0%]
- OXA-48-like: Sensitivity -, Specificity 100.0% [96.3 - 100.0%]
- IMP: Sensitivity 100.0% [67.6 - 100.0%], Specificity 97.8% [92.3 - 99.4%]
- VIM: Sensitivity 100.0% [20.7 - 100.0%], Specificity 100.0% [96.2 - 100.0%]

Predicate Device(s)

K152614

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.1640 Antimicrobial susceptibility test powder.

(a)
Identification. An antimicrobial susceptibility test powder is a device that consists of an antimicrobial drug powder packaged in vials in specified amounts and intended for use in clinical laboratories for determining in vitro susceptibility of bacterial pathogens to these therapeutic agents. Test results are used to determine the antimicrobial agent of choice in the treatment of bacterial diseases.(b)
Classification. Class II (performance standards).

Summary

0

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

May 10, 2019

GenePOC Inc. Dany Leblanc VP QA/RA 360 rue Franquet Quebec, G1P 4N3 Ca

Re: K190275

Trade/Device Name: GenePOC Carba Regulation Number: 21 CFR 866.1640 Regulation Name: Antimicrobial Susceptibility Test Powder Regulatory Class: Class II Product Code: PMY, OOI Dated: February 6, 2019 Received: February 8, 2019

Dear Dany Leblanc:

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 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

1

for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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

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

Sincerely,

for

Uwe Scherf, M.Sc., Ph.D. Director 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

2

510(K) SUMMARY 5.0

3

510K SUMMARY

510(k) Number: K190275

A. GENERAL INFORMATION

Submission Date: April 29, 2019

Submitter Information:

| Submitted By: | GenePOC Inc.
360 rue Franquet
Québec (Québec) G1P 4N3 |
|------------------------|---------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Dany Leblanc
VP Quality Assurance and Regulatory Affairs
GenePOC Inc.
Telephone: +1 418 650-3535
Email: dany.leblanc@genepoc.ca |

B. PURPOSE FOR SUBMISSION

To obtain a substantial equivalence determination for the GenePOC™ Carba assay on the revogene™ instrument for the qualitative detection and differentiation of the blaxer, blayDM, blayM, blaoxA-48-ike, and blamp gene sequences from carbapenem-non-susceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa.

C. MEASURAND

Target DNA sequence of the following genes: blakec, blandM, blaving, blaoxA-48like, and blanp

D. TYPE OF TEST

Qualitative real-time Polymerase Chain Reaction (PCR) assay

E. APPLICANT

GenePOC Inc.

F. PROPRIETARY AND ESTABLISHED NAMES

Proprietary Name: GenePOC™ Carba Common Name : GenePOC™ Carba assay

4

G. REGULATORY INFORMATION

Trade Name:	GenePOC™ Carba
Classification:	Class II
Regulation:	21 CFR 866.1640
Regulation Name:	Antimicrobial susceptibility test powder
Product Code:	PMY, OOI
Panel:	83 - Microbiology

H. INTENDED USE

1. Intended Use and Indications for Use:
  The GenePOC™ Carba assay, performed on the revogene™ instrument, is a qualitative in vitro diagnostic test designed for the detection and differentiation of the blakec. blaNDM, blackA-48-like, and blamp gene sequences associated with carbapenem-nonsusceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa, when grown on blood agar or MacConkey agar. The test utilizes automated real-time Polymerase Chain Reaction (PCR).

The GenePOC™ Carba assay should be used in conjunction with other laboratory tests including phenotypic antimicrobial susceptibility testing. A negative GenePOC™ Carba assay result does not preclude the presence of other resistance mechanisms.

The GenePOC™ Carba assay is intended as an aid for infection control in the detection of carbapenem-non-susceptible bacteria that colonize patients in healthcare settings. The identification of a bland, blaym or blaviM metallo-B-lactamase gene (i.e., the genes that encode the IMP, NDM and VIM metallo-ß-lactamases, respectively) may be used as an aid to clinicians in determining appropriate therapeutic strategies for patients with known or suspected carbapenem non-susceptible infections.

1. Special conditions for use statement(s):
  Prescription Use Only

Organisms should be identified and carbapenem non-susceptibility status should be determined prior to testing with the GenePOCTM Carba assay.

The GenePOC™ Carba assay detects blakec, blandM, blavisM, blaoxA-48-like, and blamp resistance genes from pure colonies and is not for bacterial identification.

The performance of the GenePOC™ Carba assay with bacteria other than Enterobacteriaceae, Acinetobacter baumannii or Pseudomonas aeruginosa, has not been evaluated.

5

The GenePOC™ Carba assay is not a sub-typing tool and does not report variants of the blakpc, blandm, blaym, blaoxa-48-like, and blandp

1. Special instrument requirements:
  The GenePOC™ Carba assay uses PCR technology on the revogene™ instrument platform, which extracts, amplifies, and detects the target DNA.

I. INDICATIONS FOR USE

Same as Intended Use.

J. DEVICE DESCRIPTION

The GenePOC™ Carba assay is an automated real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection and differentiation of the blaype, blayDM, blavin, blaoxA-48-ike, and blamp gene sequences in carbapenem-non-susceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa grown on blood agar or MacConkey agar. The GenePOC™ Carba assay is designed to be performed on the revogene™ instrument. The revogene™ is an instrument that automates sample homogenization, sample dilution, cells lysis, DNA amplification, and detection of the amplified PCR products.

The GenePOC™ Carba assay kit is comprised of single-use, disposable microfluidic cartridges (PIEs), Sample Buffer Tubes (SBT), and Disposable Transfer Tools (DTT). These components are used to dilute the sample, extract, amplify, and simultaneously detect blayDM, blavin, blaoxA-48-like, and blamp DNA. A Process Control (PrC) is also incorporated into each PIE to verify sample processing and amplification steps. The PrC allows for the verification of potential inhibitor substances as well as microfluidic, instrument or reagent failure.

Each GenePOC™ Carba assay kit contains 24 individual pouches. Each pouch has components for one (1) test including one (1) Carba PIE, one (1) SBT, and one (1) DTT. The single-use, multi-chambered fluidic cartridges are designed to complete sample preparation and real-time PCR for the detection of the blaker, blands, blaoxa-48-like, and blanyp gene sequences from isolates of pure cultures of carbapenem-non-susceptible gram-negative bacteria in approximately 70 minutes. User intervention is only required for preparing the standardized 0.5 McFarland bacterial suspension from characterized carbapenem-non-susceptible isolated colonies, inoculating the bacterial suspension into the Sample Buffer Tube (SBT), transferring the sample into the single-use disposable PIE, and loading/unloading the PIEs into the revogene™ carousel. Each PIE is a completely integrated closed device in which a sample is dispensed and processed through different microfluidic chambers and channels that allow for the sample processing and subsequent real-time PCR steps.

Upon completion of a run, the results are interpolated by the revogene™ from measured fluorescent signals and embedded calculation algorithms. The output results include positive, negative, indeterminate, and unresolved. Upon completion of a run, the user removes the used

6

cartridges and disposes of them in normal biological waste. The results are automatically generated at the end of the process in a report that can be viewed and printed.

K. SUBSTANTIAL EQUIVALENCE INFORMATION

1. Predicate Device Name: Xpert® Carba-R Assay
1. Predicate 510(k) Number: K152614
1. Comparison with Predicate:

The GenePOC™ Carba assay is substantially equivalent to the Xpert® Carba-R Assay (K152614). The GenePOC™ Carba and the Xpert Carba-R assays both detect target gene sequences from antibiotic-non-susceptible bacteria and use real-time PCR amplification and fluorogenic target-specific hybridization detection. The performance of the GenePOC™ Carba assay was determined in a multi-site clinical study. The table below shows the similarities and differences between the GenePOC™ Carba assay and the predicate device (Xpert® Carba-R Assay).

| Item | GenePOCT™ Carba assay | Xpert® Carba-R Assay
(Predicate Device) |
|------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| K Number | K190275 | K152614 |
| SIMILARITIES | | |
| Classification | Class II | Class II |
| Intended Use | The GenePOC™ Carba assay, performed on
the revogene™ instrument, is a qualitative
in vitro diagnostic test designed for the
detection and differentiation of the blaKPC,
blaNDM, blaVIM, blaOXA-48-like, and blaIMP
gene sequences associated with
carbapenem-non-susceptible pure colonies
of Enterobacteriaceae , Acinetobacter
baumannii , or Pseudomonas aeruginosa ,
when grown on blood agar or MacConkey
agar. The test utilizes automated real-time
Polymerase Chain Reaction (PCR).

The GenePOC™ Carba assay should be
used in conjunction with other laboratory
tests including phenotypic antimicrobial
susceptibility testing. A negative
GenePOC™ Carba assay result does not
preclude the presence of other resistance
mechanisms.

The GenePOC™ Carba assay is intended as
an aid for infection control in the detection
of carbapenem-non-susceptible bacteria that
colonize patients in healthcare settings. The
identification of a blaKPC, blaNDM or blaVIM | The Xpert® Carba-R Assay,
performed on the GeneXpert®
Instrument Systems, is a qualitative
in vitro diagnostic test for the
detection and differentiation of the
blaKPC, blaNDM, blaVIM, blaOXA-48,
and blaIMP gene sequences
associated with carbapenem-non-
susceptible pure colonies of
Enterobacteriaceae , Acinetobacter
baumannii , or Pseudomonas
aeruginosa grown on blood agar or
MacConkey agar. The test utilizes
automated real-time polymerase
chain reaction (PCR).

7

1 Note: The variants listed reflect the respective labeling of each device and the analytical studies and in silico analyses conducted at the time of 510(k) clearance.

8

The GenePOC™ Carba assay has the same general intended use as the predicate device and has the same technological characteristics as the predicate device. The differences between the GenePOC™ Carba assay and the predicate device do not raise new questions of safety and effectiveness.

L. STANDARDS/GUIDANCE DOCUMENTS REFERENCED

CLSI Guideline EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents; . Approved Guideline, 2009.
. CLSI EP28-A3c, Guideline Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline, 2008.
CLSI M02, Performance Standards for Antimicrobial Disk Susceptibility Tests; Approved . Guideline, 13th edition, 2018.
CLSI M100, Performance Standards for Antimicrobial Susceptibility Testing; Approved . Guideline, 28th edition, 2018.

M. TEST PRINICIPLE

The GenePOC™ Carba assay is a single-use test for the qualitative detection and differentiation of the blake, blaNDM, blavin, blaoxA-48-ike, and blaIMP gene sequences in carbapenem-non-susceptible pure colonies of Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa grown on blood agar or MacConkey agar that utilizes an automated sample preparation and real-time polymerase chain reaction (PCR) technology with fluorogenic detection of the amplified DNA. The GenePOC™ Carba assay utilizes the GenePOC™ Carba microfluidic cartridge (PIE) for the simultaneous detection of the blaxer, bland, blaym, blaoxa-48-like, and blamp DNA and the internal process control (PrC) DNA (to verify sample processing, amplification, and the absence of reaction inhibitors).

Well-characterized Enterobacteriaceae, Acinetobacter baumannii, or Pseudomonas aeruginosa, carbapenem-non-susceptible strain is streaked for isolation on a blood agar plate (BAP) or a MacConkey agar plate with a 10-ug meropenem disc on the first quadrant. The plate is then incubated for 18-24 hours at 35°C ± 2°C under aerobic conditions. A standardized 0.5 McFarland (McF) suspension is prepared in saline solution by collecting isolated colonies selected from the incubated agar plate with a sterile swab. Fifteen (15) uL of the 0.5 McF suspension are transferred into the Sample Buffer Tube (SBT) containing 1 mL of Sample Buffer (SB). After a 15-second vortex step at maximal speed, a volume of SB lying between the two marks of the DTT is sampled from the SBT and transferred into a GenePOC™ Carba PIE to perform the assay. The loaded GenePOC™ Carba PIE is placed into the revogene™ for further sample processing. No operator intervention is necessary once the sample is loaded onto the revogene™.

Each Carba PIE is a completely integrated and self-contained device. Each sample is sequentially transferred by centrifugation from one microfluidic chamber to the next and all reagents specific for the PCR reaction are incorporated and dried within the PCR wells. The stepwise process includes sample homogenization, lysis of cells, and sample dilution followed by the subsequent real-time PCR steps. An internal PrC is contained in the homogenization chamber and is therefore present in every test to verify critical steps of the analytical process

9

(including sample lysis, dilution and nucleic acid amplification and detection) for the presence of potential inhibitory substances as well as system or reagent failures. The amplified products are detected in real time using target-specific TagMan® chemistry-based probes. The results are interpolated by the system from measured fluorescent signals and embedded calculation algorithms. Results may be viewed, printed, transferred, and/or stored by the user.

N. PERFORMANCE CHARACTERISTICS

1. Analytical Performance

Reproducibility/Precision a.
The Reproducibility and Precision study was performed by testing a total of ten (10) positive bacterial carbapenem-non-susceptible strains, each one harboring resistance gene targeted by the GenePOC™ Carba assay (n=2 strains per resistance gene, i.e. blakec, blaNDM, blaOXA-48-like, and blamp) and one (1) negative bacterial carbapenem-non-susceptible strain that does not carry any of the resistance genes targeted by the assay from standardized 0.5 McF suspensions.

The Between-Laboratory Reproducibility study was performed by two (2) operators at three (3) sites with one (1) GenePOC™ Carba assay kit lot over a total of five (5) days (consecutive or not) per panel member tested. Two (2) runs were performed by each operator on each day. On each day of testing, four (4) panel members of each positive strain were tested on each site and by each operator for a total of 120 replicates per strain (5 days of testing x 4 panel members per strain x 3 sites x 2 operators = 120 replicates per strain). Eight (8) negative panel members were also tested on each site and by each operator, on each respective day of testing for a total of 240 replicates (5 days of testing x 8 panel members x 3 sites x 2 operators = 240 replicates).

To assess the Between-Lot Reproducibility, testing was pursued at one (1) site with two (2) additional kit lots for a total of 15 days of testing (five (5) days per kit lot, consecutive or not) per panel member tested for each kit lot. Two (2) runs per kit lot were performed by each operator on each day. On each day of testing, four (4) panel members of each positive strain were tested by each operator for a total of 120 replicates per strain (5 days of testing x 4 panel members per strain x 3 kit lots x 2 operators = 120 replicates per strain). Eight (8) negative panel members were also tested with each kit lot and by each operator, on each respective day of testing for a total of 240 replicates (5 days of testing x 8 panel members x 3 kit lots x 2 operators = 240 replicates).

The Within-Laboratory Precision for each panel member was calculated based on one (1) GenePOC™ Carba assay kit lot (kit lot 1) that was tested by two (2) operators at site 1 over a total of five (5) days (consecutive or not). A total of 40 replicates for each of the ten (10) positive bacterial carbapenem-non-susceptible strains, each one harboring one (1) resistance gene targeted by the GenePOC Carba assay (n=2 strains per resistance gene), were considered for the analysis (5 days of testing x 4 panel members per strain x 2 operators = 40 replicates). A total of 80 replicates of the

10

negative bacterial carbapenem-non-susceptible strain that does not carry any of the resistance genes targeted by the assay were also considered (5 days of testing x 8 panel members x 2 operators = 80 replicates).

For the negative samples tested, there was agreement of 98.8% (237/240; 95% CI: [96.4-99.6%], 99.6% (239/240; 95% CI: [97.7-99.9%]) and 100.0% (80/80; 95% CI: [95.4-100.0%], respectively for the Between-Laboratory Reproducibility study, the Between-Lot Reproducibility study and for the Within-Laboratory Precision study. For the positive samples, agreement of 100.0% (120/120; 95% CI: [96.9-100.0%]), 100.0% (120/120; 95% CI: [96.9-100.0%]) and 100.0% (40/40; 95% CI: [91.2-100.0%]), was achieved for each panel member in the Between-Laboratory Reproducibility study, the Between-Lot Reproducibility study and for the Within-Laboratory Precision study, respectively.

11

| | Results/Total | Agreement | |---------------------|------ | Resistance | Site 1 | Gene and
Variant | Operator A Variant | Operator A | IMP-1 | 20/20 [96.9-100.0] | IMP-1 [96.9-100.0] |
| IMP-4 | 20/20 [96.9-100.0] | IMP-4 [96.9-100.0] |
| KPC-4 | 20/20 [96.9-100.0] | KPC-4 [96.9-100.0] |
| KPC-2 | 20/20 [96.9-100.0] | KPC-2 [96.9-100.0] |
| NDM-1 | 20/20 [96.9-100.0] | NDM-1 [96.9-100.0] |
| NDM-5 | 20/20 [96.9-100.0] | NDM-5 (96.9-100.0] |
| OXA-181 | 20/20 [96.9-100.0] | OXA-181 [96.9-100.0] |
| OXA-48 | 20/20 [96.9-100.0] | OXA-48 [96.9-100.0] |
| VIM-1 | 20/20 [96.9-100.0] | VIM-1 [96.9-100.0] |
| VIM-1 | 20/20 [96.9-100.0] | VIM-1 [96.9-100.0] |
| Negative | 40/40 [96.4-99.6] | Negative [97.7-99.9] | | | | | | | | | Overall % | | Results/Total | | | | | | | | Overall %
|
---------|------------|-------|------------|------------|-------|------------|------------|-------|----------------------------------|---------------------|---------------|------------|-------|------------|------------|-------|------------|------------|------------------------|----------------------------------|
| | | Site 2 | | | Site 3 | | | Agreement | Resistance | Kit Lot 1 | | | Kit Lot 2 | | | Kit Lot 3 | | | |
| Operator B | Site | Operator A | Operator B | Site | Operator A | Operator B | Site | [95% CI] | Gene and
| Operator B | Site | Operator A | Operator B | Site | Operator A | Operator B | Site | [95% CI] |
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%)
| 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 20/20 | 20/20 | 40/40 | 120/120 (100.0%
| 40/40 | 80/80 | 39/40 | 40/40 | 79/80 | 39/40 | 39/40 | 78/80 | 237/240 (98.8%)
| 40/40 | 40/40 | 80/80 | 40/40 | 39/40 | 79/80 | 40/40 | 40/40 | 80/80 | 239/240 (99.6%)

Tested Target Results and Overall Agreement for the Between-Laboratory Reproducibility Qualitative Analysis

12

Tested Target Results and Overall Agreement for the Between-Lot Reproducibility Qualitative Analysis

13

Tested Target Results and Overall Agreement for the Within-Laboratory Precision Qualitative Analysis

| Resistance Gene
and Variant | Results/Total
Site 1 and Kit Lot 1 | | Overall % Agreement
[95% CI] |
|--------------------------------|---------------------------------------|------------|---------------------------------|
| | Operator A | Operator B | |
| IMP-1 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| IMP-4 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| KPC-4 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| KPC-2 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| NDM-1 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| NDM-5 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| OXA-181 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| OXA-48 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| VIM-1 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| VIM-1 | 20/20 | 20/20 | 40/40 (100.0%) [91.2-100.0] |
| Negative | 40/40 | 40/40 | 80/80 (100.0%) [95.4-100.0] |

14

| Resistance
Gene and
Variant | Mean Ct¹ | N | Between-
Laboratory | | Between-Operator | | Between-Day | | Repeatability | | Overall | |
|-----------------------------------|----------|-----|------------------------|-----|------------------|-----|-------------|-----|---------------|-----|---------|-----|
| | | | SD | CV% | SD | CV% | SD | CV% | SD | CV% | SD | CV% |
| IMP-1 | 29.7 | 120 | 1.2 | 4.0 | 0.6 | 2.2 | 0.0 | 0.0 | 0.9 | 3.1 | 1.6 | 5.5 |
| IMP-4 | 29.6 | 120 | 0.7 | 2.2 | 0.4 | 1.4 | 0.2 | 0.7 | 1.0 | 3.3 | 1.3 | 4.3 |
| KPC-4 | 30.5 | 120 | 1.3 | 4.1 | 0.7 | 2.3 | 0.2 | 0.8 | 1.6 | 5.1 | 2.1 | 7.0 |
| KPC-2 | 30.4 | 120 | 1.6 | 5.2 | 0.4 | 1.4 | 0.1 | 0.2 | 1.5 | 5.0 | 2.2 | 7.3 |
| NDM-1 | 30.3 | 120 | 0.3 | 0.9 | 0.1 | 0.2 | 0.0 | 0.0 | 0.9 | 3.0 | 1.0 | 3.1 |
| NDM-5 | 30.2 | 120 | 0.6 | 2.1 | 0.5 | 1.6 | 0.2 | 0.5 | 1.2 | 4.0 | 1.5 | 4.8 |
| OXA-181 | 28.5 | 120 | 0.9 | 3.2 | 0.3 | 0.9 | 0.2 | 0.8 | 1.0 | 3.4 | 1.4 | 4.8 |
| OXA-48 | 30.5 | 120 | 1.4 | 4.5 | 0.5 | 1.5 | 0.3 | 0.9 | 1.1 | 3.7 | 1.9 | 6.1 |
| VIM-1 | 27.6 | 120 | 0.3 | 1.1 | 0.4 | 1.3 | 0.0 | 0.0 | 0.9 | 3.1 | 1.0 | 3.5 |
| VIM-1 | 28.5 | 120 | 0.3 | 1.0 | 0.5 | 1.7 | 0.0 | 0.0 | 1.1 | 3.8 | 1.2 | 4.3 |
| Negative | 34.9 | 237 | 1.1 | 3.1 | 0.7 | 2.0 | 0.4 | 1.2 | 1.4 | 3.9 | 1.9 | 5.5 |

Ct Values for the Between-Laboratory Reproducibility Quantitative Analysis

N: Number; SD: Standard Deviation; CV: Coefficient of Variation

1 For resistance genes and variants, Ct values reported are for negative samples, Ct values reported are for the PC.

15

| Resistance
Gene and

Variant	Mean Ct¹	N	Between-Lot		Between-Operator		Between-Day		Repeatability		Overall
IMP-1	29.0	120	0.8	2.6	0.3	1.2	0.1	0.4	0.7	2.5	1.1	3.8
IMP-4	29.2	120	0.8	2.8	0.3	0.9	0.1	0.4	1.1	3.6	1.4	4.7
KPC-4	29.6	120	1.6	5.4	0.3	1.1	0.0	0.0	1.5	5.0	2.2	7.5
KPC-2	30.8	120	1.2	3.9	0.3	1.1	0.3	1.0	1.9	6.1	2.3	7.4
NDM-1	29.5	120	1.0	3.2	0.1	0.3	0.1	0.4	1.1	3.6	1.4	4.8
NDM-5	28.9	120	1.1	3.7	0.3	1.2	0.0	0.0	1.0	3.4	1.5	5.1
OXA-181	28.2	120	1.2	4.1	0.3	0.9	0.2	0.8	0.9	3.0	1.5	5.2
OXA-48	28.6	120	1.3	4.6	0.2	0.7	0.0	0.0	1.0	3.3	1.6	5.7
VIM-1	27.5	120	0.4	1.5	0.4	1.4	0.1	0.5	0.8	2.9	1.0	3.6
VIM-1	28.1	120	0.9	3.0	0.3	1.0	0.0	0.0	1.0	3.6	1.4	4.8
Negative	33.0	239	2.5	7.5	0.7	2.1	0.4	1.3	1.3	3.9	2.9	8.8

Ct Values for the Between-Lot Reproducibility Quantitative Analysis

N: Number; SD: Standard Deviation; CV: Coefficient of Variation

1 For resistance genes and variants, Ct values reported are for negative samples, Ct values reported are for the PC.

16

Resistance			Between-Day		Between-Run		Repeatability		Overall
Gene and
Variant	Mean Ct1	N	SD	%CV	SD	%CV	SD	%CV	SD	%CV
IMP-1	30.0	40	0.0	0.0	0.0	0.0	0.8	2.5	0.8	2.5
IMP-4	30.2	40	0.5	1.5	0.0	0.0	1.0	3.4	1.1	3.7
KPC-4	31.5	40	0.0	0.0	0.0	0.0	1.6	5.0	1.6	5.0
KPC-2	32.2	40	0.1	0.4	0.0	0.0	1.7	5.1	1.7	5.1
NDM-1	30.6	40	0.3	0.9	0.1	0.3	0.9	3.1	1.0	3.2
NDM-5	30.2	40	0.4	1.4	0.0	0.0	0.9	2.8	1.0	3.2
OXA-181	29.5	40	0.4	1.2	0.0	0.0	0.9	3.2	1.0	3.4
OXA-48	30.1	40	0.0	0.0	0.5	1.6	1.0	3.5	1.1	3.8
VIM-1	28.1	40	0.1	0.3	0.0	0.0	0.9	3.3	0.9	3.3
VIM-1	29.1	40	0.0	0.0	0.2	0.7	1.0	3.4	1.0	3.5
Negative	35.9	80	0.6	1.7	0.4	1.1	1.8	5.1	2.0	5.5

Ct Values for the Within-Laboratory Precision Quantitative Analysis

N: Number; SD: Standard Deviation; CV: Coefficient of Variation

1 For resistance genes and variants, Ct values reported are for negative samples, Ct values reported are for the PC.

b. Linearity/Assay Reportable Range

Not Applicable.

C. Traceability, Stability, Expected Values (controls, calibrators, or methods)

Internal Process Control (PrC)

Each PIE contains a PrC that controls for amplification inhibition, assay reagents, and sample processing effectiveness.

External Controls

External Controls should be prepared by the end user. These are recommended but not required to perform the GenePOC™ Carba assay. The user should select the most appropriate controls for their laboratory quality control program to comply with applicable regulations and the requirements of accrediting agencies.

17

As a Positive External Control, GenePOC recommends using a standardized 0.5 McF suspension of Klebsiella pneumoniae or Escherichia coli prepared from a commercially available strain that harbors one of the targeted carbapenemase genes (NCTC 13476 for blamp.1, CCUG 59348 for blakpc.2, ATCC® BAA-2146™ for blaNDM-1, ATCC® BAA-2523TM for blaoxA-48 and NCTC 13440 for blavIM-1).

As a Negative External Control, GenePOC recommends using a standardized 0.5 McF suspension of isolated colonies from a representative carbapenem-non-susceptible bacterial strain of Enterobacteriaceae, A. baumannii or P. aeruginosa that does not carry any of the resistance genes targeted by the GenePOC™ Carba assay.

Sample Stability in the PIE

The GenePOC™ Carba assay PIE (microfluidic cartridge) stability study was verified by testing five (5) positive carbapenem-non-susceptible bacterial strains, each carrying a different carbapenemase gene (blakpc, blayDM, blayM, blaoxA-48-like, and blamp) targeted by the GenePOC™ Carba assay. In addition, a carbapenem-non-susceptible Enterobacter cloacae strain that does not carry any of the carbapenemase resistance genes targeted by the assay was also tested.

Both positive and negative samples were tested with the GenePOC™ Carba assay from standardized 0.5 and 4 McF suspensions, respectively. The results of the study support the recommended maximum interval of one (1) hour at 25 ± 2℃ from opening of the PIE pouch and sample addition into the GenePOC™ Carba assay PIE to processing on the revogene™ instrument.

Sample Stability

Sample stability was evaluated with Sample Buffer Tube (SBT) inoculated with a standardized 0.5 McF suspension of a bacterial strain carrying one of the resistance genes targeted by the GenePOC™ Carba assay (blakec, blaNDM, blaoxA-48-like, and blaMP) and with a standardized 0.5 McF suspension of a carbapenem-nonsusceptible bacterial strain that does not carry any of the resistance genes targeted by the GenePOC™ Carba assay. The product labeling indicates that SBT inoculated with a standardized 0.5 McF suspension prepared from carbapenem non-susceptible bacterial isolates can be stored at 2-8°C for up to seven (7) days or at 25 ± 2°C for up to four (4) days prior to testing with the GenePOC™ Carba assay.

d. Detection Limit

Not applicable.

e. Analytical Reactivity

The analytical reactivity (inclusivity) of the GenePOC™ Carba assay was evaluated with 58 carbapenem-non-susceptible isolates of Enterobacteriaceae. Pseudomonas aeruginosa and Acinetobacter baumannii from various geographic origins representing

18

temporal diversity and harboring variants of the blakec, blandM, blaoxA-48-like, and blamp carbapenemase genes:

Two (2) strains with two (2) resistance genes; -
11 strains (including five [5] variants) with the blamp resistance gene; -
11 strains (including at least three [3] variants) with the blagge resistance gene; -
14 strains (including five [5] variants) with the blaNDM resistance gene; -
-11 strains (including three [3] variants) with the blaoxA-48-like resistance gene;
9 strains (including four [4] variants) with the blavM resistance gene. -

Each strain was tested from a standardized 0.5 McF bacterial suspension. Three (3) replicates per strain were tested using three (3) different GenePOC™ Carba kit lots (one (1) replicate per kit lot). The ability of the GenePOC™ Carba assay to detect multiple variants of the blaxec, blaNDM, blaoxA-48-like, and blaiMP carbapenemase genes in the 58 Enterobacteriaceae. Pseudomonas aeruginosa and Acinetobacter baumannii strains carrying these genes was demonstrated.

Carbapenem-Non-susceptible Bacterial Isolates Tested for Analytical Reactivity (Inclusivity) with the GenePOC™ Carba Assay

Species	Collection Number	Geographical and Temporal Origin	Reported Resistance Gene and Variant (When Available)
Isolates Harboring Multiple Genes
Pseudomonas
aeruginosa	ATCC® BAA-2793TM	Chile	2014 KPC-2, VIM-2
Klebsiella pneumoniae	CCRI-23061	Switzerland2	2015 OXA-232, NDM-1
KPC Isolates
Klebsiella pneumoniae	NCTC 13438	N/A1	N/A1 KPC-3
Klebsiella pneumoniae	ATCC® BAA-1705TM	USA	2007 KPC-2
Pseudomonas
aeruginosa	CCRI-21587	Canada2	2011 KPC-2
Enterobacter cloacae	CCRI-21578	Canada	2011 KPC-4
Klebsiella oxytoca	CCRI-21581	Canada	2011 KPC-3
Klebsiella pneumoniae	CCRI-19587	Canada2	2009 KPC-3
Klebsiella pneumoniae	CCRI-19570	USA	2003 KPC-2
Klebsiella pneumoniae	CCUG 59413	Sweden	2010 KPC-3
Klebsiella pneumoniae	CCUG 59348	Norway	2010 KPC-2
Klebsiella pneumoniae	CCUG 56233	Sweden	2088 KPC-2
Escherichia coli	ATCC® BAA-2340TM	USA (State Public Health
laboratories)	N/A1 KPC
NDM Isolates
Klebsiella pneumoniae	NCTC 13443	N/A1	N/A1 NDM-1
Klebsiella pneumoniae	ATCC® BAA-2146TM	USA (State Public Health
laboratories)	N/A1 NDM-1
Escherichia coli	CCRI-22255	France2	2013 NDM-1
Klebsiella pneumoniae	CCRI-21711	Canada	2011 NDM-1
Klebsiella pneumoniae	CCRI-22199	Canada	2012 NDM-1
Providencia rettgeri	CCRI-22257	France2	2013 NDM-1
Providencia stuartii	CCRI-22256	France2	2013 NDM-1
Klebsiella pneumoniae	CCRI-22254	France2	2013 NDM-4
Escherichia coli	CCRI-23064	Switzerland2	2015 NDM-5
Species	Collection Number	Geographical and Temporal Origin	Reported Resistance Gene and Variant (When Available)
Escherichia coli	CCRI-23464	Canada	2016	NDM-5
Escherichia coli	CCRI-23065	Switzerland2	2015	NDM-6
Escherichia coli	CCRI-23066	Switzerland2	2015	NDM-7
Enterobacter cloacae	ATCC® BAA-2468TM	USA (State Public Health laboratories)	N/A1	NDM-1
Klebsiella pneumoniae	CCUG 60138	Sweden	2010	NDM-1
VIM Isolates
Pseudomonas aeruginosa	NCTC 13437	N/A1	N/A1	VIM-10
Klebsiella pneumoniae	NCTC 13439	N/A1	N/A1	VIM-1
Klebsiella pneumoniae	NCTC 13440	N/A1	N/A1	VIM-1
Klebsiella pneumoniae	CCRI-19585	France	2009	VIM-1
Klebsiella pneumoniae	CCRI-22258	France2	2013	VIM-1
Pseudomonas aeruginosa	CCRI-21588	Canada2	2011	VIM-2
Serratia marcescens	CCRI-22261	France2	2013	VIM-2
Pseudomonas aeruginosa	CCRI-22720	Argentina	2014	VIM-2
Klebsiella pneumoniae	CCRI-22259	France2	2013	VIM-19
OXA-48-like Isolates
Klebsiella pneumoniae	NCTC 13442	N/A1	N/A1	OXA-48
Klebsiella pneumoniae	CCRI-22263	France2	2013	OXA-48
Escherichia coli	CCRI-22265	France2	2013	OXA-48
Enterobacter cloacae	CCRI-22266	France2	2013	OXA-48
Klebsiella pneumoniae	CCRI-22264	France2	2013	OXA-181
Providencia rettgeri	CCRI-22267	France2	2013	OXA-181
Klebsiella pneumoniae	CCRI-23060	Switzerland2	2015	OXA-204
Citrobacter freundii	CCRI-23374	Canada	2016	OXA-204
Escherichia coli	ATCC® BAA-2523TM	human clinical isolates	N/A1	OXA-48
Klebsiella pneumoniae	ATCC® BAA-2524TM	human clinical isolates	N/A1	OXA-48
Klebsiella pneumoniae	CCUG 64452	Sweden	2013	OXA-48
IMP Isolates
Escherichia coli	NCTC 13476	N/A1	N/A1	IMP-1
Acinetobacter baumannii	CCRI-19488	Canada2	2003	IMP-1
Klebsiella pneumoniae	CCRI-19569	Japan	2003	IMP-1
Klebsiella pneumoniae	CCRI-19582	Turkey	2009	IMP-1
Pseudomonas aeruginosa	CCRI-21589	Canada2	2011	IMP-1
Klebsiella pneumoniae	CCRI-19583	Taiwan	2009	IMP-4
Klebsiella pneumoniae	CCRI-19588	Taiwan	2009	IMP-4
Citrobacter youngae	CCRI-21591	Canada2	2011	IMP-4
Klebsiella pneumoniae	CCRI-19584	Taiwan	2009	IMP-8
Pseudomonas aeruginosa	CCRI-21590	China	2000	IMP-9
Serratia marcescens	CCRI-22262	France2	2013	IMP-11

19

¹ N/A = Not Available
² Isolation country

^2 Isolation country

20

An in silico analysis was performed to assess inclusivity of the primers and probes of the GenePOC™ Carba assay. For each target, all variants documented in the National Center for Biotechnology Information (NCBI) nucleotide database were analyzed on November 7th, 2018. One representative sequence of each known variant was aligned. Variants for each target, predicted to be detected, are summarized in the table below.

| Targeted
Resistance
Genes | # of
Samples | Variants
Detected | Variants
Not
Detected | Detectable1 | Potentially
Detectable2 | Not
Detectable |
|---------------------------------|-----------------|----------------------|-----------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------|
| blaKPC | 12 | 2, 3, 4 | None | 2 to 38 | None | N/A |
| blaNDM | 15 | 1, 4, 5, 6, 7 | None | 1 to 24 | None | N/A |
| blaIMP | 11 | 1, 4, 8, 9, 11 | None | 1, 2, 4 to 6, 8 to 10,
13 to 20, 23 to 26, 28
to 30, 32, 33, 37, 38,
40, 42, 45, 47 to 49,
53 to 56, 59, 60, 62,
66, 69 to 72, 74 to 79 | 3, 73, 114, 21, 22,
27, 34, 41, 43,
44, 51, 52, 58,
61, 64, 67, 68,
73 | 12, 31, 35, 63 |
| blaOXA-48-
like | 12 | 48, 181,
204, 232 | None | 48, 162, 181, 199,
204, 232, 244, 245,
2525, 370, 484, 505,
5145, 5155, 519, 5465,
5475, 566 | None | 545, 1636,
2476, 4056,
4165, 436,
4386, 4396,
517, 5355,
5385, 567 |
| blaVIM | 10 | 1, 2, 10, 19 | None | 1 to 6, 8 to 12, 14 to
20, 23 to 46, 48 to 50,
52 to 55, 57, 59, 60 | 51, 56, 58 | 7, 13, 47 |

GenePOC™ Carba Assay Target Variants Coverage

1 Based on alignments with identity and query cover ≥ 95% and E-values