(79 days)
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No
The device description and performance studies indicate a traditional immunochromatographic assay with visual interpretation, not involving AI or ML for analysis or result generation.
No.
The device is an in vitro diagnostic assay used for the detection and differentiation of carbapenemases, aiding in infection control, but explicitly states it is "not intended to guide or monitor treatment".
Yes
The device qualitatively detects and differentiates five common carbapenemases, providing an aid for infection control in healthcare settings. This involves identifying specific markers (carbapenemases) in biological samples (bacterial colonies) to understand the presence of carbapenemase-producing bacteria, which is a diagnostic function.
No
The device is described as an "in vitro rapid and visual multiplex immunochromatographic assay," which is a physical test kit that uses antibodies immobilized on a membrane to detect substances. This involves physical components and chemical reactions, not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the NG-Test CARBA 5 is an "in vitro rapid and visual multiplex immunochromatographic assay". This directly indicates that it is used outside of the body to diagnose or aid in diagnosis.
- Device Description: The description details how the assay works by detecting specific enzymes in bacterial colonies, which are samples taken from a patient (or a source related to a patient's infection). This is a characteristic of an in vitro diagnostic test.
- Performance Studies: The performance studies involve testing bacterial isolates to determine the device's accuracy (PPA, NPA, sensitivity, specificity) in detecting carbapenemases. This type of evaluation is standard for IVD devices.
- Predicate Device: The mention of a "Predicate Device(s)" with a K number (K162385; RAPIDEC CARBA NP) is a strong indicator that this device is being submitted for regulatory clearance as an IVD, as predicate devices are used for comparison in the regulatory process for new IVDs.
N/A
Intended Use / Indications for Use
NG-Test CARBA 5 is an in vitro rapid and visual multiplex immunochromatographic assay for the qualitative detection and differentiation of five common carbapenemases (KPC, OXA-48-like, VIM, IMP and NDM) from carbapenem non-susceptible pure bacterial colonies when grown on the following media:
- 5% sheep blood agar or MacConkey agar (16-24 hours) for testing Enterobacteriaceae and Pseudomonas aeruginosa
- HardyCHROM™ CRE agar (18-24 hours) for testing E. coli and KES (Klebsiella aerogenes, . Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter cloacae complex, and Serratia marcescens).
The NG-Test CARBA 5 is intended as an aid for infection control in the detection of carbapenemase-producing Enterobacteriaceae and Pseudomonas aeruginosa in healthcare settings. NG-Test CARBA 5 is not intended to guide or monitor treatment for carbapenem non-susceptible bacterial infections. A positive or negative NG-Test CARBA 5 test result does not rule out the presence of other mechanisms of antibiotic resistance. NG-Test CARBA 5 should be used in conjunction with other laboratory tests including phenotypic antimicrobial susceptibility testing.
Product codes (comma separated list FDA assigned to the subject device)
PTJ
Device Description
NG-Test CARBA 5 is an in vitro rapid and visual multiplex immunochromatographic assay that detects one or more of the five common types of carbapenemase enzymes (KPC (K), OXA-48-like (O), IMP (I), VIM (V), NDM (N)) in bacterial colonies. Liquid extraction buffer is used as a cell lysing solution when mixed with colonies. Monoclonal antibodies that individually recognize each of the five carbapenemases are immobilized on a nitrocellulose membrane. Free monoclonal antibodies are present in the sample pad and labelled with colloidal gold. Upon addition of colonies mixed with extraction buffer to the sample pad, the capillary action of the nitrocellulose draws the sample the mobile antibodies and immobile antibodies on the test strip. The immobilized control antibodies capture any mobile antibodies that run through the sample pad and nitrocellulose without binding to other test lines. A positive result occurs when a red line appears on the control region and one or more lines appear in the test regions (K. O. V. I. or N) and indicates that the sample contains one or more carbapenemases. A negative result occurs when only the control line is observed and indicates that the sample does not contain any of the five carbapenemases. If the control line does not appear, the test result is invalid. The sample pad, and nitrocellulose strip are contained within a plastic cassette. After addition of colonies in extraction buffer to the sample port, a result can be read after 15 minutes.
Mentions image processing
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Mentions AI, DNN, or ML
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Input Imaging Modality
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Anatomical Site
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Indicated Patient Age Range
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Intended User / Care Setting
Healthcare settings
Description of the training set, sample size, data source, and annotation protocol
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Description of the test set, sample size, data source, and annotation protocol
A total of 310 organisms were tested against PCR (Xpert Carba-R. Cepheid) and phenotypic tests (mCIM, eCIM, and disk diffusion). One organism did not meet enrollment criteria because it was a species of Pseudomonas (Cornell 50) other than P. aeruginosa and was therefore excluded from the analysis. Of the remaining 309 organisms tested, a total of 240 Enterobacteriaceae (which provided 244 results since four isolates co-produced two carbapenemases) and 69 P. aeruginosa isolates were tested on NG-Test CARBA 5 with concordant results obtained by phenotypic testing paired with Xpert Carba-R results. Performance was evaluated at three geographically diverse hospitals with prospectively-collected and stock bacterial isolates. The identification of carbapenemase production on NG-Test CARBA 5 was compared to another FDA-cleared device, Xpert Carba-R by Cepheid (PCR for KPC, OXA-48 or 181, IMP, VIM, NDM), modified carbapenem inactivation method (mCIM) and EDTA carbapenemase inactivation method (eCIM) as described by CLSI M100, S29, and antibiotic susceptibility testing results to ertapenem, imipenem, and meropenem. Identity and susceptibility of organisms were confirmed using FDA-cleared ID and AST systems. NG-Test CARBA 5 quality control was performed in parallel every day of testing.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Performance Study:
Performance of NG-Test CARBA 5 was evaluated at three geographically diverse hospitals with prospectively-collected and stock bacterial isolates.
A total of 310 organisms were initially considered, with 309 organisms included in the final analysis (240 Enterobacteriaceae and 69 P. aeruginosa isolates).
The identification of carbapenemase production on NG-Test CARBA 5 was compared to Xpert Carba-R by Cepheid (PCR for KPC, OXA-48 or 181, IMP, VIM, NDM), modified carbapenem inactivation method (mCIM), and EDTA carbapenemase inactivation method (eCIM) as described by CLSI M100, S29, and antibiotic susceptibility testing results to ertapenem, imipenem, and meropenem.
Key Results (PPA and NPA for primary organisms on 5% Sheep Blood or MacConkey agar):
- Enterobacteriaceae (Overall):
- Overall PPA: 100.0% (95% CI: 97.6% - 100.0%)
- Overall NPA: 95.5% (95% CI: 88.9% - 98.2%)
- After discrepant analysis for Enterobacteriaceae: PPA increased to 100.0% (97.7% - 100.0%) and NPA increased to 100.0% (95.6% - 100.0%).
- P. aeruginosa (Overall):
- Overall PPA: 100.0% (95% CI: 77.2% - 100.0%)
- Overall NPA: 94.6% (95% CI: 85.4% - 98.2%)
- After discrepant analysis for P. aeruginosa: PPA increased to 100% (80.6% - 100%) and NPA increased to 100% (93.2% - 100%).
Key Results (PPA and NPA for HardyCHROM™ CRE agar):
-
Raw Stool specimen inoculated to HardyCHROM™ CRE (Overall E. coli, KES):
- Overall PPA: 100.0% (95% CI: 97.4% - 100.0%)
- Overall NPA: 90.2% (95% CI: 77.5% - 96.1%)
- After discrepant analysis for raw stool specimen: PPA increased to 100.0% and NPA increased to 100.0% (90.6% - 100.0%).
-
C&S Cary Blair Stool specimen inoculated to HardyCHROM™ CRE (Overall E. coli, KES):
- Overall PPA: 100.0% (95% CI: 97.3% - 100.0%)
- Overall NPA: 90.2% (95% CI: 77.5% - 96.1%)
- After discrepant analysis for C&S Cary Blair stool specimen: PPA increased to 100.0% and NPA increased to 100.0%.
Analytical Reactivity Study:
Ninety-two strains characterized to have a target carbapenemase were evaluated. Each test was performed in triplicate from each media type (sheep's blood agar, MacConkey agar, HardyCHROM™ CRE agar).
- Sensitivity:
- 88/92 (95.7%) from blood agar (after mCIM analysis)
- 90/92 (97.8%) from MacConkey agar (after mCIM analysis)
- 41/41 (100%) from HardyCHROM™ CRE agar (after mCIM analysis)
Analytical Specificity Study:
81 organisms that exhibit antibiotic resistance mechanisms other than the targets, are carbapenem-susceptible, or are carbapenem non-susceptible were tested.
- Specificity:
- 81/81 (100%) of organisms tested from blood and MacConkey agar produced a negative NG-Test CARBA 5 result.
- 16/16 (100%) of organisms tested from HardyCHROM™ CRE produced a negative NG-Test CARBA 5 result.
Incubation Study:
Twenty-two strains were tested from blood and MacConkey agar every two hours from 16 to 24 hours of incubation. Fifteen of these were also tested from HardyCHROM™ CRE every two hours from 18 to 24 hours.
- All organisms tested produced the expected result on NG-Test CARBA 5 at every time point tested.
Refrigeration Storage Study:
Twelve strains were cultured and evaluated over time from refrigerated storage for up to 3 days.
- All organisms tested produced the expected result on NG-Test CARBA 5 for each day of refrigeration for up to 3 days.
Reproducibility Study:
A panel of 20 blinded isolates was tested at three distinct study sites on five work days.
- All target carbapenemase positive isolates tested (100%) were detected by NG-Test CARBA 5 on all days of the reproducibility study.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Positive Percent Agreement (PPA)
- Negative Percent Agreement (NPA)
Note: Specific values for PPA and NPA are provided in the Summary of Performance Studies section based on the tables.
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
RAPIDEC CARBA NP, K162385
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
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§ 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).
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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the seal of the Department of Health & Human Services. To the right of that is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of the word "ADMINISTRATION".
October 2, 2019
NG Biotech % Anna Klavins Lead Performance Studies Microbiologist Hardy Diagnostics 1430 West McCoy Lane Santa Maria, California 93455
Re: K191889
Trade/Device Name: NG-Test CARBA 5 Regulation Number: 21 CFR 866.1640 Regulation Name: Antimicrobial Susceptibility Test Powder Regulatory Class: Class II Product Code: PTJ Dated: July 11, 2019 Received: July 15, 2019
Dear Anna Klavins:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's
1
requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Ribhi Shawar, Ph.D. (ABMM) Chief, General Bacteriology and Antimicrobial Susceptibility Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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510(k) Summary
I. SUBMITTER
July 11th, 2019 Correspondent: Anna Klavins Lead Performance Studies Microbiologist Hardy Diagnostics 1430 W. McCoy Lane Santa Maria, CA 93455 Phone: 805-346-2766 x5752 E-mail: ClinicalTrials@hardydiagnostics.com
Applicant: Milovan Stankov-Puges CEO NG Biotech Z.A. Courbouton, secteur 1, Atelier relais le Tremplin Guipry 35480 France Phone: :+33 (0) 2 23 30 17 83 Fax: +33 (0) 9 71 70 53 10 E-mail: msp@ngbiotech.com
II. DEVICE
Name of Device: NG-Test CARBA 5 Classification Name: Antimicrobial Susceptibility Test Powder Regulatory Class: II Product Code: PTJ
III. PREDICATE DEVICE RAPIDEC CARBA NP, K162385
IV. DEVICE DESCRIPTION
NG-Test CARBA 5 is an in vitro rapid and visual multiplex immunochromatographic assay that detects one or more of the five common types of carbapenemase enzymes (KPC (K), OXA-48-like (O), IMP (I), VIM (V), NDM (N)) in bacterial colonies. Liquid extraction buffer is used as a cell lysing solution when mixed with colonies. Monoclonal antibodies that individually recognize each of the five carbapenemases are immobilized on a nitrocellulose membrane. Free monoclonal antibodies are present in the sample pad and labelled with colloidal gold. Upon addition of colonies mixed with extraction buffer to the sample pad, the capillary action of the nitrocellulose draws the sample the mobile antibodies and immobile antibodies on the test strip. The immobilized control antibodies capture any mobile antibodies that run through the sample pad and nitrocellulose without binding to other test lines. A positive result occurs when
3
a red line appears on the control region and one or more lines appear in the test regions (K. O. V. I. or N) and indicates that the sample contains one or more carbapenemases. A negative result occurs when only the control line is observed and indicates that the sample does not contain any of the five carbapenemases. If the control line does not appear, the test result is invalid. The sample pad, and nitrocellulose strip are contained within a plastic cassette. After addition of colonies in extraction buffer to the sample port, a result can be read after 15 minutes.
V. INDICATIONS FOR USE
NG-Test CARBA 5 is an in vitro rapid and visual multiplex immunochromatographic assay for the qualitative detection and differentiation of five common carbapenemases (KPC, OXA-48-like, VIM, IMP and NDM) from carbapenem non-susceptible pure bacterial colonies when grown on the following media:
- 5% sheep blood agar or MacConkey agar (16-24 hours) for testing Enterobacteriaceae and Pseudomonas aeruginosa
- HardyCHROM™ CRE agar (18-24 hours) for testing E. coli and KES (Klebsiella aerogenes, . Klebsiella oxytoca, Klebsiella pneumoniae, Enterobacter cloacae complex, and Serratia marcescens).
The NG-Test CARBA 5 is intended as an aid for infection control in the detection of carbapenemaseproducing Enterobacteriaceae and Pseudomonas aeruginosa in healthcare settings. NG-Test CARBA 5 is not intended to guide or monitor treatment for carbapenem non-susceptible bacterial infections. A positive or negative NG-Test CARBA 5 test result does not rule out the presence of other mechanisms of antibiotic resistance. NG-Test CARBA 5 should be used in conjunction with other laboratory tests including phenotypic antimicrobial susceptibility testing.
VI. PERFORMANCE DATA
Performance of NG-Test CARBA 5 was evaluated at three geographically diverse hospitals with prospectively-collected and stock bacterial isolates. The identification of carbapenemase production on NG-Test CARBA 5 was compared to another FDA-cleared device, Xpert Carba-R by Cepheid (PCR for KPC, OXA-48 or 181, IMP, VIM, NDM), modified carbapenem inactivation method (mCIM) and EDTA carbapenemase inactivation method (eCIM) as described by CLSI M100, S29, and antibiotic susceptibility testing results to ertapenem, imipenem, and meropenem. Identity and susceptibility of organisms were confirmed using FDA-cleared ID and AST systems. NG-Test CARBA 5 quality control was performed in parallel every day of testing.
A total of 310 organisms were tested against PCR (Xpert Carba-R. Cepheid) and phenotypic tests (mCIM, eCIM, and disk diffusion). One organism did not meet enrollment criteria because it was a species of Pseudomonas (Cornell 50) other than P. aeruginosa and was therefore excluded from the analysis. Of the remaining 309 organisms tested, a total of 240 Enterobacteriaceae (which provided 244 results since four isolates co-produced two carbapenemases) and 69 P. aeruginosa isolates were tested on NG-Test CARBA 5 with concordant results obtained by phenotypic testing paired with Xpert Carba-R results.
Performance was equivalent between blood and MacConkey agar. Table 1 indicates the PPA and NPA for each individual target separated out by organism group. The overall PPA for Enterobacteriaceae was 100.0% (97.6% - 100.0%) and the overall NPA was 95.5% (88.9% - 98.2%) (Table 2).
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The overall PPA for P. aeruginosa was 100.0% (77.2% - 100.0%) and the overall NPA was 94.6% (85.4% - 98.2%) (Table 3). P. aeruginosa with NDM (n=2) were evaluated analytically in the bench testing.
| Plate | Organism Group6 | Total Targets | Target | TP1 | FP3 | FN | TN | PPA | low
95%2 | high
95% | NPA | low
95% | high
95% |
|-------------------------------------|-----------------------------|---------------|-------------|-----|-----|----|-----|-------|-------------|-------------|-------|------------|-------------|
| 5% Sheep Blood or
MacConkey agar | Enterobacteriaceae
(ENT) | 244 | KPC | 84 | 0 | 0 | 160 | 100.0 | 95.6 | 100.0 | 100.0 | 97.7 | 100.0 |
| | | | OXA-48-like | 20 | 0 | 0 | 224 | 100.0 | 83.9 | 100.0 | 100.0 | 98.3 | 100.0 |
| | | | VIM | 11 | 0 | 0 | 233 | 100.0 | 74.1 | 100.0 | 100.0 | 98.4 | 100.0 |
| | | | IMP | 4 | 34 | 0 | 237 | 100.0 | 51.0 | 100.0 | 98.8 | 96.4 | 99.6 |
| | | | NDM | 37 | 14 | 0 | 206 | 100.0 | 90.6 | 100.0 | 99.5 | 97.3 | 99.9 |
| | Pseudomonas
aeruginosa | 69 | KPC | 2 | 0 | 0 | 67 | 100.0 | 34.2 | 100.0 | 100.0 | 94.6 | 100.0 |
| | | | OXA-48-like | 0 | 0 | 0 | 69 | n/a | n/a | n/a | 100.0 | 94.7 | 100.0 |
| | | | VIM | 9 | 0 | 0 | 60 | 100.0 | 70.1 | 100.0 | 100.0 | 94.0 | 100.0 |
| | | | IMP | 2 | 35 | 0 | 64 | 100.0 | 34.2 | 100.0 | 95.5 | 87.6 | 98.5 |
| | | | NDM | 0 | 0 | 0 | 69 | n/a | n/a | n/a | 100.0 | 94.7 | 100.0 |
Table 1. Performance of the Comparator Method vs. CARBA 5 for all sites combined - Analysis by Target
1No True Positive results for OXA-48-like and NDM for the P. aeruginosa organism group in multicentric clinical testing.
4 ower bounds are below 90% due to the OXA, IMP, and VIM carbapenemases. The claim of NC-Test CARBA 5 detection of OXA, IMP, and VIM carbapenemases is supported by analytical reactivity data.
For Enterobacteriaceae, three isolates were for IMP on NG-Test CARBA 5 (positive MP on NG-Test CARBA 5, positive mCM, and negative Xpert Carba-R result). One isolate was a false positive for NDM on NG-Test CARBA 5, positive mCM, negative Xpet Carba-R result). For P. aeruginou, three isolates were false positive IMP on NG-Test CARBA 5. positive InCM, and negative Xpert Carba-R result).
*All three isolates were confirmed to have the MP-3 gene, making these true positives for IMP after discrepant analysis. (IMP-8 is predicted to be detected by Xpert Carbased on in silico analysis but has not been demonstrated analytically.) One isolate was confirmed to have an NDM-1 gene making this isolate true positive for NDM after discrepant analysis. (NDM-1 is predicted to be detected by Xpert Carba-R based on in silico analysis and has been tested analytically.) After discrepant analysis, the Enterobacteriaceae overall PPA increased to 100.0% (97.7% - 100.0%) and the vereall NPA increased to 100.0% (95.6% - 100.0%).
5 MI three isolates were confirmed to have 7, IMP-15, and IMP-19) making these true positives for IMP after discrepant analysis. (IMP-7 is a known limitation of Xpert Carba R. IMP-19 is predicted by Xpert Carbed on in silico analysis but has not been tested analytically. The ability of Xpert Carba-R to detect IMP-15 is unknown.) After discrepant analysis, the P. aerwinosa overall PPA increased to 100% (80.6% - 100%) and the overall NPA increased to 100% (93.2% - 100%).
"Ertapenen disks were routinely used to maintain solated colonies of retrospective Enterobacteriaceae isolates. No selective pressure was used for isolated colonies of retrospective P. aeruginosa isolates.
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| Enterobacteriaceae | Composite Reference Method | |||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| NG-Test CARBA 5 | Positive | 156 | 41,2 | 160 |
| Negative | 0 | 84 | 84 | |
| Total | 156 | 88 | 244 | |
| Positive Percent Agreement (PPA) | 156/156 = 100% (95% CI: 97.6-100%) | |||
| Negative Percent Agreement (NPA) | 84/88 = 95.5% (95% CI: 88.9-98.2%) |
Table 2. Agreement of NG-Test CARBA 5 with the composite reference method when testing Enterobacteriaceae
1 An alternative PCR assay showed that the NDM false positive isolate harbored a blayDM-1 variant. Isolate was positive by mCIM.
2 An alternative PCR assay and bidirectional sequencing showed that the three IMP false positive isolates harbored blamp -8/-47 variant that is predicted by in silico analysis but not analytically demonstrated to be detected by the assay. Isolates were positive by mCIM.
Table 3. Agreement of NG-Test CARBA 5 with the composite reference method when testing P. aeruginosa
| Composite Reference Method | ||||
|---|---|---|---|---|
| P. aeruginosa | Positive | Negative | Total | |
| NG-Test CARBA 5 | Positive | 13 | 31 | 16 |
| Negative | 0 | 53 | 53 | |
| Total | 13 | 56 | 69 | |
| Positive Percent Agreement (PPA) | 13/13 = 100% (95% CI: 77.2-100%) | |||
| Negative Percent Agreement (NPA) | 53/56 = 94.6% (95% CI: 85.4-98.2%) |
1 An alternative PCR assay and bidirectional sequencing showed that the three IMP false positive isolates harbored blamp variants that (i) are not detected by the FDA-cleared PCR assay (blamp variant -7), (ii) are predicted by in silico analysis but not analytically demonstrated to be detected by the assay (blamp -19), or (ii) the reactivity of the assay is unknown (bla]MP variant -15). Isolates were positive by mCIM.
The bacterial isolates used to evaluate NG-Test CARBA 5 from blood and MacConkey agar were also used internally to evaluate the performance of NG-Test CARBA 5 from HardyCHROM™ CRE agar. These results were compared to Xpert Carba-R, mCIM, and eCIM as described by CLSI M100, S29, and antibiotic susceptibility testing results to ertapenem, and meropenem. Identity and susceptibility of organisms were confirmed using FDA-cleared ID and AST systems. NG-Test CARBA 5 quality control was performed in parallel every day of testing.
Of the 186 organisms enrolled, one organism was not available for testing and was excluded from the analysis. Of the 185 organisms that fell under HardyCHROM™ CRE claims, 180/185 (97.3%) organisms (184 target results) were recovered from Raw stool, and 178/185 (96.2%) organisms (182 target results) were recovered from C&S Cary Blair stool onto HardyCHROM™ CRE. Table 4 indicates the PPA and NPA for each individual target separated out by organism group. The overall PPA from raw stool specimen inoculated to HardyCHROM™ CRE was 100.0% (97.4% - 100.0%) and the overall NPA was 90.2% (77.5% - 96.1%) (Table 5). The overall PPA from C&S Cary Blair stool specimen inoculated to HardyCHROM™ CRE was 100.0% (97.3% - 100.0%) (Table 6) and the overall NPA was the same as the raw stool specimen.
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| Plate | Specimen
Type | Organism
Group | Target | TP | FP2 | FN | TN | PPA | low
95%1 | high
95% | NPA | low
95% | high
95% |
|----------------------|----------------------------|-------------------------|-------------|----|-----|----|-----|-------|-------------|-------------|-------|------------|-------------|
| HardyCHROM™ CRE agar | Raw
Stool | E. coli ,
KES | KPC | 76 | 0 | 0 | 108 | 100.0 | 95.2 | 100.0 | 100.0 | 96.6 | 100.0 |
| | | | OXA-48-like | 18 | 0 | 0 | 166 | 100.0 | 82.4 | 100.0 | 100.0 | 97.7 | 100.0 |
| | | | VIM | 9 | 0 | 0 | 175 | 100.0 | 70.1 | 100.0 | 100.0 | 97.9 | 100.0 |
| | | | IMP | 4 | 3 | 0 | 177 | 100.0 | 51.0 | 100.0 | 98.3 | 95.2 | 99.4 |
| | | | NDM | 36 | 1 | 0 | 147 | 100.0 | 90.4 | 100.0 | 99.3 | 96.3 | 99.9 |
| | C&S Cary
Blair
Stool | E. coli ,
KES | KPC | 75 | 0 | 0 | 107 | 100.0 | 95.1 | 100.0 | 100.0 | 96.5 | 100.0 |
| | | | OXA-48-like | 18 | 0 | 0 | 164 | 100.0 | 82.4 | 100.0 | 100.0 | 97.7 | 100.0 |
| | | | VIM | 8 | 0 | 0 | 174 | 100.0 | 67.6 | 100.0 | 100.0 | 97.8 | 100.0 |
| | | | IMP | 4 | 3 | 0 | 175 | 100.0 | 51.0 | 100.0 | 98.3 | 95.2 | 99.4 |
| | | | NDM | 36 | 1 | 0 | 145 | 100.0 | 90.4 | 100.0 | 99.3 | 96.2 | 99.9 |
Table 4. Performance of NG-Test CARBA 5 vs. the comparator method - Analysis by Target
'Lower bounds are below 90% due to the low prevalence of the OXA, IMP, and VIM carbapenemases. The claim of NG-Test CARBA 5 detection of OXA, IMP, and VIM carbapenemases is supported by analytical reactivity data.
"Three isolates were false positive for IMP on NG-Test CARBA 5, positive mCM, and negative Xpert Carbe-R result). All three isolates were confirmed to have the positives for IMP after discrepant analysis. (IMP-3 is predicted to be deceded by Xpert Carba-R based on in silico analysis but has not been demonstrated analytically, ) One isolate was a false positive for NDM on NG-Test CARBA 5 (positive NDM on NG-Text CARBA 5, positive mCM, negative Xpert Carba-R result). This isolate was confirmed to have an NDM-1 gene making this for NDM after discrepant analysis. (NDM-1 is predicted by Xpert Carba-R based on in silico analysis and has been tested analytically.) After discrepant analysis, the overall PPA increased to 100.0%) and the overall NPA increased to 100.0% (90.6% - 100.0%) for raw stool specimen. The overall PPA increased to 100.0% (97.4% - 100.0%) and the overall NPA increased to 100.0%) for C&S Cary Blair stool specimen.
7
| Raw Stool | Composite Reference Method | |||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| NG-Test CARBA 5 | Positive | 143 | 41,2 | 147 |
| Negative | 0 | 37 | 37 | |
| Total | 143 | 41 | 184 | |
| Positive Percent Agreement (PPA) | 143/143 = 100% (95% CI: 97.4-100%) | |||
| Negative Percent Agreement (NPA) | 37/41 = 90.2% (95% CI: 77.5-96.1%) |
Table 5. Agreement of NG-Test CARBA 5 with the composite reference method when testing bacterial growth on HardyCHROM™ CRE agar after seeded in Raw Stool
1 An alternative PCR assay showed that the NDM false positive isolate harbored a blayDM -1 variant. Isolate was positive by mCIM.
2 An alternative PCR assay and bidirectional sequencing showed that the three IMP false positive isolates harbored blamp -8/-47 variant that is predicted by in silico analysis but not analytically demonstrated to be detected by the assay. Isolates were positive by mCIM.
Table 6. Agreement of NG-Test CARBA 5 with the composite reference method when testing bacterial growth on HardyCHROM™ CRE agar after seeded in C&S Cary Blair Stool
| C&S Cary Blair Stool | Composite Reference Method | |||
|---|---|---|---|---|
| Positive | Negative | Total | ||
| Positive | 141 | 41,2 | 145 | |
| NG-Test CARBA 5 | Negative | 0 | 37 | 37 |
| Total | 141 | 41 | 182 | |
| Positive Percent Agreement (PPA) | 141/141 = 100% (95% CI: 97.3-100%) | |||
| Negative Percent Agreement (NPA) | 37/41 = 90.2% (95% CI: 77.5-96.1%) |
1 An alternative PCR assay showed that the NDM false positive isolate harbored a blayDM -1 variant. Isolate was positive by mCIM.
2 An alternative PCR assay and bidirectional sequencing showed that the three IMP false positive isolates harbored blamp -8/-47 variant that is predicted by in silico analysis but not analytically demonstrated to be detected by the assay. Isolates were positive by mCIM.
8
VII. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICE
| Attribute | Device | Predicate | Substantially
Equivalent? |
|-------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------|
| Name | NG-Test CARBA 5 | RAPIDEC CARBA NP | Yes |
| 510(k) Details | Product Code PTJ
21 CFR 866.1640
"Antimicrobial Susceptibility Test Powder"
Class II
Panel 83 Microbiology | Product Code PTJ
21 CFR 866.1640
"Antimicrobial Susceptibility Test
Powder"
Class II
Panel 83 Microbiology | Yes |
| Intended Use | NG-Test CARBA 5 is an in vitro rapid and
visual multiplex immunochromatographic
assay for the qualitative detection and
differentiation of five common
carbapenemases (KPC, OXA-48-like, VIM,
IMP and NDM) from carbapenem non-
susceptible pure bacterial colonies when
grown on the following media:
• 5% sheep blood agar or MacConkey agar
(16-24 hours) for testing
Enterobacteriaceae and Pseudomonas
aeruginosa
• HardyCHROM™ CRE agar (18-24 hours)
for testing E. coli and KES ( Klebsiella
aerogenes, Klebsiella oxytoca, Klebsiella
pneumoniae, Enterobacter cloacae
complex, and Serratia marcescens ).
The NG-Test CARBA 5 is intended as an aid
for infection control in the detection of
carbapenemase-producing
Enterobacteriaceae and Pseudomonas
aeruginosa in healthcare settings. NG-Test
CARBA 5 is not intended to guide or monitor
treatment for carbapenem non-susceptible
bacterial infections. A positive or negative
NG-Test CARBA 5 test result does not rule
out the presence of other mechanisms of
antibiotic resistance. NG-Test CARBA 5
should be used in conjunction with other
laboratory tests including phenotypic
antimicrobial susceptibility testing. | RAPIDEC CARBA NP is a
phenotypic (colorimetric) in vitro
diagnostic test for the qualitative
detection of carbapenemase enzymes
in Enterobacteriaceae and
Pseudomonas aeruginosa colonies
that have elevated MIC values to any
carbapenem. RAPIDEC CARBA NP
is performed on pure colonies grown
on non-selective sheep blood agar
culture media.
RAPIDEC CARBA NP is intended as
an aid in the prevention and control of
infection caused by carbapenemase-
producing Enterobacteriaceae and
Pseudomonas aeruginosa .
RAPIDEC CARBA NP is not
intended to guide or monitor the
treatment for these bacterial
infections. A negative result does not
preclude the presence of
carbapenemase enzymes. The ability
of RAPIDEC CARBA NP to detect
carbapenemase enzymes encoded by
genetic markers other than KPC,
NDM, OXA-48, VIM, and IMP has
not been established. RAPIDEC
CARBA NP testing should be used in
conjunction with other laboratory
tests including antimicrobial
susceptibility testing. | Yes |
| Inoculum
Preparation | By touching well-isolated colonies with a
loop | By touching well-isolated colonies
with an applicator stick | Yes |
| Sample Type | Bacterial isolates/colonial growth | Bacterial isolates/colonial growth | Yes |
| Interpretation | Visual | Visual | Yes |
| Controls | Build-in procedural control in every test strip | Build-in procedural control in every
test strip | Yes |
9
ANALYTICAL REACTIVITY
NG-Test CARBA 5 was evaluated with ninety-two strains characterized to have a target carbapenemase. Each organism was incubated aerobically for 16 hours on sheep's blood agar and MacConkey agar at 35°C or 18 hours on HardyCHROM™ CRE agar at 35°C. Each test was performed in triplicate from each type of media. NG-Test CARBA 5 test result was read 15 minutes after inoculating the buffer mixed with bacteria into the sample port. The operator was blinded to the expected result while setting up and interpreting the test. All organisms that yielded a negative NG-Test CARBA 5 result were further analyzed by modified carbapenemase inactivation method (mCIM, CLSI M100, S29). After the mCIM analysis, the final sensitivity for all target organisms evaluated was 88/92 (95.7%) from blood agar and 90/92 (97.8%) from MacConkey agar. After the mCIM analysis, the final sensitivity for all target organisms evaluated was 41/41 (100%) from HardyCHROM™ CRE agar. Two IMP-producing P. aeruginosa isolates (IMP-14 and IMP-18) were negative on NG-Test CARBA 5 but positive by mCIM. On blood agar only, two Proteus mirabilis strains resulted in false negative results.
| Organism Group | Number of
strains
tested on
Blood/
MacConkey | Detected Target | Number of
targets
tested on
Blood/
MacConkey
agar | Number
of
targets
Tested
HC
CRE | Variants Tested | Variants Not
Detected |
|---------------------------|----------------------------------------------------------|-----------------|------------------------------------------------------------------|------------------------------------------------|-----------------------------|--------------------------|
| Enterobacteriacae | 66 | KPC | 17 | 8 | 2, 3, 4, 6, 12 | |
| | | OXA-48-like | 12 | 7 | 48, 181, 163, 232 (48 type) | |
| | | VIM | 11 | 9 | 1, 4, 5, 6, 23, 27, 31 | |
| | | IMP | 8 | 7 | 4, 8/472, 261 | |
| | | NDM | 15 | 11 | 11, 5, 6, 7 | |
| | | None | 5 | 2 | | |
| | | Total | 68 | 44 | | |
| Pseudomonas
aeruginosa | 26 | KPC | 5 | | 2, 5 | |
| | | OXA-48-like | 0 | | | |
| | | VIM | 13 | | 2, 11 | |
| | | IMP | 6 | | 1, 7, 14, 18, 19, 26 | 14, 18 |
| | | NDM | 2 | | 1 | |
| | | None | 0 | | | |
| | | Total | 26 | | | |
Table 7. Analytical Reactivity Summary for Carbapenemase Producing Organisms (CPO)
'NDM-1 and IMP-26 not detected in P. mirabilis growth from blood agar, but yielded positive results from MacConkey agar.
2IMP-8 and IMP-47 were determined to be the same protein based on sequence analysis by the Beta-Lactamase Database (http://www.bldb.eu/BLDB.php?prot=B1#IMP).
10
ANALYTICAL SPECIFICTY
81 organisms that exhibit antibiotic resistance mechanisms other than the targets NG-Test CARBA 5 lateral flow assay detects, are carbapenem-susceptible, or are carbapenem nonsusceptible were tested on NG-Test CARBA 5 from blood agar and MacConkey agar. Organisms tested included Enterobacteriaceae (n=54) and P. aeruginosa (n=20), as well as other phylogenetically related organisms (n=7). HardyCHROM™ CRE was inoculated with 16 cross reactive organisms that were tested on NG-Test CARBA 5 and showed no cross reactivity. These organisms were included in the list of claimed organisms for HardyCHROM™ CRE but which do not produce one of the 5 target carbapenemases. Each organism was incubated aerobically at 35°C on sheep's blood agar and MacConkey agar for 16 hours. HardyCHROM™ CRE was incubated for 18 hours prior to testing. NG-Test CARBA 5 test result was read 15 minutes after inoculating the buffer mixed with bacteria into the sample port. The operator was blinded to the expected result while setting up and interpreting the test. 81/81 (100%) of organisms tested from blood and MacConkey agar produced a negative NG-Test CARBA 5 result. 16/16 (100%) of organisms tested from HardyCHROM™ CRE produced a negative NG-Test CARBA 5 result. Organisms with NMC-A, FRI, and GIM mechanisms were tested by the National Reference Center in France and did not cross react.
| Resistant mechanisms evaluated | ||
|---|---|---|
| Organism Group | Blood & MacConkey agar | HardyCHROM™ |
| CRE agar | ||
| Enterobacteriaceae | ACT-type, ACT-2, AmpC, CTX-M [1, 3, 8, 9, 14, 15, 22, 24, 30, 40, 55, 74, 75, 79, 124], DHA-1, ESBL, IMI, mrc-1, OmpK35, OmpK37, OXA [1, 2, 30], SHV [11(2b), 12(2be), 18, 28, 31, 89(2b), 108(u), 154, 179(u), 180(u), 182(u), OSBL(2b)], SME, SME-2, TEM [1, 1(2b), 11(2be), 63(2be), 93(2be), 210(u), OSBL(2b)], tet(A), tet(B) | ACT-2, AmpC, CTX-M [9, 14, 30], DHA-1, IMI, MIR-8, OXA, SME, TEM-129(2be), tet(A) |
| Pseudomonas | ||
| aeruginosa | aadA6, aadB, aph(3')-IIb, catB7, GES-1, GES-5(c), OXA [10, 50], PAO, PDC [1, 5, 19, 35], PER-1, strA, strB, sull, tet(c), VEB-1, inducible AmpC | N/A |
| Other | VanA | N/A |
Table 8. Resistance Mechanisms Evaluated with NG-Test CARBA 5 for Specificity
INCUBATION STUDY
In order to confirm that NG-Test CARBA 5 delivered consistent results over a range of incubation time, twenty-two strains were tested from blood and MacConkey agar every two hours from 16 to 24 hours of incubation. Fifteen of the twenty-two organisms were also tested from HardyCHROM™ CRE every two hours from 18 to 24 hours. All organisms tested produced the expected result on NG-Test CARBA 5 at every time point tested. NG-Test CARBA 5 test result was read 15 minutes after inoculating the buffer mixed with bacteria into the sample port. The operator was blinded to the expected result while setting up and interpreting the test.
11
REFRIGERATION STORAGE STUDY
In order to determine if agar media that has been stored in the refrigerator can be used with NG-Test CARBA 5, twelve strains were cultured and evaluated over time from refrigerated storage. Blood and MacConkey agar plates were inoculated directly with organism (colonies) for the fresh culture, streaking for isolation. HardyCHROM™ CRE was inoculated with organisms at 3x104 CFU/mL in raw stool and stool in C&S Cary Blair Transport Media, streaking for isolation with a 1 µL loop. Each strain was incubated aerobically on sheep's blood agar and MacConkey agar at 35°C and tests were performed after 16 to 24 hours of incubation (Day 0). Ten of the twelve organisms were also tested from HardyCHROM™ CRE after 18 to 24 hours of incubation. All organisms tested produced the expected result on NG-Test CARBA 5 for each day of refrigeration for up to 3 days. The operator was blinded to the expected result while setting up and interpreting the test.
REPRODUCIBILITY
Prior to initiating the clinical study, a panel of 20 blinded isolates provided by Hardy Diagnostics was tested at three distinct study sites on five work days to demonstrate reproducibility and to document proficiency in the performance of the test. Agreement of >95% with known test results was required before proceeding with the study. The testing was done with at least one operator and two readers, blinded to each other's results, per site. All target carbapenemase positive isolates tested (100%) were detected by NG-Test CARBA 5 on all days of the reproducibility study.
CONCLUSIONS
The analytical data presented in this submission demonstrates that NG-Test CARBA 5 is substantially equivalent to the predicate device.