(109 days)
The BioCode Gastrointestinal Pathogen Panel (GPP) is a qualitative multiplexed in vitro diagnostic test intended for use with the BioCode MDx 3000 Instrument. The BioCode GPP is capable of the simultaneous detection of nucleic acids from multiple bacteria, viruses, and parasites extracted directly from unpreserved in Cary-Blair transport medium obtained from individuals with simptoms of gastrointestinal infection. The following bacteria, parasites, and viruses are identified using the BioCode Gastrointestinal Pathogen Panel:
- . Campylobacter (C. jejuni/C. coli)
- Clostridium difficile (C. difficile) toxin A/B (Fresh samples only)
- l Salmonella spp
- Vibrio (V. parahaemolyticus/V. vulnificus/ V. cholerae), including specific identification of Vibrio parahaemolyticus .
- . Yersinia enterocolitica
- . Enteroaggregative Escherichia coli (EAEC)
- Enterotoxigenic Escherichia coli (ETEC) lt/st
- E. coli 0157 serogroup
- Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2
- Shigella/ Enteroinvasive Escherichia coli (EIEC)
- Cryptosporidium spp (C. parvum/C. hominis)
- Entamoeba histolytica
- Giardia lamblia (also known as G. intestinalis and G. duodenalis)
- . Adenovirus F 40/41
- Norovirus GI/GII ■
- . Rotavirus A
The BioCode GPP is indicated as an aid in the diagnosis of gastrointestinal illness and results are meant to be used in conjunction with other clinical, laboratory, and epidemiological data. For In Vitro Diagnostic Use Only. For Prescription Use Only.
Positive results do not rule out co-infection with organisms not included in the BioCode GPP. The agent detected may not be the definite cause of the disease. Negative results in the setting of clinical illness compatible with gastroenteriis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. Concomitant culture is necessary for organism recovery and further typing of bacterial agents. This device is not intended to monitor or guide treatment for C. difficile infection.
Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Adenovinus 40/41, Campylobacter, E. coli 0157, Shigella(EIEC, Yersinia enterocolitica, and Giardia lamblia were established additionally with retrospective clinical specimens. Performance characteristica, Giardia lamblia, Yersinia enterocolitica and Vibrio (V. parahaemolyticus, V. cholerae) were established primarily using contrived clinical specimens.
The BioCode® Gastrointestinal Pathogen Panel (GPP) is a multiplexed nucleic acid-based test designed to be used with the BioCode MDx-3000 system. The BioCode MDx-3000 is an automated system that integrates PCR amplification, target capture, signal generation and optical detection for multiple gastrointestinal pathogens from a single stool specimen, either unpreserved or in Cary Blair. Stool specimens are processed, and nucleic acids extracted with easyMAG, MagNA Pure 96, KingFisher Flex and KingFisher Apex Dx. Once the PCR plate is set up and sealed, all other operations are automated on MDx-3000. The BioCode MDx-3000 Gastrointestinal Infection Panel simultaneously tests for 17 pathogens (see table below) from unpreserved stool specimens or stool collected in Cary-Blair transport medium. Results from the BioCode Gastrointestinal Pathogen Panel (GPP) test are available within less than 4 hours.
The provided text describes the BioCode Gastrointestinal Pathogen Panel (GPP), a diagnostic test for gastrointestinal pathogens, and a study to demonstrate its substantial equivalence to a predicate device.
Here's an analysis of the acceptance criteria and study data:
1. Acceptance Criteria and Reported Device Performance
The acceptance criteria for each pathogen are implied by the reported Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) compared to the easyMAG extraction method (used with the predicate device) for each extraction system (KingFisher Flex and KingFisher Apex Dx). While explicit pre-defined acceptance criteria values are not stated, the tables present the achieved performance with 95% Confidence Intervals. Generally, a high PPA and NPA (typically >90-95%) with narrow confidence intervals are expected for substantial equivalence for diagnostic tests.
Here's a summary of the reported performance for the BioCode GPP using KingFisher Flex and KingFisher Apex Dx, focusing on the "All Archived" data as it covers a larger sample size for each target. Targets with PPA < 90% are highlighted as indicated in the document; however, the document notes that these are "Positive agreement <90%" for Archived samples only, and specifically points out individual false positives/negatives that were retested and often resolved. Targets for which "N/A" is reported for PPA/NPA indicate no positive/negative archived samples were available for comparison, and thus performance was primarily established with contrived samples (as indicated in the 'Indications for Use' section).
Table of Device Performance (Archived Samples - All Combined)
| Target (KingFisher Flex) | PPA (Agreement with easyMAG) | 95% CI | NPA (Agreement with easyMAG) | 95% CI |
|---|---|---|---|---|
| Campylobacter spp.a | 100% (43/43) | 91.8 - 100 | 99.3% (410/413) | 97.9 - 99.8 |
| Clostridium difficileb | 94.3% (66/70) | 86.2 - 97.8 | 99.5% (386/388) | 98.1 - 99.9 |
| E. coli O157c | 100% (13/13) | 77.2 - 100 | 100% (442/442) | 99.1 - 100 |
| Enteroaggregative E. coli (EAEC) d | 96.2% (50/52) | 87.0 - 98.9 | 100% (404/404) | 99.1 - 100 |
| Enterotoxigenic E. coli (ETEC) e | 100% (25/25) | 86.7 - 100 | 100% (430/430) | 99.1 - 100 |
| Shiga toxin-producing E. coli (STEC) f | 94.1% (32/34) | 80.9 - 98.4 | 99.8% (421/422) | 98.7 - 100 |
| Salmonella spp.g | 100% (41/41) | 91.4 - 100 | 99.5% (412/414) | 98.3 - 99.9 |
| Shigella/ EIEC h | 100% (31/31) | 89.0 - 100 | 99.8% (424/425) | 98.7 - 100 |
| Vibrio parahaemolyticus i | 100% (2/2) | 34.2 - 100 | 100% (453/453) | 99.2 - 100 |
| Vibrio spp. (not parahaemolyticus) j | 50% (1/2)** | 9.5 - 90.5 | 99.8% (452/453) | 98.8 - 100 |
| Yersinia enterocolitica k | 100% (6/6) | 61.0 - 100 | 99.6% (447/449) | 98.4 - 99.9 |
| Cryptosporidium spp. l | 100% (27/27) | 87.5 - 100 | 100% (430/430) | 99.1 - 100 |
| Entamoeba histolytica m | N/A | N/A | 100% (455/455) | 99.2 - 100 |
| Giardia lamblia n | 100% (15/15) | 79.6 - 100 | 100% (440/440) | 99.1 - 100 |
| Adenovirus 40/41 o | 92.3% (12/13) | 66.7 - 98.6 | 99.5% (440/442) | 98.4 - 99.9 |
| Norovirus (GI/GII) p | 95.3% (41/43) | 84.5 - 98.7 | 99.8% (413/414) | 98.6 - 100 |
| Rotavirus A q | 100% (19/19) | 83.2 - 100 | 97.7% (427/437) | 95.8 - 98.8 |
| Combined Targets | 96.1% (223/232) | 92.8 - 97.9 | 99.6% (3941/3956) | 99.4 - 99.8 |
| Target (KingFisher Apex Dx) | PPA (Agreement with easyMAG) | 95% CI | NPA (Agreement with easyMAG) | 95% CI |
|---|---|---|---|---|
| Campylobacter spp.a | 97.7% (42/43) | 87.9 – 99.6 | 99.3% (413/416) | 97.9 – 99.8 |
| Clostridium difficile b | 91.1% (51/56) | 80.7 – 96.1 | 99.5% (402/404) | 98.2 – 99.9 |
| E. coli O157 c | 92.3% (12/13) | 66.7 – 98.6 | 99.8% (445/446) | 98.7 – 100 |
| Enteroaggregative E. coli (EAEC) d | 100% (52/52) | 93.1 – 100 | 99.8% (407/408) | 98.6 – 100 |
| Enterotoxigenic E. coli (ETEC) e | 100% (25/25) | 86.7 – 100 | 99.3% (431/434) | 98.0 – 99.8 |
| Shiga toxin-producing E. coli (STEC) f | 94.1% (32/34) | 80.9 – 98.4 | 99.8% (424/425) | 98.7 – 100 |
| Salmonella spp. g | 100% (41/41) | 91.4 – 100 | 99.3% (415/418) | 97.9 – 99.8 |
| Shigella/ EIEC h | 96.8% (30/31) | 83.8 – 99.4 | 99.1% (425/429) | 97.6 – 99.6 |
| Vibrio parahaemolyticus i | 100% (2/2) | 34.2 – 100 | 99.8% (456/457) | 98.8 – 100 |
| Vibrio spp. (not parahaemolyticus) j | 50% (1/2)** | 9.5 – 90.5 | 99.8% (456/457) | 98.8 – 100 |
| Yersinia enterocolitica k | 100% (6/6) | 61.0 – 100 | 99.6% (451/453) | 98.4 – 99.9 |
| Cryptosporidium spp.l | 96.3% (26/27) | 81.7 – 99.3 | 99.8% (432/433) | 98.7 – 100 |
| Entamoeba histolytica m | N/A | N/A | 100% (459/459) | 99.1 – 100 |
| Giardia lamblia n | 93.3% (14/15) | 70.2 – 98.8 | 100% (444/444) | 99.1 – 100 |
| Adenovirus 40/41 ° | 92.3% (12/13) | 66.7 – 98.6 | 99.6% (444/446) | 98.4 – 99.9 |
| Norovirus (GI/GII) p | 97.7% (42/43) | 87.9 – 99.6 | 100% (417/417) | 99.1 – 100 |
| Rotavirus A 9 | 100% (19/19) | 83.2 – 100 | 100% (441/441) | 99.1 – 100 |
| Combined Targets | 95.4% (208/218) | 91.8 – 97.5 | 99.7% (4006/4017) | 99.5 - 99.8 |
Note:
- Asterisks (**) in the table indicate that the specific performance measure (PPA) was below 90% for archived samples. The document explicitly notes for Vibrio spp. (not parahaemolyticus) that 1 out of 2 positive concordance was observed for both KingFisher Flex and KingFisher Apex Dx, resulting in 50% PPA.
- Retesting of discordant samples often showed resolution in favor of the predicate or new device, but the summary tables reflect the initial comparison. The footnotes (a-q) provide detailed explanations for discordant cases, including instances where samples were invalid or initial false results were later identified as true negatives/positives upon retesting.
2. Sample Size and Data Provenance
- Test Set (Clinical Method Comparison Study):
- Archived Samples: 468 remnant, de-identified samples (254 frozen unpreserved stool and 214 inoculated Cary-Blair stool). These samples were prospectively collected for the clinical study that resulted in the predicate device (K180041 BioCode® GPP FDA clearance). The country of origin is not explicitly stated, but clinical studies for FDA clearance typically involve multi-center studies within the US or compliant international sites. "Mayo-GI" and "TRI" sample names suggest US clinical sites.
- Fresh Samples (for C. difficile testing): 54 freshly collected leftover samples (unpreserved).
- Contrived Samples: 120 samples (15 at 3x LoD and 15 at 6xLoD for each of three targets: Entamoeba histolytica, Yersinia enterocolitica, and Vibrio spp.).
3. Number of Experts to Establish Ground Truth for Test Set and Qualifications
The ground truth for the clinical method comparison study was established by comparing the results of the new device (BioCode GPP with KingFisher Flex and KingFisher Apex Dx extraction) against the results obtained with the easyMAG extraction, which was used with the predicate device (BioCode GPP K190585). The predicate device itself would have undergone its own clinical validation against a pre-established ground truth. Therefore, the "ground truth" for this specific study is the performance of the BioCode GPP with easyMAG extraction.
The document does not explicitly state the number or qualifications of experts used for establishing the initial ground truth of the predicate device, or for adjudicating discordant results in this study. However, the retesting notes for discordant samples mention "consensus results" were not used in the agreement calculation, implying that the retesting was performed to understand the discrepancy rather than to establish a new ground truth.
4. Adjudication Method for the Test Set
The adjudication method is described in the footnotes for Tables 11 and 16. For discordant results between the easyMAG (reference) and the KingFisher Flex/Apex Dx (new device), samples were retested twice with both original extraction methods. The outcome of these retests served to clarify the discrepancy, though the initial raw comparison was used to calculate the agreement, with retest findings noted in the footnotes. For example, a "false negative" for the new device might become a "true negative" upon retesting if the reference system also failed to detect it consistently. However, "consensus results of the discordant samples were not used in the calculation of the agreements" in the main tables but rather for detailed understanding.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done. This study focuses on a laboratory diagnostic device, not an imaging device requiring human reader interpretation. Therefore, the concept of "human readers improve with AI vs without AI assistance" is not applicable here.
6. Standalone Performance Study
Yes, a standalone performance study was done for the device in the context of the new extraction platforms. The "Nonclinical testing summary" details a Reproducibility Study and a Limit of Detection (LoD) study. These studies evaluate the algorithm's (BioCode GPP assay's) consistency and sensitivity when integrated with the new KingFisher Flex and KingFisher Apex Dx extraction systems, independent of direct human interpretative input beyond standard lab procedures.
- Reproducibility Study: Assessed intra-assay, inter-assay, day-to-day, and instrument-to-instrument (operator-to-operator) reproducibility using quantitative controls diluted in stool.
- Limit of Detection (LoD) Study: Determined the lowest concentration at which ≥95% detection was achieved in 20 replicates for each target, for both unpreserved and Cary-Blair stool, using the new extraction methods.
7. Type of Ground Truth Used
For the clinical method comparison study (test set), the ground truth was the result generated by the predicate device's established extraction method (easyMAG). This is a comparative effectiveness study where the predicate device's performance established the "truth" for evaluating the new device's substantial equivalence.
For the LoD and reproducibility studies, the ground truth was based on quantified stocks of bacteria, viruses, or parasites (except for Norovirus GI/GII which used positive clinical specimens with serial dilutions) introduced into negative stool matrices.
8. Sample Size for the Training Set
The document does not explicitly describe a "training set" for the BioCode GPP itself, as it's a multiplex nucleic acid-based test rather than a machine learning or AI algorithm in the common sense requiring explicit training data. The assay's design (probes, primers, etc.) would have been developed and optimized against a range of target and non-target organisms.
The 468 remnant clinical samples used in the method comparison study were prospectively collected for the predicate device's clinical study (K180041 BioCode® GPP FDA clearance), suggesting they were part of the validation for the predicate, not a training set for the current device's underlying assay design.
9. How the Ground Truth for the Training Set Was Established
As noted above, an explicit "training set" with ground truth in the AI/ML context isn't specified for this type of diagnostic assay. The development of such an assay involves extensive analytical validation to establish specificity, sensitivity, and inclusivity using characterized microbial strains, clinical isolates, and negative controls. The ground truth for these developmental studies would be based on molecular characterization (e.g., sequencing), culture, or other reference methods for the target organisms. The performance of the predicate device (K190585) would have been established through a clinical study where its results were compared against clinical diagnosis, culture, or other validated reference methods.
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January 10, 2025
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Applied BioCode, Inc. Alex Chang Regulatory Consultant 12130 Mora Drive Unit 2 Santa Fe Springs, California 90670
Re: K242877
Trade/Device Name: BioCode Gastrointestinal Pathogen Panel (GPP) Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal Microorganism Multiplex Nucleic Acid-Based Assay Regulatory Class: Class II Product Code: PCH Dated: September 20, 2024 Received: September 23, 2024
Dear Alex Chang:
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 (the 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 available 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.
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Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); 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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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-regulatory
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assistance/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,
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Bryan Grabias, Ph. D. Acting Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known)
K242877
Device Name
BioCode Gastrointestinal Pathogen Panel (GPP)
Indications for Use (Describe)
The BioCode Gastrointestinal Pathogen Panel (GPP) is a qualitative multiplexed in vitro diagnostic test intended for use with the BioCode MDx 3000 Instrument. The BioCode GPP is capable of the simultaneous detection of nucleic acids from multiple bacteria, viruses, and parasites extracted directly from unpreserved in Cary-Blair transport medium obtained from individuals with simptoms of gastrointestinal infection. The following bacteria, parasites, and viruses are identified using the BioCode Gastrointestinal Pathogen Panel:
- . Campylobacter (C. jejuni/C. coli)
- Clostridium difficile (C. difficile) toxin A/B (Fresh samples only)
- l Salmonella spp
- Vibrio (V. parahaemolyticus/V. vulnificus/ V. cholerae), including specific identification of Vibrio parahaemolyticus .
- . Yersinia enterocolitica
- . Enteroaggregative Escherichia coli (EAEC)
- Enterotoxigenic Escherichia coli (ETEC) lt/st
- E. coli 0157 serogroup
- Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2
- Shigella/ Enteroinvasive Escherichia coli (EIEC)
- Cryptosporidium spp (C. parvum/C. hominis)
- Entamoeba histolytica
- Giardia lamblia (also known as G. intestinalis and G. duodenalis)
- . Adenovirus F 40/41
- Norovirus GI/GII ■
- . Rotavirus A
The BioCode GPP is indicated as an aid in the diagnosis of gastrointestinal illness and results are meant to be used in conjunction with other clinical, laboratory, and epidemiological data. For In Vitro Diagnostic Use Only. For Prescription Use Only.
Positive results do not rule out co-infection with organisms not included in the BioCode GPP. The agent detected may not be the definite cause of the disease. Negative results in the setting of clinical illness compatible with gastroenteriis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. Concomitant culture is necessary for organism recovery and further typing of bacterial agents. This device is not intended to monitor or guide treatment for C. difficile infection.
Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Adenovinus 40/41, Campylobacter, E. coli 0157, Shigella(EIEC, Yersinia enterocolitica, and Giardia lamblia were established additionally with retrospective clinical specimens. Performance characteristica, Giardia lamblia, Yersinia enterocolitica and Vibrio (V. parahaemolyticus, V. cholerae) were established primarily using contrived clinical specimens.
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Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
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510(k) Summary
Prepared in accordance with the requirements of 21 CFR 807.92.
Submitter Information [807.92(a)(1)]
| Submitter/ | Applied BioCode®, Inc. | |
|---|---|---|
| Applicant | 12130 Mora Drive | |
| Santa Fe Springs, CA 90670 |
Submitter Contact: Alex Chang Phone: +1 (408) 839-5826 Email: achang@apbiocode.com
Device Information [807.92(a)(2)]
| Trade Name | BioCode Gastrointestinal Pathogen Panel (GPP) | |
|---|---|---|
| Classification | Gastrointestinal microorganism multiplex nucleic acid-based assay | |
| Name | ||
| Regulation | 21 CFR § 866.3990 |
Predicate Device [807.92(a)(3)]
BioCode Gastrointestinal Pathogen Panel (GPP) K190585
Device Description [807.92(a)(4)]
The BioCode® Gastrointestinal Pathogen Panel (GPP) is a multiplexed nucleic acid-based test designed to be used with the BioCode MDx-3000 system. The BioCode MDx-3000 is an automated system that integrates PCR amplification, target capture, signal generation and optical detection for multiple gastrointestinal pathogens from a single stool specimen, either unpreserved or in Cary Blair. Stool specimens are processed, and nucleic acids extracted with easyMAG, MagNA Pure 96, KingFisher Flex and KingFisher Apex Dx. Once the PCR plate is set up and sealed, all other operations are automated on MDx-3000. The BioCode MDx-3000 Gastrointestinal Infection Panel simultaneously tests for 17 pathogens (see table below) from unpreserved stool specimens or stool collected in Cary-Blair transport medium. Results from the BioCode Gastrointestinal Pathogen Panel (GPP) test are available within less than 4 hours.
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Bacteria. Viruses. Diarrheagenic E. coli/Shigella. and Parasites Detected by the BioCode MDx Gastrointestinal Pathogen Panel (GPP)
| Bacteria | Parasites |
|---|---|
| ■ Campylobacter spp. (C. jejuni, C. coli) | ■ Cryptosporidium spp. |
| ■ Clostridium difficile toxin A/B (Fresh samples only) | ■ Entamoeba histolytica |
| ■ Enteroaggregative E. coli (EAEC) | ■ Giardia lamblia |
| ■ Enterotoxigenic E. coli (ETEC): LT/ST | |
| ■ Shiga-toxin producing E. coli (STEC): stx1/stx2 | Viruses |
| ■ E.coli O157 | ■ Adenovirus 40/41 |
| ■ Shigella spp. /Enteroinvasive E.coli (EIEC) | ■ Norovirus GI/GII |
| ■ Salmonella spp. | ■ Rotavirus A |
| ■ Vibrio parahaemolyticus | |
| ■ Vibrio spp (not parahaemolyticus) | RNA Internal Control |
| ■ Yersinia enterocolitica |
Intended Use/ Indications for Use [807.92(a)(5)]
The BioCode Gastrointestinal Pathogen Panel (GPP) is a qualitative multiplexed nucleic acidbased in vitro diagnostic test intended for use with the BioCode MDx 3000 Instrument. The BioCode GPP is capable of the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites extracted directly from unpreserved stool samples or stool preserved in Cary-Blair transport medium obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following bacteria, parasites, and viruses are identified using the BioCode Gastrointestinal Pathogen Panel:
- Campylobacter (C. jejuni/C. coli) "
- . Clostridium difficile (C. difficile) toxin A/B (Fresh samples only)
- . Salmonella spp
- . Vibrio (V. parahaemolyticus/V. vulnificus/ V. cholerae), including specific identification of Vibrio parahaemolyticus
- Yersinia enterocolitica "
- . Enteroaggregative Escherichia coli (EAEC)
- . Enterotoxigenic Escherichia coli (ETEC) lt/st
- . E. coli 0157 serogroup
- Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2 "
- Shigella/ Enteroinvasive Escherichia coli (EIEC) .
- " Cryptosporidium spp (C. parvum/C. hominis)
- . Entamoeba histolytica
- . Giardia lamblia (also known as G. intestinalis and G. duodenalis)
- . Adenovirus F 40/41
- Norovirus GI/GII ■
- Rotavirus A .
The BioCode GPP is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness and results are meant to be used in conjunction with other clinical, laboratory, and epidemiological data. For In Vitro Diagnostic Use Only. For Prescription Use Only.
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Positive results do not rule out co-infection with organisms not included in the BioCode GPP. The agent detected may not be the definite cause of the disease. Negative results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease. Concomitant culture is necessary for organism recovery and further typing of bacterial agents. This device is not intended to monitor or guide treatment for C. difficile infection.
Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Adenovirus 40/41, Campylobacter, E. coli 0157, Shigella/EIEC, Yersinia enterocolitica, and Giardia lamblia were established additionally with retrospective clinical specimens. Performance characteristics for Entamoeba histolytica, Giardia lamblia, Yersinia enterocolitica and Vibrio (V. parahaemolyticus, V. vulnificus, and V. cholerae) were established primarily using contrived clinical specimens.
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Device Comparison [807.92(a)(6)]:
| Characteristic | Proposed Device | Predicate |
|---|---|---|
| Name | BioCode® Gastrointestinal Pathogen Panel (GPP) | BioCode® Gastrointestinal Pathogen Panel (GPP) |
| Common Name | Gastrointestinal Microorganism Multiplex Nucleic acid-based assay | Gastrointestinal Microorganism Multiplex Nucleic acid-based assay |
| 510(k) No. | K242877 | K190585 |
| Regulation | 21CFR 866.3990 | 21CFR 866.3990 |
| Product Code | PCH, OOI | PCH, OOI |
| Device Class | II | II |
| Similarities | ||
| Intended Use | The BioCode Gastrointestinal Pathogen Panel (GPP) is a qualitative, multiplexed nucleic acid-based in vitro diagnostic test intended for use with the BioCode MDx 3000 Instrument. The BioCode GPP is capable of the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites extracted directly from unpreserved stool samples or stool preserved in Cary-Blair transport medium obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following bacteria, parasites, and viruses are identified using the BioCode Gastrointestinal Pathogen Panel:Campylobacter ( C. jejuni/C. coli ) Clostridium difficile ( C. difficile ) toxin A/B (Fresh samples only) Salmonella spp Vibrio ( V. parahaemolyticus/V. vulnificus/ V. cholerae ), including specific identification of Vibrio parahaemolyticus Yersinia enterocolitica Enteroaggregative Escherichia coli (EAEC) Enterotoxigenic Escherichia coli (ETEC) lt/st E. coli O157 serogroup Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2 Shigella/ Enteroinvasive Escherichia coli (EIEC) Cryptosporidium spp ( C. parvum/C. | Same |
| Characteristic | Proposed Device | Predicate |
| Entamoeba histolytica Giardia lamblia (also known as G. intestinalis and G. duodenalis) Adenovirus F 40/41 Norovirus GI/GII Rotavirus A The BioCode® GPP is indicated as an aidin the diagnosis of specific agents ofgastrointestinal illness and results aremeant to be used in conjunction with otherclinical, laboratory, and epidemiologicaldata. For In Vitro Diagnostic Use Only.For Prescription Use Only.Positive results do not rule out co-infectionwith organisms not included in the BioCode® | ||
| GPP. The agent detected may not be thedefinite cause of the disease. Negativeresults in the setting of clinical illnesscompatible with gastroenteritis may be due toinfection by pathogens that are not detectedby this test or non-infectious causes such asulcerative colitis, irritable bowel syndrome, or | ||
| Crohn's disease. Concomitant culture isnecessary for organism recovery and furthertyping of bacterial agents. This device is notintended to monitor or guide treatment for C. difficile infection. | ||
| Instrument | Nucleic Acid Purification SystemBioCode MDx-3000 | Same |
| Sample Type | Unpreserved stool and stool in Cary BlairMedium | Same |
| Controls | Externally Sourced - Zeptometrix | Same |
| Methodology | Multiplex RT-PCR and probehybridization to biotinylated PCRproduct(s) followed by fluorescencedetection and decoding of barcodedmagnetic beads (BMB) that are coupled totarget-specific probes | Same |
| Calibrators | Internal Calibration | Same |
| Differences | ||
| Sample Extraction | easyMAG, Roche MagNA Pure 96,KingFisher Flex, KingFisher Apex Dx | easyMAG, Roche MagNA Pure96 |
Comparison of the Applied BioCode GPP with the Predicate Device
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Summary of Performance Data [807.92(b)]
All necessary testing was conducted on BioCode Gastrointestinal Pathogen Panel (GPP) to support a determination of substantial equivalence to the predicate device.
Nonclinical testing summary [807.92(b)(1)]
REPRODUCIBILITY STUDY
A study was performed to assess the Reproducibility of the BioCode® GPP using samples extracted with the KingFisher Flex and KingFisher Apex Dx. This study was designed to assess intra-assay (within run), Inter-assay (run-to-run), day-to-day and instrument-to-instrument (operator-to-operator) reproducibility. One lot of reagents was assayed at Applied BioCode on 3 instruments by 3 operators, 2 runs per day per operator for 5 days (total of 30 runs).
Organisms were diluted in Cary-Blair, and pools were constructed in Cary Blair media at 15X or 30X LoD. The prepared pools were then diluted 1:10 fold in pre-screened negative stool, frozen over night or longer, extracted in triplicate and assayed in singlicate. In total 30 sets of RP pools were run in singlicate on 3 instruments by 3 operators over 5 days.
| Organism | Concentration | % AgreementwithExpectedResult | 95% CI | MeanMFI | %CV |
|---|---|---|---|---|---|
| Salmonella enterica* | 3X LoD (6.60E+03CFU/mL) | (90/90) 100% | (95.98,100.0) | 15920 | 10.9 |
| Salmonella enterica* | 1.5X LoD(3.30E+03CFU/mL) | (89/90) 98.9% | (93.96, 99.97) | 16923 | 8.0 |
| Clostridium difficile(tcd A)* | 3X LoD (9.21E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 15693 | 19.7 |
| Clostridium difficile(tcd A)* | 1.5X LoD(4.61E+01CFU/mL) | (89/90) 98.9% | (93.96, 99.97) | 10333 | 36.3 |
| Clostridium difficile(tcd B)* | 3X LoD (9.21E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 23783 | 12.0 |
| Clostridium difficile(tcd B)* | 1.5X LoD(4.61E+01CFU/mL) | (89/90) 98.9% | (93.96, 99.97) | 18821 | 17.4 |
| Giardia lamblia | 3X LoD (5.40E+03cysts/mL) | (89/90) 98.9% | (93.96, 99.97) | 13827 | 28.3 |
| Giardia lamblia | 1.5X LoD(2.70E+03cysts/mL) | (90/90) 100% | (95.98,100.0) | 15071 | 27.6 |
| Adenovirus 40* | 3X LoD (9.90E-02TCID50/mL) | (89/90) 98.9% | (93.96, 99.97) | 13061 | 20.6 |
| Adenovirus 40* | 1.5X LoD (4.95E-02TCID50/mL) | (90/90) 100% | (95.98,100.0) | 16742 | 19.3 |
| Shigella sonnei | 3X LoD (1.32E+03CFU/mL) | (90/90) 100% | (95.98,100.0) | 19328 | 13.9 |
| Shigella sonnei | 1.5X LoD(6.60E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 19515 | 14.3 |
| Organism | Concentration | % AgreementwithExpectedResult | 95% CI | MeanMFI | %CV |
| ETEC (LT) | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98,100.0) | 13727 | 18.5 |
| ETEC (LT) | 1.5X LoD(8.42E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 13137 | 21.4 |
| ETEC (ST1a) | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98,100.0) | 23135 | 17.2 |
| ETEC (ST1a) | 1.5X LoD(8.42E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 25866 | 11.4 |
| ETEC (ST1b) | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98,100.0) | 26034 | 15.1 |
| ETEC (ST1b) | 1.5X LoD(8.42E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 27576 | 13.0 |
| Vibrio parahaemolyticus(ToxR) | 3X LoD (3.90E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 26590 | 12.1 |
| Vibrio parahaemolyticus(ToxR) | 1.5X LoD(1.95E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 21967 | 20.0 |
| Vibrio parahaemolyticus(Vib) | 3X LoD (3.90E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 10900 | 12.1 |
| Vibrio parahaemolyticus(Vib) | 1.5X LoD(1.95E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 9295 | 29.7 |
| Yersinia enterocolitica | 3X LoD (5.01E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 14645 | 22.1 |
| Yersinia enterocolitica | 1.5X LoD(2.51E+02CFU/mL) | (90/90) 100% | (95.98,100.0) | 13641 | 27.6 |
| STEC | 3X LoD (2.25E+04CFU/mL) | (90/90) 100% | (95.98,100.0) | 17774 | 15.0 |
| STEC | 1.5X LoD(1.13E+04CFU/mL) | (90/90) 100% | (95.98,100.0) | 17965 | 18.1 |
| Campylobacter coli* | 3X LoD (1.68E+02CFU/mL) | (89/90) 98.9% | (93.96, 99.97) | 22869 | 7.8 |
| Campylobacter coli* | 1.5X LoD(8.40E+01CFU/mL) | (90/90) 100% | (95.98,100.0) | 21878 | 12.9 |
| Cryptosporidium*parvum | 3X LoD (9.30E+03oocysts/mL) | (90/90) 100% | (95.98,100.0) | 23202 | 16.9 |
| Cryptosporidium*parvum | 1.5X LoD(4.65E+03oocysts/mL) | (89/90) 98.9% | (93.96, 99.97) | 22292 | 11.9 |
| Rotavirus A* | 3X LoD (6.57E+02TCID50/mL) | (90/90) 100% | (95.98,100.0) | 26666 | 24.4 |
| Rotavirus A* | 1.5X LoD(3.29E+02TCID50/mL) | (89/90) 98.9% | (93.96, 99.97) | 22240 | 14.5 |
Table 1. KingFisher Flex - Positive Agreement of Results of Reproducibility Panel.
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*False negatives became positives upon repeat from PCR, using the same extracts. The false negatives were due to sampling errors or PCR amplicon transfer during hybridization of MDx-3000.
| Target | Negative Agreement(True Negative/TrueNegative+ False Positive) | 95% CI |
|---|---|---|
| Salmonella enterica | 450/450 (100.00) | (99.15, 100.00) |
| Clostridium difficile | 450/450 (100.00) | (99.15, 100.00) |
| Giardia lamblia | 450/450 (100.00) | (99.15, 100.00) |
| Adenovirus 40 | 450/450 (100.00) | (99.15, 100.00) |
| Shigella sonnei | 450/450 (100.00) | (99.15, 100.00) |
| Vibrio parahaemolyticus* | 450/450 (100.00) | (99.15, 100.00) |
| ETEC | 450/450 (100.00) | (99.15, 100.00) |
| Yersinia enterocolitica | 450/450 (100.00) | (99.15, 100.00) |
| STEC | 450/450 (100.00) | (99.15, 100.00) |
| Campylobacter coli | 450/450 (100.00) | (99.15, 100.00) |
| Cryptosporidium parvum | 450/450 (100.00) | (99.15, 100.00) |
| Rotavirus A | 450/450 (100.00) | (99.15, 100.00) |
Table 2. KingFisher Flex – Negative Agreement of Results of Reproducibility Panel.
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| Organism | Concentration | % Agreementwith ExpectedResult | 95% CI | Mean MFI | %CV |
|---|---|---|---|---|---|
| Salmonella enterica | 3X LoD (6.60E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 16189 | 13.2 |
| 1.5X LoD (3.30E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 17748 | 18.2 | |
| Clostridium difficile(tcd A)* | 3X LoD (9.21E+01CFU/mL) | (90/90) 100% | (95.98, 100.0) | 18406 | 18.8 |
| 1.5X LoD (4.61E+01CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 12099 | 30.4 | |
| Clostridium difficile(tcd B)* | 3X LoD (9.21E+01CFU/mL) | (90/90) 100% | (95.98, 100.0) | 26553 | 9.9 |
| 1.5X LoD (4.61E+01CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 20579 | 16.5 | |
| Giardia lamblia | 3X LoD (1.80E+03cysts/mL) | (90/90) 100% | (95.98, 100.0) | 14679 | 26.5 |
| 1.5X LoD (9.00E+02cysts/mL) | (90/90) 100% | (95.98, 100.0) | 18652 | 19.5 | |
| Adenovirus 40* | 3X LoD (9.90E-02TCID50/mL) | (89/90) 98.89% | (93.96, 99.97) | 13952 | 21.4 |
| 1.5X LoD (4.95E-02TCID50/mL) | (90/90) 100% | (95.98, 100.0) | 19824 | 14.4 | |
| Shigella sonnei | 3X LoD (1.32E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 17642 | 17.0 |
| 1.5X LoD (6.60E+02CFU/mL) | (90/90) 100% | (95.98, 100.0) | 18937 | 18.5 | |
| ETEC (LT)* | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 16907 | 11.8 |
| 1.5X LoD (8.42E+02CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 15586 | 15.2 | |
| ETEC (ST1a) | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 22884 | 17.9 |
| 1.5X LoD (8.42E+02CFU/mL) | (89/90) 98.9% | (93.96, 99.97) | 27127 | 10.6 | |
| ETEC (ST1b)* | 3X LoD (1.68E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 26896 | 14.3 |
| 1.5X LoD (8.42E+02CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 29785 | 8.6 | |
| Vibrio parahaemolyticus(ToxR)* | 3X LoD (1.30E+01CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 24510 | 11.9 |
| 1.5X LoD (6.50E+00CFU/mL) | (90/90) 100% | (95.98, 100.0) | 20694 | 16.2 | |
| Vibrio parahaemolyticus(Vib)* | 3X LoD (1.30E+01CFU/mL) | (89/90) 98.89% | (93.96, 99.97) | 9362 | 16.6 |
| 1.5X LoD (6.50E+00CFU/mL) | (90/90) 100% | (95.98, 100.0) | 7110 | 24.2 | |
| Yersinia enterocolitica | 3X LoD (1.50E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 7245 | 33.6 |
| 1.5X LoD (7.52E+02CFU/mL) | (90/90) 100% | (95.98, 100.0) | 6912 | 34.0 | |
| Organism | Concentration | % Agreementwith ExpectedResult | 95% CI | Mean MFI | %CV |
| STEC | 3X LoD (2.50E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 19069 | 15.0 |
| 1.5X LoD (1.25E+03CFU/mL) | (90/90) 100% | (95.98, 100.0) | 17716 | 20.3 | |
| Campylobacter coli | 3X LoD (1.68E+02CFU/mL) | (90/90) 100% | (95.98, 100.0) | 23830 | 6.2 |
| 1.5X LoD (8.40E+01CFU/mL) | (90/90) 100% | (95.98, 100.0) | 23139 | 10.5 | |
| Cryptosporidium parvum | 3X LoD (3.09E+03oocysts/mL) | (90/90) 100% | (95.98, 100.0) | 22639 | 15.5 |
| 1.5X LoD (1.55E+03oocysts/mL) | (90/90) 100% | (95.98, 100.0) | 22527 | 12.5 | |
| Rotavirus A | 3X LoD (6.57E+02TCID50/mL) | (90/90) 100% | (95.98, 100.0) | 25647 | 20.8 |
| 1.5X LoD (3.29E+02TCID50/mL) | (90/90) 100% | (95.98, 100.0) | 25301 | 18.8 |
Table 3. KingFisher Apex Dx-Positive Agreement of Results of Reproducibility Panel.
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*False negatives became positives upon repeat from PCR, using the same extracts. The false negatives were due to sampling errors or PCR amplicon transfer during hybridization of MDx-3000.
| Table 4. KingFisher Apex Dx-Negative Agreement of Results of Reproducibility Panel. | and the country of the country of the first of the first of the first of the first of the first of the first of the first of the first of the first of the first of the first | |||
|---|---|---|---|---|
| Target | Negative Agreement(True Negative/TrueNegative+ False Positive) | 95% CI |
|---|---|---|
| Salmonella enterica | 450/450 (100.00) | (99.15, 100.00) |
| Clostridium difficile | 448/450 (99.56) | (98.39, 99.88) |
| Giardia lamblia | 450/450 (100.00) | (99.15, 100.00) |
| Adenovirus 40 | 450/450 (100.00) | (99.15, 100.00) |
| Shigella sonnei | 450/450 (100.00) | (99.15, 100.00) |
| Vibrio parahaemolyticus | 450/450 (100.00) | (99.15, 100.00) |
| ETEC | 450/450 (100.00) | (99.15, 100.00) |
| Yersinia enterocolitica | 447/450 (99.33) | (98.06, 99.77) |
| STEC | 450/450 (100.00) | (99.15, 100.00) |
| Campylobacter coli | 450/450 (100.00) | (99.15, 100.00) |
| Cryptosporidium parvum | 450/450 (100.00) | (99.15, 100.00) |
| Rotavirus A | 450/450 (100.00) | (99.15, 100.00) |
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LIMIT OF DETECTION (LoD)
A study was performed to assess the performance of the BioCode GPP on the BioCode MDx-3000 at the Limit of Detection (LoD) for both unpreserved Stool and Cary-Blair specimens. In this study the GI Panel was tested with quantified bacteria, virus or parasite stocks (except norovirus which used clinical samples). For initial screening, four replicates of each concentration (near LoD for the predicate) in negative stool and Cary-Blair were extracted on the easyMAG, the KingFisher Flex and the KingFisher Apex Dx and tested in singlet with the BioCode® GPP on the BioCode MDx-3000 system. The LoD was confirmed by extracting 20 replicates of each sample type/extraction method and testing each in singlet for a total of 20 replicates at or near presumptive LoD. LoD for each stock was defined as the lowest concentration with ≥95% detection of 20 replicates (19 out of 20), and was determined separately unpreserved stool and Cary-Blair preserved stool. The LoDs determined from the quantitated stocks or clinical samples are presented below and in the labeling. The LoDs were the same or within 3-fold for each extraction system with a few notable exceptions identified in the tables with footnotes.
| Table 5. Comparison of results for limit of detection testing for Unpreserved Stool extracted | |
|---|---|
| with the easyMAG, the KingFisher Apex Dx, KingFisher Flex and assayed with BioCode® GPP. | |
| EasyMag | KingFisher Flex | KingFisher Apex Dx | |||||
|---|---|---|---|---|---|---|---|
| Strain | Source | Unpreserved Stool LoD | Detection | Unpreserved Stool LoD | Detection | Unpreserved Stool LoD | Detection |
| Campylobactercoli | ATCC33559 | $5.60 \times 10^1$CFU/mL | 20/20 | $5.60 \times 10^1$CFU/mL | 20/20 | $5.60 \times 10^1$CFU/mL | 20/20 |
| Campylobacterjejuni spp. jejuni | ATCC33292 | $2.33 \times 10^2$CFU/mL | 20/20 | $2.33 \times 10^2$CFU/mL | 20/20 | $2.33 \times 10^2$CFU/mL | 20/20 |
| Clostridiumdifficile(toxinotype 0) | ATCC9689 | $2.11 \times 10^1$CFU/mL | 20/20 | $2.11 \times 10^1$CFU/mL | 20/20 | $2.11 \times 10^1$CFU/mL | 20/20 |
| Clostridiumdifficile(toxinotype III;Napl) | ZeptometriX0801619cf | $3.07 \times 10^1$CFU/mL | 19/20 | $3.07 \times 10^1$CFU/mL | 20/20 | $3.07 \times 10^1$CFU/mL | 20/20 |
| Enteroaggregative E. coli092:H33(EAEC) | STECTW04440 | $1.40 \times 10^3$CFU/mL | 20/20 | $1.40 \times 10^3$CFU/mL | 20/20 | $1.40 \times 10^3$CFU/mL | 20/20 |
| EnteroinvasiveE. coli O29:NM(EIEC) | ATCC43892 | $3.60 \times 10^2$CFU/mL | 20/20 | $3.60 \times 10^2$CFU/mL | 20/20 | $3.60 \times 10^2$CFU/mL | 20/20 |
| EnterotoxigenicE. coli 078:H11H10407 (ETEC) | ATCC35401 | $1.87 \times 10^2$CFU/mL | 20/20 | $5.61 \times 10^2$CFU/mL | 20/20 | $5.61 \times 10^2$CFU/mL | 20/20 |
| Salmonellabongori | SGSC4900 | $1.40 \times 10^3$CFU/mL | 20/20 | $4.67 \times 10^2$CFU/mL | 19/20 | $1.40 \times 10^3$CFU/mL | 20/20 |
| Salmonellaenterica ssp.enterica | ATCC14028 | $7.33 \times 10^2$CFU/mL | 20/20 | $2.20 \times 10^3$CFU/mL | 19/20 | $2.20 \times 10^3$CFU/mL | 20/20 |
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| EasyMag | KingFisher Flex | KingFisher Apex Dx | |||||
|---|---|---|---|---|---|---|---|
| Strain | Source | Unpreserved Stool LoD | Detection | Unpreserved Stool LoD | Detection | Unpreserved Stool LoD | Detection |
| Shiga-like toxinproducing E. coli(STEC)* | ATCCBAA-2217 | 2.50 x 103CFU/mL | 20/20 | 7.50 x103CFU/mL | 20/20 | 8.33 x102CFU/mL | 20/20 |
| E. coli 0157 | ATCC700376 | 3.30 x 103CFU/mL | 20/20 | 3.30 x 103CFU/mL | 20/20 | 3.30 x 103CFU/mL | 20/20 |
| Shigella sonnei | ATCC29930 | 4.40 x 102CFU/mL | 20/20 | 4.40 x 102CFU/mL | 20/20 | 4.40 x 102CFU/mL | 20/20 |
| Vibrio cholerae | ATCC25870 | 1.81 x 101CFU/mL | 19/20 | 1.81 x 101CFU/mL | 20/20 | 1.81 x 101CFU/mL | 20/20 |
| VibrioparahaemolyticuS | ATCC17802 | 4.33 x100CFU/mL | 19/20 | 1.31 x101CFU/mL | 20/20 | 4.33 x100CFU/mL | 20/20 |
| Yersiniaenterocolitica ** | ATCC23715 | 1.50 x103CFU/mL | 20/20 | 1.67 x102CFU/mL | 19/20 | 5.01 x102CFU/mL | 19/20 |
| Cryptosporidiumparvum | waterborneP102C | 3.10 x103oocysts/mL | 20/20 | 3.10 x103oocysts/mL | 20/20 | 1.03 x103oocysts/mL | 20/20 |
| Entamoebahistolytica HB-301:NIH | BEI NR-176 | 3.10 x10-1cysts/mL | 20/20 | 3.10 x10-1cysts/mL | 20/20 | 3.10 x10-1cysts/mL | 20/20 |
| Giardiaintestinalis (akaG. lamblia) | waterborneP101 | 6.00 x 102cysts/mL | 20/20 | 1.80 x 103cysts/mL | 20/20 | 6.00 x 102cysts/mL | 20/20 |
| Adenovirus 40(dugan) | Zeptometrix 0810084 | 3.33 x10-2TCID50/mL | 20/20 | 3.33 x10-2TCID50/mL | 19/20 | 3.33 x10-2TCID50/mL | 20/20 |
| Adenovirus 41(TAK) | Zeptometrix 0810085 | 8.46 x10-1TCID50/mL | 20/20 | 8.46 x10-1TCID50/mL | 20/20 | 2.82 x 10-1TCID50/mL | 19/20 |
| Rotavirus A | ZeptometriX0810041CF | 2.19 x102TCID50/mL | 20/20 | 2.19 x102TCID50/mL | 19/20 | 2.19 x102TCID50/mL | 20/20 |
*KingFisher Flex 9x less sensitive than KingFisher Apex Dx; **EM 9x less sensitive than KingFisher Flex
Table 6. Comparison of results for limit of detection testing for Cary-Blair Stool extracted with the easyMAG, the KingFisher Flex Apex Dx, and the KingFisher Flex and assayed with BioCode® GPP.
| EasyMag | KingFisher Flex | KingFisher Apex Dx | |||||
|---|---|---|---|---|---|---|---|
| Strain | Source | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection |
| Campylobacterr coli | ATCC33559 | $1.87 \times 10^1$CFU/mL | 20/20 | $1.87 \times 10^1$CFU/mL | 20/20 | $1.87 \times 10^1$CFU/mL | 19/20 |
| Campylobacterr jejuni spp.jejuni | ATCC33292 | $7.78 \times 10^1$CFU/mL | 19/20 | $2.33 \times 10^2$CFU/mL | 20/20 | $2.33 \times 10^2$CFU/mL | 20/20 |
| Clostridiumdifficile | ATCC 9689 | $7.04 \times 10^0$CFU/mL | 20/20 | $2.11 \times 10^1$CFU/mL | 20/20 | $6.33 \times 10^1$CFU/mL | 20/20 |
| EasyMag | KingFisher Flex | KingFisher Apex Dx | |||||
| Strain | Source | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection |
| (toxinotype0)* | |||||||
| Clostridiumdifficile(toxinotypeIII; Nap1) | Zeptometrix0801619cf | $3.07 x10^1$CFU/mL | 20/20 | $3.07 x10^1$CFU/mL | 19/20 | $3.07 x10^1$CFU/mL | 20/20 |
| Enteroaggregative E. coli092:H33(EAEC)** | STECTW04440 | $1.56 x10^2$CFU/mL | 20/20 | $1.40 x10^3$CFU/mL | 20/20 | $4.20 x10^3$CFU/mL | 20/20 |
| EnteroinvasiveE. coli029:NM(EIEC) | ATCC43892 | $3.60 x10^2$CFU/mL | 20/20 | $3.60 x10^2$CFU/mL | 20/20 | $1.20 x10^2$CFU/mL | 19/20 |
| Enterotoxigenic E. coli078:H11H10407(ETEC) | ATCC35401 | $1.87 x10^2$CFU/mL | 20/20 | $5.61 x10^2$CFU/mL | 20/20 | $1.87 x10^2$CFU/mL | 20/20 |
| Salmonellabongori | SGSC 4900 | $1.40 x10^3$CFU/mL | 20/20 | $1.40 x10^3$CFU/mL | 20/20 | $1.40 x10^3$CFU/mL | 20/20 |
| Salmonellaenterica ssp.enterica | ATCC14028 | $7.33 x10^2$CFU/mL | 20/20 | $7.33 x10^2$CFU/mL | 20/20 | $2.20 x10^3$CFU/mL | 20/20 |
| Shiga-liketoxinproducing E.coli (STEC) | ATCCBAA-2217 | $8.33 x10^2$CFU/mL | 19/20 | $2.50 x10^3$CFU/mL | 19/20 | $2.50 x10^3$CFU/mL | 20/20 |
| E. coli O157 | ATCC700376 | $1.10 x10^3$CFU/mL | 19/20 | $3.30 x10^3$CFU/mL | 20/20 | $3.30 x10^3$CFU/mL | 20/20 |
| Shigellasonnei | ATCC29930 | $1.47 x10^2$CFU/mL | 19/20 | $4.40 x10^2$CFU/mL | 20/20 | $1.47 x10^2$CFU/mL | 19/20 |
| Vibriocholerae | ATCC25870 | $1.81 x10^1$CFU/mL | 20/20 | $1.81 x10^1$CFU/mL | 20/20 | $1.81 x10^1$CFU/mL | 20/20 |
| Vibrioparahaemolyticus | ATCC17802 | $1.30 x10^1$CFU/mL | 20/20 | $1.30 x10^1$CFU/mL | 20/20 | $4.33 x10^0$CFU/mL | 20/20 |
| Yersiniaenterocolitica | ATCC23715 | $1.50 x10^3$CFU/mL | 20/20 | $5.01 x10^2$CFU/mL | 20/20 | $5.01 x10^2$CFU/mL | 20/20 |
| Cryptosporidium parvum | waterborneP102C | $3.10 x10^3$oocysts/mL | 20/20 | $1.03 x10^3$oocysts/mL | 19/20 | $1.03 x10^3$oocysts/mL | 19/20 |
| EntamoebahistolyticaHB-301:NIH | BEI NR-176 | $1.03 x10^{-1}$cysts/mL | 20/20 | $1.03 x10^{-1}$cysts/mL | 20/20 | $1.03 x10^{-1}$cysts/mL | 20/20 |
| Giardiaintestinalis(aka G.lamblia) | waterborneP101 | $6.00 x10^2$cysts/mL | 20/20 | $1.80 x10^3$cysts/mL | 20/20 | $1.80 x10^3$cysts/mL | 20/20 |
| Source | EasyMag | KingFisher Flex | KingFisher Apex Dx | ||||
| Strain | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection | Cary-BlairStool LoD | Detection | |
| Adenovirus 40(dugan) | Zeptometrix0810084 | $3.33 x10^{-2}$TCID50/mL | 19/20 | $3.33 x10^{-2}$TCID50/mL | 20/20 | $3.33 x10^{-2}$TCID50/mL | 20/20 |
| Adenovirus 41(TAK) | Zeptometrix0810085 | $8.46 x10^{-1}$TCID50/mL | 20/20 | $2.54 x10^{0}$TCID50/mL | 20/20 | $8.46 x10^{-1}$TCID50/mL | 20/20 |
| Rotavirus A | Zeptometrix0810041CF | $2.19 x10^{2}$TCID50/mL | 19/20 | $7.29 x10^{1}$TCID50/mL | 20/20 | $7.29 x10^{1}$TCID50/mL | 19/20 |
{17}------------------------------------------------
{18}------------------------------------------------
- KingFisher Apex Dx 27x less sensitive than EM; ** KingFisher Flex 9x and KingFisher Apex Dx 27x less sensitive than EM
For Norovirus GI and GII targets, positive clinical specimens were used, and serial dilutions (initial 10-fold dilution series followed by finer dilutions) were performed. Four replicates of each dilution in negative unpreserved stool and Cary-Blair stool were extracted with the easyMAG, the KingFisher Flex and KingFisher Apex Dx and tested with the BioCode® GPP on the BioCode MDx-3000 system. The LoD was confirmed by extracting 20 replicates of each sample type with each extraction method and testing at or near presumptive LoD.
For Norovirus, the LoD of the Norovirus GII in unpreserved stool was equivalent for three extraction systems. For Norovirus GI, the KingFisher Flex is 10-fold and 3-fold more sensitive than the easyMag and the KingFisher Apex Dx, respectively. For Cary-Blair stool, the LoD of the Norovirus GI was equivalent for three systems. For Norovirus GII, the KingFisher Apex Dx was 3-fold more sensitive than the EasyMag and the KingFisher Flex.
Table 7. Norovirus - Comparison of results for limit of detection testing for Unpreserved Stool extracted with easyMAG, KingFisher Flex, or KingFisher Apex Dx, and assayed with BioCode® GPP.
| Source | TargetProbe | EasyMag | KingFisher Flex | KingFisher Apex Dx | ||||
|---|---|---|---|---|---|---|---|---|
| Target | Unpreserved StoolDilution | Detection | Unpreserved StoolDilution | Detection | Unpreserved StoolDilution | Detection | ||
| Norovirus GI* | ClinicalSampleID#XTAG-24-0005574 | NoVG1 | 1:30,000 | 19/20 | 1:3,000* | 20/20 | 1:10,000 | 19/20 |
| Norovirus GII | ClinicalSampleID#02-0134 | NoVG2 | 1:30,000 | 20/20 | 1:30,000 | 19/20 | 1:30,000 | 20/20 |
*KF Flex 10-fold and 3-fold more sensitive than EM and KF Apex Dx.
{19}------------------------------------------------
Table 8. Norovirus - Comparison of results for limit of detection testing for Cary-Blair Stool extracted with the easyMAG, KingFisher Flex, or KingFisher Apex Dx, and assayed with BioCode® GPP.
| Source | TargetProbe | EasyMag | KingFisher Flex | KingFisher Apex Dx | ||||
|---|---|---|---|---|---|---|---|---|
| Target | Cary-BlairStoolDilution | Detection | Cary-BlairStoolDilution | Detection | Cary-BlairStoolDilution | Detection | ||
| NorovirusGI | ClinicalSampleID#XTAG-24-0005574 | NoVG1 | 1:30,000 | 20/20 | 1:30,000 | 20/20 | 1:30,000 | 20/20 |
| NorovirusGII | ClinicalSampleID#02-0134 | NoVG2 | 1:30,000 | 20/20 | 1:30,000 | 20/20 | 1:10,000* | 20/20 |
*KF Apex Dx 3-fold more sensitive than EM and KF Flex
Clinical testing summary [807.92(b)(2)]
METHOD COMPARISON STUDY
A clinical investigational study was performed in which a total of 468 remnant, de-identified samples (254 frozen unpreserved stool and 214 inoculated Cary-Blair stool) that were prospectively collected for the clinical study that resulted in the K180041 BioCode® GPP FDA clearance were extracted using the KingFisher Flex, KingFisher Apex Dx, and the easyMAG, and tested on the MDx-3000 system. Fifty-four (54) freshly collected leftover samples were used for the C. difficile testing. In addition, a total of 120 samples were contrived at 3x LoD and 6x LoD (15 samples at 3x LoD and 15 samples at 6xLoD for each of three targets) and tested to determine the performance characteristics for Entamoeba histolytica, Yersinia enterocolitica and Vibrio spp. (V. parahaemolyticus, V. vulnificus, and V. cholerae). The demographics of the clinical patients and the results of the clinical investigational study are as follows:
| Archived Samples | |||||
|---|---|---|---|---|---|
| Total Specimen Count | 468 | ||||
| Gender | |||||
| Male | 246/468 (52.6%) | ||||
| Female | 222/468 (47.4%) | ||||
| Age Category | |||||
| < > yrs | 87/468 (18.6%) | ||||
| 6-21 yrs | 85/468 (18.2%) | ||||
| 22-59 yrs | 197/468 (42.1%) | ||||
| 60+yrs | 99/468 (21.1%) |
Table 9. Demographic data for archived specimens (frozen unpreserved and inoculated Cary-Blair)
{20}------------------------------------------------
| Fresh Samples | |
|---|---|
| Total Specimen Count | 54 |
| Gender | |
| Male | 20/54 (37.04%) |
| Female | 34/54 (62.96%) |
| Age Category | |
| ≤ 5 yrs | 0/54 (0.00%) |
| 6-21 yrs | 2/54 (3.70%) |
| 22-59 yrs | 24/54 (44.45%) |
| 60+yrs | 28/54 (51.85%) |
Table 10. Demographic data for fresh specimens (unpreserved)
The Positive Percent Agreement (PPA) was calculated as TP/(TP+FN). TP = true positive or positive by both the EasyMag and the KingFisher Apex Dx or the KingFisher Flex; FN = false negative or negative by the KingFisher Apex Dx or the KingFisher Flex only. Negative Percent Agreement (NPA) was calculated as TN/(TN + FP). TN = true negative or negative by the EasyMag and the KingFisher Apex Dx or the KingFisher Flex; FP = false positive or positive by the KingFisher Apex Dx or the KingFisher Flex only. The exact binomial two-sided 95% confidence interval was calculated.
A. Method Comparison Study between EasyMag and KingFisher Flex
| Table 11. KingFisher Flex: Summary of Clinical Investigational Study Results (Archived | ||||||||
|---|---|---|---|---|---|---|---|---|
| Specimens) Stratified by Sample Type and Storage | ||||||||
| The Market of the country of the country of the county of the county of |
| Target | SpecimenType | (n) | Positive Agreement | Negative Agreement | ||
|---|---|---|---|---|---|---|
| Campylobacter spp.a | InoculatedCary-Blair | 209 | 18/18 (100) | 82.4 - 100 | 191/191 (100) | 98.0 - 100 |
| Campylobacter spp.a | Unpreserved(Frozen) | 247 | 25/25 (100) | 86.7 - 100 | 219/222 (98.6) | 96.1 - 99.5 |
| Campylobacter spp.a | All Archived | 456 | 43/43 (100) | 91.8 - 100 | 410/413 (99.3) | 97.9 - 99.8 |
| Clostridium difficileb | InoculatedCary-Blair | 211 | 35/37 (94.6) | 82.3 - 98.5 | 173/174 (99.4) | 96.8 - 99.9 |
| Clostridium difficileb | Unpreserved(Frozen) | 247 | 31/33 (93.9) | 80.4 - 98.3 | 213/214 (99.5) | 97.4 - 99.9 |
| Clostridium difficileb | All Archived | 458 | 66/70 (94.3) | 86.2 - 97.8 | 386/388 (99.5) | 98.1 - 99.9 |
| E. coli O157c | InoculatedCary-Blair | 209 | 5/5 (100) | 56.6 - 100 | 204/204 (100) | 98.2 - 100 |
| E. coli O157c | Unpreserved(Frozen) | 246 | 8/8 (100) | 67.6 - 100 | 238/238 (100) | 98.4 - 100 |
| E. coli O157c | All Archived | 455 | 13/13 (100) | 77.2 - 100 | 442/442 (100) | 99.1 - 100 |
| Target | Specimen Type | (n) | Positive Agreement | Negative Agreement | ||
| PPA (%) | 95% CI | NPA (%) | 95% CI | |||
| Enteroaggregative E. coli (EAEC) d | InoculatedCary-Blair | 210 | 30/30 (100) | 88.6-100 | 180/180 (100) | 97.9-100 |
| Unpreserved(Frozen) | 246 | 20/22 (90.9) | 77.2-97.5 | 224/224 (100) | 98.3-100 | |
| All Archived | 456 | 50/52 (96.2) | 87.0-98.9 | 404/404 (100) | 99.1-100 | |
| EnterotoxigenicE. coli (ETEC) e | InoculatedCary-Blair | 209 | 14/14 (100) | 78.5-100 | 195/195 (100) | 98.1-100 |
| Unpreserved(Frozen) | 246 | 11/11 (100) | 74.1-100 | 235/235 (100) | 98.4-100 | |
| All Archived | 455 | 25/25 (100) | 86.7-100 | 430/430 (100) | 99.1-100 | |
| Shiga toxin-producingE. coli (STEC) f | InoculatedCary-Blair | 209 | 12/12 (100) | 75.8-100 | 197/197 (100) | 98.1-100 |
| Unpreserved(Frozen) | 247 | 20/22 (90.9) | 72.2-97.5 | 224/225 (99.6) | 97.5-99.9 | |
| All Archived | 456 | 32/34 (94.1) | 80.9-98.4 | 421/422 (99.80) | 98.7-100 | |
| Salmonella spp.g | InoculatedCary-Blair | 209 | 20/20 (100) | 83.9-100 | 188/189 (99.4) | 97.1-99.9 |
| Unpreserved(Frozen) | 246 | 21/21 (100) | 84.5-100 | 224/225 (99.6) | 97.5-99.9 | |
| All Archived | 455 | 41/41 (100) | 91.4-100 | 412/414 (99.5) | 98.3-99.9 | |
| Shigella/ EIEC h | InoculatedCary-Blair | 210 | 13/13 (100) | 77.2-100 | 196/197 (99.5) | 97.2-99.9 |
| Unpreserved(Frozen) | 246 | 18/18 (100) | 82.4-100 | 228/228 (100) | 98.3-100 | |
| All Archived | 456 | 31/31 (100) | 89.0-100 | 424/425 (99.8) | 98.7-100 | |
| Vibrioparahaemolyticus i | InoculatedCary-Blair | 209 | 1/1 (100) | 20.7-100 | 208/208 (100) | 98.2-100 |
| Unpreserved(Frozen) | 246 | 1/1 (100) | 20.7-100 | 245/245 (100) | 98.5-100 | |
| All Archived | 455 | 2/2 (100) | 34.2-100 | 453/453 (100) | 99.2-100 | |
| Vibrio spp. (notparahaemolyticus) j | InoculatedCary-Blair | 209 | N/A | N/A | 208/209 (99.5) | 97.3-99.9 |
| Unpreserved(Frozen) | 246 | 1/2 (50%)* | 9.5–90.5 | 244/244 (100) | 98.5-100 | |
| All Archived | 455 | 1/2 (50%) | 9.5-90.5 | 452/453 (99.8) | 98.8-100 | |
| Target | SpecimenType | (n) | Positive Agreement | Negative Agreement | ||
| PPA (%) | 95% CI | NPA (%) | 95% CI | |||
| Yersiniaenterocolitica k | InoculatedCary-Blair | 209 | 3/3 (100) | 43.9 - 100 | 204/206 (99.0) | 96.5 - 99.7 |
| Unpreserved(Frozen) | 246 | 3/3 (100) | 43.9 - 100 | 243/243 (100) | 98.4 - 100 | |
| All Archived | 455 | 6/6 (100) | 61.0 - 100 | 447/449 (99.6) | 98.4 - 99.9 | |
| Cryptosporidium spp. 1 | InoculatedCary-Blair | 209 | 9/9 (100) | 70.1 - 100 | 200/200 (100) | 98.1 - 100 |
| Unpreserved(Frozen) | 248 | 18/18 (100) | 82.4 - 100 | 230/230 (100) | 98.4 - 100 | |
| All Archived | 457 | 27/27 (100) | 87.5 - 100 | 430/430 (100) | 99.1 - 100 | |
| Entamoebahistolytica m | InoculatedCary-Blair | 209 | N/A | N/A | 209/209 (100) | 98.2 - 100 |
| Unpreserved(Frozen) | 246 | N/A | N/A | 246/246 (100) | 98.5 - 100 | |
| All Archived | 455 | N/A | N/A | 455/455 (100) | 99.2 - 100 | |
| Giardia lamblia n | InoculatedCary-Blair | 209 | 4/4 (100) | 51.0 - 100 | 205/205 (100) | 98.2 - 100 |
| Unpreserved(Frozen) | 246 | 11/11 (100) | 74.1 - 100 | 235/235 (100) | 98.4 - 100 | |
| All Archived | 455 | 15/15 (100) | 79.6 - 100 | 440/440 (100) | 99.1 - 100 | |
| Adenovirus 40/41o | InoculatedCary-Blair | 209 | 5/5 (100) | 56.6 - 100 | 202/204 (99.0) | 96.5 - 99.7 |
| Unpreserved(Frozen) | 246 | 7/8 (87.5)* | 52.9 - 97.8 | 238/238 (100) | 98.4 - 100 | |
| All Archived | 455 | 12/13 (92.3) | 66.7 - 98.6 | 440/442 (99.5) | 98.4 - 99.9 | |
| Norovirus(GI/GII) p | InoculatedCary-Blair | 210 | 22/23 (95.7) | 79.0 - 99.2 | 187/187 (100) | 98.0 - 100 |
| Unpreserved(Frozen) | 247 | 19/20 (95.0) | 76.4 - 99.1 | 226/227 (99.6) | 97.5 - 99.9 | |
| All Archived | 457 | 41/43 (95.3) | 84.5 - 98.7 | 413/414 (99.8) | 98.6 - 100 | |
| Rotavirus A q | InoculatedCary-Blair | 210 | 10/10 (100) | 72.2 - 100 | 198/200 (99.0) | 96.4 - 99.7 |
| Unpreserved(Frozen) | 246 | 9/9 (100) | 70.1 - 100 | 229/237 (96.6) | 93.5 - 98.3 | |
| All Archived | 456 | 19/19 (100) | 83.2 - 100 | 427/437 (97.7) | 95.8 - 98.8 | |
| SpecimenType | (n) | Positive Agreement | Negative Agreement | |||
| Target | PPA (%) | 95% CI | NPA (%) | 95% CI | ||
| Combined Targets | InoculatedCary-Blair | 3559 | 201/204(98.5) | 95.8-99.5 | 3345/3355(99.7) | 99.5-99.8 |
| Unpreserved(Frozen) | 4188 | 223/232(96.1) | 92.8 - 97.9 | 3941/3956(99.6) | 99.4-99.8 |
{21}------------------------------------------------
{22}------------------------------------------------
{23}------------------------------------------------
*Positive agreement <90%.
Thirty-five (35) archived samples with discordant results (between the easyMAG and KingFisher Flex) retested twice with both easyMAG and KingFisher Flex.
a - Campylobacter spp. One (1) false positive retested remained false positive whereas two (2) false positives became true negatives. Twelve (12) samples were still invalid, not used in the agreement calculation.
b - Clostridium difficile. Two (2) false positives retested remained false positives. Of the four (4) false negatives retested, three (3) became true negatives whereas one (1) remained false negative. Ten (10) samples were invalid, not used in the agreement calculation.
c- E. coli O157: Thirteen (13) samples were invalid, not used in the agreement calculation.
d - EAEC: Of the two (2) false negatives retested, one (1) was true negative and one (1) became true positive. Twelve (12) samples were invalid, not used in the agreement calculation.
e- ETEC: Thirteen (13) samples were invalid, not used in the agreement calculation.
f - STEC: Two (2) false negatives retested became true negatives. One (1) false positive became false negative. Twelve (12) samples were still invalid, not used in the agreement calculation.
g - Salmonella spp. Of the two (2) false positives retested, one (1) became true negative and one (1) remained false positive. Thirteen (13) samples were invalid, not used in the agreement calculation. h - Shigella/EIEC. The false positive retested became true positive. Twelve (12) samples were invalid, not used in the agreement calculation.
i- Vibrio parahaemolyticus: Thirteen (13) samples were invalid, not used in the agreement calculation. i - Vibrio spp. One (1) false positive retested became true negative. For the false negative retested, the easyMAG had an invalid result with the 2nd run and a negative result with the 3-d run, whereas the KF Flex had negative results with both runs. Thirteen (13) samples were invalid, not used in the agreement calculation.
k – Yersinia enterocolitica: Of the (2) false positives retested, one (1) became true negative and one (1) remained false positive. Thirteen (13) samples were invalid, not used in the agreement calculation. l- Cryptosporidium spp: Eleven (11) samples were invalid, not used in the agreement calculation.
m- Entamoeba histolytica: Thirteen (13) samples were invalid, not used in the agreement calculation.
n- Giardia lamblia: Thirteen (13) samples were invalid, not used in the agreement calculation
o - Adenovirus 40/41 : The false negative and the false positive retested became true negative and true positive, respectively. Thirteen (13) samples were invalid, not used in the agreement calculation.
p - Norovirus GI/GII: The false positive and the false negative retested became true negative and true positive, respectively. Eleven (11) samples were invalid, not used in the agreement calculation. q- Rotavirus: Of the ten (10) false positives retested, six (6) became true negatives, and four (4) remained
false positives. Twelve (12) samples were invalid, not used in the agreement calculation.
{24}------------------------------------------------
The table below (Table 12) shows the results of retesting samples with discordant results between the easyMAG and the KingFisher Flex. Samples were retested twice with both easyMAG and/or KingFisher Flex. Consensus results of the discordant samples were not used in the calculation of the agreements but noted as the footnotes of Table 11 above.
| Target | SampleName | Sample Type | Extraction | Run 1(Originalrun) | Run 2 | Run 3 | ConsensusResult | |
|---|---|---|---|---|---|---|---|---|
| a | Campylobacterspp | 01-0314 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet |
| 01-0314 | Unpreserved(Frozen) | KingFisherFlex | Det | Ndet | Det | Det | ||
| 03-0153 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Det | Ndet | ||
| 03-0153 | Unpreserved(Frozen) | KingFisherFlex | Det | Ndet | Ndet | Ndet | ||
| Mayo-GI_101 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet | ||
| Mayo-GI_101 | Unpreserved(Frozen) | KingFisherFlex | Det | Det | Ndet | Det | ||
| 01-0019 | InoculatedCary-Blair | easyMAG | Det | Det | Ndet | Det | ||
| 01-0019 | InoculatedCary-Blair | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | ||
| 01-0048 | InoculatedCary-Blair | easyMAG | Det | Ndet | Ndet | Ndet | ||
| 01-0048 | InoculatedCary-Blair | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | ||
| b | Clostridiumdifficile | 01-0080 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet |
| 01-0080 | Unpreserved(Frozen) | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | ||
| 02-0009 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | ||
| 02-0009 | Unpreserved(Frozen) | KingFisherFlex | Ndet | Ndet | Det | Ndet | ||
| 02-0398 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| 02-0398 | Unpreserved(Frozen) | KingFisherFlex | Det | Ndet | Det | Det | ||
| 03-0353 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| 03-0353 | InoculatedCary-Blair | KingFisherFlex | Det | Det | Det | Det | ||
| C | EAEC | 02-0042 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet |
| EAEC | 02-0042 | Unpreserved(Frozen) | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | |
| EAEC | SP366 | Unpreserved(Frozen) | easyMAG | Det | Det | Det | Det | |
| EAEC | SP366 | Unpreserved(Frozen) | KingFisherFlex | Ndet | Det | Det | Det | |
| STEC | 02-0211 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | |
| d | STEC | 02-0211 | Unpreserved(Frozen) | KingFisherFlex | Ndet | Ndet | Ndet | Ndet |
| STEC | Mayo-GI_092 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Det | Det | |
| STEC | Mayo-GI_092 | Unpreserved(Frozen) | KingFisherFlex | Det | Ndet | Ndet | Ndet | |
| STEC | SP228 | Unpreserved | easyMAG | Det | Ndet | Ndet | Ndet | |
| Target | SampleName | Sample Type | Extraction | Run 1(Originalrun) | Run 2 | Run 3 | ConsensusResult | |
| (Frozen) | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | |||
| 01-0137 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet | ||
| e | Salmonella spp | KingFisherFlex | Det | Det | Ndet | Det | ||
| 06-0357 | Cary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherFlex | Det | Ndet | Ndet | Ndet | ||||
| 01-0267 | InoculatedCary-Blair | easyMAG | Ndet | Det | Det | Det | ||
| f | Shigella/EIEC | KingFisherFlex | Det | Det | Ndet | Det | ||
| 02-0167 | Unpreserved(Frozen) | easyMAG | Det | Invalid | Ndet | IND | ||
| Vibrio spp | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | |||
| g | 03-0396 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Invalid | ||
| KingFisherFlex | Det | Ndet | Ndet | |||||
| 06-0053 | Cary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| h | Yersinia | KingFisherFlex | Det | Ndet | Ndet | Ndet | ||
| 06-0222 | Cary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherFlex | Det | Det | Det | Det | ||||
| Adenovirus40/41 | 01-0375 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | |
| i | KingFisherFlex | Ndet | Ndet | Ndet | Ndet | |||
| SP418 | InoculatedCary-Blair | easyMAG | Ndet | Det | Det | Det | ||
| KingFisherFlex | Det | Det | Det | Det | ||||
| 01-0071 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| j | NorovirusGI/GII | KingFisherFlex | Det | Ndet | Ndet | Ndet | ||
| 02-0236 | Unpreserved(Frozen) | easyMAG | Det | Det | Det | Det | ||
| KingFisherFlex | Ndet | Det | Det | Det | ||||
| 01-0011 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherFlex | Det | Ndet | Ndet | Ndet | ||||
| 01-0023 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| k | Rotavirus | KingFisherFlex | Det | Ndet | Ndet | Ndet | ||
| 01-0062 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherFlex | Det | Ndet | Ndet | Ndet | ||||
| 01-0150 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet | ||
| KingFisherFlex | Det | Det | Det | Det | ||||
| 01-0151 | easyMAG | Ndet | Ndet | Ndet | Ndet | |||
| Target | SampleName | Sample Type | Extraction | Run 1(Originalrun) | Run 2 | Run 3 | ConsensusResult | |
| InoculatedCary-Blair | KingFisherFlex | Det | Ndet | Ndet | Ndet | |||
| CK-ST-045* | Unpreserved(Frozen) | easyMAG | Ndet | N/A | N/A | Ndet | ||
| CK-ST-045* | Unpreserved(Frozen) | KingFisherFlex | Det | N/A | N/A | Det | ||
| CK-ST-046 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| CK-ST-046 | Unpreserved(Frozen) | KingFisherFlex | Det | Det | Ndet | Det | ||
| ST-057 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| ST-057 | Unpreserved(Frozen) | KingFisherFlex | Det | Det | Det | Det | ||
| ST-058 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| ST-058 | InoculatedCary-Blair | KingFisherFlex | Det | Ndet | Ndet | Ndet | ||
| Stool_0132 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| Stool_0132 | Unpreserved(Frozen) | KingFisherFlex | Det | Ndet | Ndet | Ndet |
Table 12. KingFisher Flex: Retested Results of Discordant Specimens (Archived Specimens)
{25}------------------------------------------------
{26}------------------------------------------------
*Insufficient Volume of Sample to be retested. IND: Indeterminant; Det: Detected; Ndet: Not Detected
{27}------------------------------------------------
Twenty-seven (27) archived samples with invalid results (invalid internal control) were subjected to reflex testing per BioCode® GPP IFU. After reflex testing, fourteen (14) samples were valid, and thirteen (13) samples were still invalid, as shown in Table 13. Final results were used to calculate the agreements reported in Table 11 and/or noted in the footnote of Table 11.
| Sample Name | Sample Type | Instrument | Run 1 | Run 2 | Final Result |
|---|---|---|---|---|---|
| 01-0035 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | Valid |
| 02-0167 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | Valid |
| 03-0097 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Invalid | Invalid |
| 06-0023 | Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| 06-0119 | Cary-Blair | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 06-0223 | Cary-Blair | easyMAG | IC Invalid | IC Invalid | Invalid |
| 06-0258 | Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| 06-0331 | Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| Stool 0112 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | Valid |
| Mayo-GI_107 | Inoculated Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| Mayo-GI_117 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| Mayo-GI_123 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| Mayo-GI_126 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| SP392 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| SP392 | Inoculated Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| ST-074 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | Valid |
| ST-074 | Inoculated Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| 01-0003 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 01-0047 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 01-0335 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 02-0288 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 02-0310 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Valid | Valid |
| 03-0360 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 06-0051 | Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| 06-0137 | Cary-Blair | KingFisher Flex | IC Invalid | IC Valid | Valid |
| Mayo-GI_124 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| Stool 0036 | Unpreserved(Frozen) | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| Sample Name | Sample Type | Instrument | Run 1 | Run 2 | Final Result |
| 01-0073 | Inoculated Cary-Blair | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
| 02-0135 | Inoculated Cary-Blair | KingFisher Flex | IC Invalid | IC Invalid | Invalid |
Table 13. KingFisher Flex: Results of Reflex Testing of Invalid Samples (Archived Samples)
{28}------------------------------------------------
IC: Internal Control
| Table 14. KingFisher Flex: Summary of Clinical Investigational Study Results of Fresh | |
|---|---|
| Clostridium difficile Positives and Negatives |
| SpecimenType | (n) | Positive Agreement | Negative Agreement | |||
|---|---|---|---|---|---|---|
| Target | PPA (%) | 95% CI | NPA (%) | 95% CI | ||
| Campylobacterspp.a | Unpreserved(Fresh) | 53 | 1/1 (100.0) | 20.7 - 100.0 | 52/52 (100.0) | 93.1 - 100.0 |
| Clostridiumdifficile b | Unpreserved(Fresh) | 54 | 30/30 (100.0) | 88.6 - 100.0 | 23/24 (95.8) | 79.8 - 99.3 |
| E. coli O157 | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| EnteroaggregativeE. coli (EAEC) | Unpreserved(Fresh) | 53 | 1/1 (100.0) | 20.7 - 100.0 | 52/52 (100.0) | 93.1 - 100.0 |
| EnterotoxigenicE. coli (ETEC) | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Shiga toxin-producing E. coli(STEC) | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Salmonella spp. | Unpreserved(Fresh) | 53 | 1/1 (100.0) | 20.7 - 100.0 | 52/52 (100.0) | 93.1 - 100.0 |
| Shigella/ EIEC | Unpreserved(Fresh) | 53 | 2/2 (100.0) | 34.2 - 100.0 | 51/51 (100.0) | 93.0 - 100.0 |
| Vibrioparahaemolyticus | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Vibrio spp. (notparahaemolyticus) | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Yersiniaenterocolitica | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Cryptosporidiumspp. | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Entamoebahistolytica | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Giardia lamblia | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| Adenovirus40/41 | Unpreserved(Fresh) | 53 | 1/1 (100.0) | 20.7 - 100.0 | 52/52 (100.0) | 93.1 - 100.0 |
| Norovirus(GI/GII) | Unpreserved(Fresh) | 53 | 1/1 (100.0) | 20.7 - 100.0 | 52/52 (100.0) | 93.1 - 100.0 |
| Rotavirus A | Unpreserved(Fresh) | 53 | N/A | N/A | 53/53 (100.0) | 93.2 - 100.0 |
| CombinedTargets | Unpreserved(Fresh) | 902 | 37/37 (100.0) | 90.6 -100.0 | 864/865 (99.9) | 99.3- 100.0 |
| Target | SpecimenType | (n) | Positive Agreement | Negative Agreement | ||
| PPA (%) | 95% CI | NPA (%) | 95% CI | |||
| Vibrio parahaemolyticus | Unpreserved(Frozen) | 115 | 26/28 (92.9%) | 77.4-98.0% | 86/87 (98.9%) | 93.8-99.8% |
| Vibrio spp. (notparahaemolyticus) | Unpreserved(Frozen) | 115 | 29/30 (96.7%) | 83.3-99.4% | 82/85 (96.5%) | 90.1-98.8% |
| Yersinia enterocolitica a | Unpreserved(Frozen) | 116 | 28/28 (100%) | 87.9-100.0% | 88/88 (100%) | 95.8%-100.0% |
| Entamoeba histolytica b | Unpreserved(Frozen) | 116 | 28/28 (100%) | 87.9-100.0% | 86/88 (97.7%) | 92.1-99.4% |
| All other targets c | Unpreserved(Frozen) | 1491 | N/A | N/A | 1491/1491(100%) | 99.7-100.0% |
| Combined targets | Unpreserved(Frozen) | 1953 | 111/114(97.4%) | 92.5 -99.1% | 1833/1839(99.7%) | 99.3 -99.9% |
Note. Fifty-four (54) fresh unpreserved C. difficile positives and negatives were tested. C. difficile positive sample TRI_047A tested C. difficile positive but was invalid for all other targets (due to invalid
{29}------------------------------------------------
RNA IC, invalid results not calculated in negative agreement). Sample remained invalid after reflex testing per BioCode GPP IFU.
a. C. difficile negative sample TRI 017A had a MFI of 516 (>500 cutoff) for Campylobacter spp. The sample was Campvlobacter spp negative after being retested twice with each extraction system. b. Sample TRI 056A resulted in a false positive agreement and remained false positive after discordant analysis.
Table 15. KingFisher Flex: Summary of Contrived Specimen Results
Note. One hundred twenty (120) contrived samples were tested. Five (5) samples were invalid with the easyMAG on initial testing. When the invalid samples were tested again with PCR, the samples remained invalid.
a- Of the five (5) invalids, one (1) of the E. histolytica positives was tested E. his positive but RNA IC was invalid.
b- Of the five (5) invalids, one (1) of the Yersinia enterocolitica positives was tested Yersinia enterocolitica, but RNA IC was invalid.
c - One (1) of the negative stools being used as matrix for contriving samples was a low-positive C. difficile and erroneously enrolled as a negative sample. Because the C. difficile was not intended as part of the study, the positive agreement calculation for this target was not included.
{30}------------------------------------------------
B. Method Comparison Study between EasyMag and KingFisher Apex Dx
| Target | SpecimenType | (n) | Positive AgreementPPA (%) | 95% CI | Negative AgreementNPA (%) | 95% CI | |
|---|---|---|---|---|---|---|---|
| Campylobacter spp.a | InoculatedCary-Blair | 210 | 18/18 (100) | 82.4 – 100 | 192/192 (100) | 98.0 – 100 | |
| Unpreserved(Frozen) | 249 | 24/25 (96.0) | 80.5 – 99.3 | 221/224 (98.7) | 96.1 – 99.5 | ||
| All Archived | 459 | 42/43 (97.7) | 87.9 – 99.6 | 413/416 (99.3) | 97.9 – 99.8 | ||
| Clostridium difficile b | InoculatedCary-Blair | 211 | 34/37 (91.9) | 78.7 – 97.2 | 173/174 (99.4) | 96.8 – 99.9 | |
| Unpreserved(Frozen) | 249 | 17/19 (89.5)* | 68.6 – 97.1 | 229/230 (99.6) | 97.6 – 99.9 | ||
| All Archived | 460 | 51/56 (91.1) | 80.7 – 96.1 | 402/404 (99.5) | 98.2 – 99.9 | ||
| E. coli O157 c | InoculatedCary-Blair | 210 | 4/5 (80.0)* | 37.6 – 96.4 | 205/205 (100) | 98.2 – 100 | |
| Unpreserved(Frozen) | 249 | 8/8 (100) | 67.6 – 100 | 240/241 (99.6) | 97.7 – 99.9 | ||
| All Archived | 459 | 12/13 (92.3) | 66.7 – 98.6 | 445/446 (99.8) | 98.7 – 100 | ||
| Enteroaggregative E.coli (EAEC) d | InoculatedCary-Blair | 211 | 30/30 (100) | 88.6 – 100 | 181/181 (100) | 97.9 – 100 | |
| Unpreserved(Frozen) | 249 | 22/22 (100) | 85.1 – 100 | 226/227 (99.6) | 97.5 – 99.9 | ||
| All Archived | 460 | 52/52 (100) | 93.1 – 100 | 407/408 (99.8) | 98.6 – 100 | ||
| EnterotoxigenicE. coli (ETEC) e | InoculatedCary-Blair | 210 | 14/14 (100) | 78.5 – 100 | 194/196 (99.0) | 96.4 – 99.7 | |
| Unpreserved(Frozen) | 249 | 11/11 (100) | 74.1 – 100 | 237/238 (99.6) | 97.7 – 99.9 | ||
| All Archived | 459 | 25/25 (100) | 86.7 – 100 | 431/434 (99.3) | 98.0 – 99.8 | ||
| Shiga toxin-producingE. coli (STEC) f | InoculatedCary-Blair | 210 | 12/12 (100) | 75.8 – 100 | 198/198 (100) | 98.1 – 100 | |
| Unpreserved(Frozen) | 249 | 20/22 (90.9) | 72.2 – 97.5 | 226/227 (99.6) | 97.5 – 99.9 | ||
| All Archived | 459 | 32/34 (94.1) | 80.9 – 98.4 | 424/425 (99.8) | 98.7 – 100 | ||
| Salmonella spp. g | InoculatedCary-Blair | 210 | 20/20 (100) | 83.9 – 100 | 187/190 (98.4) | 95.5 – 99.5 | |
| Specimen | (n) | Positive Agreement | Negative Agreement | ||||
| Target | Type | PPA (%) | 95% CI | NPA (%) | 95% CI | ||
| Unpreserved(Frozen) | 249 | 21/21 (100) | 84.5 – 100 | 228/228 (100) | 98.3 – 100 | ||
| All Archived | 459 | 41/41 (100) | 91.4 – 100 | 415/418 (99.3) | 97.9 – 99.8 | ||
| InoculatedCary-Blair | 211 | 13/13 (100) | 77.2 – 100 | 194/198 (98.0) | 94.9 – 99.2 | ||
| Shigella/ EIECh | Unpreserved(Frozen) | 249 | 17/18 (94.4) | 74.2 – 99.0 | 231/231 (100) | 98.4 – 100 | |
| All Archived | 460 | 30/31 (96.8) | 83.8 – 99.4 | 425/429 (99.1) | 97.6 – 99.6 | ||
| InoculatedCary-Blair | 210 | 1/1 (100) | 20.7 – 100 | 208/209 (99.5) | 97.3 – 99.9 | ||
| Vibrio parahaemolyticus i | Unpreserved(Frozen) | 249 | 1/1 (100) | 20.7 – 100 | 248/248 (100) | 98.5 – 100 | |
| All Archived | 459 | 2/2 (100) | 34.2 – 100 | 456/457 (99.8) | 98.8 – 100 | ||
| InoculatedCary-Blair | 210 | N/A | N/A | 209/210 (99.5) | 97.4 – 99.9 | ||
| Vibrio spp.j(not parahaemolyticus) | Unpreserved(Frozen) | 249 | 1/2 (50%)* | 9.5 – 90.5 | 247/247 (100) | 98.4 – 100 | |
| All Archived | 459 | 1/2 (50%)* | 9.5 – 90.5 | 456/457 (99.8) | 98.8 – 100 | ||
| InoculatedCary-Blair | 210 | 3/3 (100) | 43.9 – 100 | 206/207 (99.5) | 97.3 – 99.9 | ||
| Yersinia enterocolitica k | Unpreserved(Frozen) | 249 | 3/3 (100) | 43.9 – 100 | 245/246 (99.6) | 97.7 – 99.9 | |
| All Archived | 459 | 6/6 (100) | 61.0 – 100 | 451/453 (99.6) | 98.4 – 99.9 | ||
| InoculatedCary-Blair | 210 | 9/9 (100) | 70.1 – 100 | 201/201 (100) | 98.1 – 100 | ||
| Cryptosporidium spp.l | Unpreserved(Frozen) | 250 | 17/18 (94.4) | 74.2 – 99.0 | 231/232 (99.6) | 97.6 – 99.9 | |
| All Archived | 460 | 26/27 (96.3) | 81.7 – 99.3 | 432/433 (99.8) | 98.7 – 100 | ||
| InoculatedCary-Blair | 210 | N/A | N/A | 210/210 (100) | 98.2 – 100 | ||
| Entamoeba histolytica m | Unpreserved(Frozen) | 249 | N/A | N/A | 249/249 (100) | 98.5 – 100 | |
| All Archived | 459 | N/A | N/A | 459/459 (100) | 99.1 – 100 | ||
| Giardia lamblia n | InoculatedCary-Blair | 210 | 4/4 (100) | 51.0 – 100 | 206/206 (100) | 98.2 – 100 | |
| Specimen | Positive Agreement | Negative Agreement | |||||
| Target | Type | (n) | PPA (%) | 95% CI | NPA (%) | 95% CI | |
| Unpreserved(Frozen) | 249 | 10/11 (90.9) | 62.3 - 98.4 | 238/238 (100) | 98.4 - 100 | ||
| All Archived | 459 | 14/15 (93.3) | 70.2 - 98.8 | 444/444 (100) | 99.1 - 100 | ||
| InoculatedCary-Blair | 210 | 5/5 (100) | 56.6 - 100 | 204/205 (99.5) | 97.3 – 99.9 | ||
| Adenovirus 40/41 ° | Unpreserved(Frozen) | 249 | 7/8 (87.5)* | 52.9 - 97.8 | 240/241 (99.6) | 97.7 - 99.9 | |
| All Archived | 459 | 12/13 (92.3) | 66.7 - 98.6 | 444/446 (99.6) | 98.4 - 99.9 | ||
| Norovirus | InoculatedCary-Blair | 210 | 22/23 (95.7) | 79.0 - 99.2 | 187/187 (100) | 98.0 - 100 | |
| (GI/GII) p | Unpreserved(Frozen) | 250 | 20/20 (100) | 83.9 - 100 | 230/230 (100) | 98.4 - 100 | |
| All Archived | 460 | 42/43 (97.7) | 87.9 – 99.6 | 417/417 (100) | 99.1 - 100 | ||
| InoculatedCary-Blair | 211 | 10/10 (100) | 72.2 - 100 | 201/201 (100) | 98.1 - 100 | ||
| Rotavirus A 9 | Unpreserved(Frozen) | 249 | 9/9 (100) | 70.1 - 100 | 240/240 (100) | 98.4 - 100 | |
| All Archived | 460 | 19/19 (100) | 83.2 - 100 | 441/441 (100) | 99.1 - 100 | ||
| Combined Targets | InoculatedCary-Blair | 3512 | 216/218(99.1) | 96.7 — 99.7 | 3280/3294(99.6) | 99.3 — 99.7 | |
| Unpreserved(Frozen) | 4235 | 208/218(95.4) | 91.8 – 97.5 | 4006/4017(99.7) | 99.5 - 99.8 |
Table 16. KingFisher Apex Dx: Summary of Clinical Investigational Study Results (Archived Specimens) Stratified by Sample Type and Storage
{31}------------------------------------------------
{32}------------------------------------------------
*Positive agreement <90%.
Forty (40) archived samples with discordant results (between the easyMAG and KingFisher Apex Dx) retested twice with both easyMAG and KingFisher Apex Dx.
a - Campylobacter spp. Two (2) false positives retested remained false positives whereas another false positive became true negative. One (1) false negative became true positive. Nine (9) samples were still invalid, not used in the agreement calculation.
b - Clostridium difficile. Of the five (5) false positives retested, three (3) became true negatives, one (1) remained false negative, and one (1) became invalid due to invalid RNA IC with the 2™ and 3ª runs with both extraction systems. Two (2) false positives became true negatives. Eight (8) samples were still invalid, not used in the agreement calculation.
c - 0157. One (1) false negative and one (1) false positive retested remained false negative and false positive.
d- EAEC: One (1) false positive retested became true negative. Eight (8) samples were still invalid, not used in the agreement calculation.
e - ETEC: Of the three (3) false positives retested, one (1) became true negative, and two (2) became true positives. Nine (9) samples were invalid, not used in the agreement calculation.
f- STEC: Of the two (2) false negatives retested, one (1) became true negative and one (1) became false positive. One (1) false positive retested remained false positive. Nine (9) samples were invalid, not used in the agreement calculation.
{33}------------------------------------------------
g - Salmonella spp: Of the three (3) false positives retested, two (2) became true negatives, and remained false positive. Nine (9) samples were invalid, not used in the agreement calculation.
h - Shigella/EIEC: Of the four (4) false positives retested, three (3) became true negatives whereas one (1) became false negative. One (1) false negative retested became true negative. Eight (8) samples were invalid, not used in the agreement calculation.
i- Vibrio parahaemolyticus: One (1) false positive retested became true negative. Nine (9) samples were invalid, not used in the agreement calculation.
i- Vibrio spp. One (1) false positive retested became true negative. For the false negative retested, the easyMAG had an invalid result with the 2nd run and a negative result with the 3rd run, whereas the KF Apex Dx had negative results with both retested runs. Nine (9) samples were invalid, not used in the agreement calculation.
k - Yersinia enterocolitica: Of the two (2) false positives retested, one (1) became true negative and one (1) remained false positive. Nine (9) samples were invalid, not used in the agreement calculation.
l - Cryptosporidium spp: One (1) false positive retested became true negative. One (1) false negative retested remained false negative. Eight (8) samples were invalid, not used in the agreement calculation. m- Entamoeba histolytica: Nine (9) samples were still invalid, not used in the agreement calculation. n - Giardia lamblia: One (1) false negative was not retested due to insufficient volume of the sample. Nine (9) samples were invalid, not used in the agreement calculation.
o - Adenovirus 40/41 : Of the wo (2) false positives retested, one (1) became true negative and one (1) became true positive. The false negative retested became true negative. Nine (9) samples were still invalid, not used in the agreement calculation.
p - Norovirus GI/GII: The false negative retested became true positive. Eight (8) samples were invalid, not used in the agreement calculation.
q- Rotavirus A: Eight (8) samples were invalid, not used in the agreement calculation.
{34}------------------------------------------------
The table below (Table 17) shows the results of retesting samples with discordant results between the easyMAG and the KingFisher Apex Dx. Samples were retested twice with both easyMag and KingFisher Apex Dx. Final results of the discordant samples were not used in the calculation of the agreements but noted as the footnotes of Table 16 above.
| Target | Sample Name | Sample Type | Extraction | Run 1(Original Run) | Run 2 | Run 3 | Final Result | |
|---|---|---|---|---|---|---|---|---|
| Campylobacter spp | 01-0237 | Unpreserved(Frozen) | easyMAG | Det | Det | Det | Det | |
| KingFisherApex Dx | Ndet | Det | Det | Det | ||||
| 03-0153 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Det | Ndet | ||
| KingFisherApex Dx | Det | Det | Ndet | Det | ||||
| Mayo-GI_101 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Det | Det | Det | ||||
| SP517 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||||
| 01-0002 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Ndet | Det | Ndet | Ndet | ||||
| 01-0019 | InoculatedCary-Blair | easyMAG | Det | Det | Ndet | Det | ||
| KingFisherApex Dx | Ndet | Ndet | Ndet | Ndet | ||||
| 01-0048 | InoculatedCary-Blair | easyMAG | Det | Ndet | Ndet | Ndet | ||
| Clostridium difficile | KingFisherApex Dx | Ndet | Ndet | Ndet | Ndet | |||
| 01-0256 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Ndet | Ndet | |||||
| 01-0373 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||||
| 02-0009 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Ndet | Ndet | Ndet | Ndet | ||||
| 02-0135 | InoculatedCary-Blair | easyMAG | Det | Invalid | Invalid | Invalid | ||
| KingFisherApex Dx | Ndet | Invalid | Invalid | Invalid | ||||
| 0157 | 06-0180 | Cary-Blair | easyMAG | Det | Det | Ndet | Det | |
| KingFisherApex Dx | Ndet | Ndet | Det | Ndet | ||||
| SP367 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Det | Ndet | ||
| KingFisherApex Dx | Det | Det | Ndet | Det | ||||
| EAEC | 01-0392 | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| Target | SampleName | SampleType | Extraction | Run 1(OriginalRun) | Run2 | Run3 | FinalResult | |
| ETEC | Unpreserved(Frozen) | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||
| e | 01-0374 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet | |
| Unpreserved(Frozen) | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | |||
| 06-0211 | Cary-Blair | easyMAG | Ndet | Det | Det | Det | ||
| KingFisherApex Dx | Det | Det | Det | Det | ||||
| 06-0305 | Cary-Blair | easyMAG | Ndet | Det | Det | Det | ||
| KingFisherApex Dx | Det | Ndet | Det | Det | ||||
| STEC | 02-0211 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | |
| f | KingFisherApex Dx | Ndet | Ndet | Ndet | ||||
| SP228 | Unpreserved(Frozen) | easyMAG | Det | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Ndet | Det | Det | Det | ||||
| SP367 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Det | Ndet | |||||
| InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | |||
| Salmonellaspp | 01-0254 | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||
| g | 01-0302 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | |
| KingFisherApex Dx | Det | Ndet | Ndet | |||||
| 06-0357 | Cary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Det | Ndet | |||||
| Shigella/EIEC | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| 01-0091 | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | |||
| h | InoculatedCary-Blair | easyMAG | Ndet | Det | Det | Det | ||
| 01-0267 | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | |||
| 01-0377 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||||
| 02-0297 | Unpreserved(Frozen) | easyMAG | Det | Det | Det | Det | ||
| KingFisherApex Dx | Ndet | Det | Det | Det | ||||
| 03-0206 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | ||
| KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||||
| Vibrioparahaemolyticus | 03-0358 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet | |
| l | KingFisherApex Dx | Det | Ndet | Ndet | ||||
| Target | SampleName | SampleType | Extraction | Run 1(OriginalRun) | Run2 | Run3 | FinalResult | |
| m | Vibrio spp | 01-0278 | InoculatedCary-Blair | easyMAG | Ndet | Ndet | Ndet | Ndet |
| 02-0167 | Unpreserved(Frozen) | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||
| 02-0167 | Unpreserved(Frozen) | easyMAG | Det | Invalid | Ndet | IND | ||
| n | Yersinia | 02-0261 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet |
| 06-0222 | Cary-Blair | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||
| o | Cryptosporidium spp | 04-047 | Unpreserved(Frozen) | easyMAG | Ndet | Det | Det | Det |
| SP399 | Unpreserved(Frozen) | KingFisherApex Dx | Det | Ndet | Det | Det | ||
| p | Giardialamblia | 03-0140* | Unpreserved(Frozen) | easyMAG | Det | N/A | N/A | Det |
| 01-0079 | Unpreserved(Frozen) | KingFisherApex Dx | Ndet | N/A | N/A | Ndet | ||
| q | Adenovirus40/41 | 01-0079 | Unpreserved(Frozen) | easyMAG | Ndet | Ndet | Ndet | Ndet |
| 01-0375 | Unpreserved(Frozen) | KingFisherApex Dx | Det | Ndet | Ndet | Ndet | ||
| r | NorovirusGI/GII | SP418 | InoculatedCary-Blair | easyMAG | Ndet | Det | Det | Det |
| 01-0290 | InoculatedCary-Blair | KingFisherApex Dx | Det | Det | Det | Det |
Table 17. KingFisher Apex Dx: Retested Results of Discordant Specimens (Archived Specimens)
{35}------------------------------------------------
{36}------------------------------------------------
- Insufficient Volume of Sample; IND: Indeterminant; Det: Detected; Ndet: Not Detected
{37}------------------------------------------------
Twenty-two (22) archived samples with invalid results (invalid internal control) were subjected to reflex testing per BioCode® GPP IFU. After reflex testing, thirteen (13) samples were valid, and nine (9) samples were still invalid, as shown in Table 18 below. Final results were used to calculate the agreement reported or noted in the footnote of Table 16.
| Sample Name | Sample Type | Instrument | Run 1 | Run 2 | Run 3 | Final Result |
|---|---|---|---|---|---|---|
| 01-0035 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| 02-0167 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| 03-0097 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Invalid | N/A | Invalid |
| 06-0023 | Cary-Blair | easyMAG | IC Invalid | IC Invalid | N/A | Invalid |
| KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid | ||
| 06-0119 | Cary-Blair | easyMAG | IC Invalid | IC Invalid | N/A | Invalid |
| KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid | ||
| 06-0223 | Cary-Blair | easyMAG | IC Invalid | IC Invalid | N/A | Invalid |
| 06-0258 | Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| 06-0331 | Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| Stool 0112 | Unpreserved(Frozen) | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| Mayo-GI_107 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid | ||
| Mayo-GI_117 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| Mayo-GI_123 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| Mayo-GI_126 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| SP392 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid | ||
| ST-074 | Inoculated Cary-Blair | easyMAG | IC Invalid | IC Valid | N/A | Valid |
| 01-0003 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid |
| 02-0288 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid |
| 02-0310 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid |
| 01-0073 | Inoculated Cary-Blair | KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid |
| Stool_0163 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid |
| 01-0238 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid |
| 01-0379 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Invalid | N/A | Invalid |
| Mayo-GI 059 | Unpreserved(Frozen) | KingFisher Apex Dx | IC Invalid | IC Valid | N/A | Valid |
Table 18. KingFisher Apex Dx: Results of Reflex Testing of Invalid Samples (Archived Samples)
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| Target | SpecimenType | (n) | Positive Agreement | Negative Agreement | ||
|---|---|---|---|---|---|---|
| PPA (%) | 95% CI | NPA (%) | 95% CI | |||
| Campylobacterspp.a | Unpreserved(Fresh) | 52 | 1/1 (100.0) | 20.7 - 100.0 | 51/51 (100.0) | 93.0 - 100.0 |
| Clostridiumdifficileb | Unpreserved(Fresh) | 54 | 29/30 (96.7) | 83.3 - 99.4 | 24/24 (100.0) | 86.2 - 100.0 |
| E. coli O157 | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Enteroaggregative E. coli (EAEC) | Unpreserved(Fresh) | 52 | 1/1 (100.0) | 20.7 - 100.0 | 50/51 (98.0) | 89.7 - 99.7 |
| EnterotoxigenicE. coli (ETEC) | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Shiga toxin-producing E. coli(STEC) | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Salmonella spp. | Unpreserved(Fresh) | 52 | 1/1 (100.0) | 20.7 - 100.0 | 51/51 (100.0) | 93.0 - 100.0 |
| Shigella/ EIEC | Unpreserved(Fresh) | 52 | 2/2 (100.0) | 34.2 - 100.0 | 50/50 (100.0) | 52.9 - 100.0 |
| Vibrioparahaemolyticus | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Vibrio spp. (notparahaemolyticus) | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Yersiniaenterocolitica | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Cryptosporidiumspp. | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Entamoebahistolytica | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Giardia lamblia | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| Adenovirus 40/41 | Unpreserved(Fresh) | 52 | 1/1 (100.0) | 20.7 - 100.0 | 51/51 (100.0) | 93.0 - 100.0 |
| Norovirus(GI/GII) | Unpreserved(Fresh) | 52 | 1/1 (100.0) | 20.7 - 100.0 | 51/51 (100.0) | 93.0 - 100.0 |
| Rotavirus A | Unpreserved(Fresh) | 52 | N/A | N/A | 52/52 (100.0) | 93.1 - 100.0 |
| CombinedTargets | Unpreserved(Fresh) | 886 | 36/37 (97.3) | 86.2 - 99.5 | 848/849 (99.9) | 99.3 - 100.0 |
| Target | SpecimenType | (n) | Positive Agreement | Negative Agreement | ||
| PPA (%) | 95% CI | NPA (%) | 95% CI | |||
| Vibrio parahaemolyticus | Unpreserved(Frozen) | 115 | 28/28 (100%) | 87.9-100.0% | 86/87 (98.9%) | 93.8-99.8% |
| Vibrio spp. (notparahaemolyticus) | Unpreserved(Frozen) | 115 | 29/30(96.7%) | 83.3-99.4% | 84/85 (98.8%) | 93.6-99.8% |
| Yersinia enterocolitica a | Unpreserved(Frozen) | 116 | 28/28 (100%) | 87.9-100.0% | 88/88 (100%) | 95.8%-100.0% |
| Entamoeba histolytica b | Unpreserved(Frozen) | 116 | 28/28 (100%) | 87.9-100.0% | 86/88 (97.7%) | 92.1-99.4% |
| All other targets c | Unpreserved(Frozen) | 1491 | N/A | N/A | 1489/1491(99.9%) | 99.5-100.0% |
| Combined Targets | Unpreserved(Frozen) | 1953 | 113/114(99.1%) | 95.2 –99.8% | 1833/1839(99.7%) | 99.3 –99.9% |
Table 19. KingFisher Apex Dx: Summary of Clinical Investigational Study Results of Fresh Clostridium difficile Positives and Negatives
Note. Fifty-four (54) fresh unpreserved samples were tested. Two (2) C. difficile positives (TRI 047A and TRI 059A) were tested C. difficile positive but invalid for all other targets (due to invalid RNA IC, invalid results not calculated in negative agreement). Samples remained invalid after reflex testing per GPP IFU.
a. C. difficile negative sample TRI 017A had a MFI of 516 (>500 cutoff) for Campylobacter spp. The sample was Campylobacter spp negative after being retested twice with each extraction system.
b. C. difficile positive sample TRI 011A initially tested as false negative resulted in a true positive after retesting. The Xpert C. difficile-Epi test results of TRI- 011A had Ct 34.8 which indicates the sample was low positive.
{39}------------------------------------------------
Table 20. KingFisher Apex Dx: Summarv of Contrived Specimen Results (Contrived at 3x LoD and 6x LoD)
Note. One hundred twenty (120) contrived samples were tested. Five (5) samples were invalid with the easyMAG on initial testing. When the invalid samples were tested again with PCR, the samples remained invalid.
a- Of the five (5) invalids, one of the E. histolytica positives was tested E. his positive but RNA IC was invalid.
b- Of the five (5) invalids, one of the Yersinia enterocolitica positives was tested Yersinia enterocolitica, but RNA IC was invalid.
c - One (1) of negative stools being used as matrix for contriving samples was a low-positive C. difficile and erroneously enrolled as a negative sample. Because the C. difficile was not intended as part of the study, the positive agreement calculation for this target was not included.
| Co-infection summary | |
|---|---|
| +1 Target | 246/466 (52.8%) |
| +2 Targets | 78/466 (16.7%) |
| +3 Targets | 13/466 (2.8%) |
Table 21. Clinical Co-infection Summary of Archived Samples
Conclusions [807.92(b)(3):
The intended use and fundamental scientific technology of the BioCode GPP using the KingFisher Flex and KingFisher Apex Dx is substantially equivalent to the predicate device. Non-clinical studies and clinical studies have established that the BioCode GPP with the KingFisher Flex and KingFisher Apex Dx is substantially equivalent to the predicate device.
§ 866.3990 Gastrointestinal microorganism multiplex nucleic acid-based assay.
(a)
Identification. A gastrointestinal microorganism multiplex nucleic acid-based assay is a qualitativein vitro diagnostic device intended to simultaneously detect and identify multiple gastrointestinal microbial nucleic acids extracted from human stool specimens. The device detects specific nucleic acid sequences for organism identification as well as for determining the presence of toxin genes. The detection and identification of a specific gastrointestinal microbial nucleic acid from individuals exhibiting signs and symptoms of gastrointestinal infection aids in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation and other laboratory findings. A gastrointestinal microorganism multiplex nucleic acid-based assay also aids in the detection and identification of acute gastroenteritis in the context of outbreaks.(b)
Classification. Class II (special controls). The special controls are set forth in FDA's guideline document entitled: “Class II Special Controls Guideline: Gastrointestinal Microorganism Multiplex Nucleic Acid-Based Assays for Detection and Identification of Microorganisms and Toxin Genes from Human Stool Specimens.” For availability of the guideline document, see § 866.1(e).