(90 days)
The BD MAX™ Extended Enteric Bacterial Panel performed on the BD MAX System, is an automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens. It is used in conjunction with the BD MAX Enteric Bacterial Panel as an optional Master Mix. The BD MAX™ Extended Enteric Bacterial Panel detects nucleic acids from
- Plesiomonas shigelloides
- Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae)
- Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable ● enterotoxin (ST) genes
- Yersinia enterocolitica ●
Testing is performed on soft to diarrheal unpreserved stool specimens or Cary-Blair preserved stool specimens from symptomatic patients with suspected acute gastroenteritis, enteritis or colitis. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of relevant gene target DNA. The test utilizes fluorogenic gene-specific hybridization probes for the detection of the amplified DNA.
This test is intended for use, in conjunction with clinical presentation, laboratory findings, and epidemiological information, as an aid in the differential diagnosis of Plesiomonas shigelloides, Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae), Enterotoxigenic Escherichia coli (ETEC) LT/ST and Yersinia enterocolitica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other organisms that are not detected by this test. and may not be the sole or definitive cause of patient illness. 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.
The BD MAX™ Extended Enteric Bacterial Panel performed on the BD MAX™ System, is an automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens. It is used in conjunction with the BD MAX Enteric Bacterial Panel as an optional Master Mix. The BD MAX Extended Enteric Bacterial Panel detects nucleic acids from
- Plesiomonas shigelloides
- . Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae)
- . Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable enterotoxin (ST) genes
- Yersinia enterocolitica
Testing is performed on unpreserved or Cary-Blair preserved soft to diarrheal stool specimens from symptomatic patients with suspected acute gastroenteritis, enteritis or colitis.
The BD MAX™ Extended Enteric Bacterial Panel (BD MAX™ xEBP) is composed of a single Master Mix that must be used in conjunction with the BD MAX™ Enteric Bacterial Panel (BD MAX EBP), performed on the BD MAX™ System. The BD MAX xEBP is performed with the Master Mix contained in the BD MAX xEBP reagent kit and the EBP reagents, including the Master Mix contained in the BD MAX™ EBP reagent kit. The BD MAX xEBP assay and accompanying Assay Definition File (ADF) were developed to allow inclusion of both BD MAX EBP and BD MAX xEBP master mix reagents in one test run.
The BD MAX™ System and the BD MAX™ Extended Enteric Bacterial Panel is run with the instrument with associated hardware and accessories, disposable microfluidic cartridges, master mixes, unitized reagent strips, extraction reagents, and sample buffer tubes. The instrument automates sample preparation including target lysis, DNA extraction and concentration, reagent rehydration, and target nucleic acid amplification and detection using real-time PCR. The assay includes a Sample Processing Control (SPC) that is present in the Extraction Tube. The SPC monitors DNA extraction steps, thermal cycling steps, reagent integrity and the presence of inhibitory substances. The BD MAX™ System software automatically interprets test result may be called as POS (Positive), NEG (Negative), or UNR (Unresolved) for each of the assay's targets, based on the amplification status of the target and of the Sample Processing Control. IND (Indeterminate) or INC (Incomplete) results are due to BD MAXTM System failure.
1. Table of Acceptance Criteria and Reported Device Performance
The acceptance criteria for each analyte are not explicitly stated in a single summarized table. However, the performance tables demonstrate the Percentage Positive Agreement (PPA) and Negative Agreement (NPA) for each target organism. A common implicit acceptance criterion for such diagnostic devices is generally a high PPA and NPA, often above 90% for positive and 95% for negative, with a lower bound of the 95% Confidence Interval (CI) also being high.
Here's a summary of the reported device performance, which implies the acceptance criteria were met given the device received clearance:
| Target Organism | Specimen Type | Specimen Origin | BD MAX Performance (PPA (95% CI)) | BD MAX Performance (NPA (95% CI)) |
|---|---|---|---|---|
| Vibrio | Cary-Blair preserved | Prospective (Fresh+Frozen) | 100 (34.2, 100) | 99.6 (99.1, 99.8) |
| Cary-Blair preserved | Retrospective (Frozen) | 100 (34.2, 100) | 100 (80.6, 100) | |
| Unpreserved | Prospective (Fresh+Frozen) | No Data for Calculation | 99.8 (99.2, 99.9) | |
| Unpreserved | Retrospective (Frozen) | 100 (34.2, 100) | 97.8 (88.7, 99.6) | |
| Cary-Blair Preserved | Contrived Samples | 100 (92.6, 100) | 100 (97.4, 100) | |
| Unpreserved | Contrived Samples | 100 (92.6, 100) | 100 (97.4, 100) | |
| Plesiomonas shigelloides | Cary-Blair preserved | Prospective | No Data for Calculation | 99.9 (99.5, 100) |
| Cary-Blair preserved | Retrospective | 100 (51, 100) | 100 (90.8, 100) | |
| Unpreserved | Prospective | No Data for Calculation | 99.9 (99.3, 100) | |
| Unpreserved | Retrospective | 100 (43.9, 100) | 97.9 (88.9, 99.6) | |
| Cary-Blair Preserved | Contrived Samples | 100 (92.6, 100) | 100 (97.4, 100) | |
| Unpreserved | Contrived Samples | 100 (92.6, 100) | 99.3 (96.2, 99.9) | |
| Yersinia enterocolitica | Cary-Blair preserved | Prospective | No Data for Calculation | 99.9 (99.6, 100) |
| Cary-Blair preserved | Retrospective | No Data for Calculation | 100 (89.3, 100) | |
| Unpreserved | Prospective | No Data for Calculation | 100 (99.6, 100) | |
| Unpreserved | Retrospective | 100 (70.1, 100) | 100 (92.4, 100) | |
| Cary-Blair Preserved | Contrived Samples | 97.9 (89.1, 99.6) | 100 (97.4, 100) | |
| Unpreserved | Contrived Samples | 100 (92.6, 100) | 100 (97.4, 100) | |
| ETEC (LT/ST) | Cary-Blair preserved | Prospective | 100 (72.2, 100) | 99.8 (99.3, 99.9) |
| Cary-Blair preserved | Retrospective | 100 (56.6, 100) | 100 (87.9, 100) | |
| Unpreserved | Prospective | 100 (80.6, 100) | 99.9 (99.3, 100) | |
| Unpreserved | Retrospective | 90 (59.6, 98.2) | 96.3 (81.7, 99.3) |
2. Sample Size Used for the Test Set and Data Provenance
- Total Clinical Specimens: 2410 specimens were enrolled.
- Prospective: 2264 specimens (882 unpreserved, 1382 Cary-Blair preserved)
- Retrospective: 146 specimens (87 unpreserved, 59 Cary-Blair preserved)
- Compliant Clinical Specimens: 2403 specimens were compliant.
- Contrived Samples:
- Vibrio: 192 (48 positive, 144 negative) for both Cary-Blair preserved and unpreserved, totaling 384 contrived samples.
- Plesiomonas shigelloides: 192 (48 positive, 144 negative) for both Cary-Blair preserved and unpreserved, totaling 384 contrived samples.
- Yersinia enterocolitica: 192 (48 positive, 144 negative) for both Cary-Blair preserved and unpreserved, totaling 384 contrived samples.
- Data Provenance: The prospective specimens were collected as part of routine patient care from six (6) geographically diverse clinical centers (multi-site investigational study), implying data from various locations. The retrospective specimens' historical results were recorded at the collection site. The study involved patients from different age groups from 0-1 month to Over 21 years old. The specific country of origin is not explicitly stated, but the submission is to the U.S. FDA, suggesting the study sites were likely within the US or compliant with US regulatory standards.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
The document does not explicitly state the number of experts or their specific qualifications (e.g., radiologist with X years of experience) used to establish the ground truth.
4. Adjudication Method (for the test set)
- For prospective fresh and prospective frozen specimens, the reference method for bacterial culture (Yersinia enterocolitica, Vibrio, and Plesiomonas shigelloides) was followed by alternate PCR assay and bi-directional sequencing on presumptive positive isolates.
- For ETEC, the comparator method was two alternate PCR assays and bidirectional sequencing performed directly from stool specimens.
- For retrospective specimens, the historical results were confirmed using an alternate PCR assay and bi-directional sequencing as part of the composite comparator method.
- Discrepant Analysis: For discrepant results (BD MAX result differed from the reference method), "discordance study" was performed, which involved retesting. The details imply that sequencing was used to resolve discrepancies.
The method can be described as a composite reference standard or adjudicated reference standard combining culture, alternative PCR, and bi-directional sequencing, with further investigation for discrepant cases. There is no explicit mention of an "N+1" method with human readers reviewing cases in the traditional sense of imaging studies.
5. If a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study involving human readers improving with AI vs. without AI assistance was not conducted. This is an IVD (In Vitro Diagnostic) device, not an imaging AI device that typically involves human interpretation. The study is focused on the device's analytical and clinical performance against a reference standard.
6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, the clinical performance study evaluated the device in a standalone manner. The BD MAX™ Extended Enteric Bacterial Panel (an automated in vitro diagnostic test) was performed, and its results were compared directly against the reference method. There is no mention of human-in-the-loop performance influencing the assay's output during the clinical evaluation. The system automates sample preparation, DNA extraction, amplification, and interpretation.
7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)
The ground truth was established using a composite reference standard:
- For Yersinia enterocolitica, Vibrio, and Plesiomonas shigelloides: Bacterial culture followed by alternate PCR assay and bi-directional sequencing on presumptive positive isolates.
- For ETEC: Two alternate PCR assays and bidirectional sequencing performed from stool specimens.
- For retrospective samples: Confirmation of historical results using alternate PCR assay and bi-directional sequencing.
This is best categorized as a laboratory-based composite reference standard combining different microbiological and molecular techniques.
8. The sample size for the training set
The document does not explicitly state the sample size used for a "training set." The performance studies described are for the analytical and clinical validation of the device, typically representing a test or validation set after development. The analytical studies (e.g., LoD, inclusivity, cross-reactivity) involve testing various strains and concentrations to characterize the device. These are often used for optimizing the assay during development, but a distinct "training set" in the context of machine learning model development is not specified.
9. How the ground truth for the training set was established
Since a dedicated "training set" is not explicitly detailed in the provided K170308 summary, the method for establishing its ground truth is also not described. The analytical studies describe preparing known positive and negative samples using quantified cultures and negative fecal matrix, which would serve a similar function to generating truth data for assay development and validation.
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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
May 2, 2017
BECTON, DICKINSON AND COMPANY LAURA STEWART REGULATORY AFFAIRS SPECIALISTS 7 LOVETON CIRCLE SPARKS MD 21152
Re: K170308
Trade/Device Name: BD Max Extended Enteric Bacterial Panel. BD Max System Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid-based assay. Regulatory Class: II Product Code: PCH Dated: January 31, 2017 Received: February 1, 2017
Dear Ms. Stewart:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours.
Kristian M. Roth -S
For : Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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| 510(k) Number (if known) |
|---|
| K170308 |
| Device Name |
| BD MAX™ Extended Enteric Bacterial Panel |
| Indications for Use (Describe) |
Indications for Use (Describe)
The BD MAX™ Extended Enteric Bacterial Panel performed on the BD MAX System, is an automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens. It is used in conjunction with the BD MAX Enteric Bacterial Panel as an optional Master Mix. The BD MAX™ Extended Enteric Bacterial Panel detects nucleic acids from
- Plesiomonas shigelloides
- Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae)
- Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable ● enterotoxin (ST) genes
- Yersinia enterocolitica ●
Testing is performed on soft to diarrheal unpreserved stool specimens or Cary-Blair preserved stool specimens from symptomatic patients with suspected acute gastroenteritis, enteritis or colitis. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of relevant gene target DNA. The test utilizes fluorogenic gene-specific hybridization probes for the detection of the amplified DNA.
This test is intended for use, in conjunction with clinical presentation, laboratory findings, and epidemiological information, as an aid in the differential diagnosis of Plesiomonas shigelloides, Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae), Enterotoxigenic Escherichia coli (ETEC) LT/ST and Yersinia enterocolitica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other organisms that are not detected by this test. and may not be the sole or definitive cause of patient illness. 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.
| Type of Use ( Select one or both, as applicable ) | |
|---|---|
| ☑Prescription Use (Part 21 CFR 801 Subpart D) | ☐Over-The Counter Use (21 CFR 801 Subpart C) |
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5. 510(k) Summary
BD MAX™ Extended Enteric Bacterial Panel (xEBP) Summary Preparation Date: 1/30/2017
Submitted by:
BD Diagnostic Systems Becton, Dickinson and Company 7 Loveton Circle Sparks, Maryland 21152
Contact:
Laura Stewart Regulatory Affairs Specialist
Tel: 410-316-4435 Fax: 410-316-4188 Email: laura stewart@bd.com
Proprietary Names:
For the instrument: BD MAX™ System For the assay: BD MAX™ Extended Enteric Bacterial Panel (xEBP)
Common Names:
For the instrument: Bench-top molecular diagnostics workstation For the assay: Gastrointestinal Bacterial Panel Multiplex Nucleic Acid-Based Assay System Enteric Bacterial Panel Enteric Bacterial Nucleic Acid Test Enteric Bacterial identification and differentiation system Enteric assay Enteric test
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Regulatory Information
Regulation section: 866.3990 - Gastrointestinal microorganism multiplex nucleic acid-based assay.
Classification: Class II
Panel: Microbiology (83)
Product Code(s):
PCI- Gastrointestinal Bacterial Panel Multiplex Nucleic Acid-Based Assay System PCH- Gastrointestinal Pathogen Panel Multiplex Nucleic Acid-Based Assay System OOI- Real Time Nucleic Acid Amplification System
Predicate Device
BioFire Diagnostics FilmArray Gastrointestinal (GI) Panel [510(k) K160459]
Device Establishment
GeneOhm Sciences Canada, Inc. (BD Diagnostics) 2555 Boul. du Parc-Technologique Quebec, QC G1P 4S5 Canada
Registration Number: 3007420875
Performance Standards
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. November 2, 2015.
Intended Use
The BD MAX™ Extended Enteric Bacterial Panel performed on the BD MAX™ System, is an automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens. It is used in conjunction with the BD MAX Enteric Bacterial Panel as an optional Master Mix. The BD MAX Extended Enteric Bacterial Panel detects nucleic acids from
- Plesiomonas shigelloides
- Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae)
- Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable enterotoxin ● (ST) genes
- Yersinia enterocolitica ●
Testing is performed on soft to diarrheal unpreserved stool specimens or Cary-Blair preserved stool specimens from symptomatic patients with suspected acute gastroenteritis or colitis. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of relevant gene target DNA. The test utilizes fluorogenic gene-specific hybridization probes for the detection of the amplified DNA.
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This test is intended for use, in conjunction with clinical presentation, laboratory findings, and epidemiological information, as an aid in the differential diagnosis of Plesiomonas shigelloides, Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae), Enterotoxigenic Escherichia coli (ETEC) LT/ST and Yersinia enterocolitica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with other organisms that are not detected by this test, and may not be the sole or definitive cause of patient illness. 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.
Special Conditions for Use Statement: For prescription use.
Special Instrument Requirements: BD MAX™ System
Device Description
The BD MAX™ Extended Enteric Bacterial Panel performed on the BD MAX™ System, is an automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens. It is used in conjunction with the BD MAX Enteric Bacterial Panel as an optional Master Mix. The BD MAX Extended Enteric Bacterial Panel detects nucleic acids from
- Plesiomonas shigelloides
- . Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae)
- . Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable enterotoxin (ST) genes
- Yersinia enterocolitica
Testing is performed on unpreserved or Cary-Blair preserved soft to diarrheal stool specimens from symptomatic patients with suspected acute gastroenteritis, enteritis or colitis.
The BD MAX™ Extended Enteric Bacterial Panel (BD MAX™ xEBP) is composed of a single Master Mix that must be used in conjunction with the BD MAX™ Enteric Bacterial Panel (BD MAX EBP), performed on the BD MAX™ System. The BD MAX xEBP is performed with the Master Mix contained in the BD MAX xEBP reagent kit and the EBP reagents, including the Master Mix contained in the BD MAX™ EBP reagent kit. The BD MAX xEBP assay and accompanying Assay Definition File (ADF) were developed to allow inclusion of both BD MAX EBP and BD MAX xEBP master mix reagents in one test run.
The BD MAX™ System and the BD MAX™ Extended Enteric Bacterial Panel is run with the instrument with associated hardware and accessories, disposable microfluidic cartridges, master mixes, unitized reagent strips, extraction reagents, and sample buffer tubes. The instrument automates sample preparation including target lysis, DNA extraction and concentration, reagent rehydration, and target nucleic acid amplification and detection using real-time PCR. The assay includes a Sample Processing Control (SPC) that is present in the Extraction Tube. The SPC monitors DNA extraction steps, thermal cycling steps, reagent integrity and the presence of inhibitory substances. The BD MAX™ System software automatically interprets test result may be called as POS (Positive), NEG (Negative), or UNR (Unresolved) for each of the assay's targets, based on the amplification status of the target and of the Sample Processing Control. IND (Indeterminate) or INC (Incomplete) results are due to BD MAXTM System failure.
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Test Principle
A stool specimen is collected and transported to the laboratory in a dry, clean container (for unpreserved specimens) or in Cary-Blair transport media. The specimen is vortexed for 15 seconds and then a 10 uL loop is used to inoculate a BD MAX™ Extended Enteric Bacterial Panel and a BD MAX™ Extended Enteric Bacterial Panel Sample Buffer Tube included in the BD MAX™ Enteric Bacterial Panel kit. The Sample Buffer Tube is closed with a septum cap, vortexed and transferred to the BD MAX™ System. A worklist is created and the Sample Buffer Tubes, the BD MAX Enteric Bacterial Panel Unitized Reagent Strip (containing both the BD MAX™ EBP and BD MAX™ xEBP master mix reagents) and the BD MAX™ PCR Cartridges are loaded onto the BD MAX™ System.
Following enzymatic bacterial cell lysis at elevated temperature, the released nucleic acids are captured on magnetic beads. The beads, with the bound nucleic acids, are washed using Wash Buffer and the nucleic acids are eluted by heat in Elution Buffer. Eluted DNA is neutralized using Neutralization Buffer and transferred to the Master Mix tubes to rehydrate the PCR reagents. After rehydration, the BD MAX™ System dispenses a fixed volume of PCR-ready solution containing extracted nucleic acids into the BD MAX™ PCR Cartridge. Microvalves in the BD MAX™ PCR Cartridge are sealed by the system prior to initiating PCR to contain the amplification mixture, thus preventing evaporation and amplicon contamination.
The amplified DNA targets are detected using hydrolysis (TaqMan®) probes labeled at one end with a fluorescent reporter dye (fluorophore) and at the other end with a quencher moiety. Probes labeled with different fluorophores are used to detect the amplicons of the enteric bacterial targets (Plesiomonas shigelloides, Vibrio (V. vulnificus, V. parahaemolyticus, and V. cholerae), heat labile and heat stabile (LT/ST) ETEC (Enterotoxigenic E. coli) and Yersinia enterocolitica) and the Sample Processing Control amplicons in five different optical channels of the BD MAX™ System. When the probes are in their native state, the fluorescence of the fluorophore is quenched due to its proximity to the quencher. However, in the presence of target DNA, the probes hybridize to their complementary sequences and are hydrolyzed by the 5'-3' exonuclease activity of the DNA polymerase as it synthesizes the nascent strand along the cDNA template. As a result, the fluorophores are separated from the quencher molecules and fluorescence is emitted. The BD MAX System monitors these signals at each cycle, and interprets the data at the end of the program to report the final results.
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Substantial Equivalence1
Table 1 shows the similarities and Table 2 shows the differences between the BD MAX™ Extended Enteric Bacterial Panel and the predicate device.
| Table 1:Similarities Comparison to Predicate Device | |
|---|---|
| --------------------------------------------------------- | -- |
| Similarities | ||
|---|---|---|
| Item | BD MAX™ xEBP | FilmArray GI Panel (K160459) |
| Intended Use | The BD MAX™ Extended Enteric Bacterial Panelperformed on the BD MAX System, is an automatedin vitro diagnostic test for the direct qualitativedetection and differentiation of enteric bacterialpathogens. It is used in conjunction with the BDMAX EntericBacterial Panel as an optional Master Mix. The BDMAX Extended Enteric Bacterial Panel detectsnucleic acids from• Plesiomonas shigelloides• Vibrio ( V. vulnificus, V. parahaemolyticus, and V.cholerae )• Enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT)/ heat-stable enterotoxin (ST)genes• Yersinia enterocoliticaTesting is performed on soft to diarrheal unpreservedstool specimens or Cary-Blair preserved stoolspecimens from symptomatic patients withsuspected acute gastroenteritis, enteritis or colitis.The test is performed directly on the specimen,utilizing real-time polymerase chain reaction (PCR)for the amplification of relevant gene target DNA.The test utilizes fluorogenic gene-specifichybridization probes for the detection of theamplified DNA.This test is intended for use, in conjunction withclinical presentation, laboratory findings, andepidemiological information, as an aidin the differential diagnosis of Plesiomonasshigelloides, Vibrio (V. vulnificus, V.parahaemolyticus, and V. cholerae) , EnterotoxigenicEscherichia coli (ETEC) LT/ST and Yersiniaenterocolitica infections. Results of this test shouldnot be used as the sole basis for diagnosis, treatment,or other patient management decisions. Positiveresults do not rule out co-infection with otherorganisms that are not detected by this test, and maynot be the sole or definitive cause of patient illness.Negative results in the setting of clinical illnesscompatible with gastroenteritis may be due toinfection by pathogens that are not detected by thistest or non-infectious causes such as ulcerativecolitis, irritable bowel syndrome, or Crohn's disease. | The FilmArray Gastrointestinal (GI) Panel is aqualitative multiplexed nucleic acid-based in vitrodiagnostic test intended for use with FilmArraysystems. The FilmArray GI Panel is capable of thesimultaneous detection and identification of nucleicacids from multiple bacteria, viruses, and parasitesdirectly from stool samples in Cary Blair transportmedia obtained from individuals with signs and/orsymptoms of gastrointestinal infection. Thefollowing bacteria (including several diarrheagenicE. coli/Shigella pathotypes), parasites, and virusesare identified using the FilmArray GI Panel:- Campylobacter (C. jejuni/C. coli/C. upsaliensis)- Clostridium difficile (C. difficile) toxin A/B- Plesiomonas shigelloides- Salmonella- Vibrio (V. parahaemolyticus/V. vulnificus/V.cholerae) , including specific identification ofVibrio cholerae- Yersinia enterocolitica- Enteroaggregative Escherichia coli (EAEC)- Enteropathogenic Escherichia coli (EPEC)- Enterotoxigenic Escherichia coli (ETEC) lt/st- Shiga-like toxin-producing Escherichia coli(STEC) stx1/stx2 (including specific identificationof the E. coli O157 serogroup within STEC)- Shigella/Enteroinvasive Escherichia coli (EIEC)- Cryptosporidium- Cyclospora cayetanensis- Entamoeba histolytica- Giardia lamblia (also known as G. intestinalisand G. duodenalis )- Adenovirus F 40/41- Astrovirus- Norovirus GI/GII- Rotavirus A- Sapovirus (Genogroups I, II, IV, and V)The FilmArray GI Panel is indicated as an aid inthe diagnosis of specific agents of gastrointestinalillness and results are meant to be used inconjunction with other clinical, laboratory, andepidemiological data. Positive results do not ruleout co-infection with organisms not included in theFilmArray GI Panel. The agent detected may not bethe definite cause of the disease.Concomitant culture is necessary for organism |
| Similarities | ||
| Item | BD MAX™ xEBP | FilmArray GI Panel (K160459) |
| recovery and further typing of bacterial agents.This device is not intended to monitor or guidetreatment for C. difficile infection.Due to the small number of positive specimenscollected for certain organisms during theprospective clinical study, performancecharacteristics for E. coli 0157, Plesiomonasshigelloides, Yersinia enterocolitica, Astrovirus,and Rotavirus A were established primarily withretrospective clinical specimens.Performance characteristics for Entamoebahistolytica, and Vibrio (V. parahaemolyticus, V.vulnificus, and Vibrio cholerae) were established | ||
| primarily using contrived clinical specimens.Negative FilmArray GI Panel results in the settingof clinical illness compatible with gastroenteritismay be due to infection by pathogens that are notdetected by this test or non-infectious causes suchas ulcerative colitis, irritable bowel syndrome, orCrohn's disease.A gastrointestinal microorganism multiplex nucleicacid-based assay also aids in the detection andidentification of acute gastroenteritis in the contextof outbreaks. | ||
| Specimen Type | Cary-Blair preserved stoolUnpreserved soft to diarrheal stool | Cary-Blair preserved stool.Not claimed (see Differences below) |
| Assay Format | Amplification: PCRDetection: fluorogenic target-specific hybridization. | Amplification: PCRDetection: non target-specific fluorescent dye |
| OrganismsDetected | • Plesiomonas shigelloides• Vibrio (V. vulnificus, V. parahaemolyticus, and V.cholerae)• Enterotoxigenic Escherichia coli heat labile andheat stabile (LT/ST) (ETEC)• Yersinia enterocolitica | • Plesiomonas shigelloides• Vibrio (V. parahaemolyticus/V. vulnificus/ V.cholerae), including specific identification ofVibrio cholerae• Enterotoxigenic Escherichia coli (ETEC) lt/st• Yersinia enterocolitica |
| Interpretation ofTest Results | Automated: BD MAX System diagnostic software | Automated |
| Analysis Platform | BD MAX™ System | Film Array Instrument |
| PCR Samplepreparation | Automated:BD MAX™ System | Automated:Film Array Instrument |
| Detection Probes | TaqMan® Probe | Fluorescent double stranded DNA binding dye (LCGreen Plus) |
| Assay Controls | Sample Processing Control (SPC) | Two controls are included in each reagent pouch tocontrol for sample processing and both stages ofPCR and melt analysis. |
| Differences | ||
| Item | BD MAX TM xEBP | FilmArray GI Panel |
| Specimen Type | Unpreserved soft to diarrheal stool | Not claimed |
| OrganismsDetected | Listed in device Similarities above. | Other organisms detected: |
| • Campylobacter ( C. jejuni/C. coli/C. upsaliensis ) | ||
| • Clostridium difficile ( C. difficile ) toxin A/B | ||
| • Salmonella | ||
| • Enteroaggregative Escherichia coli (EAEC) | ||
| • Enteropathogenic Escherichia coli (EPEC) | ||
| • Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2(including specific identification of the E. coli O157serogroup within STEC) | ||
| • Shigella/ Enteroinvasive Escherichia coli (EIEC) | ||
| • Cryptosporidium | ||
| • Cyclospora cayetanensis | ||
| • Entamoeba histolytica | ||
| • Giardia lamblia (also known as G. intestinalis and G. duodenalis ) | ||
| • Adenovirus F 40/41 | ||
| • Astrovirus | ||
| • Norovirus GI/GII | ||
| • Rotavirus A• Sapovirus (Genogroups I, II, IV, and V) |
The term "substantial equivalence" as used in this 510(k) notification is limited to the definition of substantial equivalence as found in the Federal Food, Drug and Cosmetic Act, as amended and as applied under which a device can be marketed without pre-market approval or reclassification. A determination of substantial equivalency under this not intended to have any bearing whatsoever on the resolution of patent infringement suits or any other patent matters. No statements related to, or in support of substantial equivalence herein shall be construed as an admission against interest under the US Patent Laws or the courts.
{8}------------------------------------------------
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Table 2: Differences Comparison to Predicate Device
Analytical Performance
Precision
Within-laboratory precision was evaluated for the BD MAX™ Extended Enteric Bacterial Panel at one (1) internal site. Testing was performed over 12 days, with two runs per day (one each by 2 operators), for a total of 24 runs. The Precision panel members were divided into four (4) concentration categories, based upon organism concentration relative to the LoDs established for each of the assay targets and expected correct percent positive . The panel members contained Vibrio cholerae, Plesiomonas shigelloides, ETEC and Yersinia enterocolitica. The following values were used as spike levels for the target organisms contained in each panel member:
- Moderate Positive (MP): ≥2 to ≤3x LoD; positive approximately 95% of the time .
- Low Positive (LP): ≥1 to <2x LoD; positive approximately 95% of the time ●
- High Negative (HN): C20-80 LoD: negative between 20 and 80% of the time .
- True Negative (TN): No target; negative 100% of the time ●
Each panel member was spiked with negative unpreserved stool matrix. True negative samples contained no target. High negative samples were spiked with target organisms below the analytical LoD of the assay; however, the HN samples were expected to yield a positive result in approximately 20% to 80% of the replicates due the inherent sensitivity of the PCR assays. Results are summarized by target and concentration in Table 3.
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| Category | Agreement with Expected Results | Vibrio(95% CI) | P. shigelloides(95% CI) | Y. enterocolitica(95% CI) | ETEC(95% CI) |
|---|---|---|---|---|---|
| TNa | 10096/96(96.2, 100) | 10096/96(96.2, 100) | 10096/96(96.2, 100) | 10096/96(96.2, 100) | |
| HNa | 58.328/48(44.3, 71.2) | 45.822/48(32.6, 59.7) | 41.720/48(28.8, 55.7) | 47.923/48(34.5, 61.7) | |
| LP | 10048/48(92.6, 100) | 10048/48(92.6, 100) | 97.947/48(89.1, 99.6) | 10048/48(92.6, 100) | |
| MP | 10048/48(92.6, 100) | 10048/48(92.6, 100) | 10048/48(92.6, 100) | 97.947/48(89.1, 99.6) |
Table 3: Precision study result Using One Lot of the BD MAX Extended Enteric Bacterial Panel
4 For the True Negative (TN) and High Negative (HN) categories, the expected assay result was deemed to be negative. Therefore, percent agreement was calculated for negative results.
Reproducibility
The Site-to-Site reproducibility study was performed at three (3) clinical sites using one (1) reagent lot. Two (2) operators performed 2 runs per day, over five (5) distinct days (consecutive or not), for a total of 30 runs. The panels used were the same as described under the Precision heading, above.
The overall site-to-site reproducibility percent agreement was 100% for the TN category for all targets, and ranged from 30.0 to 48.9%, 97.8 to 100 % and 98.9 to 100% for the HN, LP and MP categories, respectively. Results are summarized in Table 4. The quantitative reproducibility results across sites by target are presented in Table 6. Ct. Score is an internal criterion used to determine final assay results and was selected as a means of assessing quantitative assay reproducibility. Mean Ct. Score and the mean Cycle EP values with variance components (SD and % CV) are shown in Table 5 and Table 6.
| Table 4: | Site-to-Site Reproducibility Results Using One Lot of the BD MAX Extended Enteric Bacterial Panel |
|---|---|
| ----------------- | --------------------------------------------------------------------------------------------------- |
| Category | Agreement with Expected Results | |||
|---|---|---|---|---|
| Vibrio(95% CI) | P. shigelloides(95% CI) | Y. enterocolitica(95% CI) | ETEC(95% CI) | |
| TNa | 100180/180(97.9, 100) | 100180/180(97.9, 100) | 100180/180(97.9, 100) | 100180/180(97.9, 100) |
| HNa | 48.944/90(38.8, 59.0) | 30.027/90(21.5, 40.1) | 35.632/90(26.4, 45.8) | 46.742/90(36.7, 56.9) |
| LP | 10090/90(95.9, 100) | 98.989/90(94.0, 99.8) | 10090/90(95.9, 100) | 97.888/90(92.3, 99.4) |
| MP | 10090/90(95.9, 100) | 98.989/90(94.0, 99.8) | 98.989/90(94.0, 99.8) | 10090/90(95.9, 100) |
4 For the True Negative (TN) and High Negative (HN) categories, the expected assay result was deemed to be negative. Therefore, percent agreement was calculated for negative results.
{11}------------------------------------------------
| Target | PCRMetric | Category | N | Mean | Within Run | Between RunWithin Day | Between DayWithin Site | Between Site | Total | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |||||
| Vibrio | Ct.Score | HN | 46 | 36.9 | 1.11 | 3.0 | 0.00 | 0.0 | 0.00 | 0.0 | 0.40 | 1.1 | 1.18 | 3.2 |
| Score | LP | 90 | 32.1 | 0.43 | 1.3 | 0.00 | 0.0 | 0.00 | 0.0 | 0.13 | 0.4 | 0.45 | 1.4 | |
| MP | 90 | 31.7 | 0.55 | 1.7 | 0.00 | 0.0 | 0.00 | 0.0 | 0.13 | 0.4 | 0.56 | 1.8 | ||
| CycleEndPoint | HN | 46 | 805.1 | 334.64 | 41.6 | 0.00 | 0.0 | 0.00 | 0.0 | 103.89 | 12.9 | 350.40 | 43.5 | |
| LP | 90 | 2725.7 | 398.88 | 14.6 | 303.13 | 11.1 | 250.16 | 9.2 | 588.42 | 21.6 | 812.29 | 29.8 | ||
| MP | 90 | 3076.6 | 459.04 | 14.9 | 127.59 | 4.1 | 92.07 | 3.0 | 0.00 | 0.0 | 485.26 | 15.8 | ||
| Plesiomonasshigelloides | Ct.Score | HN | 63 | 36.0 | 1.44 | 4.0 | 0.05 | 0.1 | 0.00 | 0.0 | 0.43 | 1.2 | 1.50 | 4.2 |
| LP | 89 | 31.5 | 0.73 | 2.3 | 0.00 | 0.0 | 0.02 | 0.1 | 0.52 | 1.7 | 0.90 | 2.9 | ||
| MP | 89 | 30.7 | 0.73 | 2.4 | 0.00 | 0.0 | 0.00 | 0.0 | 0.16 | 0.5 | 0.74 | 2.4 | ||
| CycleEndPoint | HN | 63 | 596.5 | 284.02 | 47.6 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 284.02 | 47.6 | |
| LP | 89 | 2349.7 | 332.60 | 14.2 | 0.00 | 0.0 | 0.00 | 0.0 | 250.45 | 10.7 | 416.36 | 17.7 | ||
| MP | 89 | 2320.9 | 342.77 | 14.8 | 313.96 | 13.5 | 86.03 | 3.7 | 416.63 | 18.0 | 630.12 | 27.1 | ||
| Yersiniaenterocolitica | Ct.Score | HN | 58 | 36.3 | 0.99 | 2.7 | 0.35 | 1.0 | 0.13 | 0.4 | 0.00 | 0.0 | 1.06 | 2.9 |
| LP | 90 | 32.9 | 0.69 | 2.1 | 0.00 | 0.0 | 0.00 | 0.0 | 0.37 | 1.1 | 0.78 | 2.4 | ||
| MP | 89 | 32.7 | 0.58 | 1.8 | 0.14 | 0.4 | 0.06 | 0.2 | 0.23 | 0.7 | 0.64 | 2.0 | ||
| EndPoint | HN | 58 | 599.6 | 232.01 | 38.7 | 139.57 | 23.3 | 0.00 | 0.0 | 121.25 | 20.2 | 296.66 | 49.5 | |
| LP | 90 | 1191.1 | 272.51 | 22.9 | 107.96 | 9.1 | 0.00 | 0.0 | 121.52 | 10.2 | 317.31 | 26.6 | ||
| MP | 89 | 1264.2 | 363.00 | 28.7 | 0.00 | 0.0 | 0.00 | 0.0 | 139.76 | 11.1 | 388.98 | 30.8 | ||
| ETEC | Ct.Score | HN | 48 | 36.8 | 1.37 | 3.7 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 1.37 | 3.7 |
| LP | 88 | 33.7 | 0.74 | 2.2 | 0.00 | 0.0 | 0.09 | 0.3 | 0.29 | 0.9 | 0.80 | 2.4 | ||
| MP | 90 | 32.4 | 0.67 | 2.1 | 0.00 | 0.0 | 0.00 | 0.0 | 0.26 | 0.8 | 0.72 | 2.2 | ||
| CycleEndPoint | HN | 48 | 976.0 | 577.46 | 59.2 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 577.46 | 59.2 | |
| LP | 88 | 2049.6 | 511.41 | 25.0 | 61.48 | 3.0 | 0.00 | 0.0 | 121.96 | 6.0 | 529.33 | 25.8 | ||
| MP | 90 | 2640.0 | 491.69 | 18.6 | 217.12 | 8.2 | 0.00 | 0.0 | 263.58 | 10.0 | 598.65 | 22.7 | ||
| SPC | Ct.Score | TN | 90 | 27.2 | 0.35 | 1.3 | 0.00 | 0.0 | 0.00 | 0.0 | 0.18 | 0.7 | 0.40 | 1.5 |
| CycleEndPoint | TN | 90 | 6020.7 | 672.52 | 11.2 | 0.00 | 0.0 | 0.00 | 0.0 | 148.61 | 2.5 | 688.74 | 11.4 |
Table 5: Quantitative Site-to-Site Reproducibility for all Targets and Sample Processing Control
Table 6: Quantitative Site-to-Site Reproducibility Results Summary
| PCRMetric | Parameter | Vibrio | P. shigelloides | Y. enterocolitica | ETEC | SPC | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HN | LP | MP | HN | LP | MP | HN | LP | MP | HN | LP | MP | TN | ||
| N | 46 | 90 | 90 | 63 | 89 | 89 | 58 | 90 | 89 | 48 | 88 | 90 | 90 | |
| Ct.Score | Mean | 36.9 | 32.1 | 31.7 | 36.0 | 31.5 | 30.7 | 36.3 | 32.9 | 32.7 | 36.8 | 33.7 | 32.4 | 27.2 |
| SD | 1.18 | 0.45 | 0.56 | 1.50 | 0.90 | 0.74 | 1.06 | 0.78 | 0.64 | 1.37 | 0.80 | 0.72 | 0.40 | |
| %CV | 3.2 | 1.4 | 1.8 | 4.2 | 2.9 | 2.4 | 2.9 | 2.4 | 2.0 | 3.7 | 2.4 | 2.2 | 1.5 | |
| N | 46 | 90 | 90 | 63 | 89 | 89 | 58 | 90 | 89 | 48 | 88 | 90 | 90 | |
| CycleEP | Mean | 805.1 | 2725.7 | 3076.6 | 596.5 | 2349.7 | 2320.9 | 599.6 | 1191.1 | 1264.2 | 976.0 | 2049.6 | 2640.0 | 6020.7 |
| SD | 350.40 | 812.29 | 485.26 | 284.02 | 416.36 | 630.12 | 296.66 | 317.31 | 388.98 | 577.46 | 529.33 | 598.65 | 688.74 | |
| %CV | 43.5 | 29.8 | 15.8 | 47.6 | 17.7 | 27.1 | 49.5 | 26.6 | 30.8 | 59.2 | 25.8 | 22.7 | 11.4 |
The Lot-to-lot reproducibility study was performed at one (1) site using three (3) reagent lots. Two (2) operators performed 2 runs per day, over five (5) distinct days (consecutive or not), for a total of 30 runs. The panels used were the same as described under the Precision heading, above. Results from 5 days of the accuracy and precision study were used to comprise data for one lot of reagents for the Lot-to-Lot study.
The overall Lot-to-lot reproducibility percent agreements were 100% for TN, and ranged from 23.3 to 41.1%, 97.8 to 100 % and 98.9 to 100% for the HN, LP and MP respectively. Results are summarized in Table 7. The quantitative results across lots and by target are presented in Table 8 and Table 9. Ct. Score and the Cycle EP, an internal criteria used to determine a final assay result, was
{12}------------------------------------------------
selected as a means of assessing quantitative assay reproducibility. Mean Ct. Score and the mean Cycle EP values with variance components (SD and % CV) are shown in Table 8 and Table 9.
| Category | Agreement with Expected Results | |||
|---|---|---|---|---|
| Vibrio(95% CI) | P. shigelloides(95% CI) | Y. enterocolitica(95% CI) | ETEC(95% CI) | |
| TNa | 100180/180(97.9, 100) | 100180/180(97.9, 100) | 100180/180(97.9, 100) | 100180/180(97.9, 100) |
| HNa | 41.137/90(31.5, 51.4) | 28.926/90(20.5, 39.0) | 23.321/90(15.8, 33.1) | 41.137/90(31.5, 51.4) |
| LP | 10090/90(95.9, 100) | 10090/90(95.9, 100) | 10090/90(95.9, 100) | 97.888/90(92.3, 99.4) |
| MP | 10090/90(95.9, 100) | 10090/90(95.9, 100) | 98.989/90(94.0, 99.8) | 10090/90(95.9, 100) |
Table 7: Lot-to-lot Reproducibility Results for BD MAX Extended Enteric Bacterial Panel
4 For the True Negative (TN) and High Negative (HN) categories, the expected assay result was deemed to be negative. Therefore, percent agreement was calculated for negative results.
| Target | PCRMetric | Category | N | Mean | Within Run | Between RunWithin Day | Between DayWithin Site | Between Site | Total | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | |||||
| Vibrio | Ct.Score | HN | 53 | 36.6 | 1.18 | 3.2 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 1.18 | 3.2 |
| LP | 90 | 32.2 | 0.42 | 1.3 | 0.18 | 0.6 | 0.00 | 0.0 | 0.23 | 0.7 | 0.51 | 1.6 | ||
| MP | 90 | 31.9 | 0.61 | 1.9 | 0.00 | 0.0 | 0.00 | 0.0 | 0.20 | 0.6 | 0.64 | 2.0 | ||
| CycleEndPoint | HN | 53 | 931.1 | 369.81 | 39.7 | 0.00 | 0.0 | 0.00 | 0.0 | 0.00 | 0.0 | 369.81 | 39.7 | |
| LP | 90 | 3124.7 | 189.01 | 6.0 | 0.00 | 0.0 | 70.08 | 2.2 | 135.26 | 4.3 | 242.75 | 7.8 | ||
| MP | 90 | 3033.5 | 207.01 | 6.8 | 0.00 | 0.0 | 53.48 | 1.8 | 101.18 | 3.3 | 236.53 | 7.8 | ||
| Plesiomonasshigelloides | Ct.Score | HN | 64 | 35.8 | 1.41 | 3.9 | 0.20 | 0.6 | 0.00 | 0.0 | 0.00 | 0.0 | 1.42 | 4.0 |
| LP | 90 | 31.3 | 0.52 | 1.7 | 0.05 | 0.2 | 0.00 | 0.0 | 0.31 | 1.0 | 0.61 | 2.0 | ||
| MP | 90 | 30.7 | 0.68 | 2.2 | 0.29 | 0.9 | 0.00 | 0.0 | 0.52 | 1.7 | 0.90 | 2.9 | ||
| CycleEndPoint | HN | 64 | 686.9 | 320.86 | 46.7 | 0.00 | 0.0 | 0.00 | 0.0 | 104.23 | 15.2 | 337.36 | 49.1 | |
| LP | 90 | 2444.1 | 411.42 | 16.8 | 0.00 | 0.0 | 0.00 | 0.0 | 269.42 | 11.0 | 491.79 | 20.1 | ||
| MP | 90 | 2428.7 | 170.72 | 7.0 | 140.04 | 5.8 | 0.00 | 0.0 | 353.06 | 14.5 | 416.43 | 17.1 | ||
| Yersiniaenterocolitica | Ct.Score | HN | 69 | 36.1 | 1.08 | 3.0 | 0.70 | 1.9 | 0.00 | 0.0 | 0.00 | 0.0 | 1.29 | 3.6 |
| LP | 90 | 32.4 | 0.73 | 2.3 | 0.03 | 0.1 | 0.00 | 0.0 | 0.58 | 1.8 | 0.94 | 2.9 | ||
| MP | 89 | 32.3 | 0.87 | 2.7 | 0.18 | 0.6 | 0.00 | 0.0 | 0.25 | 0.8 | 0.93 | 2.9 | ||
| EndPoint | HN | 69 | 731.3 | 252.85 | 34.6 | 154.25 | 21.1 | 0.00 | 0.0 | 12.15 | 1.7 | 296.44 | 40.5 | |
| LP | 90 | 1339.7 | 140.87 | 10.5 | 0.00 | 0.0 | 0.00 | 0.0 | 72.45 | 5.4 | 158.41 | 11.8 | ||
| MP | 89 | 1370.3 | 162.58 | 11.9 | 25.73 | 1.9 | 0.00 | 0.0 | 19.29 | 1.4 | 165.73 | 12.1 | ||
| ETEC | Ct.Score | HN | 53 | 37.0 | 1.06 | 2.9 | 0.13 | 0.3 | 0.00 | 0.0 | 0.00 | 0.0 | 1.06 | 2.9 |
| LP | 88 | 33.6 | 0.65 | 1.9 | 0.00 | 0.0 | 0.00 | 0.0 | 0.43 | 1.3 | 0.78 | 2.3 | ||
| MP | 90 | 32.4 | 0.74 | 2.3 | 0.00 | 0.0 | 0.00 | 0.0 | 0.46 | 1.4 | 0.87 | 2.7 | ||
| CycleEndPoint | HN | 53 | 838.3 | 308.54 | 36.8 | 105.56 | 12.6 | 0.00 | 0.0 | 64.26 | 7.7 | 332.37 | 39.6 | |
| LP | 88 | 2056.8 | 267.41 | 13.0 | 0.00 | 0.0 | 0.00 | 0.0 | 323.86 | 15.7 | 419.99 | 20.4 | ||
| MP | 90 | 2244.2 | 315.85 | 14.1 | 0.00 | 0.0 | 0.00 | 0.0 | 332.48 | 14.8 | 458.59 | 20.4 | ||
| SPC | Ct.Score | TN | 90 | 27.2 | 0.30 | 1.1 | 0.00 | 0.0 | 0.00 | 0.0 | 0.06 | 0.2 | 0.30 | 1.1 |
| CycleEndPoint | TN | 90 | 5544.7 | 567.21 | 10.2 | 0.00 | 0.0 | 0.00 | 0.0 | 282.58 | 5.1 | 633.70 | 11.4 |
Table 8: Quantitative Lot-to-lot for all Targets and SPC
{13}------------------------------------------------
| PCRMetric | Parameter | Vibrio | P. shigelloides | Y. enterocolitica | ETEC | SPC | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| HN | LP | MP | HN | LP | MP | HN | LP | MP | HN | LP | MP | TN | ||
| N | 53 | 90 | 90 | 64 | 90 | 90 | 69 | 90 | 89 | 53 | 88 | 90 | 90 | |
| Ct.Score | Mean | 36.6 | 32.2 | 31.9 | 35.8 | 31.3 | 30.7 | 36.1 | 32.4 | 32.3 | 37.0 | 33.6 | 32.4 | 27.2 |
| SD | 1.18 | 0.51 | 0.64 | 1.42 | 0.61 | 0.90 | 1.29 | 0.94 | 0.93 | 1.06 | 0.78 | 0.87 | 0.30 | |
| %CV | 3.2 | 1.6 | 2.0 | 4.0 | 2.0 | 2.9 | 3.6 | 2.9 | 2.9 | 2.9 | 2.3 | 2.7 | 1.1 | |
| N | 53 | 90 | 90 | 64 | 90 | 90 | 69 | 90 | 89 | 53 | 88 | 90 | 90 | |
| CycleEP | Mean | 931.1 | 3124.7 | 3033.5 | 686.9 | 2444.1 | 2428.7 | 731.3 | 1339.7 | 1370.3 | 838.3 | 2056.8 | 2244.2 | 5544.7 |
| SD | 369.81 | 242.75 | 236.53 | 337.36 | 491.79 | 416.43 | 296.44 | 158.41 | 165.73 | 332.37 | 419.99 | 458.59 | 633.70 | |
| %CV | 39.7 | 7.8 | 7.8 | 49.1 | 20.1 | 17.1 | 40.5 | 11.8 | 12.1 | 39.6 | 20.4 | 20.4 | 11.4 |
Table 9: Ouantitative Lot-to-lot Reproducibility Results Summary
Storage and Stability
- . Collected specimens, either unpreserved stool or stool stored in 15 mL Cary-Blair transport media should be kept between 2 ℃ and 25 ℃ during transport. Protect against exposure to excessive heat.
- Specimen can be stored for up to 120 hours (5 days) at 2-8 °C or for up to 24 hours at 2-25 °C before testing.
- BD MAX Extended Enteric Bacterial Panel Master Mix is stable at 2-25 ℃ through the stated expiration date. Do not use expired components.
- BD MAX Extended Enteric Bacterial Master Mix Tubes are provided in sealed pouches. To . protect product from humidity, immediately re-seal after opening. Master Mix tubes are stable for up to 14 days at 2-25 °C after initial opening and re-sealing of the pouch.
- Unreconstituted Master Mix tubes are stable for up to 24 hours at 2-25 °C after being removed . from their protective pouch.
Controls
External Control materials are not provided by BD. External Positive and Negative Controls are not used by the BD MAX System software for the purpose of sample test result interpretation. External Controls are treated as if they were patient samples. However, Quality Control strains and procedures are included in the package insert. Various types of External Controls are recommended to allow the user to select the most appropriate for their laboratory quality control program:
- External Negative Control: Commercially available control material or a previously characterized a. sample known to be negative. BD recommends that the External negative Control be prepared prior to the External Positive Control in order to reduce the potential for contamination as a result of control preparation.
- External Positive Control: Commercially available control materials, such as the ATCC strains b. listed below, or previously characterized samples known to be positive:
- . Yersinia enterocolitica (ATCC 9610)
- Vibrio cholerae (ATCC 14033)
- Vibrio parahaemolyticus (ATCC 17802)
- ETEC (ATCC 35401)
- Plesiomonas shigelloides (ATCC 14029) ●
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The assay includes a Specimen Processing Control (SPC) that is present in the Extraction Tube. The Sample Processing Control monitors the efficiency of DNA capture, washing and elution during the sample processing steps, as well as the efficiency of DNA target amplification and detection during PCR analysis.
Analytical Sensitivity
The analytical sensitivity (Limit of Detection or LoD) for the BD MAX Extended Enteric Bacterial Panel was determined as follows: Each target organism was prepared and quantified from culture prior to inclusion in this study. Individual inoculating loops were dipped into each organism preparation and were then transferred to a Sample Buffer Tube already containing fecal matrix (preserved or unpreserved) that was pre-determined to be negative for all the targets detected by the BD MAX Extended Enteric Bacterial Panel. Each organism was tested with a minimum of 24 replicates per sample type (preserved or unpreserved), by 2 operators, using 3 different production lots of the BD MAX Extended Enteric Bacterial Panel. The LoD for a specific organism was confirmed by testing at least 24 additional replicates at the determined LoD concentration. Analytical sensitivity (LoD), defined as the lowest concentration at which greater than 95% of all replicates are expected to test positive, ranged from 34 to 539 CFU/SBT and 3,434 to 53,852 CFU/mL (in stool) for unpreserved specimens for both strains and 79 to 257 CFU/SBT and 7,860 to 25,712 CFU/mL (in stool) for preserved specimens (Table 10).
| Unpreserved (1st strain) | Unpreserved (2nd strain) | Cary-Blair Preserved (1st strain) | ||||
|---|---|---|---|---|---|---|
| Target organism | LoDa(CFU/SBT) | LoDa(CFU/mL in stool) | LoDa(CFU/SBT) | LoDa(CFU/mL in stool) | LoDa(CFU/SBT) | LoDa(CFU/mL in stool) |
| PlesiomonasshigelloidesATCC 14029b;ATCC 14030c | 458 | 45,752 | 155 | 15,481 | 257 | 25,712 |
| YersiniaenterocoliticaATCC 9610b;CCUG 4588c | 209 | 20,900 | 311 | 31,099 | 227 | 22,723 |
| ETEC ST/LTATCC 35401b;CCUG 47194c | 34 | 3,434 | 539 | 53,852 | 137 | 13,706 |
| Vibrio choleraeATCC 14033b;ENF 9786c | 149 | 14,942 | 43 | 4,344 | 252 | 25,238 |
| VibrioparahaemolyticusATCC17802b;ENF 5887c | 207 | 20,708 | 80 | 8,031 | 124 | 12,424 |
| Vibrio vulnificusATCC 27562b;ENF 10727c | 131 | 13,093 | 80 | 7,959 | 79 | 7,860 |
BD MAX™ Extended Enteric Bacterial Panel Limit of Detection for Individual Target Table 10•
LoD concentrations are expressed in CFU/SBT and CFU/mL, except for Vibrio, which is expressed in cells/SBT and cells/mL 1st strain
C2nd strain
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Analytical Inclusivity
A variety of BD MAX Extended Enteric Bacterial Panel assay target strains were included in this study. Strain selection criteria included prevalence, serotype and geographic location, where appropriate. Sixty-nine (69) strains were tested, including strains from public collections and well-characterized clinical isolates.
Inclusivity testing included 10 strains of Plesiomonas shigelloides, 10 strains of Yersinia enterocolitica, 36 strains of Vibrio (cholerae, parahaemolyticus and vulnificus) and 13 strains of ETEC LT/ST. The strains were tested at < 3 x LoD (Limit of Detection) of the corresponding strain in unpreserved stool matrix. The BD MAX Extended Enteric Bacterial Panel correctly identified 68 of the 69 strains tested upon initial testing. One strain of ETEC ST/LT (CCUG 38088) did not meet acceptance criteria and was further evaluated. This strain was titrated and tested to determine the minimum concentration sufficient for detection. Upon repeat, the CCUG 38088 strain of ETEC ST/LT was detected at 10 x LoD.
Analytical Specificity (Cross-Reactivity and Exclusivity)
The BD MAX Extended Enteric Bacterial Panel was performed on samples containing phylogenetically related species and other organisms (bacteria, viruses, parasites and yeast) likely to be found in stool specimens. The bacterial cells, yeasts, parasites and viruses were tested in the Sample Buffer Tube at ≥ 10° CFU, cells or genome equivalents/mL in stool, or ≥ 10° PFU/mL in stool or TCID50/mL in stool or equivalent amount of RNA/DNA/PCR reaction. Overall, 184 organisms were tested.
- . Most bacterial strains, yeast, parasites and viruses tested produced negative results with the BD MAX Extended Enteric Bacterial Panel.
- Two (2) strains of Vibrio mimicus, associated with human disease, produced positive results with the BD MAX Extended Enteric Bacterial Panel. However, no positive result was recorded at ≤ 1.0 x 10' cells/mL in stool with these two strains.
- . The following 8 Vibrio species, NOT associated with infections in humans and therefore unlikely to be encountered in human stool, were detected by the BD MAX Extended Enteric Bacterial Panel: V. brasiliensis, V. campbellii, V. harveyi, V. hispanicus, V. nereis, V. pacini, V. rotiferianus and V. tubiashii.
Based upon in silico analysis, the following Vibrio species could be detected by the BD MAX Extended Enteric Bacterial Panel:
- 2 Vibrio species, NOT associated with infections in humans and therefore unlikely to be encountered in human stool: V. coralliilyticus and Moritella marina (formerly known as Vibrio marinus).
- One (1) uncategorized Vibrio species associated with infections in humans, Vibrio HENC. ●
Interfering Substances
Nineteen (19) biological and chemical substances that may occasionally be present in stool specimens were evaluated for potential interference with the BD MAX Extended Enteric Bacterial Panel. Included in this study was an Antibiotics Mixture, which consisted of a combination of 8 different antibiotics tested simultaneously (each antibiotic at a concentration that may be found in a stool sample). Nystatin cream was found to interfere at levels above 3.1 mg/mL. Spermicidal lubricant and hydrocortisone cream were found to interfere at levels above 2.5 mg/mL. Vagisil was found to interfere at levels above 0.92 mg/mL. Results demonstrated no reportable interference with any other substance tested (Table 11).
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In addition, microorganisms that may be endogenously present in stool specimens were evaluated for potential interference with the BD MAX Extended Enteric Bacterial Panel. Ten (10) organisms were tested at high concentration (> 2 x 10° CFU/mL of stool). Results demonstrated no reportable interference with any microorganism tested (Table 12).
Endogenous and Commercial Exogenous Substances Tested with the BD MAX Extended Table 11: Enteric Bacterial Panel
| Brand Name of Description | Result | Brand Name of Description | Result |
|---|---|---|---|
| Fecal Fat | NI | Spermicial Lubricant | P |
| Human DNA | NI | Diaper Rash Cream | NI |
| Mucus | NI | Vagisil® | P |
| Whole Human Blood | NI | Laxatives | NI |
| Hydrocortisone Cream | P | Anti-Diarrheal (liquid) | NI |
| Antiseptic Towelettes | NI | Anti-Diarrheal (pill) | NI |
| Enema | NI | Antibiotics Mixture | NI |
| Hemorrhoidal Gel | NI | Antacids | NI |
| Nystatin Cream | P | Non-Steroidal Anti-Inflammatory(NSAID) | NI |
| Topical Antibiotic | NI |
P: Potential interference with the BD MAX Extended Enteric Bacterial Panel at high concentrations. NI: No reportable interference with the BD MAX Extended Enteric Bacterial Panel.
Table 12: Microorganisms Tested for Interference with the BD MAX Extended Enteric Bacterial Panel
| Microorganism | Result |
|---|---|
| Salmonella typhimurium | NI |
| Shigella sonnei | NI |
| Campylobacter coli | NI |
| Escherichia coli (stx1/stx2) | NI |
| Citrobacter amalonaticus | NI |
| Proteus vulgaris | NI |
| Bacteroides thetaiotaomicron | NI |
| Ruminococcus bromii | NI |
| Enterococcus faecalis | NI |
| Peptostreptococcus anaerobius | NI |
NI: No reportable interference with the BD MAX Extended Enteric Bacterial Panel.
Carryover/Cross-Contamination
A study was conducted to investigate within-run carryover and between-run carryover while processing samples with high bacterial load of analytes in the BD MAX™ Extended Enteric Bacterial Panel. A panel made of one high positive member containing one target organism and one negative member was used to prepare numerous samples. The enterotoxigenic Escherichia coli strain was used to represent the high positive panel member (~1 x 10 CFU/mL of SBT). The negative member did not contain any target analyte. Twelve (12) replicates of the high positive panel member and 12 replicates of the negative panel member were tested in each run by alternating negative and positive samples. Three (3) operators performed 3 consecutive runs across 3 BD MAX instruments for a total of nine (9) runs containing 24 samples. Of the 108 negative samples tested in this study, one (1) sample produced a positive ETEC ST/LT result.
Mixed Infection/Competitive Interference
The mixed infection/competitive interference study was designed to evaluate the ability of the BD MAX Extended Enteric Bacterial Panel to detect low positive results in the presence of other targets at high concentrations. A mix of three (3) out of four (4) organisms (Plesiomonas shigelloides, Yersinia enterocolitica, ETEC ST/LT, and Vibrio cholerae) were prepared at < 2x their respective LoD to serve as
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low targets in the BD MAX Extended Enteric Bacterial Panel Sample Buffer Tube. The fourth BD MAX Enteric Bacterial Panel analyte (absent from the low targets mix) was spiked into the Sample Buffer Tube at a concentration ≥ 1 x 10° CFU/mL (representing a high load target) along with 10 uL of unpreserved stool and tested to simulate mixed infections. In the presence of high loads of Plesiomonas shigelloides (> 1 x 10° CFU/mL) and Vibrio cholerae (> 1 x 10° cells/mL), all three (3) organisms corresponding to their respective simulated mixed infection preparations were successfully detected by the BD MAX Extended Enteric Bacterial Panel. Successful detection of all three (3) low target organisms by the BD MAX Extended Enteric Bacterial Panel was achieved in the presence of Yersinia enterocolitica at 1.0 x 10 CFU/mL and ETEC ST/LT at 9.44 x 102 CFU/mL.
Clinical Performance Studies
Clinical performance characteristics of the BD MAX Extended Enteric Bacterial Panel were determined in a multi-site investigational study. The study involved a total of six (6) geographically diverse clinical centers where stools specimens were collected as part of routine patient care, enrolled into the trial, and tested with the BD MAX Extended Enteric Bacterial Panel. Specimens were obtained from pediatric or adult patients suspected of acute bacterial gastroenteritis, enteritis or colitis, for which stool culture had been ordered by healthcare provider. The reference method for both prospective fresh and prospective frozen specimens, was a combination of bacterial culture followed by alternate PCR assay and bi-directional sequencing on presumptive positive isolates for Yersinia enterocolitica, Vibrio and Plesiomonas shigelloides. For ETEC, the comparator method was two alternate PCR assays and bidirectional sequencing performed from the stool specimens. For retrospective specimens, the historical results were recorded at the collection site. The historical results were confirmed using an alternate PCR assay and bi-directional sequencing as part of the composite comparator method in order to confirm the presence of the target DNA.
A total of 2264 prospective specimens (882 unpreserved and 1382 Cary-Blair preserved) and 146 retrospective specimens (87 unpreserved and 59 Cary-Blair preserved) were enrolled in the clinical evaluation for a total of 2410 specimens enrolled. All test results from the BD MAX Exteric Bacterial Panel and the comparator method were single results, no coinfections were detected. Table 13 describes the number of compliant specimens enrolled by patient age and specimen type with a total of 2403 compliant specimens overall. Table 14 through Table 22 describe the performance characteristics of the BD MAX Extended Enteric Bacterial Panel that were observed during the clinical trial.
| Age Group | Cary-Blairpreserved | Unpreserved | Combined |
|---|---|---|---|
| 0-1 month | 6 | 0 | 6 |
| 1 month to 2 years | 250 | 66 | 316 |
| 2-12 | 311 | 164 | 475 |
| 13-18 | 141 | 85 | 226 |
| 19-21 | 44 | 23 | 67 |
| Over 21 | 671 | 621 | 1292 |
| Unknown | 16 | 5 | 21 |
| Total | 1439 | 964 | 2403 |
Compliant Clinical Trial Enrollment Summary by Age Group and Specimen Type Table 13:
Vibrio Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 100% and 99.6% of the Vibrio prospective and negative specimens, respectively, and 100% of the retrospective positive and negative specimens. For the unpreserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 99.8% of the Vibrio negative specimens. No unpreserved prospective positive was found for Vibrio, therefore no performance can be estimated. 100% and 97.8% of the retrospective positive and negative specimens respectively was identified (Table 14).
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As Vibrio prevalence is low, an evaluation of contrived samples was performed to supplement data collected in the study. These were prepared by spiking four (4) different strains for each species of Vibrio detected by BD MAX Extended Enteric Bacterial Panel in negative stool matrix. Strains were spiked at various clinically relevant loads and randomly distributed among three (3) clinical sites for BD MAX Extended Enteric Bacterial Panel testing. A positive agreement of 100% was obtained across the tested loads. Results are shown in Table 15.
| Specimen Type | Specimen Origin | BD MAX | Reference Method | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|---|
| P | N | Total | |||||
| Cary-Blair preserved | Prospective(Fresh+Frozen) | P | 2 | 5 | 7 | 100(34.2, 100) | 99.6(99.1, 99.8) |
| N | 0 | 1351 | 1351 | ||||
| Total | 2 | 1356 | 1358 | ||||
| Cary-Blair preserved | Retrospective(Frozen) | P | 2 | 0 | 2 | 100(34.2, 100) | 100(80.6, 100) |
| N | 0 | 16 | 16 | ||||
| Total | 2 | 16 | 18 | ||||
| Unpreserved | Prospective(Fresh+Frozen) | P | 0 | 2 | 2 | No Data forCalculation | 99.8(99.2, 99.9) |
| N | 0 | 866 | 866 | ||||
| Total | 0 | 868 | 868 | ||||
| Unpreserved | Retrospective(Frozen) | P | 2 | 1 | 3 | 100(34.2, 100) | 97.8(88.7, 99.6) |
| N | 0 | 45 | 45 | ||||
| Total | 2 | 46 | 48 |
Vibrio – Overall Performance Table 14:
While the BD MAX Extended Enteric Bacterial Panel is designed to detect V. paraheamoliticus, V. cholerae and V. vulnificus, the panel does not report results to the species level. The Vibrio species identification was obtained by sequencing of the alternate PCR performed from the reference method presumptive positive isolate. During the study, equal number of V. cholerae and V. parahaemoliticus were obtained and no V. vulnificus was obtained. Three (3) V. parahaemoliticus and three (3) V. cholerae were obtained.
| Specimen Type | BD MAX | Expected Result | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|
| Positive | Negative | Total | ||||
| Cary-Blair Preserved | Positive | 48 | 0 | 48 | 100(92.6, 100) | 100(97.4, 100) |
| Negative | 0 | 144 | 144 | |||
| Total | 48 | 144 | 192 | |||
| Unpreserved | Positive | 48 | 0 | 48 | 100(92.6, 100) | 100(97.4, 100) |
| Negative | 0 | 144 | 144 | |||
| Total | 48 | 144 | 192 |
Table 15: Vibrio Contrived Samples Results per Specimen Type
Plesiomonas shigelloides Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 99.9% of the Plesiomonas shigelloides prospective specimens, and 100% of the retrospective positive and negative specimens. No Cary-Blair prospective positive P. shigelloides were identified: therefore no performance can be estimated. For the unpreserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 99.9% of the P. shigelloides negative specimens. No prospective positive P. shigelloides were identified; therefore no performance can be estimated. 100% and 97.9% of the retrospective positive and negative specimens respectively was identified (Table 16).
As P. shigelloides prevalence is low, an evaluation of contrived samples was performed to supplement data collected in the study. These were prepared by spiking 12 different strains of
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P. shigelloides in negative stool matrix. Strains were spiked at various clinically relevant loads and randomly distributed among three (3) clinical sites for BD MAX Extended Enteric Bacterial Panel testing. A positive agreement of 100% was obtained across the tested loads. Results are shown in Table 17.
| Specimen Type | Specimen Origin | BD MAX | Reference Method | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|---|
| Positive | Negative | Total | |||||
| Cary-Blairpreserved | Prospective | Positive | 0 | 2 | 2 | No Data forCalculation | 99.9(99.5, 100) |
| Negative | 0 | 1355 | 1355 | ||||
| Total | 0 | 1357 | 1357 | ||||
| Cary-Blairpreserved | Retrospective | Positive | 4 | 0 | 4 | 100(51, 100) | 100(90.8, 100) |
| Negative | 0 | 38 | 38 | ||||
| Total | 4 | 38 | 42 | ||||
| Unpreserved | Prospective | Positive | 0 | 1 | 1 | No Data forCalculation | 99.9(99.3, 100) |
| Negative | 0 | 863 | 863 | ||||
| Total | 0 | 864 | 864 | ||||
| Unpreserved | Retrospective | Positive | 3 | 1 | 4 | 100(43.9, 100) | 97.9(88.9, 99.6) |
| Negative | 0 | 46 | 46 | ||||
| Total | 3 | 47 | 50 |
Plesiomonas shigelloides - Overall Performance Table 16:
| Table 17: Plesiomonas shigelloides Contrived Samples Results per Specimen Type |
|---|
| [Table Content Placeholder] |
| Specimen Type | BD MAX | Expected Result | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|
| Positive | Negative | Total | ||||
| Cary-Blair Preserved | Positive | 48 | 0 | 48 | 100 | 100 |
| Negative | 0 | 144 | 144 | (92.6, 100) | (97.4, 100) | |
| Total | 48 | 144 | 192 | |||
| Unpreserved | Positive | 48 | 1a | 49 | 100 | 99.3 |
| Negative | 0 | 143 | 143 | (92.6, 100) | (96.2, 99.9) | |
| Total | 48 | 144 | 192 |
4 Sample XW0007C was initially found positive for Plesiomonas shigelloides, but found negative for this target once retested from the SBT (Discrepant analysis).
Yersinia enterocolitica Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 99.9% of the Yersinia enterocolitica prospective negative specimens and 100% of the retrospective negative specimens. No Cary-Blair prospective and retrospective positive Y. enterocolitica were identified, therefore no performance can be estimated. For the unpreserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 100% of the Y. enterocolitica negative prospective specimens. No prospective Y. enterocolitica were identified, therefore no performance can be estimated. 100% of the retrospective positive and negative specimens was identified (Table 18).
As Y. enterocolitica prevalence is low, an evaluation of contrived samples was performed to supplement data collected in the study. These were prepared by spiking twelve (12) different strains of Y. enterocolitica in negative stool matrix. Strains were spiked at various clinically relevant loads and randomly distributed among three (3) clinical sites for BD MAX Extended Enteric Bacterial Panel testing. A positive agreement of 100% was obtained across the tested loads, except for one Cary-Blair preserved sample type at clinical load which the agreement obtained is 97.9% (Results are shown in Table 19).
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| Specimen Type | Specimen Origin | BD MAX | Reference Method | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|---|
| Positive | Negative | Total | |||||
| Cary-Blairpreserved | Prospective | Positive | 0 | 1 | 1 | No Data forCalculation | 99.9(99.6, 100) |
| Negative | 0 | 1341 | 1341 | ||||
| Total | 0 | 1342 | 1342 | ||||
| Cary-Blairpreserved | Retrospective | Positive | 0 | 0 | 0 | No Data forCalculation | 100(89.3, 100) |
| Negative | 0 | 32 | 32 | ||||
| Total | 0 | 32 | 32 | ||||
| Unpreserved | Prospective | Positive | 0 | 0 | 0 | No Data forCalculation | 100(99.6, 100) |
| Negative | 0 | 863 | 863 | ||||
| Total | 0 | 863 | 863 | ||||
| Unpreserved | Retrospective | Positive | 9 | 0 | 9 | 100% (70.1%,100%) | 100(92.4, 100) |
| Negative | 0 | 47 | 47 | ||||
| Total | 9 | 47 | 56 |
Table 18: Yersinia enterocolitica – Overall Performance
Table 19: Yersinia enterocolitica Contrived Samples Results per Specimen Type
| Specimen Type | BD MAX | Expected Result | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|
| Positive | Negative | Total | ||||
| Cary-Blair Preserved | Positive | 47 | 0 | 47 | 100(97.4, 100) | |
| Negative | 1b | 144 | 145 | 97.9(89.1, 99.6) | ||
| Total | 48 | 144 | 192 | |||
| Unpreserved | Positive | 48 | 0 | 48 | ||
| Negative | 0 | 144 | 144 | 100(92.6, 100) | 100(97.4, 100) | |
| Total | 48 | 144 | 192 |
8 Sample XW0351C was initially found negative for Yersinia enterocolitica, but found positive for this target once retested from the SBT (Discrepant analysis).
Enterotoxigenic E. coli (ETEC LT/ST) Performance Results
For the Cary-Blair preserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 100% and 99.8% of the ETEC prospective and negative specimens, respectively, and 100% of the retrospective positive and negative specimens. For the unpreserved specimen type, the BD MAX Extended Enteric Bacterial Panel identified 100% and 99.9% of the ETEC prospective positive and negative specimens, respectively, and 90% and 96.3% of the retrospective positive and negative specimens respectively was identified (Table 20).
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| Specimen Type | Specimen Origin | BD MAX | Comparator Method | PPA (%) | NPA (%) | ||
|---|---|---|---|---|---|---|---|
| Positive | Negative | Total | |||||
| Cary-Blairpreserved | Prospective | Positive | 10 | 3 | 13 | 100(72.2, 100) | 99.8(99.3, 99.9) |
| Negative | 0 | 1348 | 1348 | ||||
| Total | 10 | 1351 | 1361 | ||||
| Retrospective | Positive | 5 | 0 | 5 | 100(56.6, 100) | 100(87.9, 100) | |
| Negative | 0 | 28 | 28 | ||||
| Total | 5 | 28 | 33 | ||||
| Unpreserved | Prospective | Positive | 16 | 1 | 17 | 100(80.6, 100) | 99.9(99.3, 100) |
| Negative | 0 | 818 | 818 | ||||
| Total | 16 | 819 | 835 | ||||
| Retrospective | Positive | 9 | 1 | 10 | 90(59.6, 98.2) | 96.3(81.7, 99.3) | |
| Negative | 1 | 26 | 27 | ||||
| Total | 10 | 27 | 37 |
Table 20: Enterotoxigenic E. coli (ETEC LT/ST) - Overall Performance
The ETEC toxin identification was obtained by sequencing of the alternate PCR performed from stool specimens (Table 21). While the BD MAX Extended Enteric Bacterial Panel is designed to detect the toxin types described below, the panel does not report results to the species or toxin level.
| ETEC | LT | ST | Unknownc | |||
|---|---|---|---|---|---|---|
| PPA | STPPA | STpPPA | SThPPA | PPA | ||
| SpecimenType | Specimen Origin | EstimatePercent(95% CI) | EstimatePercent(95% CI) | EstimatePercent(95% CI) | EstimatePercent(95% CI) | EstimatePercent(95% CI) |
| Cary-BlairPreserved | Prospective | 100(4/4)(51.0, 100) | 100(5/5)(56.6, 100) | 100(2/2)(34.2, 100) | 100(1/1)(20.7, 100) | 100(3/3)(43.9, 100) |
| Retrospective | 100(4/4)(51.0, 100) | 100(5/5)(56.6, 100) | 100(4/4)(51.0, 100) | 0 | 0 | |
| Unpreserved | Prospective | 100(4/4)(51.0, 100) | 100(7/7)(64.6, 100) | 100(3/3)(43.9, 100) | 100(2/2)(34.2, 100) | 100(7/7)(64.6, 100) |
| Retrospective | 100(7/7)(64.6, 100) | 80.0(4/5)(37.6, 96.4) | 100(2/2)(34.2, 100) | 100(2/2)(34.2, 100) | 0 |
Table 21: ETEC Performance per Toxin Observed During the Clinical Trial
C ETEC toxin was detected but the specific species not identified.
Discrepant Results
There were nineteen (19) discrepant results in the clinical trial. Eighteen (18) falsepositive (FP) and one (1) false-negative (FN) result. Three (3) retrospective specimens, one (1) ETEC FN, one (1) ETEC FP and one (1) Vibrio FP, were not tested in the discrepant analysis, due to limited specimen volume. Of the sixteen (16) FP discrepant specimens tested, there were seven (7) FP Vibrio, four (4) P. shigelloides, one (1) Y. enterocolitica and four (4) ETEC. Five (5) out of the seven (7) Vibrio discrepant specimens were negative for both BD MAX Extended Enteric Bacterial Panel and the comparator method. The two (2) remaining Vibrio FP specimens, obtained a positive status in the BD MAX Extended Enteric Bacterial Panel coupled with a negative status in the comparator method testing in the discordance study.
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Out of the four (4) P. shigelloides discrepant results, one (1) was negative for both BD MAX Extended Enteric Bacterial Panel and the comparator method. Two (2) specimens obtained a positive status on the BD MAX Extended Enteric Bacterial Panel coupled with a negative status in the comparator method testing in the discordance study. They were also both positive for another target on the BD MAX Extended Enteric Bacterial Panel, ETEC and Y. enterocolitica, respectively. One (1) specimen, a Retrospective specimen, was Vibrio true positive in the study and P. shigelloides FP. In discrepant analysis this specimen was P. shigelloides negative for the comparator method and positive for Vibrio and P. shigelloides (1/3). The Y. enterocolitica FP specimen was negative for both BD MAX Extended Enteric Bacterial Panel and the comparator method in discrepant analysis. One (1) of the six (6) ETEC FP was negative for both BD MAX Extended Enteric Bacterial Panel and the comparator method. Two (2) ETEC specimens obtained a positive status on the BD MAX Extended Enteric Bacterial Panel coupled with an ETEC positive status in the RM testing in the discordance study. One (1) ETEC specimen obtained a positive status on the BD MAX Extended Enteric Bacterial Panel coupled with a negative status in the comparator method testing in the discordance study.
Non-Reportable
The initial unresolved rate was calculated when considering the unresolved rate of both BD MAX Enteric Bacterial Panel and BD MAX Extended Enteric Bacterial Panel assays. Of the 2264 prospective specimens initially evaluated, 3.1% of the Cary-Blair preserved and 3.9% of the unpreserved specimens initially reported as unresolved rate following a valid repeat test was calculated when considering BD MAX Extended Enteric Bacterial Panel only; 0.1% of the prospective Cary-Blair preserved specimens and 0.3% of the prospective unpreserved specimens remained unresolved. Of all the specimens initially evaluated with both BD MAX Enteric Bacterial Panel and BD MAX Extended Enteric Bacterial Panel assays, 1.0% of the Cary-Blair preserved and 1.5% of the unpreserved initially reported as Indeterminate. Following a valid repeat test, 0.1% of the Cary-Blair preserved and none of the unpreserved remained Indeterminate. No incompletes were reported during this study (Table 22).
| Unresolved Rate | Indeterminate Rate | Incomplete Rate | ||||
|---|---|---|---|---|---|---|
| SpecimenType | InitialxEBP(95% CI) | InitialEBP+xEBP(95% CI) | FinalxEBP(95% CI) | InitialEBP+xEBP(95% CI) | FinalEBP+xEBP(95% CI) | InitialEBP+xEBP(95% CI) |
| Cary-Blairpreserved | 2.435/1430(1.8, 3.4) | 3.144/1430(2.3, 4.1) | 0.11/1427(0.0, 0.4) | 1.015/1430(0.6, 1.7) | 0.11/1427(0.0, 0.4) | 00/1430(0.0, 0.3) |
| Unpreserved | 2.221/958(1.4, 3.3) | 3.937/958(2.8, 5.3) | 0.33/958(0.1, 0.9) | 1.514/958(0.9, 2.4) | 0.0/958(0.0, 0.4) | 00/958(0.0, 0.4) |
Table 22: Non-reportable Rates
§ 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).