Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description focuses on automated PCR and detection based on predefined criteria, with no mention of AI or ML algorithms for interpretation or analysis.

Therapeutic

No
The device is an in vitro diagnostic test used to detect and differentiate bacterial pathogens, aiding in diagnosis, not providing therapy.

Diagnostic

Yes
The "Intended Use / Indications for Use" section explicitly states that the device is an "automated in vitro diagnostic test for the direct qualitative detection and differentiation of enteric bacterial pathogens" and is intended "as an aid in the differential diagnosis."

SaMD (Software as a Medical Device)

No

The device description explicitly states that the BD MAX™ Extended Enteric Bacterial Panel is performed on the BD MAX™ System, which includes "instrument with associated hardware and accessories, disposable microfluidic cartridges, master mixes, unitized reagent strips, extraction reagents, and sample buffer tubes." This indicates the device is a system that includes both hardware and software components, not a software-only medical device.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the BD MAX™ Extended Enteric Bacterial Panel is an "automated in vitro diagnostic test".
Purpose: The test is designed for the "direct qualitative detection and differentiation of enteric bacterial pathogens" from patient specimens (stool). This is a core function of an in vitro diagnostic device.
Methodology: It utilizes real-time PCR and fluorogenic gene-specific hybridization probes to detect nucleic acids from specific pathogens, which are common techniques used in IVD tests.
Clinical Context: The test is intended to be used "in conjunction with clinical presentation, laboratory findings, and epidemiological information, as an aid in the differential diagnosis" of specific infections. This clearly indicates its role in the diagnostic process.
Specimen Type: The test is performed on "unpreserved or Cary-Blair preserved soft to diarrheal stool specimens from symptomatic patients". Testing on human specimens is characteristic of IVDs.
Device Description: The "Device Description" further reinforces that it is an "automated in vitro diagnostic test".

All of these points align with the definition and purpose of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for 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, 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.

Product codes

PCH, PCI, OOI

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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Stool specimens

Indicated Patient Age Range

Pediatric or adult patients

Intended User / Care Setting

Not Found

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

Not Found

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

The 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.

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

Precision Study:

Study Type: Within-laboratory precision
Sample Size: Each panel member (Vibrio cholerae, Plesiomonas shigelloides, ETEC, Yersinia enterocolitica) was tested with a minimum of 24 replicates per concentration category.
Key Results:
- True Negative (TN): 100% agreement with expected negative results for all targets (96/96).
- High Negative (HN): Agreement with expected negative varied by target, ranging from 41.7% to 58.3%.
- Low Positive (LP): High agreement with expected positive results; 100% for Vibrio, P. shigelloides, and ETEC, 97.9% for Y. enterocolitica.
- Moderate Positive (MP): High agreement with expected positive results; 100% for Vibrio, P. shigelloides, and Y. enterocolitica, 97.9% for ETEC.

Reproducibility Study (Site-to-Site):

Study Type: Multi-site reproducibility
Sample Size: 3 clinical sites, 2 operators per site, 2 runs per day for 5 days, totaling 30 runs. Panels were the same as in the Precision study.
Key Results:
- Overall TN (True Negative) agreement: 100% for all targets.
- HN (High Negative) agreement: Ranged from 30.0% to 48.9%.
- LP (Low Positive) agreement: Ranged from 97.8% to 100%.
- MP (Moderate Positive) agreement: Ranged from 98.9% to 100%.
- Quantitative Ct. Score and Cycle End Point values and their variability (SD and %CV) were also reported.

Reproducibility Study (Lot-to-Lot):

Study Type: Lot-to-lot reproducibility
Sample Size: 1 site, 2 operators, 2 runs per day for 5 days using 3 reagent lots, totaling 30 runs. Panels were the same as in the Precision study.
Key Results:
- Overall TN (True Negative) agreement: 100%.
- HN (High Negative) agreement: Ranged from 23.3% to 41.1%.
- LP (Low Positive) agreement: Ranged from 97.8% to 100%.
- MP (Moderate Positive) agreement: Ranged from 98.9% to 100%.
- Quantitative Ct. Score and Cycle End Point values and their variability (SD and %CV) were also reported.

Analytical Sensitivity (Limit of Detection or LoD):

Study Type: LoD determination
Sample Size: Each target organism tested with a minimum of 24 replicates per sample type (preserved or unpreserved), by 2 operators, using 3 different production lots. An additional 24 replicates were tested at the determined LoD concentration.
Key Results:
- LoD ranged from 34 to 539 CFU/SBT (Stool Buffer Tube) and 3,434 to 53,852 CFU/mL (in stool) for unpreserved specimens.
- LoD ranged from 79 to 257 CFU/SBT and 7,860 to 25,712 CFU/mL (in stool) for preserved specimens.

Analytical Inclusivity:

Study Type: Evaluation of assay target strains detection
Sample Size: 69 strains tested (Plesiomonas shigelloides (10), Yersinia enterocolitica (10), Vibrio (36), ETEC LT/ST (13)).
Key Results: 68 of 69 strains were correctly identified upon initial testing. One ETEC ST/LT strain (CCUG 38088) was detected at 10x LoD.

Analytical Specificity (Cross-Reactivity and Exclusivity):

Study Type: Evaluation of potential cross-reactivity with phylogenetically related species and other organisms.
Sample Size: 184 organisms (bacterial cells, yeasts, parasites, viruses) tested.
Key Results:
- Most organisms tested produced negative results.
- Two Vibrio mimicus strains produced positive results, but not at concentrations ≤ 1.0 x 10^4 cells/mL.
- Eight Vibrio species not associated with human infections were detected.
- In silico analysis suggested detection of two more Vibrio species (V. coralliilyticus, Moritella marina) and Vibrio HENC.

Interfering Substances:

Study Type: Evaluation of potential interference from biological and chemical substances.
Sample Size: 19 biological and chemical substances, plus 10 microorganisms.
Key Results:
- Nystatin cream (>3.1 mg/mL), Spermicidal lubricant (>2.5 mg/mL), Hydrocortisone cream (>2.5 mg/mL), and Vagisil (>0.92 mg/mL) caused interference.
- No reportable interference from other substances or any of the 10 tested microorganisms.

Carryover/Cross-Contamination:

Study Type: Evaluation of within-run and between-run carryover.
Sample Size: 108 negative samples alongside high positive samples in 9 runs.
Key Results: One out of 108 negative samples produced a positive ETEC ST/LT result.

Mixed Infection/Competitive Interference:

Study Type: Evaluation of detection in the presence of other high-concentration targets.
Key Results: All three low target organisms were successfully detected in the presence of high loads of Plesiomonas shigelloides, Vibrio cholerae, Yersinia enterocolitica, and ETEC ST/LT in simulated mixed infections.

Clinical Performance Studies:

Study Type: Multi-site investigational study.
Sample Size: 2264 prospective specimens (882 unpreserved, 1382 Cary-Blair preserved) and 146 retrospective specimens (87 unpreserved, 59 Cary-Blair preserved), totaling 2410 specimens. 2403 compliant specimens in total.
Key Results:
- Vibrio Performance:
  - Cary-Blair preserved (Prospective): PPA 100% (2/2), NPA 99.6% (1351/1356).
  - Cary-Blair preserved (Retrospective): PPA 100% (2/2), NPA 100% (16/16).
  - Unpreserved (Prospective): No Data for Calculation (0/0 positive), NPA 99.8% (866/868).
  - Unpreserved (Retrospective): PPA 100% (2/2), NPA 97.8% (45/46).
  - Contrived samples (Vibrio): PPA 100% (48/48) for both Cary-Blair and Unpreserved types.
- Plesiomonas shigelloides Performance:
  - Cary-Blair preserved (Prospective): No Data for Calculation (0/0 positive), NPA 99.9% (1355/1357).
  - Cary-Blair preserved (Retrospective): PPA 100% (4/4), NPA 100% (38/38).
  - Unpreserved (Prospective): No Data for Calculation (0/0 positive), NPA 99.9% (863/864).
  - Unpreserved (Retrospective): PPA 100% (3/3), NPA 97.9% (46/47).
  - Contrived samples (P. shigelloides): PPA 100% for Cary-Blair preserved (48/48) and Unpreserved (48/48).
- Yersinia enterocolitica Performance:
  - Cary-Blair preserved (Prospective): No Data for Calculation (0/0 positive), NPA 99.9% (1341/1342).
  - Cary-Blair preserved (Retrospective): No Data for Calculation (0/0 positive), NPA 100% (32/32).
  - Unpreserved (Prospective): No Data for Calculation (0/0 positive), NPA 100% (863/863).
  - Unpreserved (Retrospective): PPA 100% (9/9), NPA 100% (47/47).
  - Contrived samples (Y. enterocolitica): PPA 97.9% for Cary-Blair preserved (47/48) and 100% for Unpreserved (48/48).
- Enterotoxigenic E. coli (ETEC LT/ST) Performance:
  - Cary-Blair preserved (Prospective): PPA 100% (10/10), NPA 99.8% (1348/1351).
  - Cary-Blair preserved (Retrospective): PPA 100% (5/5), NPA 100% (28/28).
  - Unpreserved (Prospective): PPA 100% (16/16), NPA 99.9% (818/819).
  - Unpreserved (Retrospective): PPA 90% (9/10), NPA 96.3% (26/27).
- Discrepant Results: 19 discrepant results (18 false positives, 1 false negative). 3 retrospective samples not retested due to volume. Of 16 FP retested, 7 Vibrio, 4 P. shigelloides, 1 Y. enterocolitica, 4 ETEC.
- Non-Reportable Rates:
  - Initial Unresolved Rate (xEBP only): 2.4% (Cary-Blair), 2.2% (Unpreserved).
  - Final Unresolved Rate (xEBP only): 0.1% (Cary-Blair), 0.3% (Unpreserved).
  - Initial Indeterminate Rate (EBP+xEBP): 1.0% (Cary-Blair), 1.5% (Unpreserved).
  - Final Indeterminate Rate (EBP+xEBP): 0.1% (Cary-Blair), 0% (Unpreserved).
  - No Incomplete results reported.

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

Key metrics reported are:

Percent Agreement (Agreement with Expected Results) for Precision and Reproducibility studies.
- True Negative (TN), High Negative (HN), Low Positive (LP), Moderate Positive (MP).
Limit of Detection (LoD) for Analytical Sensitivity.
Positive Percent Agreement (PPA) for Clinical Performance.
Negative Percent Agreement (NPA) for Clinical Performance.
Quantitative metrics like Mean Ct. Score and Mean Cycle End Point with Standard Deviation (SD) and % Coefficient of Variation (CV) for reproducibility studies.
Non-reportable rates (Unresolved, Indeterminate, Incomplete).

Predicate Device(s)

K160459

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 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).

Summary

0

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, stacked one behind the other.

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.

1

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.

7

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 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 FilmArray Gastrointestinal (GI) Panel is a
qualitative multiplexed nucleic acid-based in vitro
diagnostic test intended for use with FilmArray
systems. The FilmArray GI Panel is capable of the
simultaneous detection and identification of nucleic
acids from multiple bacteria, viruses, and parasites
directly from stool samples in Cary Blair transport
media obtained from individuals with signs and/or
symptoms of gastrointestinal infection. The
following bacteria (including several diarrheagenic
E. coli/Shigella pathotypes), parasites, and viruses
are 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 of
Vibrio 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 identification
of the E. coli O157 serogroup 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 FilmArray GI Panel 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. Positive results do not rule
out co-infection with organisms not included in the
FilmArray GI Panel. The agent detected may not be
the 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 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 E. coli 0157, Plesiomonas
shigelloides, Yersinia enterocolitica, Astrovirus,
and Rotavirus A were established primarily with
retrospective clinical specimens.
Performance characteristics for Entamoeba
histolytica, and Vibrio (V. parahaemolyticus, V.
vulnificus, and Vibrio cholerae) were established |
| | | primarily using contrived clinical specimens.
Negative FilmArray GI Panel 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.
A gastrointestinal microorganism multiplex nucleic
acid-based assay also aids in the detection and
identification of acute gastroenteritis in the context
of outbreaks. |
| Specimen Type | Cary-Blair preserved stool
Unpreserved soft to diarrheal stool | Cary-Blair preserved stool.
Not claimed (see Differences below) |
| Assay Format | Amplification: PCR
Detection: fluorogenic target-specific hybridization. | Amplification: PCR
Detection: non target-specific fluorescent dye |
| Organisms
Detected | • Plesiomonas shigelloides
• Vibrio (V. vulnificus, V. parahaemolyticus, and V.
cholerae)
• Enterotoxigenic Escherichia coli heat labile and
heat stabile (LT/ST) (ETEC)
• Yersinia enterocolitica | • Plesiomonas shigelloides
• Vibrio (V. parahaemolyticus/V. vulnificus/ V.
cholerae), including specific identification of
Vibrio cholerae
• Enterotoxigenic Escherichia coli (ETEC) lt/st
• Yersinia enterocolitica |
| Interpretation of
Test Results | Automated: BD MAX System diagnostic software | Automated |
| Analysis Platform | BD MAX™ System | Film Array Instrument |
| PCR Sample
preparation | Automated:
BD MAX™ System | Automated:
Film Array Instrument |
| Detection Probes | TaqMan® Probe | Fluorescent double stranded DNA binding dye (LC
Green Plus) |
| Assay Controls | Sample Processing Control (SPC) | Two controls are included in each reagent pouch to
control for sample processing and both stages of
PCR and melt analysis. |
| Differences | | |
| Item | BD MAX TM xEBP | FilmArray GI Panel |
| Specimen Type | Unpreserved soft to diarrheal stool | Not claimed |
| Organisms
Detected | 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 O157
serogroup 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

9

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 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	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)
HNa	48.9
44/90
(38.8, 59.0)	30.0
27/90
(21.5, 40.1)	35.6
32/90
(26.4, 45.8)	46.7
42/90
(36.7, 56.9)
LP	100
90/90
(95.9, 100)	98.9
89/90
(94.0, 99.8)	100
90/90
(95.9, 100)	97.8
88/90
(92.3, 99.4)
MP	100
90/90
(95.9, 100)	98.9
89/90
(94.0, 99.8)	98.9
89/90
(94.0, 99.8)	100
90/90
(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 Metric | Category | N | Mean Within Day | | Between Day
Within Site | | Between Site | |-----------------------------| | | | Vibrio Score | HN | | | | End
Point | HN | 46 | 805.1 | 334.64 | | | | | Plesiomonas
shigelloides | Ct.
Score | HN | | | | | End
Point | HN | 63 | 596.5 | 284.02 | | | | | Yersinia
enterocolitica | Ct.
Score | HN | | | | | Point | HN | | | | | ETEC Score | HN | | | | | End
Point | HN | 48 | 976.0 | 577.46 | | | | | SPC Score | TN | End
Point | TN | 90 | 6020.7 | 672.52 | PCR
| Within Run | | Between Run
| Total | |
-----------------------|----------|----|--------|------------|------|---------------------------|------|----------------------------|-----|--------------|------|--------|------|
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Ct.
| 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 |
| Cycle
| 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 |
| 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 |
| Cycle
| 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 |
| 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 |
| End
| 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 |
| Ct.
| 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 |
| Cycle
| 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 |
| Ct.
| 90 | 27.2 | 0.35 | 1.3 | 0.00 | 0.0 | 0.00 | 0.0 | 0.18 | 0.7 | 0.40 | 1.5 |
| Cycle
| 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

| PCR

Metric	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
Cycle
EP	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	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)	100
180/180
(97.9, 100)
HNa	41.1
37/90
(31.5, 51.4)	28.9
26/90
(20.5, 39.0)	23.3
21/90
(15.8, 33.1)	41.1
37/90
(31.5, 51.4)
LP	100
90/90
(95.9, 100)	100
90/90
(95.9, 100)	100
90/90
(95.9, 100)	97.8
88/90
(92.3, 99.4)
MP	100
90/90
(95.9, 100)	100
90/90
(95.9, 100)	98.9
89/90
(94.0, 99.8)	100
90/90
(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 | PCR
Metric | Category | N | Mean | Within Run | | Between Run
Within Day | | Between Day
Within 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 |
| | Cycle
End
Point | 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 |
| Plesiomonas
shigelloides | 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 |
| | Cycle
End
Point | 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 |
| Yersinia
enterocolitica | 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 |
| | End
Point | 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 |
| | Cycle
End
Point | 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 |
| | Cycle
End
Point | 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

| PCR

Metric	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
Cycle
EP	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) ●

14

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)
Plesiomonas
shigelloides
ATCC 14029b;
ATCC 14030c	458	45,752	155	15,481	257	25,712
Yersinia
enterocolitica
ATCC 9610b;
CCUG 4588c	209	20,900	311	31,099	227	22,723
ETEC ST/LT
ATCC 35401b;
CCUG 47194c	34	3,434	539	53,852	137	13,706
Vibrio cholerae
ATCC 14033b;
ENF 9786c	149	14,942	43	4,344	252	25,238
Vibrio
parahaemolyticus
ATCC17802b;
ENF 5887c	207	20,708	80	8,031	124	12,424
Vibrio vulnificus
ATCC 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

15

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 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 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-Blair
preserved | 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
			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 for
Calculation	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 (%)
		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

19

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 (%)
			Positive	Negative	Total
Cary-Blair
preserved	Prospective	Positive	0	2	2	No Data for
Calculation	99.9
(99.5, 100)
		Negative	0	1355	1355
		Total	0	1357	1357
Cary-Blair
preserved	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 for
Calculation	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).

20

Specimen Type	Specimen Origin	BD MAX	Reference Method			PPA (%)
			Positive	Negative	Total
Cary-Blair
preserved	Prospective	Positive	0	1	1	No Data for
Calculation	99.9
(99.6, 100)
		Negative	0	1341	1341
		Total	0	1342	1342
Cary-Blair
preserved	Retrospective	Positive	0	0	0	No Data for
Calculation	100
(89.3, 100)
		Negative	0	32	32
		Total	0	32	32
Unpreserved	Prospective	Positive	0	0	0	No Data for
Calculation	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).

21

Specimen Type	Specimen Origin	BD MAX	Comparator Method			PPA (%)
			Positive	Negative	Total
Cary-Blair
preserved	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
		PPA	ST
PPA	STp
PPA	STh
PPA	PPA
Specimen
Type	Specimen Origin	Estimate
Percent
(95% CI)	Estimate
Percent
(95% CI)	Estimate
Percent
(95% CI)	Estimate
Percent
(95% CI)	Estimate
Percent
(95% CI)
Cary-Blair
Preserved	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.

22

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
Specimen
Type	Initial
xEBP
(95% CI)	Initial
EBP+xEBP
(95% CI)	Final
xEBP
(95% CI)	Initial
EBP+xEBP
(95% CI)	Final
EBP+xEBP
(95% CI)	Initial
EBP+xEBP
(95% CI)
Cary-Blair
preserved	2.4
35/1430
(1.8, 3.4)	3.1
44/1430
(2.3, 4.1)	0.1
1/1427
(0.0, 0.4)	1.0
15/1430
(0.6, 1.7)	0.1
1/1427
(0.0, 0.4)	0
0/1430
(0.0, 0.3)
Unpreserved	2.2
21/958
(1.4, 3.3)	3.9
37/958
(2.8, 5.3)	0.3
3/958
(0.1, 0.9)	1.5
14/958
(0.9, 2.4)	0.
0/958
(0.0, 0.4)	0
0/958
(0.0, 0.4)

Table 22: Non-reportable Rates