K Number
K143648
Date Cleared
2015-08-25

(246 days)

Product Code
Regulation Number
866.3990
Reference & Predicate Devices
Predicate For
AI/MLSaMDIVD (In Vitro Diagnostic)TherapeuticDiagnosticis PCCP AuthorizedThirdpartyExpeditedreview
Intended Use

The BD MAX™ Enteric Parasite Panel performed on the BD MAX™ System is an automated in vitro diagnostic test for the direct qualitative detection of enteric pathogens. The BD MAX Enteric Parasite Panel detects nucleic acids from:
• Giardia lamblia

  • · Cryptosporidium (C. hominis and C. parvum only)
  • Entamoeba histolytica
    Testing is performed on unpreserved or 10% formalin-fixed stool specimens from symptomatic patients with suspected gastroenteritis, enteritis or colitis. The assay is intended to aid in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation and other laboratory findings. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of specific targets. The test utilizes fluorogenic gene-specific hybridization probes for 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 Giardia lamblia, Cryptosporidium hominis and C. parvum, as well as Entamoeba histolytica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decision. 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 ss compatible with gastroenteritis and/or colitis 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.

Device Description

The BD MAX™ System and the BD MAX™ Enteric Parasite Panel are comprised of an 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 results. A test result may be called as POS, NEG or UNR 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 MAX™ System failure.

AI/ML Overview

The provided document describes the analytical and clinical performance of the BD MAX™ Enteric Parasite Panel, an in vitro diagnostic test. The acceptance criteria and device performance are outlined in the "Clinical Performance Studies" section.

Here's a breakdown of the requested information:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state pre-defined acceptance criteria for the clinical performance endpoints in a single, clear table. However, the reported performance metrics (Sensitivity, Specificity, PPA, NPA) for each target and specimen type are presented. For the purpose of this response, I will interpret the reported performance values themselves as the "device performance" and highlight the key metrics that would typically serve as acceptance criteria in such a submission.

Target & Specimen TypePerformance Metric (Acceptance Criteria Implied)Reported Device Performance
Giardia lamblia
10% Formalin-Fixed (Prospective)Sensitivity95.5% (95% CI: 78.2%, 99.2%)
Specificity99.7% (95% CI: 99.1%, 99.9%)
10% Formalin-Fixed (Retrospective)Positive Percent Agreement (PPA)100.0% (95% CI: 93.5%, 100.0%)
Negative Percent Agreement (NPA)100.0% (95% CI: 94.9%, 100.0%)
Unpreserved (Prospective)Sensitivity94.4% (95% CI: 74.2%, 99.0%)
Specificity100.0% (95% CI: 99.4%, 100.0%)
Unpreserved (Retrospective)PPA98.6% (95% CI: 92.6%, 99.8%)
NPA94.9% (95% CI: 89.8%, 97.5%)
Cryptosporidium parvum/hominis
10% Formalin-Fixed (Prospective)Sensitivity90.3% (95% CI: 80.5%, 95.5%)
Specificity99.8% (95% CI: 99.2%, 99.9%)
10% Formalin-Fixed (Retrospective)PPA93% (95% CI: 81.4%, 97.6%)
NPA100% (95% CI: 95.3%, 100%)
Unpreserved (Prospective)Sensitivity100% (95% CI: 90.1%, 100%)
Specificity99.5% (95% CI: 98.6%, 99.8%)
Unpreserved (Retrospective)PPA97.7% (95% CI: 88.2%, 99.6%)
NPA98.4% (95% CI: 95.3%, 99.4%)
Entamoeba histolytica
10% Formalin-Fixed (Prospective)Specificity100% (95% CI: 99.5%, 100%)
10% Formalin-Fixed (Retrospective)NPA100% (95% CI: 93.4%, 100%)
Unpreserved (Prospective)Specificity100% (95% CI: 99.3%, 100%)
Unpreserved (Retrospective)PPA100% (95% CI: 74.1%, 100%)
NPA100% (95% CI: 98.0%, 100%)
Entamoeba histolytica (Contrived)PPA100% (95% CI: 92.9%, 100%)
NPA100% (95% CI: 92.9%, 100%)
Unresolved Rates (Final)Max Unresolved Rate (10% FF)0.0% (0/1084-0/146)
Max Unresolved Rate (Unpreserved)1.5% (11/747-0/244)
Indeterminate Rates (Final)Max Indeterminate Rate (all types)0.0%
Incomplete Rates (Final)Max Incomplete Rate (all types)0.0%

2. Sample Size Used for the Test Set and Data Provenance

  • Total Compliant Specimens: 2495 (1273 10% formalin-fixed, 1222 unpreserved).
  • Split by Type:
    • Prospective: 2204 specimens (1128 10% formalin-fixed, 1058 unpreserved, 18 noncompliant - excluded).
    • Retrospective: 411 specimens (148 10% formalin-fixed, 251 unpreserved, 12 noncompliant - excluded).
    • Excluded from Performance Calculations: 128 retrospective specimens (historical results not confirmed by alternate PCR and bi-directional sequencing).
  • Adjusted Compliant Specimens for Performance:
    • Giardia lamblia: 1021 (10% Formalin-Fixed Prospective), 126 (10% Formalin-Fixed Retrospective), 673 (Unpreserved Prospective), 209 (Unpreserved Retrospective)
    • Cryptosporidium parvum/hominis: 1015 (10% Formalin-Fixed Prospective), 121 (10% Formalin-Fixed Retrospective), 663 (Unpreserved Prospective), 228 (Unpreserved Retrospective)
    • Entamoeba histolytica: 827 (10% Formalin-Fixed Prospective), 54 (10% Formalin-Fixed Retrospective), 577 (Unpreserved Prospective), 202 (Unpreserved Retrospective)
    • Entamoeba histolytica (Contrived): 100 (50 10% formalin-fixed, 50 unpreserved)
  • Data Provenance: The study was a multicenter clinical study with specimens collected as part of routine patient care. It involved five (5) clinical sites and seven (7) collection sites. An internal site also performed testing on specimens from collection centers. The data includes both prospective and retrospective specimens. The country of origin of the data is not explicitly stated, but given the FDA submission, it is likely primarily from the USA, though multi-site studies can include international sites.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

The document describes a "reference method" for establishing ground truth, but does not specify the number or qualifications of experts (e.g., medical technologists, pathologists, or specific specialists) involved in establishing the ground truth.

The "reference method" included:

  • Standard microscopic examination (possibly O&P, though not explicitly named as the sole method).
  • For confirmed positive specimens, an "alternate PCR and bi-directional sequencing" was used.
  • For Giardia lamblia and Cryptosporidium, DFA (Direct Fluorescent Antibody) was also part of the reference method.
  • Discrepant analysis involved repeat testing with the alternate PCR and bi-directional sequencing, DFA, EIA (Enzyme Immunoassay), and repeat testing with the BD MAX™ Enteric Panel.

4. Adjudication Method for the Test Set

The document details a discrepant resolution process for the clinical study:

  • For specimens where the BD MAX Enteric Parasite Panel result differed from the initial reference method, "discrepant repeat testing" was performed.
  • This repeat testing included:
    • Alternate PCR and bi-directional sequencing.
    • DFA (for Giardia and Cryptosporidium).
    • EIA (for Giardia).
    • Repeat testing with the BD MAX™ Enteric Panel (often in multiple replicates, e.g., 6 or 12 replicates).

This indicates an adjudication method that goes beyond a simple "x+1" or "x+y" rule, involving multiple different assays and follow-up analysis to establish a confirmed ground truth, particularly for discrepant results. It's a method driven by discrepancy rather than a fixed "consensus" rule initially.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study comparing human readers with and without AI assistance was not mentioned or performed. This device is an automated molecular diagnostic test and does not involve human "readers" in the interpretation of results in the way an imaging AI device would. The BD MAX System itself automates the interpretation of test results.

6. Standalone (Algorithm Only Without Human-in-the-Loop Performance) Study

Yes, this entire study can be considered a standalone performance evaluation. The BD MAX™ Enteric Parasite Panel is an automated in vitro diagnostic test, and its performance was evaluated against a reference method (ground truth) without a human-in-the-loop component for the device's operation or result interpretation. The "Test Principle" section explicitly states, "The BD MAX™ System software automatically interprets test results. A test result may be called as POS, NEG or UNR for each of the assay's targets..."

7. Type of Ground Truth Used

The ground truth for the clinical studies was established using a composite reference method consisting of:

  • Microscopic examination (implied, given the nature of parasite detection tests, though not explicitly detailed for all cases).
  • Alternate PCR and bi-directional sequencing: This is a highly specific molecular method for confirming the presence and identity of the target pathogens.
  • Direct Fluorescent Antibody (DFA): Used for Giardia and Cryptosporidium.
  • Enzyme Immunoassay (EIA): Used in discrepant analysis for Giardia.

For Entamoeba histolytica, given its rarity, contrived specimens (spiked residual negative clinical specimens) were also used to supplement clinical data for evaluating positive percent agreement.

8. Sample Size for the Training Set

The document does not explicitly state a sample size for a "training set" in the context of machine learning or algorithm development. This device is a real-time PCR assay, and its performance is based on the specificity of its primers and probes and the robustness of the extraction and amplification process, rather than a machine learning model that requires a distinct training set. The "Analytical Inclusivity" and "Analytical Specificity" sections describe testing against known strains and related organisms, which could be considered akin to "training" or "development" data for assay design, but not "training set" in the common AI/ML sense.

9. How the Ground Truth for the Training Set Was Established

As there's no explicitly defined "training set" in the AI/ML sense, there's no ground truth established in that context. However, for the analytical studies (e.g., Limit of Detection, Inclusivity, Specificity), the ground truth was established by:

  • Known concentrations of purified organisms: For LoD, organisms were inoculated at specific concentrations into negative stool matrix.
  • Known strains/isolates: For inclusivity, specific ATCC strains or known clinical isolates of the target organisms were used.
  • Known phylogenetically related species and other organisms: For specificity, a wide range of known bacteria, viruses, parasites, and yeast were tested at specified concentrations.

This involves laboratory-controlled methods, where the "ground truth" (presence/absence and concentration of a specific organism) is inherently known by design.

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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 an abstract symbol that resembles three human profiles facing to the right, stacked on top of each other.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

August 27, 2015

BECTON, DICKINSON AND COMPANY PAUL SWIFT REGULATORY AFFAIRS MANAGER 7 LOVETON CIR. SPARKS MD 21152

Re: K143648

Trade/Device Name: BD Max Enteric Parasite Panel, BD Max Instrument Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid based assay Regulatory Class: II Product Code: PCH, OOI Dated: July 30, 2015 Received: July 31, 2015

Dear Mr. Swift:

This letter corrects our substantially equivalent letter of August 25, 2015.

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

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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.

If you desire specific advice for your device on our labeling regulation (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/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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,

Ribhi Shawar -S

For Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health (OIR) Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K143648

Device Name BD MAX™ Enteric Parasite Panel

Indications for Use (Describe)

The BD MAX™ Enteric Parasite Panel performed on the BD MAX™ System is an automated in vitro diagnostic test for the direct qualitative detection of enteric pathogens. The BD MAX Enteric Parasite Panel detects nucleic acids from:

• Giardia lamblia

  • · Cryptosporidium (C. hominis and C. parvum only)
  • Entamoeba histolytica

Testing is performed on unpreserved or 10% formalin-fixed stool specimens from symptomatic patients with suspected gastroenteritis, enteritis or colitis. The assay is intended to aid in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation and other laboratory findings. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of specific targets. The test utilizes fluorogenic gene-specific hybridization probes for 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 Giardia lamblia, Cryptosporidium hominis and C. parvum, as well as Entamoeba histolytica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decision. 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 ss compatible with gastroenteritis and/or colitis 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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510(k) Summary

BD MAX Enteric Parasite Panel (EPP)

Summary Preparation Date:

August 18, 2015

Submitted by:

BD Diagnostic Systems Becton, Dickinson and Company 7 Loveton Circle Sparks, Maryland 21152

Contact:

Paul Swift, RAC Regulatory Affairs Manager

Tel: 410-316-4905 Fax: 410-316-4188 Email: Paul_Swift@bd.com

Proprietary Names:

For the instrument:

BD MAX™M

For the assay:

BD MAX™ Enteric Parasite Panel (EPP)

Common Names:

For the instrument: Bench-top molecular diagnostics workstation

For the assay:

Enteric Parasite Nucleic Acid Test Enteric Parasite identification and differentiation system Enteric assay Enteric test

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Regulatory Information

Regulation section:

  1. 3990 - Gastrointestinal microorganism multiplex nucleic acid-based assay

Classification:

Class II

Panel:

Microbiology (83)

Product Code(s):

PCH Gastrointestinal Pathogen Panel Multiplex Nucleic Acid-Based Assay System

Predicate Device

FilmArray Gastrointestinal (GI) Panel Kit [K140407 (May 2, 2014)], BioFire Diagnostics, LLC

Device Establishment

GeneOhm Sciences Canada, Inc. (BD Diagnostics) 2555 Boul. du Parc-Technologique Quebec, QC G1P 4S5 Canada

Registration Number: 3007420875

Performance standards

No performance standards have been developed under Section 514 of the Food, Drug and Cosmetic Act.

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Intended Use

The BD MAX™ Enteric Parasite Panel performed on the BD MAX™ System is an automated in vitro diagnostic test for the direct qualitative detection of enteric pathogens. The BD MAX Enteric Parasite Panel detects nucleic acids from:

  • Giardia lamblia
  • Cryptosporidium (C. hominis and C. parvum only)
  • . Entamoeba histolytica

Testing is performed on unpreserved or 10% formalin-fixed stool specimens from symptomatic patients with suspected gastroenteritis or colitis. The assay is intended to aid in the diagnosis of gastrointestinal infection when used in conjunction with clinical evaluation and other laboratory findings. The test is performed directly on the specimen, utilizing real-time polymerase chain reaction (PCR) for the amplification of specific targets. The test utilizes fluorogenic gene-specific hybridization probes for 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 Giardia lamblia. Cryptosporidium hominis and C. parvum, as well as Entamoeba histolytica infections. Results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decision. 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 and/or colitis may be due to infection by pathogens that are not detected by this test or noninfectious 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™ System and the BD MAX™ Enteric Parasite Panel are comprised of an 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 results. A test result may be called as POS, NEG or UNR 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 MAX™ System failure.

Test Principle

Stool specimens are collected from patients and transported to the laboratory either unpreserved in a clean container or 10% formalin-fixed. The specimen is vortexed and a 10 uL loop is inserted to the depth of the loop into the specimen, and expressed using a swirling motion into a BD MAX Sample Buffer Tube (SBT). The SBT is closed with a septum cap and then heated on the BD Pre-Warm Heater to facilitate

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lysis of the parasite organisms. The SBT is then vortexed and transferred to the BD MAX System. Once the work list is generated and the clinical sample is loaded on the BD MAX instrument with a EPP Unitized Reagent Strip (URS) and PCR Cartridge, the run is started and no further operator intervention is required. The BD MAX System automates sample preparation, including target organism lysis, DNA extraction and concentration, reagent rehydration, target nucleic acid sequence amplification and detection using real-time PCR. The interpretation of the signal is performed automatically by the BD MAX System. The assay also includes a Sample Processing Control (SPC) that is provided in the Extraction Tube and subjected to extraction, concentration and amplification steps. The SPC is incorporated into the lysis, extraction and amplification steps to monitor for the presence of potential inhibitory substances as well as system or reagent failures.

Following enzymatic cell lysis at an elevated temperature, the released nucleic acids are captured on magnetic affinity beads. The beads, with the bound nucleic acids, are washed and the nucleic acids are eluted by heat and high pH in Elution Buffer. Eluted DNA is pH neutralized and transferred to the Master Mix Tube to rehydrate the PCR reagents. After rehydration, the BD MAX System dispenses a fixed volume of DNA eluent 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 heat evaporation and contamination. The amplified DNA targets are detected using hydrolysis (TagMan") 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 amplicons for enteric parasite targets and the Sample Processing Control in four different optical channels of the BD MAX System: Giardia lamblia target amplicons are detected in the FAM channel, Cryptosporidium parvum/hominis target amplicons are detected in the ROX channel, Entamoeba histolytica target amplicons are detected in the VIC channel and Sample Processing Control amplicons are detected in the Cy5.5 channel.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 DNA 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 differences between the BD MAX Enteric Parasite Panel and the predicate device.

Table 1: Comparison to Predicate Device
ITEMBD MAX Enteric Parasite PanelFilmArray Gastrointestinal (GI) Panel Kit[K140407 (May 2, 2014)]BioFire Diagnostics, LLC
Specimen typeUnpreserved and 10% formalin-fixed stool.Human stool samples collected in Cary Blair transport media.
Assay FormatAmplification: PCRDetection: fluorogenic target-specific hybridization.Nested multiplex RT-PCR followed by high resolution melting analysis to confirm identity of amplified product.
Interpretation of Test ResultsAutomated (BD MAX System diagnostic software)Automated test interpretation and report generation by the FilmArray™ Instrument.
Analysis PlatformBD MAX SystemFilmArray™ Instrument
PCR Sample PreparationAutomated by the BD MAX SystemAutomated by the FilmArray™ Instrument
Detection ProbesTaqMan® ProbeUnknown - information not available publicly
Assay ControlsSample Processing Control (SPC)Two controls are included in each reagent pouch to control for sample processing and both stages of PCR and melt analysis

Таы» 1. С. to Predicate Da

1 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 21 CFR 807, Subpart E under which a device can be marketed without pre-market approval or reclassification. A determination of substantial equivalency under this notification is 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 their application by the courts.

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Analytical Performance

Precision

Within-laboratory precision was evaluated for the BD MAX™ Enteric Bacterial Panel at one (1) site. The Precision panel consisted of 4 sample categories near the LoD. Each specimen contained negative stool matrix. Target strains were tested as follows:

  • For moderate positives (MP): overall correct percentage of approximately 100% with 95% CI
  • . For low positives (LP): overall correct percentage of approximately 95% with 95% CI
  • . For true negatives (TN): overall correct percentage of approximately 100% with 95% CI
  • For high negatives (HN): overall correct percentage between 20 and 80% ●

Testing was performed in triplicate, over 12 days, with 2 runs per day, by 2 different technologists. Precision study results are summarized below in Table 2.

Panel Member LevelSpecimen/TargetPercentage Correct (%)
High Negative (HN)G. lamblia48.6
C. parvum41.7
E. histolytica37.5
Low Positive (LP)G. lamblia98.6
C. parvum98.6
E. histolytica98.6
Moderate Positive (MP)ALL100
True Negative (TN)ALL100

Table 2: Within-laboratory Precision Testing

Reproducibility

For the Site-to-Site reproducibility study, three (3) clinical sites were provided with a total of sixteen (16) panels, each consisting of 12 tubes. The panels used were the same as described under the Precision heading, above. Each site was asked to perform the study on eight (8) distinct days (consecutive or not), wherein each day, two (2) panels were tested, one (1) for each of two (2) technologists.

The overall Site-to-Site Reproducibility percent agreement was 100% for the TN and MP categories for all targets, and ranged from 38.2% to 48.6% and 97.2% to 98.6% for the HN and LP categories, respectively (Table 3). The qualitative and quantitative reproducibility across sites and by target is presented below in Tables 4 through 9. Ct.Score is an internal criterion used to determine final assay results and was selected as an additional means of assessing assay

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reproducibility. Overall mean Ct.Score values with variance components (SD and %CV) are shown in Tables 5, 7 and 9.

CategoryGiardia lambliaCryptosporidium parvumEntamoeba histolytica
TN100.0% (144/144)(97.4%, 100.0%)100.0% (144/144)(97.4%, 100.0%)100.0% (144/144)(97.4%, 100.0%)
HN48.6% (70/144)(40.6%, 56.7%)38.2% (55/144)(30.7%, 46.3%)47.2% (68/144)(39.2%, 55.3%)
LP97.2% (140/144)(93.1%, 98.9%)97.2% (140/144)(93.1%, 98.9%)98.6% (142/144)(95.1%, 99.6%)
MP100.0% (144/144)(97.4%, 100.0%)100.0% (144/144)(97.4%, 100.0%)100.0% (144/144)(97.4%, 100.0%)

Table 3: Site-to-Site Reproducibility Study Results using one lot of the BD MAX Enteric Parasite Panel

Table 4: G. lamblia Site-to-Site Qualitative Reproducibility Across Sites with Pooled Days, Runs and Replicates

CategoryX LoDSITETotal
CINCLEBD NCCorrectIncorrect
CorrectIncorrectCorrectIncorrectCorrectIncorrectn%n%
TN048100.00048100.00048100.000144100.000
HN0.1-0.52347.92552.12245.82654.22552.12347.97048.67451.4
LP0.8-1.24695.824.24797.912.14797.912.114097.242.8
MP3-548100.00048100.00048100.000144100.000

Table 5: G. lamblia Site-to-Site Quantitative Reproducibility Across Sites, Days, Runs and Within Run

Ct.scoreWithin RunBetween Run within DayBetween Day within SiteBetween SiteOverall
CategorynMeanSD%CVSD%CVSD%CVSD%CVSD%CV
HN7431.21.173.7%0.551.8%0.561.8%0.000.0%1.414.5%
LP14028.70.782.7%0.000.0%0.000.0%0.371.3%0.863.0%
MP14426.80.441.6%0.150.6%0.000.0%0.150.6%0.491.8%

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Table 6:C. parvum Site-to-Site Qualitative Reproducibility Across Siteswith Pooled Days, Runs, and Replicates
----------------------------------------------------------------------------------------------------------------------------
X LoDSITE
CINCLEBD NCTotal
CategoryCorrectIncorrectCorrectIncorrectCorrectIncorrectCorrectIncorrect
n%n%n%n%n%n%n%n%
TN048100.00048 100.00048100.000144100.000
HN0.1-0.51735.43164.6 1837.53062.52041.72858.3રેર38.28961.8
LP0.8-1.24695.824.24797.912.14797.912.114097.22.8
MP3-248100.00048 100.00048 100.000144100.000

Table 7: C. parvum Site-to-Site Quantitative Reproducibility Across Sites, Days, Runs and Within Run

Ct.scoreWithinRunBetween Run withinDayBetween Day withinSiteBetween SiteOverall
CategorynMeanSD%CVSD%CVSD%CVSD%CVSD%CV
HN8935.71.534.3%0.541.5%0.601.7%0.431.2%1.785.0%
LP14031.41.073.4%0.411.3%0.000.0%0.110.3%1.153.7%
MP14430.10.742.4%0.441.5%0.000.0%0.170.6%0.882.9%

Table 8: E. histolytica Site-to-Site Qualitative Reproducibility Across Sites with Pooled Days, Runs and Replicates

CategoryX LoDSITETotal
CINCLEBD NCCorrectIncorrect
CorrectIncorrectCorrectIncorrectCorrectIncorrectn%n%
TN048100.00048100.00048100.000144100.000
HN0.1-0.52245.82654.22654.22245.82041.72858.36847.27652.8
LP0.8-1.24797.912.14797.912.148100.00014298.621.4
MP3-548100.00048100.00048100.000144100.000

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Ct.scoreWithin RunBetween Run within DayBetween Day within SiteBetween SiteOverall
CategorynMeanSD%CVSD%CVSD%CVSD%CV
HN7630.42.347.7%1.354.4%0.662.2%0.321.0%2.809.2%
LP14225.50.883.4%0.261.0%0.000.0%0.000.0%0.913.6%
MP14423.50.582.5%0.371.6%0.000.0%0.090.4%0.692.9%

Table 9: E. histolytica Site-to-Site Quantitative Reproducibility Across Sites, Days, Runs and Within Run

For the Lot-to-Lot reproducibility study, two users each completed a single run of 12 panel members on a single instrument for each of two lots of reagents over a 5-day period. 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 agreement was 100% for the TN and MP categories for all targets, and ranged from 48.9% to 55.6% and 97.8% to 98.9% for the HN, and :LP categories, respectively (Table 10).

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95% CI
TargetLevelCorrectTotal% CorrectLowerCIUpperCI
TN**9090100.0%95.9%100.0%
G. lambliaHN*449048.9%38.8%59.0%
LP889097.8%92.3%99.4%
MP9090100.0%95.9%100.0%
C. parvumTN9090100.0%95.9%100.0%
HN509055.6%45.3%65.4%
LP889097.8%92.3%99.4%
MP9090100.0%95.9%100.0%
E. histolyticaTN9090100.0%95.9%100.0%
HNવર્સ9050.0%39.9%60.1%
LP899098.9%94.0%99.8%
MP9090100.0%95.9%100.0%

Table 10: Lot-to-Lot Reproducibility - Study Results using three lots of the BD MAX Enteric Parasite Panel

  • HNs are dilutions of the LoD designed to produce results that are negative for 20% to 80% of replicates. As such, "% Correct" correlates to the percent of negative results.

** TNs were blanks, therefore, "% Correct" correlates to the percent of negative results.

Sample Storage

Collected specimens, either unpreserved or 10% formalin fixed stool, should be kept between 2 °C and 25 °C during transport. Protect against freezing or exposure to excessive heat.

Specimens, either unpreserved or 10% formalin fixed stool, can be stored for up to 120 hours (5 days) at 2-8 ℃ or for a maximum of 48 hours at 2-25 ℃ before testing.

Prior to or following pre-warm, inoculated Sample Buffer Tubes can be stored for testing) for a total of up to:

TimeTemperature
120 hours (5 days)2 - 8 °C
48 hours2 - 25 °C

Controls

External Control materials are not provided by BD; 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:

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  • Commercially available positive control materials
  • Giardia intestinalis (Lambl) (ATCC 3088D)
  • Cryptosporidium parvum (ATCC PRA-67D) —
  • Entamoeba histolytica (ATCC 30459D)
  • External negative control —
  • Use a non-inoculated BD MAX™ Enteric Parasite Panel SBT —

The assay includes a Specimen 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.

Analytical Sensitivity

The analytical sensitivity (Limit of Detection or LoD) for the BD MAX Enteric Parasite Panel was determined for each assay target individually. Purified organisms were used to prepare targets that were inoculated into the Sample Buffer Tube along with pooled negative stool matrix (both unpreserved and 10% formalin fixed were evaluated, separately). The pooled negative stool matrix was created from stool specimens obtained from patients that were characterized by the BD MAX Enteric Parasite Panel. The organism concentrations were used to simulate positive samples with a wide range of organism loads. The LoD was determined for each organism tested with both unpreserved and 10% formalin fixed targetnegative stool matrix. The results from the LoD study can be found below in Table 11.

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T

Unpreserved[95% Confidence Interval]10% formalin fixed[95% Confidence Interval]
Cryptosporidium parvum
LoD(organism/mL in SBT)[95% confidence interval]160.17 [96.93 - 264.22]154.25 [100.46 - 236.57]
LoD(organism/mL in stool)[95% confidence interval]24,026 [14,540 - 39,633]92,550 [60,276 - 141,942]
Giardia lamblia
LoD(organism/mL in SBT)[95% confidence interval]10.67 [5.69 - 15.66]10.04 [4.83 - 15.25]
LoD(organism/mL in stool)[95% confidence interval]1,601 [854 - 2,349]6,024 [2,898 - 9,150]
Entamoeba histolytica
LoD(organism/mL in SBT)[95% confidence interval]16.79 [11.97 - 23.42]15.52 [10.99 - 21.76]
LoD(organism/mL in stool)[95% confidence interval]2,519 [1,796 - 3,570]9,300 [6,600 - 13,080]

Table 11: BD MAX Enteric Parasite Panel Target Limits of Detection

Analytical Inclusivity

The objective of this study was to demonstrate that the BD MAX Enteric Parasite Panel is able to detect clinically relevant and geographically diverse serovars/strains/ subspecies for each of the BD MAX™ Enteric Parasite Panel targets found in various geographical origins. The study was designed to validate the functional performance of the BD MAX™ Enteric Parasite Panel by verifying the specificity of the assay's primers and probes for the targeted bacterial enteric analytes.

In total, eleven (11) distinct strains/isolates of whole organisms of Giardia lamblia/intestinalis, ten (10) distinct strains/isolates of whole organisms of Entamoeba histolytica and one (1) strain of Cryptosporidium hominis whole organism were screened with the BD MAX Enteric Parasite Panel at 2X the point estimate of the 95% LoD (Table 12). One genomic DNA preparation, representative of each BD MAX Enteric Parasite Panel target, was screened at 2X LoD (Table 13).

Due to the lack of availability of Cryptosporidium parvum/hominis organisms, commercially available genomic DNA libraries, prepared from nine (9) distinct Cryptosporidium parvum strains/isolates, were also evaluated as supplementary information.

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An additional study was performed for Cryptosporidium due to the lack of commercially available organisms. DNA extracts from thirteen (13) clinical stool specimens with known Cryptosporidium spp. infection status, were obtained (Table 14). Because it is not possible to discriminate human or other DNA from parasite DNA by spectrophotometry, quantification of the DNA extracts was not performed. Instead, Sample Buffer Tubes were spiked with 10 uL of negative, pooled stool matrix and with 5 uL of the clinical specimen DNA extract, in duplicate.

OrganismATCC IDOrganismATCC ID
Giardia lambliaPRA-242Entamoeba histolyticaPRA-358
Giardia lambliaPRA-244Entamoeba histolytica50007
Giardia lambliaPRA-247Entamoeba histolytica30890
Giardia lambliaPRA-249Entamoeba histolytica30889
Giardia lamblia30888Entamoeba histolytica30458
Giardia intestinalis30957Entamoeba histolytica30459
Giardia intestinalis50114Entamoeba histolyticaPRA-357
Giardia intestinalis50137Entamoeba histolytica50738
Giardia intestinalis50581Entamoeba histolytica30190
Giardia intestinalis50584Entamoeba histolytica50541
Giardia intestinalis50585Cryptosporidium parvum502 (Tufts)
Table 12: Inclusivity Organisms
-----------------------------------------

Table 13: Target Genomic DNA

OrganismGenomic DNAATCC ID
Cryptosporidium parvumPRA-67D
Giardia lamblia30888D
Entamoeba histolytica30459D

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OrganismSubtypeOrganismSubtype
Cryptosporidium hominisIbA10G2Cryptosporidium parvumUnknown
Cryptosporidium hominisIbA10G2Cryptosporidium parvumUnknown
Cryptosporidium hominisUnknownCryptosporidium parvumUnknown
Cryptosporidium hominisIbA10G2Cryptosporidium hominisIeA11G3T3
Cryptosporidium hominisIbA10G2Cryptosporidium hominisIbA10G2
Cryptosporidium hominisIbA10G2Cryptosporidium ubiquitum*Unknown
Cryptosporidium hominisUnknown
Table 14: Clinical Specimen DNA Extracts: Cryptosporidium spp. Positive
-----------------------------------------------------------------------------

Analytical Specificity

The BD MAX Enteric Parasite Panel was performed on samples containing phylogenetically related species and other organisms (bacteria, viruses, parasites and yeast) likely to be found in stool specimens. Potentially cross-reacting organisms are summarized in Tables 15 - 17.

  • Six (6) out of 6 Entamoeba species other than E. histolytica) produced negative ● results with the BD MAX Enteric Parasite Panel. The organisms were tested directly from stock at a 1:10 dilution to obtain the highest possible concentration in the Sample Buffer Tube, with concentrations ranging from 4.00x103 organisms/mL to 2.90x103 organisms/mL in the Sample Buffer Tube. E. barretti (ATCC 30996) was provided as non-titered stock.
  • . One (1) out of 1 Cryptosporidium meleagridis strain tested at a concentration ≥ 1x10 ° cycts/mL in the Sample Buffer Tube, produced positive results with the BD MAX Enteric Parasite Panel. Cryptosporidium meleagridis has been documented in symptomatic human infection.
  • . One hundred thirteen (113) out of 113 bacterial strains, tested at a concentration > 1x10 CFU/mL in the Sample Buffer Tube, produced negative results with the BD MAX Enteric Parasite Panel.
  • Fifteen (15) out of 15 viruses, produced negative results with the BD MAX Enteric Parasite . Panel. Thirteen (13) were tested directly from stock at a 1:10 dilution to obtain the highest possible concentration in the Sample Buffer Tube, with concentrations ranging from 1.6x100 TCID50 - 8.9x10' TCID50 Human Papillomavirus was tested as plasmid in Escherichia coli and Rotavirus was tested as a high titer qualitative standard.
  • . Five (5) out of 5 phylogenetically unrelated parasites, tested at a concentration ≥ 1x10 organisms/mL in the Sample Buffer Tube, produced negative results with the BD MAX Enteric Parasite Panel.
  • Two (2) out of 2 Candida spp. tested at a concentration > 1x10° organisms/mL in the Sample Buffer Tube, produced negative results with the BD MAX Enteric Parasite Panel.
  • Three (3) enteric organisms representing each target of the BD MAX Enteric Parasite Panel were tested, with results as follows:
    • One (1) of 1 Cryptosporidium spp.; Cryptosporidium parvum tested at a concentration ≥ O 1x10 cycts/mL in the Sample Buffer Tube, produced positive results for Cryptosporidium and negative results for all other targets with the BD MAX Enteric Parasite Panel.

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  • o One (1) of 1 Giardia spp.; Giardia lamblia tested at a concentration ≥ 1x10° cycts/mL in the Sample Buffer Tube, produced positive results for Giardia and negative results for all other targets with the BD MAX Enteric Parasite Panel.
  • One (1) of 1 Entamoeba spp.; Entamoeba histolytica tested at a concentration ≥ 1x10 O cycts/mL in the Sample Buffer Tube, produced positive results for Entamoeba and negative results for all other targets with the BD MAX Enteric Parasite Panel.

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Table 15:

Potential Cross-Reactant Bacteria and Yeast

GenusSpeciesStrain
AbiotrophiadefectivaATCC 49176
AcinetobacterbaumanniiATCC 19606
IwoffiiATCC 17925
AeromonashydrophilaATCC 49847
Alcaligenesfaecalis subsp. faecalisATCC 8750
AnaerococcustetradiusATCC 35098
ArcobacterbutzleriATCC 49616
ArcobactercryaerophilusATCC 43157
BacilluscereusATCC 49064
BacteroidescaccaeATCC 43185
merdaeATCC 43184
stercorisATCC 43183
BifidobacteriumadolescentisATCC 15706
longumATCC 15707
coliATCC 43134
CamplylobacterconcisusCCUG 17580
curvusCCUG 47528
fetus subsp. fetusATCC 27374
fetus subsp. venerealisATCC 19438
gracilisATCC 33236
hominisATCC BAA-381
jejuniATCC 43429
CamplylobacterlariATCC 43675
CamplylobacterrectusATCC 33238
CamplylobacterupsaliensisATCC 49815
CandidaalbicansATCC 24433
catenulateATCC 18821
CedeceadavisaeATCC 33431
ChlamydiatrachomatisATCC VR-879
CitrobacteramalonaticusATCC 25405
fruendiibATCC 33128
koseriATCC 27156
sedlakiiATCC 51115
ClostridiumdifficileATCC 17858
ATCC 43598
CCUG 8864- ATCC 9689
ATCC 43255
ATCC BAA-1805
ATCC 43593
ClostridiumperfringensATCC 10543
CollinsellaaerofaciensATCC 35085
CorynebacteriumgenitaliumATCC 33030
DesulfovibriopigerATCC 29098
EdwardsiellatardaATCC 15947
EggerthellalentaATCC 25559
EnterobacteraerogenesATCC 13048
cloacaebATCC 35030
EnterobactercasseliflavusATCC 49605
EnterobactercecorumATCC 43198
EnterobacterdisparATCC 51266
EnterobacterfaecalisATCC 29212
EnterobactergallinarumATCC 49573
EnterobacterhiraeATCC 49612
EnterobacterraffinosusATCC 49427
EscherichiacoliATCC 25922
coli O157 stx 1BD RD012313-01
Escherichiacoli O157 stx 2coli O157 stx 2BD K092012-01BD RD092612-01
GenusSpeciesStrain
EscherichiacoliATCC 12014ATCC 8739ATCC 10536ATCC 33605
FusobacteriumfergusoniiATCC 35469
hermanniiATCC 33650
vulnerisATCC 33821
GardnerellavariumATCC 27725
GemellavaginalisATCC 14019
HafniamorbillorumATCC 27824
HelicobacteralveiATCC 11604
fennelliaeATCC 35683
KlebsiellapyloriATCC 43504
oxytocaATCC 13182
LactobacilluspneumoniaeATCC 33495
acidophilusATCC 4355
LactococcusreuteriATCC 23272
lactisATCC 15346ATCC 49032
LeminorellagrimontiiATCC 33999
grayiATCC 19120
ListeriainnocuaATCC 33090
monocytogenesATCC 19115
morganiiATCC 25830
MorganellaasaccharolyticusATCC 14963
PeptoniphilusanaerobiusATCC 27337
PeptostreptococcusshigelloidesATCC 14029
PlesiomonasasaccharolyticaATCC 25260
PorphyromonasmelaninogenicaATCC 25845
ProteusmirabilisATCC 29906
penneriATCC 35198
vulgarisATCC 13315
ProvidenciaalcalifaciensATCC 27971
rettgeriATCC 29944
stuartiiATCC 33672
PseudomonasaeruginosaATCC 27853
fluorescensATCC 13525
RuminococcusbromiiATCC 27255
SalmonellatyphimuriumATCC 14028
enteriditisATCC 13076
SerratialiquefaciensATCC 35551
marcescensATCC 13880
ShigellasonneiBD ENF 7142
flexneriATCC 700930
StaphylococcusaureusATCC 25923
epidermidisATCC 12228
StenotrophomonasmaltophiliaATCC 13637
StreptococcusagalactiaeATCC 13813
dysgalactiaeATCC 43078
intermediusATCC 27335
TrabulsiellauberisATCC 19436
VeillonellaguamensisATCC 49490
VibrioparvulaATCC 10790
choleraeBD ENF 13503
YersiniaparahaemolyticusbercovieriATCC 17802ATCC 43970
enterocoliticaATCC 9610
rohdeiATCC 43380

{19}------------------------------------------------

VirusStrain
Adenovirus type 2ATCC VR-680
Adenovirus type 14ATCC VR-15
Adenovirus type 40ATCC VR-931
Adenovirus type 41ATCC VR-930
Coxsackie A9ATCC VR-186
Coxsackie B1ATCC VR-687
HHV-5 CytomegalovirusATCC VR-538
Enterovirus type 69ATCC VR-785
Human Papillomavirus Type 16ATCC 45113
Human Papillomavirus Type 18ATCC 45152
Herpes Simplex Virus IATCC VR-539
Herpes Simplex Virus IIATCC VR-734
Norovirus I0810086CF
Norovirus II0810087CF
RotavirusNATROTA

Table 16: Potential Cross-Reactant Viruses

Potential Cross-Reactant Parasites Table 171:

ParasiteStrain
Blastocystis hominisATCC 50608
Encephalitozoon intestinalisATCC 50651
Encephalitozoon hellumATCC 50504
Encephalitozoon cuniculiATCC 50602
Pentatrichomonas hominisATCC 30098
Entamoeba barrettiATCC 30996
Entamoeba disparATCC PRA-260
Entamoeba gigivalisATCC 30927
Entamoeba invadensATCC 30994
Entamoeba moshkovskiiATCC 30041
Entamobea ranarumATCC 50389
Cryptosporidium meleagridis1867

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Interfering Substances

Twenty-two (22) biological and chemical substances occasionally used or found in stool specimens were evaluated for potential interference with the BD MAX Enteric Parasite Panel near the LOD for each particular target. Included in this study was an Antibiotics Mixture, which consisted of a combination of 8 different antibiotics tested simultaneously, with each antibiotic at a concentration that may be excreted in a stool specimen. Three of the substances tested exhibited potential interference with the BD MAX Enteric Parasite Panel (refer to Table 18). Vagisil cream demonstrated potential interference at concentrations greater than 9% in stool. Whole human blood demonstrated potential interference at concentrations greater than 25% in stool. Additional testing with grossly bloody clinical stool specimens showed potential interference in 1 out of a total of 12 specimens tested. Substances that demonstrated interference may result in potential unresolved, indeterminate or false negative results.

Brand Name or DescriptionResultBrand Name or DescriptionResult
Fecal FatNISpermicidal LubricantNI
Human DNANIDiaper Rash CreamNI
MucusNIVagisil (>9%)I
Whole Human Blood (>25%)ILaxativesNI
Hydrocortisone CreamNIAnti-Diarrheal (liquid)NI
Antiseptic TowelettesNIAnti-Diarrheal (pill)NI
EnemaNIAntibiotics MixtureNI
Hemorrhoidal GelNIAntacidsNI
Nystatin CreamNINon-Steroidal Anti-Inflammatory (NSAID)NI
10% Buffered FormalinNITopical AntibioticNI
20% Buffered FormalinNIGrossly Bloody StoolI

Table 18: Endogenous and Commercial Exogenous Substances Tested with the BD MAX Enteric Parasite Panel

I: Interference with the BD MAX Enteric Parasite Panel.

NI: No reportable interference with the BD MAX Enteric Parasite Panel.

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Carryover/Cross-Contamination

A study was conducted to investigate within-run carryover and between-run carryover while processing specimens with a high load of Giardia lamblia. Cryptosporidium parvum and Entamoeba histolytica in the BD MAX Enteric Parasite Panel made of one high positive member containing the three target organisms and one negative member was used to prepare numerous samples. Isolates of Giardia lamblia, Entamoeba histolytica and Cryptosporidium parvum were used for the high positive panel member (1 x 10 cysts/trophozoites per mL). 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. Two (2) operators performed a total of 15 runs with each run containing 24 samples.

Carryover contamination was assessed for each target in the BD MAX Enteric Parasite Panel. A total of 180 Sample Buffer Tubes, each containing the 3 BD MAX Enteric Parasite Panel targets, were assessed in the carvover contamination study. All of the 180 spiked Sample Buffer Tubes produced the expected positive results for all 3 target organisms. A total of 180 Sample Buffer Tubes, each negative for all 3 BD MAX Enteric Parasite Panel targets, were also assessed in the carryover contamination study. One hundred and seventy-eight (178) of the 180 spiked Sample Buffer Tubes produced the expected negative results for all 3 target organisms. One expected negative for Giardia lamblia and the second was positive for both Giardia lamblia and Entamoeba histolytica in a single Sample Buffer Tube.

Mixed Infection/Competitive Interference

The mixed infection/competitive interference study was designed to evaluate the ability of the BD MAX Enteric Panel to detect low positive results in the presence of other targets at high concentrations. Four (4) organisms (Giardia lamblia, Cryptosporidium parvum and two preparations of Entamoeba histolytica) were individually prepared at 2X their respective LOD to serve as a low target in the BD MAX Enteric Parasite Panel Sample Buffer Tube. A high target mix comprised of the organisms representative of the other two BD MAX Enteric Parasite Panel analytes at a concentration of > 1x10 organisms/mL in the Sample Buffer Tube was spiked into the Sample Buffer Tube along with 10 µL of unpreserved stool and tested to simulate mixed infections. The second target mix for Entamoeba histolytica also contained Entamoeba dispar prepared in the same manner as just described. All four low target organisms were successfully detected by the BD MAX Enteric Parasite Panel when combined with their respective simulated high target concentration mixed infection preparations.

Clinical Performance Studies

A clinical study was designed to assess the performance of the BD MAX Enteric Parasite Panel for the identification of Giardia lamblia, Cryptosporidium parvum/hominis and Entamoeba histolytica, from unpreserved or 10% formalin-fixed stool specimens from symptomatic patients with suspected gastroenteritis, enteritis and/or colitis. This multicenter study evaluated results obtained with the BD MAX Enteric Parasite Panel compared to those obtained with the reference method. Clinical centers were employed to collect and test patient specimens; whereas collection centers were employed to collect patient specimens, with the reference method and EPP testing being performed off-site.

The study involved a total of five (5) clinical sites where specimens were collected as part of routine patient care, enrolled into the trial, and tested on the BD MAX Enteric Parasite Panel as well as seven (7) collection sites. Only excess, de-identified patient specimens were used. Additionally, an internal site was involved as a clinical center to perform BD MAX Enteric Parasite Panel testing on specimens supplied by

{22}------------------------------------------------

collection centers. Samples tested at BD were obtained from all collection sites. Specimens collected at the sites consisted of a mix of 10% formalin-fixed and unpreserved specimens as well as a mix of prospective and retrospective specimens.

Clinical centers were selected for the clinical study based on a number of criteria, such as investigator and site personnel availability, number of specimens of interest tested for each target, prevalence, and familiarity with PCR methodology. The clinical centers were also selected according to the specimen types that they routinely collect.

A total of 2204 prospective specimens (1128 10% formalin-fixed, 1058 unpreserved and 18 noncompliant) and 411 retrospective specimens (148 10% formalin-fixed, 251 unpreserved and 12 noncompliant) were enrolled in the clinical evaluation. Table 19 describes the number of compliant specimens enrolled by patient age and specimen type. A total of 128 retrospective specimens were not included in the performance calculations below as the historical results were not confirmed by an alternate PCR and bi-directional sequencing.

Age GroupSpecimen TypeFormalin 10%Specimen TypeUnpreservedSpecimen TypeCombined
0-1 month101
1 month to 2 years11151162
2-12 years21876294
13-18 years12177198
19-21 years373471
Over 21 years7237821505
Unknown62202264
Total127312222495

Table 19: Compliant clinical trial enrollment summary by age group and specimen type

For the 10% formalin fixed specimen type, the BD MAX Enteric Parasite Panel identified 95.5% and 99.7% of the Giardia lamblia prospective and negative specimens, respectively, and 100% and 100% of the retrospective positive and negative specimens, respectively. For the unpreserved specimen type, the BD MAX Enteric Parasite Panel identified 94.4% and 100% of the Giardia lamblia prospective positive and negative specimens, respectively, and 98.6% and 94.9% of the retrospective and negative specimens, respectively (Table 20).

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SpecimenTypeSpecimenOriginBD MAXEntericParasitePanelRMTotal
PN
10% FormalinFixedProspectiveP213224
N11996997
Total229991021
SENSITIVITY (95% CI): 95.5% (78.2%, 99.2%)SPECIFICITY (95% CI): 99.7% (99.1%, 99.9%)
RetrospectiveP55055
N07171
Total5571126
PPA (95% CI): 100.0% (93.5%, 100.0%)NPA (95% CI): 100.0% (94.9%, 100.0%)
UnpreservedProspectiveP17017
N13655656
Total18655673
SENSITIVITY (95% CI): 94.4% (74.2%, 99.0%)SPECIFICITY (95% CI): 100.0% (99.4%, 100.0%)
RetrospectiveP7275,679
N14129130
Total73136209
PPA (95% CI): 98.6% (92.6%, 99.8%)NPA (95% CI): 94.9% (89.8%, 97.5%)

Table 20: Giardia lamblia – Clinical Performance

  • l The alternate PCR and bi-directional sequencing component of the reference method was negative for this specimen. The DFA component of the reference method was positive. Discrepant repeat testing with an alternate PCR and bi-directional sequencing was performed and gave a negative result. Discrepant repeat testing with DFA was done a negative result. Discrepant repeat testing with the BD MAX™ Enteric Panel was performed in twelve (12) replicates and gave all negative results (012). The specimen was also tested using a Giardia antigen EIA as discrepant testing and gave a negative result.
  • 2 One (1) specimen was tested using a Giardia antisen detecting EIA as discrepant testing and gave a negative result. Discrepant repeat testing with the BD MAX™ Enteric Panel was performed in six (6) replicates of this specimen and gave 1/6 positive result. No discrepant testing was performed for the other two (2) specimens.
  • 3 The alternate PCR and bi-directional sequencing component of the reference method was negative for this specimen. The DFA segment of the reference method was positive. Discrepant repeat testing with the alternate PCR and bi-directional sequencing was performed and gave a negative result. Discrepant repeat testing with DFA was performed and gave a positive result. Discrepant repeat testing with the BD MAX™ Enteric Parasite Panel was performed in six (6) replicates and gave all negative results (0/6). This specimen was also tested using an antigen detecting EIA as discrepant testing and gave a negative result.
  • 4 No discrepant testing was performed for this specimen.
  • 5 One (1) specimen was tested using an antigen detecting EIA and a commercially-available molecular assay as discrepant testing and gave a positive result for both. Discrepant repeat testing with the BD MAX™ Enteric Parasite Parasite in six (6) replicates of this specimen and gave all positive results (6/6).
  • 6 One (1) specimen was tested using an EIA and a commercially-available molecular assay as discrepant testing. The EIA gave a positive result and the molecular assay gave a negative result. Discrepant repeat testing with the BD MAX™ Enteric Panel was done in eleven (11) replicates of this specimen and gave 5 positive results (5/11). No discrepant testing was performed for the other five (5) specimens.

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For the 10% formalin fixed specimen type, the BD MAX Enteric Parasite Panel identified 90.3% of the Cryptosporidium parvum/hominis prospective positive and negative specimens, respectively, and 93% and 100% of the retrospective positive and negative specimens, respectively. For the unpreserved specimen type, the BD MAX Enteric Parasite Panel identified 100% and 99.5% of the Cryptosporidium parvum/hominis prospective positive and negative specimens, respectively, and 97.7% and 98.4% of the retrospective positive and negative specimens, respectively (Table 21). Because DFA identifies Cryptosporidium to the genus level, DFA-positive specimens identified by bi-directional sequencing as other than C. hominis or C. parvum were considered reference method negative.

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SpecimenTypeSpecimenOriginBD MAXTMEntericParasite PanelRMTotal
PN
10% FormalinFixedProspectiveP562358
N61, 29514957
Total629531015
SENSITIVITY: 90.3% (80.5%, 95.5%)SPECIFICITY: 99.8% (99.2%, 99.9%)
RetrospectiveP40040
N357881
Total4378121
PPA: 93% (81.4%, 97.6%)NPA: 100% (95.3%, 100%)
UnpreservedProspectiveP353638
N0625625
Total35628663
SENSITIVITY: 100% (90.1%, 100%)SPECIFICITY: 99.5% (98.6%, 99.8%)
RetrospectiveP433746
N1181182
Total44184228
PPA: 97.7% (88.2%, 99.6%)NPA: 98.4% (95.3%, 99.4%)

Table 21: Cryptosporidium parvum/hominis - Clinical Performance

  • All six (6) specimens were positive by the DFA composite reference method. One (1) specimen sequenced as C. 1 parvum, three (3) specimens were negative and two (2) were non-reportable by the alternate PCR and bi-directional sequencing components of the composite reference method.
  • 2 Discrepant repeat testing with the alternate PCR and bi-directional sequencing was performed on all six (6) specimens. One (1) specimen sequenced as Cryptosporidium parvum, one (1) specimen sequenced as Cryptosporidium felis and the remaining four (4) were PCR negative by discrepant repeat testing was also performed using an antigen detecting EIA that does not distinguish between Cryptosporidium and Giardia. Two specimens were EIA negative and four specimens were EIA positive, of which two were positive for Giardia by other test methods.
  • 3 One DFA-positive specimen was classified as reference method negative based on alternate PCR and bi-directional sequencing results that identified Cryptosporidium meleagridis.
  • 4 Six DFA-positive specimens were classified as reference method negative based on alternate PCR and bi-directional sequencing results that identified (4) Cryptosporidium canis, (1) C. meleagridis and (1) Cryptosporidium spp. (undefined).
  • ર Discrepant repeat testing was performed with BD MAX Enteric Parasite Panel in twelve (12) replicates per specimen. One specimen was positive for five (5) of twelve (12) replicates and one specimen was positive for two (2) of twelve (12) replicates.
  • Discrepant repeat testing was performed with BD MAX Enteric Panel in six (6) replicates per specimen. One specimen was 6 positive for five (5) of six (6) replicates and one specimen was positive for three (3) of six (6) replicates. A third specimen was negative in six (6) of six (6) replicates.
  • 7 Discrepant repeat testing was performed with BD MAX Enteric Panel in six (6) replicates per specimen. Two specimens were negative for six (6) of six (6) replicates.

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For the 10% formalin fixed specimen type, the BD MAX Enteric Parasite Panel identified 100% of the Entamoeba histolytica negative specimens for both the prospective and retrospective specimens. There were no prospective or retrospective 10% formalin fixed Entamoeba histolytica positive specimens found during the clinical evaluation. For the unpreserved specimen type, the BD MAX Enteric Parasite Panel identified 100% Entamoeba histolytica prospective negative specimens and 100% of the retrospective positive and negative specimens, respectively (Table 22). There were no prospective unpreserved Entamoeba histolytica positive specimens found during the clinical evaluation.

Specimen TypeSpecimen OriginBD MAXEntericParasite PanelRMTotal
PN
10% FormalinFixedProspectiveP000
N0827827
Total0827827
SENSITIVITY (95% CI): No data for calculationSPECIFICITY (95% CI): 100% (99.5%, 100%)
RetrospectiveP000
N05454
Total05454
PPA (95% CI): No data for calculationNPA (95% CI): 100% (93.4%, 100%)
UnpreservedProspectiveP000
N0577577
Total0577577
SENSITIVITY (95% CI): No data for calculationSPECIFICITY (95% CI): 100% (99.3%, 100%)
RetrospectiveP11011
N0191191
Total11191202
PPA (95% CI): 100% (74.1%, 100%)NPA (95% CI): 100% (98.0%, 100%)

Table 22: Entamoeba histolytica - Clinical Performance

As Entamoeba histolytica is a rare analyte, both prospective testing efforts were unable to demonstrate the positive percent agreement of the BD MAX Enteric Parasite Panel. To supplement the prospective and retrospective data, an evaluation of contrived specimens was performed. Surrogate clinical specimens were prepared using residual specimens that had previously tested negative for all BD MAX Enteric Parasite Panel targets. Specimens were spiked at clinically relevant levels at various concentrations of the limit of detection for each specimen type. Users analyzing the specimens were blinded as to the specimen status.

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For both the 10% formalin fixed and unpreserved specimen types, the BD MAX Enteric Parasite Panel correctly identified 100% of both the positive and negative specimens. The contrived study results obtained with the BD MAX Enteric Parasite Panel were compared to the expected results and are summarized in Table 23.

Specimen TypeBD MAXEnteric Parasite PanelExpected ResultTotal
PN
Formalin 10%P50050
N05050
Total5050100
PPA (95% CI): 100% (92.9%, 100%)NPA (95% CI): 100% (92.9%, 100%)
UnpreservedP50050
N05050
Total5050100
PPA (95% CI): 100% (92.9%, 100%)NPA (95% CI): 100% (92.9%, 100%)
Table 23: Entamoeba histolytica - Contrived Specimen Performance
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Performance of the BD MAX Enteric Parasite Panel by Cryptosporidium hominis and Cryptosporidium parvum species identification as observed during the clinical trial is presented below in Table 24. The species identification was obtained from the alternate PCR and bi-directional sequencing segment of the composite reference method. While the BD MAX Enteric Parasite Panel is designed to detect the species described below, the panel does not report results to the species level.

Table 24: Cryptosporidium PPA per species observed during the clinical trial

CryptosporidiumPPA
Specimen TypeSpecimen Origin SpeciesEstimate95% CI
10% Formalin FixedProspectivehominis100.0% (17/17)(81.6%, 100.0%)
parvum97.4% (37/38)(86.5%, 99.5%)
Retrospectivehominis95.0% (19/20)(76.4%. 99.1%)
parvum91.3% (21/23)(73.2%. 97.6%)
Unpreservedhominis100.0% (22/22)(85.1%, 100.0%)
Prospectiveparvum100.0% (11/11)(74.1%, 100.0%)
Retrospectivehominis96.2% (25/26)(81.1%, 99.3%)
parvum100.0% (18/18)(82.4%. 100.0%)

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There were twenty-three (23) co-infections detected by the BD MAX Enteric Parasite Panel. Table 25 below shows the co-infections detected by the BD MAX Enteric Parasite Panel during the clinical trial.

Co-InfectionsNumber of Co-InfectionsObservedNumber ofDiscrepantCo-InfectionsDiscrepant TargetUnavailableReference MethodResult for Comparison
Cryptosporidiumparvum/hominisEntamoebahistolytica31Cryptosporidiumhominis/parvum1 Cryptosporidiumparvum/hominis
Giardia lambliaCryptosporidiumparvum/hominis112Cryptosporidiumparvum/hominis3 Giardia lamblia and4 Cryptosporidiumparvum/hominis
Giardia lambliaEntamoebahistolytica941Giardia lamblia1 Giardia lamblia and3 Entamoeba histolytica

Table 25: Co-infections observed during the BD MAX Enteric Parasite Panel clinical trial

1 All four (4) are retrospective specimens with unconfirmed historical results.

Of the 2226 specimens initially evaluated with the BD MAX Enteric Parasite Panel, 1.5% of the 10% formalin fixed and 4.7% of the unpreserved specimens were initially reported as Unresolved. Following a valid repeat test, 0% of the 10% formalin fixed and 1.2% of the unpreserved specimens remained Unresolved. The total numbers provided in Table 26 are based on compliant specimens and BD MAX Enteric Parasite Panel results.

Table 26: Unresolved Rates

Initial Unresolved Rate with95% CIFinal Unresolved Rate with Valid Repeatwith 95% CI
SpecimenTypeSpecimenOriginPercent95% CIPercent95% CI
Formalin 10%Prospective1.5% (16/1084)(0.9%, 2.4%)0.0% (0/1084)(0.0%, 0.4%)
Formalin 10%Retrospective1.4% (2/146)(0.4%, 4.9%)0.0% (0/146)(0.0%, 2.6%)
UnpreservedProspective5.6% (42/752)(4.2%, 7.5%)1.5% (11/747)(0.8%, 2.6%)
UnpreservedRetrospective2.0% (5/244)(0.9%, 4.7%)0.4% (1/244)(0.1%, 2.3%)

Of the 2226 specimens initially evaluated with the BD MAX Enteric Parasite Panel, 0.3% of the 10% formalin fixed and 0.1% of the unpreserved specimens were initially reported as Indeterminate. Following a valid repeat test, 0% of both the 10% formalin fixed and the unpreserved specimens remained Indeterminate. The total numbers provided in Table 27 are based on compliant specimens and BD MAX Enteric Parasite Panel results.

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Initial Indeterminate Rate with95% CIFinal Indeterminate Rate with ValidRepeat with 95% CI
SpecimenTypeSpecimenOriginPercent95% CIPercent95% CI
Formalin 10%Prospective0.4% (4/1084)(0.1%, 0.9%)0.0% (0/1084)(0.0%, 0.4%)
Retrospective0.0% (0/146)(0.0%, 2.6%)0.0% (0/146)(0.0%, 2.6%)
UnpreservedProspective0.1% (1/752)(0.0%, 0.7%)0.0% (0/747)(0.0%, 0.5%)
Retrospective0.0% (0/244)(0.0%, 1.5%)0.0% (0/244)(0.0%, 1.5%)

Table 27: Indeterminate Rates

Of the 2226 specimens initially evaluated with the BD MAX Enteric Parasite Panel, 0.6% of the 10% formalin fixed and 0.4% of the unpreserved specimens were initially reported as Incomplete. Following a valid repeat test, 0% of both the 10% formalin fixed and the unpreserved specimens remained Incomplete. The total numbers provided in Table 28 are based on compliant specimens and BD MAX Enteric Parasite Panel results.

Table 28: Incomplete Rates

Initial Incomplete Rate with95% CIFinal Incomplete Rate withValid Repeat with 95% CI
SpecimenTypeSpecimenOriginPercent95% CIPercent95% CI
Formalin10%Prospective0.6% (6/1084)(0.3%, 1.2%)0.0% (0/1084)(0.0%, 0.4%)
Retrospective0.7% (1/146)(0.1%, 3.8%)0.0% (0/146)(0.0%, 2.6%)
UnpreservedProspective0.0% (0/752)(0.0%, 0.5%)0.0% (0/747)(0.0%, 0.5%)
Retrospective1.6% (4/244)(0.6%, 4.1%)0.0% (0/244)(0.0%, 1.5%)

§ 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).