Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a multiplexed nucleic acid test using PCR technology for the detection of pathogens. There is no mention of AI or ML in the device description, intended use, or performance studies. The analysis focuses on analytical and clinical performance metrics typical of molecular diagnostic assays.

Therapeutic

No
The device is an in vitro diagnostic test for detecting infectious agents from stool samples, which aids in diagnosis rather than providing therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the device "is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness".

SaMD (Software as a Medical Device)

No

The device is a multiplexed nucleic acid test that requires a physical analyzer (QIAstat-Dx Analyzer 1.0) and single-test cartridges with pre-packaged reagents to perform sample preparation and detection. It is not solely software.

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 QIAstat-Dx Gastrointestinal Panel 2 is a "multiplexed nucleic acid test intended for use with the QIAstat-Dx Analyzer 1.0. for the simultaneous in vitro qualitative detection of nucleic acids from multiple viruses, bacteria. and parasites directly from preserved stool samples...". The phrase "in vitro" is a key indicator of an IVD, meaning it's used to test samples outside of the body.
Sample Type: The test is performed on "preserved stool samples," which are biological specimens taken from the body.
Purpose: The test is intended as "an aid in the diagnosis of specific agents of gastrointestinal illness," which is a diagnostic purpose.
Device Description: The description details a system that processes a sample within a cartridge using pre-packaged reagents, which is typical of an in vitro diagnostic assay.

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

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The QIAstat-Dx Gastrointestinal Panel 2 is a multiplexed nucleic acid test intended for use with the QIAstat-Dx Analyzer 1.0. for the simultaneous in vitro qualitative detection of nucleic acids from multiple viruses, bacteria. and parasites directly from preserved stool samples (Para-Pak C&S or FecalSwab) obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following viruses, bacteria (including several diarrheagenic E. col/ Shigella pathotypes), and parasites are identified with the QIAstat-Dx Gastrointestinal Panel 2 :

Pathogen	Para-Pak C&S	FecalSwab
• Adenovirus F40/F41	Reported	Reported
• Astrovirus	Reported	Reported
• Norovirus GI/GII	Reported	Reported
• Rotavirus A	Reported	Reported
• Campylobacter (C. jejuni, C. coli and C. upsaliensis)	Reported	Reported
• Shigella/Enteroinvasive Escherichia coli (EIEC)	Reported	Reported
• Enteropathogenic Escherichia coli (EPEC)	Reported	Not Reported
• Enterotoxigenic Escherichia coli (ETEC) lt/st	Reported	Reported
• Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2(including specific identification of E. coli O157 serogroup within STEC)	Reported	Not Reported
• Salmonella	Reported	Reported
• Plesiomonas shigelloides	Reported	Reported
• Yersinia enterocolitica	Reported	Reported
• Cryptosporidium	Reported	Reported
• Cyclospora cayetanensis	Reported	Reported
• Entamoeba histolytica	Reported	Reported
• Giardia lamblia*	Reported	Reported

*(Also known as Giardia intestinalis and Giardia duodenalis)

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

The QIAstat-Dx Gastrointestinal Panel 2 is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule-out coinfection with organisms not detected by the QIAstat-Dx Gastrointestinal Panel 2. The organisms detected may not be the sole or definitive cause of the disease.

Negative QIAstat-Dx Gastrointestinal Panel 2 results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this assay test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease.

Product codes

PCH

Device Description

QIAstat-Dx is based on single-test cartridges with pre-packaged reagents including both wet and dry chemistry to handle the sample preparation and detection steps for the presence of a range of selected analytes by PCR technology. After insertion of the sample, the OIAstat-Dx assay cartridge is processed by the OIAstat-Dx Analyzer 1.0.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Gastrointestinal (via stool samples)

Indicated Patient Age Range

Not Found (Age groups are presented in expected values summary, but no specific age range for indication is given).

Intended User / Care Setting

Clinical diagnostics laboratories (hospital-associated or independent)

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 of QIAstat-Dx Gastrointestinal Panel 2 was established during a multi-center international prospective study conducted at thirteen clinical settings representative of different geographical areas within USA and Europe (9 sites in USA and 4 sites in Europe) between May and July 2021. All study sites were hospital-associated or independent clinical diagnostics laboratories that perform routine diagnostics of GI infections. 1939 eligible specimens (stool preserved in Para-Pak C&S (Meridian Bioscience) or FecalSwab (COPAN)) were collected and tested.

Additional prospective archived samples were collected for Norovirus GI/GII (81 samples) and STEC (18 samples). These were prospectively collected samples from four different collection sites (3 US and 1 EU), where only those positive for the pathogen by standard of care method were archived for analysis alongside 20 negative specimens.

In addition, to supplement the results of the prospective clinical studies, a total of 750 preselected archived frozen (retrospective) specimens were also evaluated. These specimens served to increase the sample size for analytes that showed low prevalence in the clinical prospective study or that were less represented in a particular sample type (Para-Pak C&S or FecalSwab).

Total: 2808 specimens (1939 prospective, 119 prospective archived and 750 retrospective) were evaluated in the clinical study. These specimens were collected using Para-Pak C&S (1217) or FecalSwab (1591).

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

Study Type: Non-Clinical Studies (Analytical Sensitivity (LoD), Analytical Reactivity (Inclusivity), Analytical Specificity (Cross-Reactivity and Exclusivity), Interfering Substances, Microbial Interference, Competitive Interference, Carryover, Sample Stability, Reproducibility, Repeatability) and Clinical Studies (Clinical Performance, Failure Rates, Co-infections, Contrived Specimens Testing).

Analytical Sensitivity (Limit of Detection (LoD)):

Sample Size: 36 pathogen strains, analyzed in serial dilutions in human stool matrix (Para-Pak C&S or FecalSwab).
Key Results: LoD for each target ensured >95% detection rate. Individual LoD values expressed in molecular and microbiological units are provided in Table 5.2.

Analytical Reactivity (Inclusivity):

Sample Size: 114 pathogen strains tested in vitro.
Key Results: QIAstat-Dx Gastrointestinal Panel 2 is inclusive for 100% (114 out of 114) of the pathogen strains tested in vitro. Most strains detected at ≤3-fold of corresponding LoD. Some strains (ETEC, EIEC/Shigella, Rotavirus, STEC, Adenovirus, Norovirus) showed

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

May 31, 2024

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Food and Drug Administration (FDA). On the left is the Department of Health and Human Services logo. To the right of that is a blue square with the letters FDA in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

QIAGEN GmbH % Selina Salthouse Regulatory Affairs Manager QIAGEN Manchester Ltd CityLabs 2.0. 200 Hathersage Road Manchester, Manchester M13 0BH, United Kingdom

Re: K220062

Trade/Device Name: QIAstat-Dx Gastrointestinal Panel 2 Regulation Number: 21 CFR 866.3990 Regulation Name: Gastrointestinal microorganism multiplex nucleic acid-based assay Regulatory Class: Class II Product Code: PCH Dated: April 4, 2023 Received: April 4, 2023

Dear Selina Salthouse:

We have reviewed your section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (the Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

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Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Noel J. Gerald -S

Noel J. Gerald, Ph.D. Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

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Indications for Use

510(k) Number (if known) K220062

Device Name QIAstat-Dx Gastrointestinal Panel 2

Indications for Use (Describe)

The QIAstat-Dx Gastrointestinal Panel 2 is a multiplexed nucleic acid test intended for use with the QIAstat-Dx Analyzer 1.0. for the simultaneous in vitro qualitative detection of nucleic acids from multiple viruses, bacteria. and parasites directly from preserved stool samples (Para-Pak C&S or FecalSwab) obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following viruses, bacteria (including several diarrheagenic E. col/ Shigella pathotypes), and parasites are identified with the QIAstat-Dx Gastrointestinal Panel 2 :

Pathogen	Para-Pak C&S	FecalSwab
• Adenovirus F40/F41	Reported	Reported
• Astrovirus	Reported	Reported
• Norovirus GI/GII	Reported	Reported
• Rotavirus A	Reported	Reported
• Campylobacter (C. jejuni, C. coli and C. upsaliensis)	Reported	Reported
• Shigella/Enteroinvasive Escherichia coli (EIEC)	Reported	Reported
• Enteropathogenic Escherichia coli (EPEC)	Reported	Not Reported
• Enterotoxigenic Escherichia coli (ETEC) lt/st	Reported	Reported
• Shiga-like toxin-producing Escherichia coli (STEC) stx1/stx2
(including specific identification of E. coli O157 serogroup within STEC)	Reported	Not Reported
• Salmonella	Reported	Reported
• Plesiomonas shigelloides	Reported	Reported
• Yersinia enterocolitica	Reported	Reported
• Cryptosporidium	Reported	Reported
• Cyclospora cayetanensis	Reported	Reported
• Entamoeba histolytica	Reported	Reported
• Giardia lamblia*	Reported	Reported

*(Also known as Giardia intestinalis and Giardia duodenalis)

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

The QIAstat-Dx Gastrointestinal Panel 2 is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule-out coinfection with organisms not detected by the QIAstat-Dx Gastrointestinal Panel 2. The organisms detected may not be the sole or definitive cause of the disease.

Negative QIAstat-Dx Gastrointestinal Panel 2 results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this assay 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)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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5.0 510(k) SUMMARY

General Information

| Submitted by: | QIAGEN GmbH
Qiagen Strasse 1
Hilden, 40724,Germany |
|-----------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact Person: | Selina Salthouse
Manager, Regulatory Affairs
QIAGEN Manchester Ltd
Citylabs 2.0, 200 Hathersage Road
Manchester, M13 0BH, UK
Phone: +44 7823 862 321
Fax: 44 161 204 1200
Email: selina.salthouse@qiagen.com |
| Date Prepared: | May 30, 2024 |
| Device Name: | QIAstat-Dx® Gastrointestinal Panel 2 |
| Trade Name: | QIAstat-Dx® Gastrointestinal Panel 2 |
| Common Name: | QIAstat-Dx® Gastrointestinal Panel 2 |
| Classification: | 21 CFR 866.3990 - Gastrointestinal Pathogen Panel Multiplex
Nucleic Acid-Based Assay System |
| Product Code: | PCH |

Predicate Device

Manufacturer	Product Name	510(k) No.
BioFire Diagnostics LLC	FilmArray®
Gastrointestinal (GI) Panel	K140407

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Device Description

QIAstat-Dx is based on single-test cartridges with pre-packaged reagents including both wet and dry chemistry to handle the sample preparation and detection steps for the presence of a range of selected analytes by PCR technology. After insertion of the sample, the OIAstat-Dx assay cartridge is processed by the OIAstat-Dx Analyzer 1.0.

Principle of Operation

The QIAstat-Dx Gastrointestinal Panel 2 is part of the QIAstat-Dx system and works with the QIAstat-Dx Analyzer 1.0.

The OIAstat-Dx Gastrointestinal Panel 2 is intended to be used with stool samples in Para-Pak C&S or FecalSwab transport media.

Once the cartridge has been inserted into the instrument, the test starts automatically and runs for approximately 78 minutes. When the test is finished, the cartridge is removed by the user and discarded. The QIAstat-Dx Analyzer 1.0 automatically interprets test results and displays a summary on the analyzer display screen. The results can be printed using a connected printer if needed. The detected analytes are displayed in red. For other analytes tested, they are displayed in green if not detected or in gray if not applicable or invalid. The analyzer will report if an error occurs during processing, in which case the test will fail and no results will be provided (screen will show "FAIL").

Sample Pre-treatment for PCR Inhibitors removal

Following insertion of the cartridge, the buffer located in Reservoir 1 is added inside the lysis chamber and homogenized with the sample using a rotor in the presence of silica beads. This enables separation of commonly found inhibitory substances in stool from the DNA/RNA by chemical means.

Resuspension of Internal Control (IC) and Proteinase K

Following sample pre-treatment, the IC and Proteinase K is resuspended with the buffer located in Reservoir 2 (resuspension buffer). The buffer from Reservoir 2 is added to the interconnected IC cavity and Proteinase K cavity and transferred repeatedly between the Transfer Chamber and the cavities to ensure resuspension. The resuspended IC and Proteinase K are transferred to the sample cavity.

Cell Lysis

Primary lysis of the cells and analytes present in a stool sample and IC occurs by a combination of chemical and mechanical processes using a rotor inside the lysis chamber in the presence of silica beads and a buffer that acts as a chemical agent in aiding the mechanical process. The fast movement of the rotor in the presence of the silica beads results in sample agitation, which creates turbulence and shear forces that favor the lysis of the cell wall.

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After mechanical lysis is completed, the primary lysate is transferred to the purification chamber through a frit with 2-5 um pore size. The second lysis buffer (from Reservoir 3) is added to the primary lysate to complete chemical lysis.

Purification

Binding reagent (from Reservoir 4) is added to the lysate in the purification chamber, and the mix is passed through the silica membrane. In this process, the DNA/RNA molecules stick to the membrane, and the remaining components of the lysate are delivered to the waste chamber. Then the membrane is washed with a first washing buffer (from Reservoir 5) to wash away proteins. This is followed by a second washing step with a second washing buffer (from Reservoir 6) to remove any remaining substances other than the nucleic acids. A subsequent drying step eliminates volatile substances from the silica membrane. Prior to the elution step, the Transfer Chamber is rinsed with the rinsing buffer (from Reservoir 7) in order to remove any potential inhibitors from previous processing steps. At the end of the process, the nucleic acids are released from the membrane using an elution buffer (from Reservoir 8). The eluate is collected in the Transfer Chamber.

Rehydration of Master Mix

A defined volume (135 µL) of the eluate is delivered to the reservoir of the Dry Chemistry Container (DCC) to rehydrate the Master Mix. Any remaining eluate is transferred to the Reservoir 7. The eluate/Master Mix solution is mixed by repeated transfer between the Transfer Chamber and the DCC.

Aliquoting and PCR

Defined aliquots (15 µL) of mixed eluate/Master Mix are sequentially transferred from the Transfer Chamber to each of eight Reaction Chambers containing the specified, air dried primers and probes.

Within each Reaction Chamber, a reverse-transcription step followed by real time, multiplex PCR ("rtPCR") is performed. Increase in fluorescence (indicative of detection of each target analyte) is detected directly within each Reaction Chamber.

The rtPCR process is conducted by two submodules of the QIAstat-Dx Analyzer 1.0: the Thermal Cycler and the qPCR Sensor.

Components Description

OIAstat-Dx Gastrointestinal Panel 2 Cartridge:

The QIAstat-Dx Gastrointestinal Panel 2 cartridge is a disposable plastic device that allows performing fully automated molecular assays. The main features of the OIAstat-Dx Gastrointestinal Panel 2 cartridge for the Gastrointestinal assay include the ability to test liquid samples and the capacity to store all necessary reagents within the cartridge needed for such testing. All sample preparation and assay steps will be performed within the cartridge.

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All the reagents required for the complete execution of the test are pre-loaded and selfcontained in the OIAstat-Dx Gastrointestinal Panel 2. The user does not need to manipulate any reagents. During the test, reagents are handled by pneumatically-operated microfluidics without any direct contact with the user or the analyzer actuators. This eliminates any possibility of exposure of the user or the analyzer to chemicals contained in the cartridge during the test and up to the disposal of used cartridges.

Reagents may be found in three different physical forms: liquid, air-dried on surfaces or lyophilized powder cake.

Within the cartridge, multiple steps are automatically performed in sequence by using pneumatic pressure and a multiport valve to transfer sample and fluids via the Transfer Chamber to their intended destinations.

QIAstat-Dx Analyzer 1.0

The QIAstat-Dx Analyzer 1.0 is the unit that hosts a cartridge and, on command from the user, is able to run predefined assay protocols. The software specific to this test is preloaded on the QIAstat-Dx Analyzer 1.0.

Other Materials

Each QIAstat-Dx Gastrointestinal Panel 2 cartridge will be used with a transfer pipette (provided with device). The stool sample from the patient will be collected in Para-Pak C&S or FecalSwab transport medium following the manufacturer's instructions for use (not provided with device).

QIAstat-Dx Analyzer 1.0 - the QIAstat-Dx Gastrointestinal Panel 2 cartridge can only be run on the QIAstat-Dx Analyzer 1.0.

Calibrators and/or Controls

Blank controls are not applicable to the device because it is a single test disposable cartridge. Negative and positive external controls are recommended by the company but not provided with the OIAstat-Dx Gastrointestinal Panel 2.

QIAGEN provides an IC within the QIAstat-Dx Gastrointestinal Panel 2 cartridge which provides a full process control covering lysis, nucleic acid purification, reverse transcription and DNA amplification. The IC is Schizosaccharomyces pombe. The IC is located in the IC cavity and is mixed with the sample during sample preparation and the eluate is mixed with the Master Mix, then aliquoted in all Reaction Chambers. The results screen displays a message indicating that the Internal Control "Passed" when the test is run successfully. An IC message of "Failed" indicates that the internal control was not amplified; 'Positive' test results are then reported as POSITIVE* (positive with warning), all 'Negative' results are invalid.

The OIAstat-Dx Analyzer 1.0 is provided factory calibrated and does not require user calibration. The OIAstat-Dx Analyzer 1.0 includes self-check controls to verify the performance of all sensors and actuators and will alert the user in case of failure.

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The RCA will provide the results to the Application Software (SW). The Application SW will store the information related to a given result in the database and will display a summary of detected analytes and the result for the IC. All POSITIVE analytes will be listed as "DETECTED PATHOGENS". The screen will also display the complete list of all "TESTED PATHOGENS", including positive, not applicable or invalid analytes. All results for tests executed on each QIAstat-Dx Analyzer 1.0 are stored and can be reviewed in a specific archive on each QIAstat-Dx Analyzer 1.0.

Specimen collection and transport materials

Stool samples are collected and resuspended in Para-Pak C&S or FecalSwab transport medium following the manufacturer's instructions (Para-Pak C&S (Meridian Bioscience) or FecalSwab (COPAN)).

Intended Use

The QIAstat-Dx® Gastrointestinal Panel 2 is a multiplexed nucleic acid test intended for use with the OIAstat-Dx Analyzer 1.0 for the simultaneous in vitro qualitative detection and identification of nucleic acids from multiple viruses, bacteria, and parasites directly from preserved stool samples (Para-Pak® C&S or FecalSwab™) obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following viruses, bacteria (including several diarrheagenic E.coli/Shigella pathotypes), and parasites are identified with the QIAstat-Dx® Gastrointestinal Panel 2:

Pathogen	Para-Pak C&S	FecalSwab
Adenovirus F40/F41	✓	✓
Astrovirus	✓	✓
Norovirus GI/GII	✓	✓
Rotavirus A	✓	✓
Campylobacter (C. jejuni, C. coli and C. upsaliensis)	✓	✓
Shigella/Enteroinvasive Escherichia coli (EIEC)	✓	✓
Enteropathogenic Escherichia coli (EPEC)	✓	Not reported
Enterotoxigenic Escherichia coli (ETEC) lt/st	✓	✓
Shiga-like toxin-producing Escherichia coli (STEC)
stx1/stx2 (including specific identification of E. coli
O157 serogroup within STEC)	✓	Not reported
Salmonella	✓	✓
Plesiomonas shigelloides	✓	✓
Yersinia enterocolitica	✓	✓
Cryptosporidium	✓	✓
Cyclospora cayetanensis	✓	✓
Entamoeba histolytica	✓	✓
Giardia lamblia*	✓	✓

Also known as Giardia intestinalis and Giardia duodenalis

Concomitant culture is necessary for organism recovery and further typing of bacterial agents.

9

The QIAstat-Dx® Gastrointestinal Panel 2 is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness, in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule-out co-infection with organisms not detected by the QIAstat-Dx® Gastrointestinal Panel 2. The organisms detected may not be the sole or definitive cause of the disease.

Negative QIAstat-Dx® Gastrointestinal Panel 2 results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this assay test or non-infectious causes such as ulcerative colitis, irritable bowel syndrome, or Crohn's disease.

Comparison of the QIAstat-Dx® Gastrointestinal Panel 2 and the Predicate Device

The QIAstat-Dx Gastrointestinal Panel 2 is substantially equivalent to the predicate device:

K140407: BioFire Diagnostics LLC FilmArray® Gastrointestinal (GI) Panel
Similarities and differences between the QIAstat-Dx Gastrointestinal Panel 2 and the predicate device are shown in Table 5.1.

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Table 5.1: Comparison of the QIAstat-Dx Gastrointestinal Panel 2 with the predicate device

Characteristic	Device	Predicate
Name	QIAstat-Dx Gastrointestinal Panel 2	BioFire Diagnostics, LLC.
FilmArray Gastrointestinal (GI) Panel
510(k) No.	K220062	K140407
Regulation	21 CFR 866.3990	21 CFR 866.3990
Product Code	PCH	PCH
Device Class	Class II	Class II
Similarities
Intended Use	The QIAstat-Dx® Gastrointestinal Panel 2 is a multiplexed nucleic acid test intended for use with the QIAstat-Dx Analyzer 1.0 for the simultaneous qualitative detection and identification of nucleic acids from multiple viruses, bacteria, and parasites directly from preserved stool samples (Para-Pak® C&S or FecalSwabTM) obtained from individuals with signs and/or symptoms of gastrointestinal infection. The following viruses, bacteria (including several diarrheagenic E.coli/Shigella pathotypes), and parasites are identified with the QIAstat-Dx® Gastrointestinal Panel 2: Adenovirus F40/F41 Astrovirus Norovirus GI/GII Rotavirus A Campylobacter ( C. jejuni, C. coli and C. upsaliensis ) Shigella/Enteroinvasive Escherichia coli (EIEC) Enteropathogenic Escherichia coli (EPEC) (only with Para-Pak C&S, not reported for FecalSwab) Enterotoxigenic Escherichia coli (ETEC) lt/st	The FilmArray® Gastrointestinal (GI) Panel is a qualitative multiplexed nucleic acid-based test intended for use with the FilmArray® Instrument for the simultaneous detection and identification of nucleic acids from multiple bacteria, viruses, and parasites directly from stool samples in Cary Blair medium, 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
Characteristic	Device	Predicate
	Shiga-like toxin-producing
Escherichia coli (STEC) stx1/stx2 (including specific identification of E. coli O157 serogroup within STEC) (only with Para-Pak C&S, not reported for FecalSwab) Salmonella Plesiomonas shigelloides Yersinia enterocolitica Cryptosporidium Cyclospora cayetanensis Entamoeba histolytica Giardia lamblia (Also known as Giardia intestinalis and Giardia duodenalis ) Concomitant culture is necessary for organism recovery and further typing of bacterial agents.
The QIAstat-Dx® Gastrointestinal Panel 2 is indicated as an aid in the diagnosis of specific agents of gastrointestinal illness in conjunction with other clinical, laboratory, and epidemiological data. Positive results do not rule-out co-infection with organisms not detected by the QIAstat-Dx® Gastrointestinal Panel 2. The organisms detected may not be the sole or definitive cause of the disease.
Negative QIAstat-Dx® Gastrointestinal Panel 2 results in the setting of clinical illness compatible with gastroenteritis may be due to infection by pathogens that are not detected by this assay test or non-infectious causes such as ulcerative colitis,	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® Gastrointestinal 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.
Characteristic	Device	Predicate
	irritable bowel syndrome, or
Crohn's disease.	Concomitant culture is
necessary for organism
recovery and further typing of
bacterial agents.
		This device is not intended to
monitor or guide treatment for
C. difficile infection.
		Due to the small number of
positive specimens collected
for certain organisms during
the prospective clinical study,
performance characteristics for
E. coli O157, 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
Characteristic	Device	Predicate
		identification of acute
gastroenteritis in the context of
outbreaks.
Specimen Type	Preserved stool in Para-Pak C&S
or FecalSwab transport media	Stool in Cary-Blair Medium
Analyte Detected	RNA/DNA	RNA/DNA
Organism Detected	See above	See above
Amplification and
Detection
Technology	PCR	PCR
Assay Controls	One internal control in each
cartridge to control for sample
processing that is subjected to all
nucleic acid extraction and
amplification steps similar to
patient samples. Labeling
recommends use of negative and
positive external controls
regularly. Use transport media as
the external Negative Control,
and previously characterized
positive samples or negative
sample spiked with well
characterized target organisms as
external positive controls.	Two controls are included in
each reagent pouch to control
for sample processing and both
stages of PCR and melt
analysis. Labeling
recommends the use of
external positive and negative
controls regularly. Enteric
transport media can be used as
an external negative control,
and previously characterized
positive samples or negative
samples spiked with well
characterized organisms as
external positive controls.
Differences
Assay Targets	No additional targets to those
listed above which are in
common with the predicate
device.	The FilmArray®
Gastrointestinal Panel has four
additional targets: a generic
Vibrio target which
differentiates Vibrio cholerae ,
Clostridium difficile ( C. difficile ) toxin A/B,
Enteroaggregative Escherichia coli (EAEC) and Sapovirus.
	Nucleic Acid
Extraction	Extraction of nucleic acids using
silica membrane	Extraction of nucleic acids
using magnetic beads
	Characteristic	Device	Predicate
	Technology	QIAstat-Dx Gastrointestinal
Panel 2 detection of amplified
targets uses an increase in
fluorescence due to specific probe
binding to generate the assay
results.	The FilmArray®
Gastrointestinal Panel uses
melting curve analysis to
confirm the identity of
amplified targets to generate
assay results
	Operational	The sample is loaded straight into
the cartridge.	The FilmArray®
Gastrointestinal Panel has to
rehydrate the cartridge before
use
Amplification and
Detection Instrument
System	QIAstat-Dx Analyzer 1.0	FilmArray Instrument

11

Page 8 of 58

12

13

14

Performance Characteristics - Non-Clinical Studies

Sensitivity (Limit of Detection)

The Analytical Sensitivity, or Limit of Detection (LoD), is defined as the lowest concentration at which >95% of the tested samples generate a positive call.

The LoD for each of the QIAstat-Dx Gastrointestinal Panel 2 target pathogenic organisms was assessed, using in total 36 pathogens strains, by analyzing serial dilutions of analytical samples prepared from culture isolates from commercial suppliers (e.g., ZeptoMetrix® and ATCC®) or clinical samples positive for target analytes commercially unavailable. Each sample tested was prepared in human stool matrix, which consists of a pool of previously tested negative clinical stool specimens resuspended in Para-Pak C&S (Meridian Biosciences) transport medium following the manufacturer's instructions collection device. A matrix equivalency study between Para-Pak C&S and FecalSwab transport media was conducted to support the conclusions in the section, except for EPEC, STEC stx1/stx2 and STEC O157 that are only applicable for Para-Pak C&S resuspended samples.

Individual LoD values for each QIAstat-Dx Gastrointestinal Panel 2 target is shown in Table 5.2.

Table 5.2: LoD concentration details for the 36 pathogen strains tested with QIAstat-Dx Gastrointestinal Panel 2

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen
Strain | Supplier &
Catalog ID | Dilution
from
the
stock | Detection
Rate | Concentration
(molecular
units) # | Concentration
(microbiological
units) |
|---------------------------------------------------------------|---------------------------------------------------------------------|------------------------------------------|----------------------------------|-----------------------------------------------|-----------------------------------------|---------------------------------------------|
| Campylobacter | Campylobacte
r coli 76-GA2
[LMG 21266] | ATCC
43478 | 1.00E-
06 | 20/20 | 5802
copies/mL | 1.2 CFU/mL |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen
Strain | Supplier &
Catalog ID | Dilution
from
the
stock | Detection
Rate | Concentration
(molecular
units) # | Concentration
(microbiological
units) |
| | Campylobacte
r coli CIP
7080 | ATCC
33559 | 1.00E-
05 | 20/20 | 8941
copies/mL | 0.6 CFU/mL |
| | Campylobacte
r jejuni Z086 | ZeptoMetrix
0801650 | 1.00E-
06 | 20/20 | 14491
copies/mL | 1660 CFU/mL |
| | Campylobacte
r jejuni subsp.
Jejuni
RM3193 | ATCC
BAA-1234 | 1.00E-
06 | 19/20 | 7210
copies/mL | 110 CFU/mL |
| | Campylobacte
r upsaliensis
NCTC 11541 | ZeptoMetrix
0801999 | 3.16E-
06 | 20/20 | 56165
copies/mL | 2259.4 CFU/mL |
| | Campylobacte
r upsaliensis
RM3195 | ATCC
BAA-1059 | 1.00E-
06 | 19/20 | 7631
copies/mL | 35 CFU/mL |
| Plesiomonas | Bader | ATCC
14029 | 3.16E-
07 | 19/20 | 116
copies/mL | 2.7 CFU/mL |
| shigelloides | Z130 | ZeptoMetrix
0801899 | 3.16E-
07 | 20/20 | 481
copies/mL | 2291 CFU/mL |
| Salmonella | Salmonella
enterica
Serovar
Typhimurium
Z005 | ZeptoMetrix
0801437 | 3.16E-
07 | 20/20 | 1441
copies/mL | 4518.8 CFU/mL |
| | Salmonella
enterica
Serovar
choleraseus | ATCC
13312 | 3.16E-
07 | 20/20 | 647
copies/mL | 91.6 CFU/mL |
| Yersinia
enterocolitica | subsp.
enterocolitica
NTCC 11175,
Biotype 4,
serotype 3 | ATCC
700822 | 3.16E-
07 | 20/20 | 2496
copies/mL | 120.1 CFU/mL |
| | Z036 | ZeptoMetrix
0801734 | 1.00E-
07 | 20/20 | 719
copies/mL | 2070 CFU/mL |
| | Escherichia
coli O111:NM
(EPEC) | ZeptoMetrix
0801747 | 3.16E-
07 | 20/20 | 1817
copies/mL | 2581.7 CFU/mL |
| Enteropathogenic
E. coli (EPEC)* | Escherichia
coli 7.1493;
EPEC;
084:H28 | ZeptoMetrix
0801938 | 1.00E-
06 | 20/20 | 29021
copies/mL | 1190 CFU/mL |
| Enterotoxigenic
E. coli (ETEC)
lt/st | Escherichia
coli ETEC;
ST+, LT+ | ZeptoMetrix
0801624 | 1.00E-
07 | 20/20 | 855
copies/mL | 567 CFU/mL |
| | Escherichia
coli H10407,
078:H11 | ATCC
35401 | 3.16E-
08 | 19/20 | 367
copies/mL | 10.1 CFU/mL |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen
Strain | Supplier &
Catalog ID | Dilution
from
the
stock | Detection
Rate | Concentration
(molecular
units) # | Concentration
(microbiological
units) |
| Shigella/
Enteroinvasive
E. coli (EIEC) | Escherichia
coli CDC EDL
1282,
029:NM | ATCC
43892 | 1.00E-
08 | 20/20 | 1431
copies/mL | 41.3 CFU/mL |
| | Shigella
sonnei NCDC
1120-66 | ATCC
25931 | 3.16E-
08 | 20/20 | 488
copies/mL | 0.2 CFU/mL |
| Shiga-like toxin-
producing
E.coli (STEC)
stx1/stx2* | Escherichia
coli O26:H4 | ZeptoMetrix
0801748 | 3.16E-
07 | STEC
stx1/stx2:
20/20 | 2012
copies/mL | 726.8 CFU/mL |
| Shiga-like toxin-
producing E.
coli (STEC)
0157* | Escherichia
coli O157:H7;
EDL933 | ZeptoMetrix
0801622 | 3.16E-
07 | STEC
stx1/stx2:
19/20
0157:
19/20 | 1217
copies/mL | 2281.5 CFU/mL |
| Cryptosporidium | Cryptosporidi
um parvum —
Iowa isolate | Waterborne
® P102C | 1.00E-
05 | 20/20 | 661
copies/mL | n/a |
| | Cryptosporidi
um hominis | Public
Health
Wales UKM
84 | 3.16E-
03 | 20/20 | 357
copies/mL | n/a |
| Cyclospora
cayetanensis | N/A | LACNY -
Clinical
sample
LAC2825 | 1.00E-
04 | 19/20 | 53 copies/mL | n/a |
| | N/A | LACNY
Clinical
sample
LAC2827 | 3.16E-
04 | 20/20 | 137
copies/mL | n/a |
| Entamoeba
histolytica | HM-1:IMSS
(Mexico City
1967) | ATCC
30459 | 1.00E-
06 | 20/20 | 7 copies/mL | 0.2 cells/mL |
| | HK-9 (Korea) | ATCC
30015 | 1.00E-
07 | 19/20 | 1 copy/mL | 0.13 cells/mL |
| Giardia lamblia | WB
(Bethesda) | ATCC
30957 | 3.16E-
05 | 19/20 | 11850
copies/mL | 790 cells/mL |
| | Portland-1 | ATCC
30888 | 1.00E-
04 | 20/20 | 14500
copies/mL | 635 cells/mL |
| Adenovirus
F40/F41 | Type 40
(Dugan) | ZeptoMetrix
0810084CF | 1.00E-
06 | 20/20 | 11726
copies/mL | 0.1 TCID50/mL |
| | Type 41 (Tak) | ZeptoMetrix
0810085CF | 1.00E-
07 | 19/20 | 979
copies/mL | 0.05 TCID50/mL |
| Astrovirus | ERE IID 2371
(type 8) | Zeptometrix
0810277CF | 1.00E-
04 | 20/20 | 11586371
copies/mL | 11.7 TCID50/mL |
| | ERE IID 2868
(type 4) | Zeptometrix
0810276CF | 3.16E-
06 | 19/20 | 52184
copies/mL | 1.3 TCID50/mL |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen
Strain | Supplier &
Catalog ID | Dilution
from
the
stock | Detection
Rate | Concentration
(molecular
units) # | Concentration
(microbiological
units) |
| Norovirus
GI/GII | GI.1
(recombinant) | ZeptoMetrix
0810086CF | 3.16E-
05 | 19/20 | 24629
copies/mL | 891.1 TCID50/mL |
| | GII.4
(recombinant) | ZeptoMetrix
0810087CF | 1.00E-
05 | 20/20 | 8998
copies/mL | 10.5 TCID50/mL |
| Rotavirus A | Wa | ZeptoMetrix
0810041CF | 1.00E-
04 | 19/20 | 5201
copies/mL | 14.1 TCID50/mL |
| | 69M | ZeptoMetrix
0810280CF | 3.16E-
05 | 19/20 | 5787
copies/mL | 436.1 TCID50/mL |

15

16

17

Molecular unit titers were determined using in-house developed and validated qPCR assays.

Target applicable for Para-Pak C&S samples only.

Analytical Reactivity (Inclusivity)

Analytical Reactivity (Inclusivity) was evaluated with gastrointestinal pathogen isolates/strains that were selected based on clinical relevance and genetic, temporal and geographical diversity. Based on in vitro (wet) testing and in silico analysis, the QIAstat-Dx Gastrointestinal Panel 2 primers and probes are specific and inclusive for clinically prevalent and relevant strains for each pathogen tested.

In vitro testing

QIAstat-Dx Gastrointestinal Panel 2 is inclusive for 100% (114 out of 114) of the pathogen strains tested in vitro. Most pathogen strains evaluated in laboratory testing were detected at ≤ 3-fold (104/114) of the corresponding LoD reference strain which is written in bold (refer to Table 5.3 to Error! Reference source not found.). Less than 100% detection was observed for one strain each of ETEC, EIEC/Shigella and Rotavirus and two strains each of STEC (one STEC O157), Adenovirus and Norovirus at 3x LoD. Testing of these strains at 10x LoD generated the expected positive resulted for all replicates.

Table 5.3: Inclusivity test results for Campylobacter strains

18

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
|--------------------------------------------------|---------------------------------------|----------------------------------|-----------------|---------------|---------------------------|
| | Campylobacter coli | 76-GA2
[LMG
21266] | ATCC | 43478* | 1x
LoD |
| | Campylobacter coli | Z293 | ZeptoMe
trix | 804272 | 1x
LoD |
| Campylobacter | Campylobacter coli | CIP 7080
[1407, CIP
70.80] | ATCC | 33559* | 3x
LoD |
| | Campylobacter jejuni | Z086 | ZeptoM
etrix | 080165
0* | 1x
LoD |
| | Campylobacter jejuni | subsp. jejuni
RM3193 | ATCC | BAA-
1234* | 0.1x
LoD |
| | Campylobacter jejuni
subsp. jejuni | O:19 HL7;
D3180 | ATCC | BAA-
218 | 0.1x
LoD |
| | Campylobacter jejuni
subsp. jejuni | AS-83-79 | ATCC | 33291 | 0.1x
LoD |
| | Campylobacter jejuni
subsp. doylei | NCTC
11951 | ATCC | 49349 | 0.1x
LoD |
| | Campylobacter
upsaliensis | NCTC
11541 | ZeptoM
etrix | 080199
9* | 1x
LoD |
| | Campylobacter
upsaliensis | RM 3195
(1994) | ATCC | BAA-
1059* | 0.3x
LoD |
| | Campylobacter
upsaliensis | NCTC
11541
[C2311 | ATCC | 43954 | 1x
LoD |

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
|--------------------------------------------------|-----------------------------|------------------------------------------------------------------------|-----------------|---------------|---------------------------|
| Plesiomonas
shigelloides | Plesiomonas
shigelloides | Z130 | ZeptoM
etrix | 080189
9* | 1x
LoD |
| | Plesiomonas
shigelloides | GNI 14 | ATCC | 51903 | 1x
LoD |
| | Plesiomonas
shigelloides | CDC 3085-
55 [Bader
M51, NCIB
9242, NCTC
10360, RH
798] | ATCC | 14029* | 0.3x
LoD |

				Table 5.4: Inclusivity test results for Plesiomonas shigelloides strains
--	--	--	--	--------------------------------------------------------------------------	--	--	--

19

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
|--------------------------------------------------|---------------------|-------------------------------------------------------------------------------|-------------|---------------|---------------------------|
| Salmonella | Salmonella enterica | Serovar
Typhimurium Z005 | ZeptoMetrix | 080143
7* | 1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Bareilly | NCTC | NC057
45 | 1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
typhi, Z152 | ZeptoMetrix | 080193
3 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Enteridis,
CDC K-1891
[ATCC
25928] | ATCC | 13076 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Infantis,
MZ1479
[SARB27] | ATCC | BAA-
1675 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Montevideo,
G4639 | ATCC | BAA-
710 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Javiana | NCTC | NC064
95 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Thompson | NCTC | NC084
96 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica, | ATCC | 9712 | 0.1x
LoD |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
| | | serovar
Saintpaul | | | |
| | Salmonella enterica | Subsp.
Enterica,
serovar Berta | NCTC | NC057
70 | 0.1x
LoD |
| | Salmonella enterica | Subps.
Salame, II
NCTC
10310
[JT945,
SS140/61] | ATCC | 700151 | 0.1x
LoD |
| | Salmonella enterica | Subps.
diarizonae
IIIb, 62 | ATCC | 29934 | 0.1x
LoD |
| | Salmonella enterica | Subps.
houtenae IV,
CIP 82.32
[264.66] | ATCC | 43974 | 0.1x
LoD |
| | Salmonella enterica | Subps.
Indica VI,
CIP 102501
[F.
Kauffmann
1240] | ATCC | 43976 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Agona, CDC
873 [CDC
1111-61] | ATCC | 51957 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Muenchen,
54 | ATCC | 8388 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Oranienburg,
E1093 | ATCC | 9239 | 0.1x
LoD |
| | Salmonella enterica | Subsp.
Enterica, | ATCC | 51962 | 0.1x
LoD |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
| | | serovar
Paratyphi B
var. Java,
CDC 5 | | | |
| | Salmonella bongori | CIP 82.33
[1224.72] | ATCC | 43975 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Choleraesius
,NCTC 5735
[1348, K.34] | ATCC | 13312* | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Newport,
C487-69 | ATCC | 27869 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica, 4,
5, 12:7:-,
serovar
Typhimurium
m | NCTC | NC139
52 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Braenderup | ATCC | 700136 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Anatum | NCTC | NC057
79 | 0.3x
LoD |
| | Salmonella enterica | Subps.
arizonae IIIa,
NCTC 7311
[CDAI 426] | ATCC | 700156 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica,
serovar
Heidelberg,
[16] | ATCC | 8326 | 0.3x
LoD |
| | Salmonella enterica | Subsp.
Enterica, | ATCC | BAA-
2739 | 0.3x
LoD |
| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detected |
| | | serovar
Mississippi,
CDC 2012K-
0487 | | | |

Table 5.5: Inclusivity test results for Salmonella strains

20

21

22

Table 5.6: Inclusivity test results for Yersinia enterocolitica strains

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catal
og ID | x-fold
LoD
detected |
|--------------------------------------------------|----------------------------|---------------------------------------------------------------------------------------------------------------|-----------------|----------------|---------------------------|
| Yersinia
enterocolitica | Yersinia
enterocolitica | Z036 | ZeptoMet
rix | 08017
34* | 1x LoD |
| | Yersinia
enterocolitica | NTCC 11175,
Biotype 4,
serotype 3
(0:3) | ATCC | 70082
2* | 1x LoD |
| | Yersinia
enterocolitica | 33114 [CCUG
11291, CCUG
12369, CIP
80.27, DSM
4780, LMG
7899, NCTC
12982], Biovar
1, 0:8 | ATCC | 9610 | 1x LoD |
| | Yersinia
enterocolitica | 0:9 | ATCC | 55075 | 3x LoD |

Strain tested during LoD verification study

Table 5.7: Inclusivity test results for Enteropathogenic E. coli (EPEC) strains. Only applicable for Para-Pak C&S samples.

| QIAstat-Dx
Gastrointestinal
Panel 2 Target | Pathogen | Strain | Supplier | Catalog
ID | x-fold
LoD
detecte
d |
|--------------------------------------------------|------------------------------------|--------------------------------|-----------------|---------------|-------------------------------|
| Enteropathogenic E.
coli (EPEC) | Enteropathogenic E. coli
(EPEC) | O111:NM | ZeptoMetri
x | 080174
7* | 1x
LoD |
| | Enteropathogenic E. coli
(EPEC) | 7.1493,O84:H28 | ZeptoMetrix | 080193
8* | 1x
LoD |
| | Enteropathogenic E. coli
(EPEC) | Stoke
W,O111:K58(B
4):H- | ATCC | 33780 | 1x
LoD |

23

Table 5.8: Inclusivity test results for Enterotoxigenic E. coli (ETEC) strains
QIAstat-Dx
Gastrointestinal
Panel 2 Target	Pathogen	Strain	Supplier	Catalog
ID	x-fold
LoD
detected
Enterotoxigenic E.
coli (ETEC) lt/st	Enterotoxigenic E. coli
(ETEC) lt/st	ST+, LT+	ZeptoMet
rix	0801624

| 1x LoD |
| | Enterotoxigenic E. coli
(ETEC) lt/st | H10407,O78:H1
1,LT(+) / ctx
A11(+) | ATCC | 35401* | 0.3x
LoD |
| | Enterotoxigenic E. coli
(ETEC) lt/st | O27:H7,ST (+)/
LT (-) | SSI
Diagnostic
a | 82173 | 0.1x
LoD |
| | Enterotoxigenic E. coli
(ETEC) lt/st | O115:H15,ST
(+)/ LT (-) | SSI
Diagnostic
a | 82174 | 3x LoD |
| | Enterotoxigenic E. coli
(ETEC) lt/st | O169:H-,ST (-) /
LT (+) | SSI
Diagnostic
a | 82172 | 10x
LoD# |

Table 5.8: Inclusivity test results for Enterotoxigenic E. coli (ETEC) strains

Strain tested during LoD verification study

Testing at a lower concentration resulted in a detection rate of Shigella boydii 1,2,3,

1Note that these potential cross-reactions identified by in-silico analysis reflects sequences which can be acquired between species by horizontal gene transfer.

2Rare or less common eae intimin carrier organisms

3On-panel target.

41n vitro testing of Campylobacter lari and Campylobacter helveticus strains at high concentration confirmed potential cross-reactivity of these Campylobacter species with the QIAstat-Dx Gastrointestinal Panel 2 assay. 5 Rare or less common Stx toxins producers

6 E. coli 0157 reported by the QIAstrointestinal Panel will only be called when there is a positive amplification for the E. coli (STEC) design according to the calling algorithm. An infrequent case of an E. coli (STEC)

and an E. coli 0157 co-infection they will not be differentiated from a single infection caused by an STEC 0157:H7 strain.

7 Target only applicable for Para-Pak C&S samples.

Interfering Substances

The effect of potentially interfering substances on the detectability of the QIAstat-Dx Gastrointestinal Panel 2 organisms was evaluated. Thirty-four (34) potentially interfering substances were spiked into the sample mixes at a level predicted to be above the concentration of the substance likely to be found in stool specimens. Each organism was tested at 3x LoD and testing was performed in triplicates. Endogenous substances such as human whole blood, human genomic DNA and several pathogens were tested alongside exogenous substances like antibiotics, other gastrointestinal-related medications and different technique-specific substances.

For the vast majority of substances tested, no inhibition was observed, with the exceptions of mucin, calcium carbonate, nonoxynol-9 and Rotavirus reassortants, that may cause inhibition at high concentration.

Mucin at 5% w/v was found to generate false positives results for the Yersinia target. These signals were investigated by testing the interfering substance with and FDA-cleared method and they were confirmed to be present in the endogenous substance.

36

Calcium carbonate was found to interfere with the detection of all the QIAstat-Dx Gastrointestinal Panel 2 targets at concentrations above 0.5% w/v.

Nonoxynol-9 was found to interfere with the detection of Entamoeba histolytica at concentrations above 0.02% v/v.

As predicted, Rotavirus reassortants WC3:2-5, R574(9) and W179-4,9 used in Rotavirus A vaccines generated positive results for Rotavirus A in the QIAstat-Dx Gastrointestinal Panel 2. Final concentrations without interference (i.e. no false positive results for Rotavirus) for WC3:2-5, R574(9) and W179-4,9 were 8.89x1055 TCID50/m1 and 1.10 PFU/ml, respectively (see Table 5.24 for other concentrations tested).

Results from the 34 interfering substances that could be present or introduced in a stool specimen are provided in Table 5.24.

				Table 5.24: Final highest concentrations per potentially interfering substance
without observable interference on the QIAstat-Dx Gastrointestinal Panel 2

Substance tested	Concentration tested	Result
Endogenous substances
Bovine and ovine bile	12% w/v	No Interference
Cholesterol	1.5% w/v	No Interference
Fatty acids (palmitic acid)	0.2% w/v	No Interference
Fatty acids (stearic acid)	0.4% w/v	No Interference
Human genomic DNA	20 µg/mL	No Interference
	10 µg/mL	No Interference
	5 µg/mL	No Interference
Human stool (overfill of Cary Blair vial)	300 mg/mL	No Interference
Human urine	50% v/v	No Interference
Human whole blood with Na Citrate	40% v/v	No Interference
Mucin from bovine submaxillary	5% w/v	Interference#
	2.5% w/v	No Interference
Triglycerides	5% w/v	No Interference
Exogenous substances
Bacitracin	250U/mL	No Interference
Bisacodyl	0.3% w/v	No Interference
	0.15% w/v	No Interference
Bismuth subsalicylate	0.35% w/v	No Interference
Calcium carbonate (TUMS® Extra
Strength 750)	5% w/v	Interference
Docusate sodium	0.5% w/v	No Interference
Docusate sodium	2.5% w/v	No Interference
Doxycycline hydrochloride	0.05% w/v	No Interference
Glycerin	50% v/v	No Interference
Hydrocortisone	0.5% w/v	No Interference
Loperamide hydrochloride	0.078% w/v	No Interference

37

Substance tested	Concentration tested	Result
Magnesium hydroxide	0.1% w/v	No Interference
Metronidazole	1.5% w/v	No Interference
Mineral oil	50% v/v	No Interference
Naproxen sodium	0.7% w/v	No Interference
Nonoxynol-9	1.2% v/v	Interference
	0.6% v/v	Interference
	0.3% v/v	Interference
	0.15% v/v	Interference
	0.075% v/v	Interference
	0.02% v/v	No Interference
Nystatin	10000 USP units/mL	No Interference
Phenylephrine hydrochloride	0.075% w/v	No Interference
Sodium phosphate	5% w/v	No Interference
Vaccine components
Rotavirus reassortant WC3:2-5, R574(9) -
VR 2195	$8.89 \times 10^{-3}$ TCID50/mL	Interference
	$8.89 \times 10^{-4}$ TCID50/mL	Interference
	$8.89 \times 10^{-5}$ TCID50/mL	No Interference
Rotavirus reassortant WI79-4,9 - VR 2415	$1.10 \times 10^{2}$ pfu/mL	Interference
	$1.10 \times 10^{1}$ pfu/mL	Interference
	1.10 pfu/mL	No Interference
Technique-specific Substances, Transport Media
Bleach	0.5% v/v	No Interference
Ethanol	0.2% v/v	No Interference
Puritan Fecal Opti-Swab Collection &
Transport System with Cary-Blair
Medium*	100%	No Interference
PurSafe DNA/RNA Preservative*	100%	No Interference
Sigma Fecal Transwab*	1 swab/2mL Cary Blair	No Interference

*Performance not stablished for this transport media

This substance was tested by another FDA-cleared that also detected Yersinia positive signals.

38

Microbial Interference

A microbial interference study was conducted to assess the inhibitory effects of select nontarget organisms on the ability to detect the QIAstat-Dx Gastrointestinal Panel 2 targets. Clinically relevant and challenging concentrations of non-target organisms were individually mixed with negative clinical stool matrix with spiked targeted pathogens at 3x LoD. Testing was performed in triplicate. All combinations and replicates successfully detected all the QIAstat-Dx Gastrointestinal Panel 2 targets. See Table 5.25 for a list of the non-target organisms tested and the result summary.

Substance tested	Concentration tested	Result
Non-target microorganisms
Aeromonas hydrophila	1 x 106 units/mL	No Interference
Bacteroides vulgatus	1 x 106 units/mL	No Interference
Bifidobacterium bifidum	1 x 106 units/mL	No Interference
Enterovirus Specie D, Serotype EV-D68	1 x 105 units/mL	No Interference
Non-pathogenic E. coli	1 x 106 units/mL	No Interference
Helicobacter pylori	1 x 106 units/mL	No Interference
Saccharomyces cerevisiae (deposited as S.
boulardii)	1 x 105 units/mL	No Interference

Table 5.25: Final highest concentration without observable inhibitory effect

Competitive Interference

Competitive interference was tested in a subset of pathogens. No interference was observed when evaluating competitive interference by target pathogens when two QIAstat-Dx Gastrointestinal Panel target pathogens were tested by spiking samples with one pathogen target at 3x LoD and one at 50x LoD. Results from the pathogen targets tested are provided in Table 5.26.

39

| Mix | Pathogen | Final
concentration
(molecular
units)* | Final
concentration
tested xLoD | Detection
rate | Target detected |
|--------------------|-----------------------|-------------------------------------------------|---------------------------------------|-------------------|-----------------|
| Norovirus
50x - | Norovirus
GI/GII | 4.5E+05
copies/mL | 50x | 3/3 | Yes |
| Rotavirus
3x | Rotavirus A | 1.7E+04
copies/mL | 3x | 3/3 | Yes |
| Norovirus
3x - | Norovirus
GI/GII | 2.7E+04
copies/mL | 3x | 3/3 | Yes |
| Rotavirus
50x | Rotavirus A | 2.9E+05
copies/mL | 50x | 3/3 | Yes |
| Giardia 50x | Giardia
lamblia | 7.2 E+05
copies/mL | 50x | 3/3 | Yes |
| Adenovirus
3x | Adenovirus
F40/F41 | 2.9E+03
copies/mL | 3x | 3/3 | Yes |

Table 5.26: QIAstat-Dx Gastrointestinal Panel 2 Detection Rate Results for Competitive Interference

Molecular unit titers were determined using in house developed and validated qPCR assays.

Carryover

A carryover study was performed to evaluate the potential occurrence of crosscontamination between consecutive runs when using the QIAstat-Dx Gastrointestinal Panel 2 on the QIAstat-Dx Analyzer 1.0.

Pathogen samples of stool sample matrix, with alternating high-positive (10-10% organism/mL) and negative samples, were conducted on two QIAstat-Dx Analyzer 1.0.

No carryover between samples was observed in the QIAstat-Dx Gastrointestinal Panel 2, demonstrating that the system design and recommended sample handling and testing practices are effective in preventing false positive results due to carryover or crosscontamination between samples.

40

Sample Stability

Sample stability testing demonstrated that the OIAstat-Dx Gastrointestinal Panel 2 assay is capable of processing samples which are stored prior to analysis under conditions typically utilized for stool specimens according to the intended use.

The results of this study support the following recommendations for storage of stool resuspended in Para-Pak C&S and FecalSwab transport media before testing:

Room temperature up to 4 days at 15–25°C
Refrigerated up to 4 days at 2-8°C ●

Reproducibility

Reproducibility testing of contrived samples was performed at three test sites including one internal site (Site A) and two external sites (Site B and Site C). The study incorporated a range of potential variation introduced by sites, days, replicates, cartridge lots, operators, and QIAstat-Dx analyzers. For each site, testing was performed across 5 non-consecutive days with 6 replicates per day (leading to a total of 30 replicates per target, concentration and site), 4 OIAstat-Dx Analyzers (2 analyzers per operator and per site), and at least 2 operators on each testing day. A total of 5 sample mixes (two combined samples at 1x LoD and 3x LoD plus one negative sample) were prepared. For each mix, 6 replicates were tested and evaluated. shows the detection rate per target and concentration for each site of the Reproducibility study. In addition, data obtained at all three sites have been compiled to calculate the exact 2-sided 95% Confidence Interval by target and concentration.

Table 5.27 shows the detection rate per target and concentration for each site of the Reproducibility study. In addition, data obtained at all three sites have been compiled to calculate the exact 2-sided 95% Confidence Interval by target and concentration.

41

Table 5.27: Detection rate per target and concentration for each site of the Reproducibility study and exact 2-sided 95% Confidence Interval by target and concentration

| Pathogen Tested | Concentration
Tested | Expected
Result | % Agreement with Expected Result | | | All Sites
(95%
Confidence
Interval) |
|-----------------------------------------------------------------|-------------------------|----------------------------------|----------------------------------|-----------------|-----------------|----------------------------------------------|
| | | | Site A | Site B | Site C | |
| Adenovirus F41
ZeptoMetrix
0810085CF | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | None | Not
Detected | 29/30
96.67% | 29/30
96.67% | 29/30
96.67% | 87/90a
96.67%
(90.98-
98.9%) |
| Campylobacter
ZeptoMetrix
0801650 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30b
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| Pathogen Tested | Concentration Tested | Expected Result | % Agreement with Expected Result | | | All Sites (95% Confidence Interval) |
| | | | Site A | Site B | Site C | |
| Escherichia coli EPECd
ZeptoMetrix 0801747 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| | 1x LoD | Detected | 30/30
100% | 29/30
96.67% | 30/30
100% | 89/90
98.89%
(93.96 – 99.97%) |
| | None | Not Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| Entamoeba histolytica
ATCC 30459 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30
100% | 29/30
96.67% | 89/90
98.89%
(93.96 – 99.97%) |
| | None | Not Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| Giardia lambliab
ATCC 30888 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| | None | Not Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 - 100.00%) |
| | | | % Agreement with Expected Result | | | |
| Pathogen Tested | Concentration
Tested | Expected
Result | Site A | Site B | Site C | All Sites
(95%
Confidence
Interval) |
| Norovirus GII
ZeptoMetrix
0810087CF | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 29/30
96.67% | 30/30
100% | 30/30
100% | 89/90
98.89%
(93.96 -
99.97%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| Rotavirus Ac
ZeptoMetrix
0810280CF | 3x LoD | Detected | 29/30
96.67% | 29/30
96.67% | 30/30
100% | 88/90
97.8%
(92.20 -
99.73%) |
| | 1x LoD | Detected | 23/30
76.67% | 26/30
86.67% | 12/12
100% | 61/72
84.7%
(74.31 -
92.12%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| Escherichia coli
(STEC) O157:H7d
ZeptoMetrix
0801622 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30
100% | 29/30
96.67% | 89/90
98.89%
(93.96 -
99.97%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00% ) |
| | | % Agreement with Expected Result | | | | |
| Pathogen Tested | Concentration
Tested | Expected
Result | Site A | Site B | Site C | All Sites
(95%
Confidence
Interval) |
| Escherichia coli
(STEC) stx1/stx2d
ZeptoMetrix
0801622 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| Salmonella
enterica
ZeptoMetrix
0801437 | 3x LoD | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 -
100.00%) |
| | 1x LoD | Detected | 30/30
100% | 29/30
96.67% | 29/30
96.67% | 88/90
97.78%
(92.20 –
99.73%) |
| | None | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(95.98 –
100.00%) |

42

43

44

45

			% Agreement with Expected Result
Pathogen Tested	Concentration
Tested	Expected
Result	Site A	Site B	Site C	All Sites
(95%
Confidence
Interval)
Yersinia
enterocolitica
ZeptoMetrix
0801734	3x LoD	Detected	30/30
100%	30/30
100%	30/30
100%	90/90
100%
(95.98 -
100.00% )
	1x LoD	Detected	30/30
100%	30/30
100%	30/30
100%	90/90
100%
(95.98 -
100.00%)
	None	Not
Detected	30/30
100%	30/30
100%	30/30
100%	90/90
100%
(95.98 -
100.00%)

4 Three (3) Adenovirus F40/41 false positives were observed when testing negative sample. Retesting of the three samples resulted in the expected negative results.

b One (1) Giardia lamblia false positive was observed when testing a positive sample not containing the pathogen. Repeat testing of this sample resulted in the expected negative result.

6 The Reproducibility study was fully re-tested for Rotavirus A with a new sample set due to an unexpected number of false negatives for Rotavirus A at the 1x LoD concentration. This was observed during an interim data evaluation (61/72, 84.7%) and was attributed to the sample manufacture while unrelated to the study workflow variables (operator, lot, day, instrument and site). Test runs derived from Rotavirus A new sample set resulted in 90/90 (100%; 95.98-100% CI) for the 3x LoD and 89/90 (98.89%; 93.96-99.97% CI) for the 1x LoD. During this testing, one (1) Campylobacter false positive was observed. Retesting of this sample resulted in the expected negative result.

d Target only applicable for Para-Pak C&S samples.

During the reproducibility study, potential variation introduced by sites, days, replicates, cartridge lots, operators, and QIAstat-Dx analyzers was analyzed showing no significant contribution to variability (Standard Deviation and Coefficient of Variation values below 1 and 5%, respectively) caused by any of the assessed variables (Table 5.28).

46

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Table 5.28: Variability attributable to each variable according to ANOVA model for QIAstat-Dx Gastrointestinal Panel 2.

Pathogen	Concentration tested	Positive (n)	Negative (n)	Mean	Variance component (SD, %CV*)					Total**
					Between Site	Between Lot	Between Day	Between Operator	Between Instrument	Between Replicate
Adenovirus	1x LoD	90	0	34.15	(0.2979, 0.87%)	(0.1630, 0.48%)	(0.1135, 0.33%)	(0.0000, 0.00%)	(0.1883, 0.55%)	(0.6153, 1.80%)	(0.7067, 2.07%)
	3x LoD	90	0	32.14	(0.0000, 0.00%)	(0.0741, 0.23%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.3716, 1.16%)	(0.3766, 1.17%)
Campylobacter	1x LoD	90	0	32.64	(0.1289, 0.39%)	(0.0000, 0.00%)	(0.0200, 0.06%)	(0.0000, 0.00%)	(0.0661, 0.20%)	(0.4232, 1.30%)	(0.4411, 1.35%)
	3x LoD	90	0	31.00	(0.0397, 0.13%)	(0.1446, 0.47%)	(0.0806, 0.26%)	(0.0000, 0.00%)	(0.1488, 0.48%)	(0.3420, 1.10%)	(0.3855, 1.24%)
EPEC	1x LoD	89	1	34.24	(0.4422, 1.29%)	(0.2000, 0.58%)	(0.0000, 0.00%)	(0.1599, 0.47%)	(0.1044, 0.30%)	(0.6699, 1.96%)	(0.7927, 2.31%)
	3x LoD	90	0	32.49	(0.1053, 0.32%)	(0.1429, 0.44%)	(0.0000, 0.00%)	(0.2081, 0.64%)	(0.0000, 0.00%)	(0.5238, 1.61%)	(0.5710, 1.76%)
Entamoeba	1x LoD	89	1	33.01	(0.0000, 0.00%)	(0.1349, 0.41%)	(0.0000, 0.00%)	(0.1175, 0.36%)	(0.0000, 0.00%)	(0.3182, 0.96%)	(0.3456, 1.05%)
	3x LoD	90	0	31.34	(0.1826, 0.58%)	(0.1265, 0.40%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.0906, 0.29%)	(0.4000, 1.28%)	(0.4449, 1.42%)
Giardia	1x LoD	90	0	28.65	(0.2341, 0.82%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.1737, 0.61%)	(0.5386, 1.88%)	(0.5953, 2.08%)
	3x LoD	90	0	26.68	(0.0840, 0.31%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.1025, 0.38%)	(0.0857, 0.32%)	(0.3531, 1.32%)	(0.3798, 1.42%)
Norovirus GII	1x LoD	89	1	33.53	(0.1346, 0.40%)	(0.3090, 0.92%)	(0.1111, 0.33%)	(0.0000, 0.00%)	(0.0431, 0.13%)	(0.4102, 1.22%)	(0.5083, 1.52%)
	3x LoD	90	0	32.42	(0.1742, 0.54%)	(0.3405, 1.05%)	(0.2895, 0.89%)	(0.0632, 0.19%)	(0.1972, 0.61%)	(0.4966, 1.53%)	(0.6721, 2.07%)
Rotavirus ***	1x LoD	61	11	36.71	(0.0000, 0.00%)	(0.1487, 0.41%)	(0.0000, 0.00%)	(0.1824, 0.50%)	(0.0000, 0.00%)	(0.9172, 2.50%)	(0.9356, 2.55%)
	3x LoD	88	2	35.95	(0.3468, 0.96%)	(0.4463, 1.24%)	(0.1231, 0.34%)	(0.1295, 0.36%)	(0.1132, 0.31%)	(1.2073, 3.36%)	(1.3014, 3.62%)
STEC_0157	1x LoD	89	1	33.83	(0.2218, 0.66%)	(0.3083, 0.91%)	(0.0000, 0.00%)	(0.0204, 0.06%)	(0.0000, 0.00%)	(0.5455, 1.61%)	(0.6295, 1.86%)
	3x LoD	90	0	32.22	(0.2431, 0.75%)	(0.2146, 0.67%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.2051, 0.64%)	(0.4625, 1.44%)	(0.5633, 1.75%)
STEC_stx1	1x LoD	90	0	34.07	(0.2297, 0.67%)	(0.1214, 0.36%)	(0.1281, 0.38%)	(0.0000, 0.00%)	(0.1170, 0.34%)	(0.4206, 1.23%)	(0.4991, 1.46%)
	3x LoD	90	0	32.54	(0.2380, 0.73%)	(0.2222, 0.68%)	(0.0802, 0.25%)	(0.0000, 0.00%)	(0.2000, 0.61%)	(0.4919, 1.51%)	(0.5887, 1.81%)
STEC_stx2	1x LoD	90	0	34.11	(0.0620, 0.18%)	(0.0979, 0.29%)	(0.0310, 0.09%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.3987, 1.17%)	(0.4109, 1.20%)
	3x LoD	90	0	32.69	(0.1173, 0.36%)	(0.2599, 0.80%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.1316, 0.40%)	(0.4131, 1.26%)	(0.4821, 1.47%)
Salmonella	1x LoD	88	2	34.53	(0.2888, 0.84%)	(0.2631, 0.76%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.0000, 0.00%)	(0.4880, 1.41%)	(0.5857, 1.70%)
	3x LoD	90	0	33.44	(0.0279, 0.08%)	(0.2100, 0.63%)	(0.0000, 0.00%)	(0.1814, 0.54%)	(0.1177, 0.35%)	(0.4088, 1.22%)	(0.4715, 1.41%)
Yersinia	1x LoD	90	0	34.53	(0.1616, 0.47%)	(0.1688, 0.49%)	(0.0564, 0.16%)	(0.0000, 0.00%)	(0.1036, 0.30%)	(0.5172, 1.50%)	(0.5637, 1.63%)
	3x LoD	90	0	33.02	(0.0503, 0.15%)	(0.2129, 0.64%)	(0.0370, 0.11%)	(0.0000, 0.00%)	(0.0801, 0.24%)	(0.3434, 1.04%)	(0.3951, 1.20%)

SD is first figure presented, subsequent figure is CV (Coefficient of variation presented as a percentage (%)
** TOTAL means total variance observed across the study

*** Results in table correspond to the original sample set analyzed. Results from the new dataset were:

					Variance component (SD, %CV*)
Pathogen	Concentration
tested	Positive
(n)	Negative
(n)	Mean	Between
Site	Between
Lot	Between
Day	Between
Operator	Between
Instrument	Between
Replicate	Total**
Rotavirus	1x LoD	89	1	34.86	(0.1706,
0.49%)	(0.0000,
0.00%)	(0.3458,
0.99%)	(0.1207,
0.35%)	(0.3224,
0.92%)	(0.9814,
2.82%)	(1.0891,
3.12%)
	3x LoD	90	0	32.42	(0.1742,
0.54%)	(0.3405,
1.05%)	(0.2895,
0.89%)	(0.0632,
0.19%)	(0.1972,
0.61%)	(0.4966,
1.53%)	(0.6721,
2.07%)

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Repeatability

A repeatability study was conducted on the OIAstat-Dx Analyzer 1.0 instruments using a set of samples composed of low-concentrated analytes spiked into stool matrix (3x LoD and 1x LoD) and negative stool samples. OIAstat-Dx Gastrointestinal Panel 2 detected pathogens included in the positive samples were Adenovirus, Campylobacter, Enteropathogenic E. coli (EPEC), Entamoeba histolytica, Giardia lamblia, Norovirus GII, Rotavirus, E. coli 0157, STEC stx1/stx2, Salmonella enterica, and Yersinia enterocolitica. Each sample was tested with the same instrument over 12 days. In total. 60 replicates of 1x LoD and 60 replicates of 3x LoD per each of the tested targets and 60 replicates of negative samples were run. Overall results showed a 93.33-100.00-% and 95.00-100.00% detection rate for 1x LoD and 3x LoD samples, respectively. Negative samples showed 100% of negative calls for all panel analytes.

Performance Characteristics - Clinical Studies

Expected values

In the prospective clinical performance evaluation of the QIAstat-Dx Gastrointestinal Panel 2, 1939 eligible specimens (stool preserved in Para-Pak C&S (Meridian Bioscience) or FecalSwab (COPAN)) were collected and tested at 13 clinical sites across 5 countries (4 sites in Europe and 9 sites in USA) from May 2021. The number and percentage of positive results as determined by the QIAstat-Dx Gastrointestinal Panel 2, stratified by age group, are presented in Table 5.29. Overall, the QIAstat-Dx Gastrointestinal Panel 2 detected at least 1 organism in in 17.4% (213/1222) and 23.8% (171/717) of the prospectively collected stool specimens in FecalSwab and Para-Pak C&S, respectively (Table 5.29).

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Table 5.29: Expected Values Summary by Age Group for the Prospective Clinical study as determined by the QIAstat-Dx Gastrointestinal Panel 2

| Pathogen | Medium Brand | Overall | 0-5 years | 6-21 years | 22-49 years | 50+ years | Not
Reported |
|----------------------------------------------|--------------|-----------|------------|------------|-------------|-----------|-----------------|
| Viruses | | | | | | | |
| Adenovirus F40/F41 | FecalSwab | 5 (0.4%) | 3 (1.7%) | 2 (1.7%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 2 (0.3%) | 1 (3.2%) | 0 (0.0%) | 0 (0.0%) | 1 (0.2%) | 0 (0.0%) |
| Astrovirus | FecalSwab | 3 (0.2%) | 3 (1.6%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 6 (0.8%) | 2 (6.5%) | 0 (0.0%) | 3 (1.4%) | 1 (0.2%) | 0 (0.0%) |
| Norovirus GI/GII | FecalSwab | 43 (3.5%) | 22 (12.1%) | 1 (0.8%) | 14 (4.8%) | 6 (1.0%) | 0 (0.0%) |
| | Para-Pak C&S | 16 (2.3%) | 3 (9.7%) | 1 (2.8%) | 3 (1.4%) | 9 (2.2%) | 0 (0.0%) |
| Rotavirus A | FecalSwab | 23 (1.9%) | 13 (7.1%) | 2 (1.7%) | 7 (2.4%) | 1 (0.2%) | 0 (0.0%) |
| | Para-Pak C&S | 4 (0.6%) | 2 (6.5%) | 0 (0.0%) | 0 (0.0%) | 2 (0.5%) | 0 (0.0%) |
| Bacteria | | | | | | | |
| Campylobacter | FecalSwab | 69 (5.6%) | 25 (13.7%) | 7 (5.8%) | 17 (5.9%) | 20 (3.2%) | 0 (0.0%) |
| | Para-Pak C&S | 30 (4.2%) | 2 (6.5%) | 0 (0.0%) | 10 (4.7%) | 18 (4.3%) | 0 (0.0%) |
| Plesiomonas shigelloides | FecalSwab | 2 (0.2%) | 0 (0.0%) | 0 (0.0%) | 2 (0.7%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 7 (1.0%) | 1 (3.2%) | 0 (0.0%) | 4 (1.9%) | 2 (0.5%) | 0 (0.0%) |
| Salmonella | FecalSwab | 14 (1.1%) | 5 (2.7%) | 4 (3.3%) | 3 (1.0%) | 2 (0.3%) | 0 (0.0%) |
| | Para-Pak C&S | 17 (2.4%) | 4 (12.9%) | 0 (0.0%) | 3 (1.4%) | 10 (2.4%) | 0 (0.0%) |
| Yersinia enterocolitica | FecalSwab | 22 (1.8%) | 3 (1.6%) | 2 (1.7%) | 9 (3.1%) | 8 (1.3%) | 0 (0.0%) |
| | Para-Pak C&S | 8 (1.1%) | 0 (0.0%) | 0 (0.0%) | 4 (1.9%) | 4 (1.0%) | 0 (0.0%) |
| Diarrheagenic E. coli/Shigella | | | | | | | |
| Enteropathogenic E. coli
(EPEC) | Para-Pak C&S | 56 (7.9%) | 9 (29.0%) | 2 (5.6%) | 18 (8.4%) | 27 (6.5%) | 0 (0.0%) |
| Enterotoxigenic E. coli | FecalSwab | 18 (1.5%) | 2 (1.1%) | 2 (1.7%) | 11 (3.8%) | 3 (0.5%) | 0 (0.0%) |
| (ETEC) lt/st | Para-Pak C&S | 17 (2.4%) | 1 (3.2%) | 0 (0.0%) | 7 (3.3%) | 9 (2.2%) | 0 (0.0%) |
| Shiga-like toxin E. coli
(STEC) stx1/stx2 | Para-Pak C&S | 9 (1.3%) | 0 (0.0%) | 0 (0.0%) | 6 (2.8%) | 3 (0.7%) | 0 (0.0%) |
| E. coli 0157 | Para-Pak C&S | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |
| Shigella/Enteroinvasive | FecalSwab | 10 (0.8%) | 1 (0.5%) | 0 (0.0%) | 6 (2.1%) | 3 (0.5%) | 0 (0.0%) |
| E. coli (EIEC) | Para-Pak C&S | 3 (0.4%) | 0 (0.0%) | 0 (0.0%) | 1 (0.5%) | 2 (0.5%) | 0 (0.0%) |
| Parasites | | | | | | | |
| Cryptosporidium | FecalSwab | 2 (0.2%) | 0 (0.0%) | 1 (0.8%) | 1 (0.3%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 7 (1.0%) | 0 (0.0%) | 1 (2.8%) | 4 (1.9%) | 2 (0.5%) | 0 (0.0%) |
| Cyclospora cayetanensis | FecalSwab | 3 (0.2%) | 0 (0.0%) | 1 (0.8%) | 2 (0.7%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 18 (2.5%) | 0 (0.0%) | 0 (0.0%) | 6 (2.8%) | 12 (2.9%) | 0 (0.0%) |
| Giardia lamblia | FecalSwab | 15 (1.2%) | 3 (1.6%) | 1 (0.8%) | 7 (2.4%) | 4 (0.6%) | 0 (0.0%) |
| | Para-Pak C&S | 1 (0.1%) | 1 (3.2%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |
| Entamoeba histolytica | FecalSwab | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |
| | Para-Pak C&S | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) |

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Clinical Performance

The clinical performance of QIAstat-Dx Gastrointestinal Panel 2 was established during a multi-center international prospective study conducted at thirteen clinical settings representative of different geographical areas within USA and Europe (9 sites in USA and 4 sites in Europe) between May and July 2021. All study sites were hospital-associated or independent clinical diagnostics laboratories that perform routine diagnostics of GI infections. Table 5.30 provides a summary of prospective specimen's distribution across all study sites.

	No. of prospective specimens
Site/Country	FecalSwab	Para-Pak C&S	Total
Germany	293	46	339
Denmark	293	0	293
Spain	247	0	247
France	63	0	63
USA site 1	0	186	186
USA site 2	0	43	43
USA site 3	282	0	282
USA site 4	0	177	177
USA site 5	44	0	44
USA site 6	0	39	39
USA site 7	0	0	0 a
USA site 8	0	131	131
USA site 9	0	95	95
Total	1222	717	1939

Table 5.30: Prospective Specimens Distribution Across the study sites

4 the specimens from this site (148) were excluded from the analysis because they were collected with another device different to Para-Pak C&S or FecalSwab

Table 5.31: provides all demographic information for the 1,939 specimens evaluated in the prospective study.

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Table 5.31: Demographic data for prospective evaluated specimens
Demographic data	FecalSwab	Para-Pak C&S
Gender
Female	628	442	22.8
Male	594	275	14.2
Age Group
0-5 years	182	31	1.6
6-21 years	121	38	2.0
22-49 years	290	215	11.1
50+ years	629	426	22.0
Not Reported	0	7	0.4
Patient population
Emergency room	46	29	1.5
Hospitalized	342	143	7.4
Immunocompromised	3	0	0.0
Outpatient	491	325	16.8
No information available	340	220	11.3
No. Days between Symptom Onset and QIAstat-Dx Testing
> 7 days	89	0	0.0
≤ 7 days	146	16	0.8
Not Reported	987	701	36.2
	32.4
	30.6
	9.4
	6.2
	15.0
	32.4
	0.0
	2.4
	17.6
	0.2
	25.3
	17.5
	4.6
	7.5
	50.9

Table 5 31. Demographic data for prospective evaluated specimens

The performance of the QIAstat-Dx Gastrointestinal Panel 2 was evaluated for each panel test result using one FDA-cleared test as comparator for the most analytes. A composite comparator consisting of either three independent FDA-cleared test methods or two independent FDA-cleared tests methods and two validated PCR assays followed by bidirectional sequencing was used for Norovirus GI/GII, ETEC, STEC and Giardia lamblia. (Table 5.32: ). The composite endpoint was determined as the majority of the three results.

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QIAstat-Dx GI Panel 2 Test Result	Comparator Method
Adenovirus F40/41
Astrovirus
Rotavirus A
Sapovirus
Campylobacter
Plesiomonas shigelloides
Salmonella	One FDA-cleared test method
Yersinia enterocolitica
E. coli O157 a
Enteropathogenic E. coli (EPEC) a
Shigella/Enteroinvasive E. coli (EIEC)
Cryptosporidium
Cyclospora cayetanensis
Entamoeba histolytica
Norovirus GI/GII	Composite of three FDA-cleared test
methods
Enterotoxigenic E. coli (ETEC) lt/st	Composite of three FDA-cleared test
methods
Shiga-like toxin- E. coli (STEC) stx1/stx2 a	Composite of three FDA-cleared test
methods
Giardia lamblia	Composite of two FDA-cleared test methods
and two validated PCR tests followed by bi-
directional sequencing b

Table 5.32: OIAstat-Dx Gastrointestinal Panel 2 Clinical studies comparator method

a Targets evaluated in Para-Pak C&S specimens only.

b Each of the two PCR assays used were well-characterized and validated nucleic acid amplification tests (NAAT) followed by bi-directional sequencing analysis. Both assays were designed to amplify different sequences than those targeted by the OIAstat-Dx Gastrointestinal Panel 2. Positive results reguired to generate sequences from bi-directional sequencing with at least 200 bases of adequate quality that by BLAST analyses matched a sequence of the expected organism or gene from NCBI GenBank database with at least 95% query coverage and at least 95% identity compared to the reference.

Additional prospective archived samples were collected for Norovirus GI/GII (81 samples) and STEC (18 samples). These were prospectively collected samples from four different collection sites (3 US and 1 EU), where only those positive for the pathogen by standard of care method were archived for analysis alongside 20 negative specimens.

In addition, to supplement the results of the prospective clinical studies, a total of 750 preselected archived frozen (retrospective) specimens were also evaluated. These specimens served to increase the sample size for analytes that showed low prevalence in the clinical prospective study or that were less represented in a particular sample type (Para-Pak C&S or FecalSwab). The same Comparator Methods detailed in Table 5.31 were used as confirmatory testing for the presence of the nucleic acids from the expected.

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In total 2808 specimens (1939 prospective, 119 prospective archived and 750 retrospective) were evaluated in the clinical study. These specimens were collected using Para-Pak C&S (1217) or FecalSwab (1591).

The positive percentage agreement (PPA) and the negative percentage agreement (NPA) were calculated for the prospective and retrospective studies and for each sample type (Para-Pak C&S and FecalSwab) separately.

The PPA was calculated as 100% x (TP / (TP + FN)). True positive (TP) indicates that both the QIAstat-Dx Gastrointestinal Panel 2 and comparator method showed a positive result for this specific target, and false negative (FN) indicates that the OIAstat-Dx Gastrointestinal Panel 2 result was negative while the comparator method result was positive. The NPA was calculated as 100% x (TN / (TN + FP)). True negative (TN) indicates that both the QIAstat-Dx Gastrointestinal Panel 2 and the comparator method showed negative results, and a false positive (FP) indicates that the QIAstat-Dx Gastrointestinal Panel 2 result was positive, but the comparator method result was negative. The PPA and NPA exact binomial two-sided 95% confidence interval was calculated.

Where a composite comparator was used (Table 5.32), the result was determined as the majority of the three individual test results (i.e., a positive composite comparator result is based on positive results for at least two comparator tests and a negative composite comparator result is based on negative results for at least two comparator tests). If insufficient pathogen positive sample was available to obtain a majority test result a worstcase model was applied in the PPA calculation. In this model the PPA was calculated including all observed true positive and false negative samples between QIAstat-Dx and the composite comparator while for the samples where it was not possible to conduct testing with the complete comparator due to insufficient sample volume, the following was done:

· samples that were negative in QIAstat-Dx and positive for one comparator assay, negative (or insufficient volume) for a second comparator and insufficient volume for a third comparator were included in the calculations as worst-case false negatives,
samples that were positive in QIAstat-Dx and positive in one comparator test, negative o (or insufficient volume) for a second comparator and insufficient volume for the third comparator, were considered as worst-case false positives and therefore, excluded in the PPA calculations.

The results of the clinical performance of the prospective, prospective archived and retrospective studies are summarized in Tables: Table 5.34 and Table 5.35. respectively.

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Discrepancies between the QIAstat-Dx Gastrointestinal Panel 2 and the comparator methods were investigated for the analytes that the QIAstat-Dx Gastrointestinal Panel 2 test result was compared to one FDA-cleared method. Discrepancies analyses are footnoted on each clinical performance summary Table below (Table 5.33, and Table 5.35).

		Positive Percent Agreement			Negative Percent Agreement
Analyte	Medium
Brand	TP/TP+FN	0/0	95% CI	TN/TN+FP	0/0	95% CI
			Viruses
Adenovirus F40/F41	FecalSwab	5/6 a	83.3	43.7-97.0	1214 / 1214	100.0	99.7-100.0
	Para-Pak
C&S	1 / 2 b	50.0	9.5-90.6	703 / 704 b	99.9	99.2-100.0
Astrovirus	FecalSwab	313	100.0	43.9-100.0	1219 / 1219	100.0	99.7-100.0
	Para-Pak
C&S	6 / 6	100.0	61.0-100.0	700 / 700	100.0	99.5-100.0
Norovirus GI/GII	FecalSwab	31 / 33 €	93.9	80.4-98.3	493 / 495 €	99.6	98.6-100.0
	Para-Pak
C&S	14 / 18 d	77.8	54.8-91.0	399 / 399 d	100.0	99.1-100.0
Rotavirus A	FecalSwab	21 / 23 €	91.3	73.2-97.6	1197 / 1199
e	99.8	99.4-100.0
	Para-Pak
C&S	313	100.0	43.9-100.0	702 / 703 f	99.9	99.2-100.0
			Bacteria
Campylobacter	FecalSwab	65 / 67 8	97.0	89.8-99.2	1151 / 1155
g	99.7	99.1-99.9
	Para-Pak
C&S	30 / 31 h	96.8	83.8-99.4	675 / 677 h	99.7	98.9-99.9
Plesiomonas
shigelloides	FecalSwab	0 / 0	N/A	N/A	1220 / 1222	99.8	99.4-100.0
	Para-Pak
C&S	5/6 !	83.3	43.7-97.0	698 / 700 i	99.7	99.0-99.9
Salmonella	FecalSwab	14 / 16 k	87.5	64.0-96.5	1206 / 1206	100.0	99.7-100.0
	Para-Pak
C&S	19 / 201	95.0	76.4-99.1	688 / 688	100.0	99.4-100.0
Yersinia enterocolitica	FecalSwab	15 / 16 m	93.8	71.7-99.0	1199 / 1206
m	99.4	98.8-99.7
	Para-Pak
C&S	313	100.0	43.9-100.0	698 / 703 "	99.3	98.4-99.7
				Diarrheagenic E. coli/Shigella
Enteropathogenic E.
coli (EPEC)	Para-Pak
C&S	57 / 65	87.7	77.6-93.6	632 / 632	100.0	99.4-100.0
Enterotoxigenic E. coli	FecalSwab	9 / 100	90.0	59.6-99.2	427 / 430 °	99.3	98.0-99.8
(ETEC) lt/st	Para-Pak
C&S	9 / 10 P	90.0	59.6-99.2	390 / 395 P	98.7	97.1-99.5
Shiga-like toxin E. coli
(STEC) stx1/stx2	Para-Pak
C&S	ર / વિત	83.3	43.6-97.0	397 / 400 9	99.3	97.8-99.7
E. coli 0157	Para-Pak
C&S	0 / 0	N/A	N/A	ર્ / 5	100.0	56.6-100.0
Shigella/Enteroinvasive	FecalSwab	10 / 10	100.0	72.3-100.0	1212 / 1212	100.0	99.7-100.0
E. coli (EIEC)	Para-Pak
C&S	2/2	100.0	34.2-100.0	703 / 704 ﺗ	99.9	99.2-100.0
			Parasites
Cryptosporidium	FecalSwab	2/4	50.0	15.0-85.0	1218 / 1218	100.0	99.7-100.0
	Para-Pak
C&S	6 / 6	100.0	61.0-100.0	699 / 700 ×	99.9	99.2-100.0
Cyclospora	FecalSwab	313	100.00	43.9-100.0	1219 / 1219	100.0	99.7-100.0
cayetanensis	Para-Pak
C&S	18 / 19 t	94.7	75.4-99.1	687 / 687	100.0	99.4-100.0
		Positive Percent Agreement			Negative Percent Agreement
Analyte	Medium Brand	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Entamoeba histolytica	FecalSwab	0 / 0	N/A	N/A	1222 / 1222	100.0	99.7-100.0
	Para-Pak C&S	0 / 0	N/A	N/A	706 / 706	100.0	99.5-100.0
Giardia lamblia	FecalSwab	6 / 8 u	75.0	40.9-92.9	434 / 441 u	98.4	96.8-99.2
	Para-Pak C&S	1 / 1	100.0	20.7-100.0	406 / 406 v	100.0	99.1-100.0

Table 5.33: Clinical Performance in the Prospective study

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a Adenovirus F40/41 was not detected in the single false negative specimen (0/1) in FecalSwab using a different FDA-cleared test method

b Adenovirus F40/41 was not detected in the single false negative specimen (0/1) and in the single false positive specimen (0/1) in Para-Pak C&S using a different FDA-cleared test method

s ten (10) FecalSwab samples positive for Norovirus GI/GII in both QIAstat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for composite comparator testing. The sample size for NPA is smaller for Norovirus GVGII as only a portion of the samples with a negative result in Q1Astat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

4 two (2) Para-Pak C&S samples positive for Norovirus GI/GII in both QIAstat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for comparator testing. One (1) Para-Pak C&S sample negative in QIAstat-Dx and positive with insufficient volume for complete composite comparator testing were classed as false negative in the sample size for NPA is smaller for Norovirus GI/GII as only a portion of the samples with a negative result in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

® Rotavirus A was detected in one of the two false specimens (1/2) and was not detected in the two false positive specimens (0/2) in FecalSwab using a different FDA-cleared test method.

f Rotavirus A was not detected in the single specimen (0/1) in Para-Pak C&S using a different FDA-cleared test method. s Campylobacter was not detected in the two false negative specimens (0/2) and was detected in three of the four false positive specimens (3/4) in FecalSwab using a different FDA-cleared test method.

A Campylobacter was not detected in the single false negative specimens (0/1) and was detected in one of the two false positive specimens (1/2) in Para-Pak C&S using a different FDA-cleared test method.

i Plesiomonas shigelloides was not detected in the two false positive specimens (0/2) in FecalSwab using a different FDA-cleared test method

i Plesiomonas shigelloides was not detected in the single false negative specimen (0/1) and was not detected in the two false positive specimens (0/2) in Para-Pak C&S using a different FDA-cleared test method

Salmonella was not detected in the two false negative specimens (0/2) in FecalSwab using a different FDA-cleared test method. 1 Salmonella was not detected in the single specimen (0/1) in Para-Pak C&S using a different FDA-cleared test method. ™ Yersinia enterocolitica was not detected in the single specimen (0/1) and was not detected in the seven false positive specimens (0/7) in FecalSwab using a different FDA-cleared test method.

11 Yersinia enterocolitica was not detected in the five specimens (0/5) using a different FDA-cleared test method. 9 six (6) FecalSwab samples positive for ETEC in both QIAstat-Dx and the primary FDA-cleared comparator were excluded from the

PPA calculations because the samples did not have sufficient volume for comparator testing. The sample size for NPA is smaller for ETEC as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

P three (3) Para-Pak C&S samples positive for ETEC in both QIAstat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for comparator testing. The sample size for NPA is smaller for ETEC as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

9 one (1) FecalSwab sample positive for STEC in both OIAstat-Dx and one FDA-cleared comparator was excluded from the PPA calculations because the samples did not have sufficient volume for comparator testing. The sample size for NPA is smaller for STEC as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

™ Shigella/ EIEC was detected in the single specimen (1/1) in Para-Pak C&S using a different FDA-cleared test method. S Cryptosporidium was not detected in the single false positive specimen (0/1) in Para-Pak C&S using PCR followed by bi-directional sequence analysis.

t For Cyclospora cayetanensis there was one (1) false negative specimen in Para-Pak C&S that was not further investigated by discrepant analyses.

4 two (2) FecalSwab samples positive for Giardia lamblia in both Q1Astat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for complete comparator testing. Two (2) FecalSwab samples negative in Q1Astat-Dx and positive with one FDA-cleared comparator with insufficient volume for complete composite comparator testing were classed as false negative in the PPA calculations. The sample size for Giardia lamblia as only a portion of the samples with a negative result in one FDA-cleared comparator was tested with the complete composite comparator in the prospective study.

v the sample size for NPA is smaller for Giardia as only a portion of the samples with a negative result in QIAstat-Dr and in one FDA-cleared comparator was tested with the complete comparator in the prospective study.

55

		Positive Percent Agreement		Negative Percent Agreement
Analyte	Medium Brand	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Norovirus
GI/GII	FecalSwab	48 / 49a	98.0	89.3 - 99.6	2 / 4a	50.0	15.0-85.0
GI/GII	Para-Pak C&S	26 / 28 a,b	92.9	77.4 - 98.0	37 / 38 a	97.4	86.5-99.5
Shiga-like
toxin E. coli
(STEC)
stx1/stx2	Para-Pak C&S	12 / 13 c,d	92.3	66.7 - 98.6	51 / 52 c	98.1	89.9-99.7

Table 5.34: Clinical Performance in the Prospective Archived study

4 For Norovirus GUGII/GII four out of the eighty-one (4/81) positive prospectively archived samples (positive by standard of care) were negative by the composite comparator and therefore included as negative samples in the NPA calculations.

b One (1) Para-Pak C&S sample negative in QIAstat-Dx and positive for Norovirus GI/GII with one FDA-cleared comparator with insufficient volume for complete comparator testing was classed as false negative in the PPA calculations.

· For STEC five out of the eighten (5/18) positived samples (positive by standard of care) were negative by the composite comparator and therefore included as negative samples in the NPA calculations.

4 One (1) Para-Pak C&S sample positive for STEC in both Q1Astat-Dx and one FDA-cleared comparator was excluded from the PPA calculations because the samples did not have sufficient volume for complete comparator testing.

56

		Positive Percent Agreement			Negative Percent Agreement
	Medium
Brand	TP/TP+FN	%	95% CI	TN/TN+FP	%	95% CI
Viruses
Adenovirus F40/F41	FecalSwab	23 / 26 a	88.5	71.0-96.0	203 / 203	100.0	98.2-100.0
	Para-Pak
C&S	29 / 29	100.0	88.3-100.0	39 / 39	100.0	91.0-100.0
Astrovirus	FecalSwab	2/3 b	66.7	20.8-93.9	191 / 191	100.0	98.0-100.0
	Para-Pak
C&S	0 / 0	N/A	N/A	14 / 14	100.0	78.5-100.0
Norovirus GI/GII	FecalSwab	28 / 32 c	87.5	71.9-95.0	74 / 75 c	98.7	92.8-100.0
	Para-Pak
C&S	27 / 29	93.1	78.0-98.1	86 / 86 d	100.0	95.8-100.0
Rotavirus A	FecalSwab	8/9e	88.9	56.5-98.0	185 / 185	100.0	98.0-100.0
	Para-Pak
C&S	2/2	100.0	34.2-100.0	12 / 12	100.0	75.8-100.0
				Bacteria				Campylobacter	FecalSwab	31 / 31	100.0	89.0-100.0	161 / 163 f	98.8	95.6-99.7		Para-Pak
C&S	3/3	100.0	43.9-100.0	11 / 11	100.0	74.1-100.0	Plesiomonas
shigelloides	FecalSwab	2/2	100.0	34.2-100.0	192 / 192	100.0	98.0-100.0		Para-Pak
C&S	33 / 36 g	91.7	78.2-97.1	117 / 117	100.0	96.8-100.0	Salmonella	FecalSwab	30 / 31 h	96.8	83.8-99.4	161 / 163 h	98.8	95.6-99.7		Para-Pak
C&S	1 / 1	100.0	20.7-100.0	13 / 13	100.0	77.2-100.0	Yersinia enterocolitica	FecalSwab	32 / 34 i	94.1	80.9-98.4	160 / 160	100.0	97.7-100.0		Para-Pak
C&S	1 / 1	100.0	20.7-100.0	14 / 14	100.0	78.5-100.0					Diarrheagenic E. coli/Shigella				Enteropathogenic E.
coli (EPEC)	Para-Pak
C&S	60 / 65 j	92.3	83.2-96.7	42 / 42	100.0	91.6-100.0	Enterotoxigenic E. coli
(ETEC) lt/st	FecalSwab	22 / 24 k	91.7	74.2-97.7	85 / 86 k	98.8	93.7-99.8		Para-Pak
C&S	23 / 24	95.8	79.8-99.3	61 / 61 l	100.0	94.1-100.0	Shiga-like toxin E. coli
(STEC) stx1/stx2	Para-Pak
C&S	60 / 64	93.8	85.0-97.5	44 / 44 m	100.0	92.0-100.0	E. coli 0157	Para-Pak
C&S	39 / 42 n	92.9%	80.1-99.4	16 / 16	100.0	80.6-100.0	Shigella/Enteroinvasive
E. coli (EIEC)	FecalSwab	22 / 24 o	91.7	73.0-99.0	170 / 170	100.0	97.8-100.0		Para-Pak
C&S	0 / 0	N/A	N/A	14 / 14	100.0	78.5-100.0					Parasites				Cryptosporidium	FecalSwab	6 / 6	100.0	54.1-100.0	186 / 188 p	98.9	96.2-99.9	Para-Pak
C&S	26 / 26	100.0	86.8-100.0	117 / 117	100.0	96.9-100.0	Cyclospora
cayetanensis	FecalSwab	1 / 1	100.0	20.7-100.0	193 / 193	100.0	98.1-100.0		Para-Pak
C&S	1 / 1	100.0	20.7-100.0	13 / 13	100.0	77.2-100.0	Entamoeba histolytica	FecalSwab	0 / 0	N/A	N/A	194 / 194	100.0	98.1-100.0		Para-Pak
C&S	0 / 0	N/A	N/A	14 / 14	100.0	76.5-100.0	Giardia lamblia	FecalSwab	29 / 31 q	93.6	79.3-98.2	46 / 48 q	95.8	86.0-98.9		Para-Pak
C&S	27 / 28 r	96.4	82.3-99.4	92 / 92 r	100.0	96.0-100.0
				Bacteria
Campylobacter	FecalSwab	31 / 31	100.0	89.0-100.0	161 / 163 f	98.8	95.6-99.7
	Para-Pak
C&S	3/3	100.0	43.9-100.0	11 / 11	100.0	74.1-100.0
Plesiomonas
shigelloides	FecalSwab	2/2	100.0	34.2-100.0	192 / 192	100.0	98.0-100.0
	Para-Pak
C&S	33 / 36 g	91.7	78.2-97.1	117 / 117	100.0	96.8-100.0
Salmonella	FecalSwab	30 / 31 h	96.8	83.8-99.4	161 / 163 h	98.8	95.6-99.7
	Para-Pak
C&S	1 / 1	100.0	20.7-100.0	13 / 13	100.0	77.2-100.0
Yersinia enterocolitica	FecalSwab	32 / 34 i	94.1	80.9-98.4	160 / 160	100.0	97.7-100.0
	Para-Pak
C&S	1 / 1	100.0	20.7-100.0	14 / 14	100.0	78.5-100.0
				Diarrheagenic E. coli/Shigella
Enteropathogenic E.
coli (EPEC)	Para-Pak
C&S	60 / 65 j	92.3	83.2-96.7	42 / 42	100.0	91.6-100.0
Enterotoxigenic E. coli
(ETEC) lt/st	FecalSwab	22 / 24 k	91.7	74.2-97.7	85 / 86 k	98.8	93.7-99.8
	Para-Pak
C&S	23 / 24	95.8	79.8-99.3	61 / 61 l	100.0	94.1-100.0
Shiga-like toxin E. coli
(STEC) stx1/stx2	Para-Pak
C&S	60 / 64	93.8	85.0-97.5	44 / 44 m	100.0	92.0-100.0
E. coli 0157	Para-Pak
C&S	39 / 42 n	92.9%	80.1-99.4	16 / 16	100.0	80.6-100.0
Shigella/Enteroinvasive
E. coli (EIEC)	FecalSwab	22 / 24 o	91.7	73.0-99.0	170 / 170	100.0	97.8-100.0
	Para-Pak
C&S	0 / 0	N/A	N/A	14 / 14	100.0	78.5-100.0
				Parasites
Cryptosporidium	FecalSwab	6 / 6	100.0	54.1-100.0	186 / 188 p	98.9	96.2-99.9
	Para-Pak
C&S	26 / 26	100.0	86.8-100.0	117 / 117	100.0	96.9-100.0
Cyclospora
cayetanensis	FecalSwab	1 / 1	100.0	20.7-100.0	193 / 193	100.0	98.1-100.0
	Para-Pak
C&S	1 / 1	100.0	20.7-100.0	13 / 13	100.0	77.2-100.0
Entamoeba histolytica	FecalSwab	0 / 0	N/A	N/A	194 / 194	100.0	98.1-100.0
	Para-Pak
C&S	0 / 0	N/A	N/A	14 / 14	100.0	76.5-100.0
Giardia lamblia	FecalSwab	29 / 31 q	93.6	79.3-98.2	46 / 48 q	95.8	86.0-98.9
	Para-Pak
C&S	27 / 28 r	96.4	82.3-99.4	92 / 92 r	100.0	96.0-100.0

Table 5.35: Clinical Performance in the Retrospective study

4 Adenovirus F40/41 was detected in one of the three false negatives (1/3) in FecalSwab using a different FDA-cleared test method b Astrovirus was detected in the single specimen (1/1) in FecalSwab using a different FDA-cleared test method. 6 two (2) FecalSwab samples positive for Norovirus GI/GII in both Q1Astat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for composite comparator testing. The sample size for NPA is smaller for Norovirus GVGII as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the retrospective study.

4 the sample size for NPA is smaller for Norovirus GVGII in Para-Pak C&S as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the composite comparator in the retrospective study · Rotavirus A was detected in the single false negative specimen (1/1) in FecalSwab using a different FDA-cleared test method.

57

f Campylobacter was detected in one of the two false positive specimens (1/2) in FecalSwab using a different FDA-cleared test method

s Plesiomonas shigelloides was detected in one of the three false negative specimens (1/3) in Para-Pak C&S using a different FDAcleared test method.

4 Salmonella was not detected in the single false negative specimen (0/1) and was not detected in the two false positive specimens (0/2) in FecalSwab using a different FDA-cleared test method

1 Yersinia enterocolitica was not detected in the two false negative specimens (0/2) in FecalSwab using a different FDA-cleared test method.

I Enteropathogenic E. coli (EPEC) was detected in all three false negative specimens (3/3) in Para-Pak C&S using PCR followed by bi-directional sequence analysis. There were two (2) other false negative specimens that were not further investigated by discrepant analyses

ten (10) FecalSwab samples positive for ETEC in both OIAstat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for composite comparator testing. One (1) FecalSwab sample negative in QIAstat-Dx and positive with one FDA-cleared comparator with insufficient volume for composite comparator testing was classed as false negative in the PPA calculations. The sample size for ETEC as only a portion of the samples with a negative result in QIAstat-Dx and one FDA-cleared comparator was tested with the composite comparator in the retrospective study.

1 The sample size for NPA is smaller for ETEC in Para-Pak CS&S as only a portion of the samples with a negative result in QIAstat-Dx and one FDA-cleared comparator was tested with the composite comparator in the retrospective study).

" The sample size for NPA is smaller for STEC in Para-Pak C&S as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the composite comparator in the retrospective study

" E. coli O157 was not detected in two false negativens (0/2) in Para-Pak C&S using a different FDA-cleared test method. There was one (1) false negative specimen in Para-Pak C&S that was not further investigated by discrepant analyses.

° Shigella/ EIEC was detected in one of the two false negative specimens (1/2) in FecalSwab using a different FDA-cleared test method.

P Cryptosporidium was not detected in the two false positive specimens (0/2) in FecalSwab using PCR followed by bi-directional sequence analysis

9 four (4) samples positive for Giardia lambia in both QIAstat-Dx and one FDA-cleared comparator were excluded from the PPA calculations because the samples did not have sufficient volume for composite comparator testing. Two (2) FecalSwab samples negative in QIAstat-Dx and positive with one FDA-cleared comparator with insufficient volume for composite comparator testing was classed as false negative in the PPA calculations. The sample size for NPA is smaller for Giardia as only a portion of the samples with a negative result in QIAstat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the retrospective study

r One (1) Para-Pak C&S samples positive for Giardia lamblia in both QIAstat-Dx and primary FDA-cleared comparator (were excluded from the PPA calculations because the samples did not have sufficient volume for composite comparator testing. One (1) Para-Pak C&S sample negative in QIAstat-Dx and positive with one FDA-cleared comparator with insufficient volume for complete comparator testing was classed as false negative in the PPA calculations. The sample size for NPA is smaller for Giardia lamblia as only a portion of the samples with a negative result in Q1Astat-Dx and in one FDA-cleared comparator was tested with the complete composite comparator in the retrospective study.

58

The proportion of failed runs on initial attempt, and following repeats are summarized in Table 5.36. The error breakdown due to instrument, invalid results, 'sample too concentrated' failures and other run failures are summarized in Table 5.37.

| Transport

Media	Study	Initial Runs			Final Runs
		N / Total	%	95% CI	N / Total	%	95% CI
FecalSwab	Prospective	16 / 1227	1.3	0.7-2.1	3 / 1227	0.2	0.1-0.7
	Prospective
Archived	0 / 53	0.0	0.0-6.7	0 / 53	0.0	0.0-6.7
	Retrospective	11 / 366	3.0	1.5-5.3	5 / 366	1.4	0.4-3.2
	Total	27 / 1646	1.6	1.1 - 2.4	8 / 1646	0.5	0.2 - 1.0
Para-Pak	Prospective	66 / 740	8.9	7.0-11.2	21 / 740	2.8	1.8-4.3
	Prospective
Archived	3 / 66	4.5	0.9-12.7	0 / 66	0.0	0.0-5.4
	Retrospective	46 / 454	10.1	7.5 - 13.3	25 / 454	5.5	3.6-8.0
	Total	115 / 1260	9.1	7.6-10.9	46 / 1260	3.7	2.7-4.8

Table 5.36 Failure Rates Summary

Table 5.37 Failure Types Breakdown

| Transport
Media | Study N / Total | % | Final Runs
N / Total | % |
|--------------------| | FecalSwab | | | | | | | | Prospective
Archived | | | | | | | | | | | | | | Para-Pak C&S | | | | | | | | Prospective
Archived | | | | | | | Archived
Frozen | Instrument | | | | | | | Failure Reason | Initial Runs
-------------------------------------|---------------------------|---------------------------|-----|-------------------------|-----|
| Prospective | Instrument | 0 / 1227 | 0.0 | 0 / 1227 | 0.0 |
| Invalid a | 0 / 1227 | 0.0 | 0 / 1227 | 0.0 |
| Sample too Concentrated b | 5 / 1227 | 0.4 | 0 / 1227 | 0.0 |
| Other c | 11 / 1227 | 0.9 | 3 / 1227 | 0.2 |
| Instrument | 0 / 53 | 0.0 | 0 / 53 | 0.0 |
| Invalid | 0 / 53 | 0.0 | 0 / 53 | 0.0 |
| Sample too Concentrated | 0 / 53 | 0.0 | 0 / 53 | 0.0 |
| Other | 0 / 53 | 0.0 | 0 / 53 | 0.0 |
| Retrospective | Instrument | 1 / 366 | 0.3 | 0 / 366 | 0.0 |
| Invalid | 1 / 366 | 0.3 | 0 / 366 | 0.0 |
| Sample too Concentrated | 0 / 366 | 0.0 | 0 / 366 | 0.0 |
| Other | 9 / 366 | 2.5 | 5 / 366 | 1.4 |
| Prospective | Instrument | 9 / 740 | 1.2 | 2 / 740 | 0.3 |
| Invalid | 5 / 740 | 0.7 | 5 / 740 | 0.7 |
| Sample too Concentrated | 35 / 740 | 4.7 | 7 / 740 | 0.9 |
| Other | 17 / 740 | 2.3 | 7 / 740 | 0.9 |
| Instrument | 0 / 66 | 0.0 | 0 / 66 | 0.0 |
| Invalid | 1 / 66 | 1.5 | 0 / 66 | 0.0 |
| Sample too Concentrated | 1 / 66 | 1.5 | 0 / 66 | 0.0 |
| Other | 1 / 66 | 1.5 | 0 / 66 | 0.0 |
| Retrospective
| 1 / 454 | 0.2 | 0 / 454 | 0.0 |
| Invalid | 10 / 454 | 2.2 | 6 / 454 | 1.3 |
| Sample too Concentrated | 10 / 454 | 2.2 | 2 / 454 | 0.4 |
| Other | 25 / 454 | 5.5 | 17 / 454 | 3.7 |

a Internal Control failures with at least one analyte detected and the other analytes reported as 'invalid'

b Run failures related to 'sample concentration too high'. These specimens were repeated with 100 microliters as detailed in Appendix C.

C Run failures related to workflow checkpoints.

59

Co-infections

The QIAstat-Dx Gastrointestinal Panel 2 reported multiple organism detections (i.e., coinfections) for a total of 15 and 29 prospective specimens in FecalSwab and Para-Pak C&S, respectively. This represents 7.0% of positive specimens (15/213) in FecalSwab and 17.0% of positive specimens (29/171) in Para-Pak C&S. Most multiple detections in FecalSwab specimens (14/15; 93%) contained two organisms, while 6.7% (1/15) contained three organisms. In Para-Pak C&S specimens, most multiple detections (22/29; 75.9%) contained two organisms, while 24.1% (7/29) contained three organisms. The most common multiple infections are shown in Table 5.38 and Table 5.39 below.

Table 5.38: Most Prevalent Multiple Detection Combinations (≥2 instances) as Determined by the QIAstat-Dx Gastrointestinal Panel 2 in FecalSwab specimens

| Multiple Detection Combination | Number of FecalSwab
Specimens |
|---------------------------------------------------------|----------------------------------|
| Campylobacter + Norovirus GI/GII | 2 |
| Enterotoxigenic E. coli (ETEC) lt/st + Norovirus GI/GII | 3 |
| Campylobacter + Rotavirus A | 4 |

Table 5.39: Most Prevalent Multiple Detection Combinations (≥2 instances) as Determined by the QIAstat-Dx Gastrointestinal Panel 2 in Para-Pak C&S specimens

| Multiple Detection Combination | Number of Para-Pak C&S
Specimens |
|-----------------------------------------------------------------------------------------|-------------------------------------|
| Campylobacter + Enteropathogenic E. coli (EPEC) | 3 |
| Enteropathogenic E. coli (EPEC) + Salmonella | 3 |
| Enteropathogenic E. coli (EPEC) + Enterotoxigenic E. coli (ETEC)
lt/st | 4 |

The analytes most commonly found in mixed infections in the FecalSwab specimens were Campylobacter (9), Norovirus GI/GII (7), Rotavirus (4) and ETEC (3) as shown in Table 5.4040 while the analytes most commonly found in mixed infections in the Para-Pak C&S specimens were EPEC (17), ETEC (8), Campylobacter (7), Norovirus GI/GII (5), Rotavirus (4) and STEC (5) as shown in Table 5.4141 and Table 5.4242.

60

Table 5.4040: Prevalence of Analytes in Mixed Infections in FecalSwab specimens as
determined by the QIAstat-Dx Gastrointestinal Panel 2

Analyte	N	%
Adenovirus F40/F41	1	3.2
Astrovirus	1	3.2
Campylobacter	9	29.0
Enterotoxigenic E. coli (ETEC) lt/st	3	9.7
Giardia lamblia	2	6.5
Norovirus GI/GII	7	22.6
Plesiomonas shigelloides	1	3.2
Rotavirus A	4	12.9
Shigella/Enteroinvasive E. coli (EIEC)	2	6.5
Yersinia enterocolitica	1	3.2

Prevalence of Analytes in Mixed Infections in Para-Pak C&S Table 5.4141: specimens as determined by the QIAstat-Dx Gastrointestinal Panel 2

Analyte	N	%
Adenovirus F40/F41	1	1.5
Astrovirus	1	1.5
Campylobacter	7	10.8
Cryptosporidium	2	3.1
Cyclospora cayetanensis	2	3.1
Enteropathogenic E. coli (EPEC)	17	26.2
Enterotoxigenic E. coli (ETEC) lt/st	8	12.3
Giardia lamblia	1	1.5
Norovirus GI/GII	5	7.7
Plesiomonas shigelloides	7	10.8
Rotavirus A	1	1.5
Salmonella	4	6.2
Shiga-like toxin E. coli (STEC) stx1/stx2	5	7.7
Shigella/Enteroinvasive E. coli (EIEC)	3	4.6
Yersinia enterocolitica	1	1.5

Contrived Specimens Testing

Several analytes, such as Entamoeba histolytica are so rare that both prospective and archived testing efforts were insufficient to demonstrate system performance. To

61

supplement the prospective and archived specimens' test results, an evaluation of contrived specimens was performed. Contrived specimens were prepared using negative residual specimens that had previously tested negative by QIAstat-Dx Gastrointestinal Panel 2 and comparator methods. At least, 50% of these specimens were spiked at concentrations at concentrations slightly above the Limit of Detection (2x LoD) and the rest at 5x and 10x LoD, using quantified strains for each pathogen. A minimum of 50 contrived specimens were tested for each evaluated analyte. The analyte status of each contrived specimen was blinded to the users analyzing the specimens. Results are summarized in Table 5.4242 Table 5.42.

QIAstat-Dx GI2 Target	Medium Brand	Fraction	Percentage	95% CI
Astrovirus	FecalSwab	33 / 34	97.1	85.1-99.5
	Para-Pak C&S	34 / 34	100.0	89.8-100.0
Rotavirus A	FecalSwab	35 / 35	100.0	90.1-100.0
	Para-Pak C&S	34 / 35	97.1	85.5-99.5
Plesiomonas shigelloides	FecalSwab	33 / 33	100.0	89.6-100.0
	Para-Pak C&S	34 / 35	97.1	85.5-99.5
Yersinia enterocolitica	FecalSwab	34 / 34	100.0	89.8-100.0
	Para-Pak C&S	34 / 35	97.1	85.5-99.5
Shigella/EIEC	FecalSwab	35 / 35	100.0	90.1-100.0
	Para-Pak C&S	34 / 34	100.0	89.8-100.0
Cryptosporidium	FecalSwab	27 / 27	100.0	87.5-100.0
	Para-Pak C&S	31 / 31	100.0	89.0-100.0
Cyclospora cayetanensis	FecalSwab	26 / 26	100.0	87.1-100.0
	Para-Pak C&S	30 / 30	100.0	88.6-100.0
Entamoeba histolytica	FecalSwab	35 / 35	100.0	90.1-100.0
	Para-Pak C&S	34 / 35	97.1	85.5-99.5

Table 5.4242: Test Results Summary for Contrived Specimens
-------------------------------------------------------------	--	--	--	--

Conclusions

The QIAstat-Dx Gastrointestinal Panel 2 is substantially equivalent to the legally marketed: BioFire Diagnostics LLC FilmArray Gastrointestinal (GI) Panel 510(k) # K140407.