BioFire Global Fever Special Pathogens Panel; BIOFIRE SHIELD Control Kit for the BioFire Global Fever Special Pathogens Panel

K213362 · Biofire Defense, LLC · QVR · Mar 22, 2023 · Immunology

Device Facts

• Record ID: K213362
• Device Name: BioFire Global Fever Special Pathogens Panel; BIOFIRE SHIELD Control Kit for the BioFire Global Fever Special Pathogens Panel
• Applicant: Biofire Defense, LLC
• Product Code: QVR · Immunology
• Decision Date: Mar 22, 2023
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.4000
• Device Class: Class 2

Indications for Use

The BioFire® Global Fever Special Pathogens Panel is a qualitative, multiplexed, nucleic acid-based test intended for use with BioFire® FilmArray® 2.0 and BioFire® FilmArray® Torch Systems. The BioFire Global Fever Special Pathogens Panel is for the simultaneous qualitative detection and identification of multiple bacterial, viral, and protozoan nucleic acids directly from EDTA whole blood collected from individuals with signs and/or symptoms of acute febrile illness or recent acute febrile illness and known or suspected exposure to the target pathogens described below. Pathogens identified: Chikungunya virus, Dengue virus (serotypes 1, 2, 3 and 4), Leishmania spp. that cause visceral leishmaniasis (e.g., L. donovani and L. infantum), Leptospira spp., Plasmodium spp. (including species differentiation of Plasmodium falciparum and Plasmodium vivax/ovale), West Nile virus. Pathogens presumptively identified: Bacillus anthracis, Crimean-Congo hemorrhagic fever virus, Ebolavirus spp., Francisella tularensis, Lassa virus, Marburgvirus, Yellow fever virus, Yersinia pestis. Pathogens for which the panel provides presumptive identification results require additional testing and confirmation procedures in consultation with the appropriate public health authorities for whom reports may be necessary. Positive results do not rule out co-infections with pathogens not included on the BioFire Global Fever Special Pathogens Panel. Not all pathogens that cause acute febrile illness are detected by this test, and negative results do not preclude infection with the pathogens targeted by the device and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Evaluation for more common causes of acute febrile illness (e.g., infections of the upper and lower respiratory tract or gastroenteritis, as well as non-infectious causes) should be considered prior to evaluation with this panel. In the United States, patient travel history, exposure risk, and consultation of the CDC Yellow Book should be considered prior to use of the BioFire Global Fever Special Pathogens Panel as some pathogens are more common in certain geographical locations. Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities. The BioFire Global Fever Special Pathogens Panel is indicated for use in laboratories having appropriate biosafety equipment, personal protective equipment (PPE), containment facilities and personnel trained in the safe handling of diagnostic clinical specimens potentially containing any of the pathogens detected by this panel. The BioFire Global Fever Special Pathogens Panel is indicated for use in laboratories that follow public health guidelines that address appropriate biosafety conditions, interpretation of test results, and coordination of findings with public health authorities. For In Vitro Diagnostic Use.

Device Story

Multiplexed nucleic acid-based test for BioFire FilmArray 2.0/Torch systems; detects bacterial, viral, and protozoan pathogens from EDTA whole blood. Pouch contains freeze-dried reagents for nucleic acid purification and nested, multiplexed PCR with DNA melt analysis. User loads hydration solution and sample buffer/blood mixture into pouch; instrument automates liquid movement via bladders/pistons and thermal cycling via Peltier devices. Digital camera captures fluorescent images of PCR2 reactions; software interprets melt curves to identify pathogens. Results available in ~1 hour. Used in clinical laboratories by trained personnel. Output provides qualitative identification or presumptive identification of pathogens. Results support clinical decision-making in conjunction with other data; aids in managing patients with acute febrile illness and potential exposure to biothreat/special pathogens.

Clinical Evidence

Prospective clinical study (n=2139) evaluated PPA/NPA against comparator methods. Results showed high sensitivity/specificity for identified pathogens. Archived specimen study (n=416) and contrived specimen study (n=50 replicates per analyte) supplemented performance data for rare pathogens. Performance metrics (PPA/NPA) and 95% CIs reported for all analytes.

Technological Characteristics

Multiplexed nested nucleic acid amplification test with melt analysis. Uses BioFire FilmArray 2.0/Torch instruments. Reagents freeze-dried in pouch. Magnetic bead-based extraction. Fluorescent DNA binding dye (LC Green Plus). Automated software interpretation. Room temperature storage.

Indications for Use

Indicated for individuals with signs/symptoms of acute febrile illness and known/suspected exposure to target pathogens. For use in laboratories with appropriate biosafety equipment, PPE, containment facilities, and trained personnel.

Regulatory Classification

Identification

A multiplex nucleic acid detection system for biothreat agents is a qualitative in vitro diagnostic test for detection and identification of nucleic acids from biothreat pathogens in human clinical specimens

Predicate Devices

• BioFire Global Fever Panel (DEN200043)
• FilmArray NGDS Warrior Panel (K170883)

Submission Summary (Full Text)

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March 22, 2023

Image /page/0/Picture/1 description: The image shows the logo of 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 the FDA logo, which consists of a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG" in blue, and the word "ADMINISTRATION" in a smaller font size below that.

BioFire Defense, LLC David Rabiger Associate Director of Regulatory and Clinical Affairs 79 W 4500 S. Suite 14 Salt Lake City, Utah 84107

### Re: K213362

Trade/Device Name: BioFire Global Fever Special Pathogens Panel; BIOFIRE SHIELD Control Kit for the BioFire Global Fever Special Pathogens Panel Regulation Number: 21 CFR 866.4000 Regulation Name: Device To Detect And Identify Biothreat Microbial Agents In Human Clinical Specimens. Regulatory Class: Class II Product Code: QVR, QMV, PMN Dated: October 8, 2021 Received: October 12, 2021

Dear David Rabiger:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. 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 located 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.

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

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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 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-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 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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

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

Enclosure

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# Indications for Use

510(k) Number (if known)

K213362

Device Name

BioFire Global Fever Special Pathogens Panel

### Indications for Use (Describe)

The BioFire® Global Fever Special Pathogens Panel is a qualitative, mucleic acid-based test intended for use with BioFire® FilmArray® 2.0 and BioFire® FilmArray® Torch Systems. The BioFire Global Fever Special Pathogens Panel is for the simultaneous qualitative detection and identification of multiple bacterial, viral, and protozoan nucleic acids directly from EDTA whole collected from individuals with signs and or symptoms of acute febrile illness and known or suspected exposure to the target pathogens described below.

### Pathogens identified:

Chikungunya virus Dengue virus (serotypes 1, 2, 3 and 4) Leishmania spp. that cause visceral leishmaniasis (e.g., L. donovani and L. infantum) Leptospira spp. Plasmodium spp. (including species differentiation of Plasmodium falciparum and Plasmodium vivax/ovale) West Nile virus

### Pathogens presumptively identified:

Bacillus anthracis Crimean-Congo hemorrhagic fever virus Ebolavirus spp. Francisella tularensis Lassa virus Marburgvirus Yellow fever virus Yersinia pestis

Pathogens for which the panel provides presumptive identification results resting and confirmation procedures in consultation with the appropriate public health authorities for whom reports may be necessary.

Positive results do not rule out co-infections with pathogens not included on the BioFire Global Fever Special Pathogens Panel. Not all pathogens that cause acute febrile illness are detected by this test, and nectude infection with the pathogens targeted by the device and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.

Evaluation for more common causes of acute illness (e.g., infections of the upper and lower respiratory tract or gastroenteritis, as well as non-infectious causes) should be considered prith this panel. In the United States, patient travel history, exposure risk, and consultation of the CDC Yellow Book should be considered prior to use of the BioFire Global Fever Special Pathogens Panel as some pathogens are more common in certain geographical locations. Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guided by the relevant public health authorities.

The BioFire Global Fever Special Pathogens Panel is indicatories having appropriate biosafety equipment, personal protective equipment (PPE), contamment facilities and persomel trained in the safe handling of diagnostic clinical specimens potentially containing any of the pathogens detected by this panel.

The BioFire Global Fever Special Pathogens Panel is indicated for use in laboratories that follow public health guidelines that address appropriate biosafety conditions, interpretation of test results, and coordination of findings with public health authorities.

For In Vitro Diagnostic Use.

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)

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# 510(k) Summary

#### Submitter I.

BioFire Defense, LLC 79 West 4500 South, Suite 14 Salt Lake City, UT 84107 Phone: [phone] Fax: [phone]

Contact Person: David Rabiger Date Prepared: 2022-Oct-07

#### II. Device

Name of Device: BioFire® Global Fever Special Pathogens Panel Common or Usual Name: Same Product Code: QMV Regulation: 21 CFR 866.3966 Classification Name: Device to detect and identify selected microbial agents that cause acute febrile illness Regulatory Class: Class II (Special Controls) Panel: Microbiology - 83

Additional Product Code: QVR Regulation: 21 CFR 866.4000 Classification Name: A multiplex nucleic acid detection system for biothreat agents Regulatory Class: Class II (Special Controls) Panel: Microbiology - 83

#### Predicate Devices III.

BioFire® Global Fever Panel (BioFire Defense, LLC) [DEN200043] This predicate has not been subject to a design-related recall.

FilmArray® NGDS Warrior Panel (BioFire Defense, LLC) [K170883] This predicate has not been subject to a design-related recall.

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#### IV. Device Description

The BioFire® Global Fever Special Pathogens Panel is a multiplexed nucleic acid-based test designed to be used with BioFire® FilmArray® Systems (BioFire® FilmArray® 2.0 or BioFire® FilmArray® Torch). The BioFire Global Fever Special Pathogens Panel pouch contains freezedried reagents to perform nucleic acid purification and nested, multiplexed polymerase chain reaction (PCR) with DNA melt analysis. The BioFire Global Fever Special Pathogens Panel conducts tests for the identification of bacterial, viral, and protozoan pathogens from whole blood specimens collected in EDTA tubes (Table 1). Results from the BioFire Global Fever Special Pathogens Panel are available in about 1 hour.

| Type | Disease | Pathogen |
|-----------|---------------------------------|------------------------------------------------------------------|
| Bacterial | Anthrax | Bacillus anthracis 1 |
| | Leptospirosis | Leptospira spp. |
| | Plague | Yersinia pestis 1 |
| | Tularemia | Francisella tularensis 1 |
| Viral | Crimean-Congo hemorrhagic fever | Crimean-Congo hemorrhagic fever virus 1 |
| | Chikungunya fever | Chikungunya virus |
| | Dengue fever | Dengue virus (serotypes 1, 2, 3 and 4) |
| | Lassa fever | Lassa virus 1 |
| | Ebola virus disease | Ebolavirus spp. (Bundibugyo, Reston, Sudan, Taï Forest, Zaire) 1 |
| | Marburg virus disease | Marburgvirus 1 |
| | West Nile fever | West Nile virus |
| | Yellow fever | Yellow fever virus 2 |
| Protozoan | Malaria | Plasmodium spp. |
| | | Plasmodium falciparum |
| | | Plasmodium vivax/ovale |
| | Visceral Leishmaniasis | Leishmania spp. including L. donovani and L. infantum |

Table 1. Pathogens Detected by the BioFire Global Fever Special Pathogens Panel

1 Select agents are subject to additional requirements. Definitive identification requires confirmatory testing.

2 Definitive identification requires confirmatory testing.

A test is initiated by loading Hydration Solution into one port of the pouch and a whole blood specimen mixed with the provided Sample Buffer into the other port of the pouch. The pouch contains all the reagents required for specimen testing and analysis in a freeze-dried format; the addition of Hydration Solution and the Sample Buffer rehydrates the reagents. After the pouch is prepared, the BioFire® FilmArray® Software guides the user through the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, selecting the appropriate protocol, and initiating the run on the BioFire FilmArray system.

The BioFire FilmArray instruments contain a coordinated system of inflatable bladders and seal points, which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting

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channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically controlled pneumatic pistons are positioned over multiple plungers to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and subsequent melt.

Nucleic acid extraction occurs within the BioFire FilmArray pouch using mechanical and chemical lysis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplexed PCR is executed in two stages. During the first stage, a single, large volume, multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The second stage PCR and melt, or nested singleplex PCR, is performed in each well of the array. At the end of the second stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR2 reactions and software interprets the data.

Materials provided in each BioFire Global Fever Special Pathogens Panel Kit:

- Individually packaged BioFire Global Fever Special Pathogens Panel Pouches ●
- Single-use (1.0 mL) BioFire® FilmArray® Sample Buffer Tubes ●
- Single-use pre-filled (1.5 mL) BioFire® FilmArray® Hydration Injection Vials ●
- . Individually packaged BioFire® FilmArray® Sample Injection Vials
- Individually packaged Transfer Pipettes
- . BioFire Global Fever Special Pathogens Panel – Ouick Guide
- Instructions available online at: www.biofiredefense.com 0 BioFire Global Fever Special Pathogens Panel – Instructions for Use

Materials required but not provided:

- BioFire FilmArray System including: ●
- BioFire® FilmArray® 2.0 or BioFire® FilmArray® Torch Instrument System o including accompanying platform-specific core software
- BioFire® FilmArray® Pouch Loading Station O
- BioFire® Global Fever Special Pathogens Pouch Module Software is required to o run the BioFire Global Fever Special Pathogens Panel and is available by request at www.biofiredefense.com if not already installed on the instrument system.
- 10% bleach solution or a similar disinfectant ●

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#### v. Intended Use

The BioFire® Global Fever Special Pathogens Panel is a qualitative, multiplexed, nucleic acid-based test intended for use with BioFire® FilmArray® 2.0 and BioFire® FilmArray® Torch Systems. The BioFire Global Fever Special Pathogens Panel is for the simultaneous qualitative detection and identification of multiple bacterial, viral, and protozoan nucleic acids directly from EDTA whole blood collected from individuals with signs and/or symptoms of acute febrile illness or recent acute febrile illness and known or suspected exposure to the target pathogens described below.

### Pathogens identified:

Chikungunya virus Dengue virus (serotypes 1, 2, 3 and 4) Leishmania spp. that cause visceral leishmaniasis (e.g., L. donovani and L. infantum) Leptospira spp. Plasmodium spp. (including species differentiation of Plasmodium falciparum and Plasmodium vivax/ovale) West Nile virus

# Pathogens presumptively identified:

Bacillus anthracis Crimean-Congo hemorrhagic fever virus Ebolavirus spp. Francisella tularensis Lassa virus Marburgvirus Yellow fever virus Yersinia pestis

Pathogens for which the panel provides presumptive identification results require additional testing and confirmation procedures in consultation with the appropriate public health authorities for whom reports may be necessary.

Positive results do not rule out co-infections with pathogens not included on the BioFire Global Fever Special Pathogens Panel. Not all pathogens that cause acute febrile illness are detected by this test, and negative results do not preclude infection with the pathogens targeted by the device and should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.

Evaluation for more common causes of acute febrile illness (e.g., infections of the upper and lower respiratory tract or gastroenteritis, as well as non-infectious causes) should be considered prior to evaluation with this panel. In the United States, patient travel history, exposure risk, and consultation of the CDC Yellow Book should be considered prior to use of the BioFire Global Fever Special Pathogens Panel as some pathogens are more common in certain geographical locations. Results are meant to be used in conjunction with other clinical, epidemiologic, and laboratory data, in accordance with the guidelines provided by the relevant public health authorities.

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The BioFire Global Fever Special Pathogens Panel is indicated for use in laboratories having appropriate biosafety equipment, personal protective equipment (PPE), containment facilities and personnel trained in the safe handling of diagnostic clinical specimens potentially containing any of the pathogens detected by this panel.

The BioFire Global Fever Special Pathogens Panel is indicated for use in laboratories that follow public health guidelines that address appropriate biosafety conditions, interpretation of test results, and coordination of findings with public health authorities.

### For In Vitro Diagnostic Use.

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#### Substantial Equivalence VI.

The BioFire Global Fever Special Pathogens Panel is substantially equivalent to the BioFire Global Fever Panel (previously known as the FilmArray Global Fever Panel) and the FilmArray NGDS Warrior Panel. The BioFire Global Fever Panel was granted De Novo classification on November 20, 2020 [DEN200043] and was categorized as a Class II device. The FilmArray NGDS Warrior Panel was cleared on June 22, 2017 and was also categorized as a Class II device [K170883].

In addition, the BioFire Global Fever Special Pathogens Panel is substantially equivalent to the FilmArray NGDS Warrior Panel in that they both have the ability to detect select agents in whole blood specimens. While there are differences between these panels regarding pouch chemistry, pouch protocols, and pouch module software, both panels were developed using the same FilmArray technology and design principles.

Table 2 outlines the similarities and differences between the BioFire Global Fever Special Pathogens Panel and the predicate devices.

| Element | Subject Device:<br>BioFire Global Fever Special<br>Pathogens Panel | Predicate:<br>BioFire Global Fever Panel<br>[DEN200043] | Predicate:<br>FilmArray NGDS Warrior Panel<br>[K170883] |
|-----------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Specimen Type | Whole Blood (collected in EDTA<br>tube) | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Whole Blood (collected in EDTA<br>tube), positive blood culture, or<br>sputum |
| Intended Use<br>Setting | Individuals with signs and/or<br>symptoms of acute febrile<br>illness or recent acute febrile<br>illness and known or suspected<br>exposure to pathogens on the<br>panel. | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Individuals with signs and<br>symptoms of infection from<br>biothreat agents and/or<br>individuals who are at risk for<br>exposure or may have been<br>exposed to pathogens tested by<br>the panel. |
| Element | Subject Device:<br>BioFire Global Fever Special<br>Pathogens Panel | Predicate:<br>BioFire Global Fever Panel<br>[DEN200043] | Predicate:<br>FilmArray NGDS Warrior Panel<br>[K170883] |
| Pathogens Detected | Identification:<br>Chikungunya virus, Dengue<br>virus (serotypes 1, 2, 3 and 4),<br>Leishmania spp., Leptospira<br>spp., Plasmodium spp.<br>(including species<br>differentiation of Plasmodium<br>falciparum and Plasmodium<br>vivax/ovale),<br>West Nile virus<br>Presumptive Identification:<br>Bacillus anthracis, Crimean-<br>Congo hemorrhagic fever virus,<br>Ebolavirus spp., Francisella<br>tularensis, Lassa virus,<br>Marburgvirus, Yellow fever<br>virus, Yersinia pestis | Identification:<br>Chikungunya virus, Dengue<br>virus (serotypes 1,2,3, and<br>4), Leptospira spp.,<br>Plasmodium spp. (including<br>species differentiation of<br>Plasmodium falciparum<br>and Plasmodium<br>vivax/ovale) | Presumptive Identification in<br>whole blood EDTA:<br>Bacillus anthracis, Coxiella<br>burnetii, Ebolavirus spp.,<br>Francisella tularensis,<br>Marburgvirus, Yersinia Pestis<br>Presumptive Identification in<br>positive blood culture:<br>Bacillus anthracis, Yersinia pestis<br>Presumptive Identification in<br>sputum:<br>Francisella tularensis, Yersinia<br>pestis |
| Analyte | RNA/DNA | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| Technological<br>Principles | Highly multiplexed nested<br>nucleic acid amplification test<br>with melt analysis | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| Instrumentation1 | BioFire FilmArray 2.0 and<br>BioFire FilmArray Torch | BioFire FilmArray 2.0 | BioFire FilmArray 2.0 |
| Time to result | About 1 hour | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| Reagent Storage | Room temperature | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| Test Interpretation | Automated test interpretation<br>and report generation. User<br>cannot access raw data. | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| Controls | Two controls are included in<br>each reagent pouch to control<br>for sample processing and both<br>stages of PCR and melt<br>analysis. | Same as BioFire Global<br>Fever Special Pathogens<br>Panel | Same as BioFire Global Fever<br>Special Pathogens Panel |
| User Complexity | High | Moderate | Moderate |

Table 2. Comparison of the BioFire Global Fever Special Pathogens Panel and Predicate Devices

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1 The BioFire FilmArray 2.0 [K143178] and BioFire FilmArray Torch [K160068] instruments are based on the same technological principles.

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#### VII. Summary of Performance Data

# Clinical Performance

### Prospective Clinical Study

The prospective clinical study was designed to evaluate the sensitivity/positive percent agreement (PPA) and specificity/negative percent agreement (NPA) for each analyte of the BioFire Global Fever Special Pathogens Panel when testing prospectively collected whole blood specimens in the intended use setting.

Between March 2018 and March 2021, 11 sites contributed 2139 prospectively collected whole blood specimens from individuals who had a recorded or self-reported fever within the past two days. A summary of demographic information for the 2139 specimens included in the prospective study is given in Table 3.

| | Demographic | Overall |
|-------|-------------|--------------|
| Sex | Female | 1095 (51.2%) |
| | Male | 1044 (48.8%) |
| Age | <5 | 178 (8.3%) |
| | 5 to 21 | 822 (38.4%) |
| | 22 to 50 | 779 (36.4%) |
| | >50 | 360 (16.8%) |
| Total | | 2139 |

Table 3. Demographic Summary for Prospective BioFire Global Fever Special Pathogens Panel Clinical Evaluation

PPA for each analyte was calculated as 100% × (TP + FN)). True positive (TP) indicates that both the BioFire Global Fever Special Pathogens Panel and the comparator method had a positive result for this specific analyte, and false negative (FN) indicates that the BioFire Global Fever Special Pathogens Panel result was negative while the comparator result was positive. NPA was calculated as 100% × (TN / (TN + FP)). True negative (TN) indicates that both the BioFire Global Fever Special Pathogens Panel and the comparator method had negative results and a false positive (FP) indicates that the BioFire Global Fever Special Pathogens Panel result was positive, but the comparator result was negative. Results are summarized in Tables 4 (viruses), 5 (bacteria), and 6 (protozoa).

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| BioFire Global Fever | | PPA | | | NPA | | |
|---------------------------------------------|------------------|-----------------|-------|----------------|-----------------|-------|-----------|
| Special Pathogens<br>Panel Detected Result | Number<br>Tested | TP/(TP +<br>FN) | % | 95% CI | TN/(TN +<br>FP) | % | 95% CI |
| Chikungunya virusa | 1875c | 25/25 | 100% | 86.7-<br>100% | 1848/1850 | 99.9% | 99.6-100% |
| Crimean-Congo<br>hemorrhagic<br>fever virus | 2139 | 1/1 | 100% | 20.7-<br>100% | 2138/2138 | 100% | 99.8-100% |
| Dengue virusb | 1875c | 266/283 | 94.0% | 90.6-<br>96.2% | 1592/1592 | 100% | 99.8-100% |
| Ebola virus | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |
| Lassa virus | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |
| Marburg virus | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |
| West Nile virus | 2139 | 1/1 | 100% | 20.7-<br>100% | 2138/2138 | 100% | 99.8-100% |
| Yellow fever virus | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |

Table 4. BioFire Global Fever Special Pathogens Panel Clinical Performance Summary - Viruses

ª Evidence of Chikungunya virus was found in 2/2 FP specimens by additional PCR.

ֿ Evidence of Dengue virus was found in 15/17 FN specimens were positive upon BioFire Global Fever Special Pathogens Panel retest and by additional PCR, two were positive Global Fever Special Pathogens Panel retest, and eight were detected only by additional PCR.

^ Comparator analysis was not performed for Chikungunya virus on specimens collected after September 2019.

| BioFire Global Fever Special<br>Pathogens Panel Detected<br>Result | Number<br>Tested | TP/(TP + FN) | PPA<br>% | 95% CI | TN/(TN + FP) | NPA<br>% | 95% CI |
|--------------------------------------------------------------------|------------------|--------------|----------|----------------|--------------|----------|------------|
| Bacillus anthracis | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |
| Francisella tularensis | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |
| Leptospira spp.a | 1875b | 15/16 | 93.8% | 71.7-<br>98.9% | 1855/1859 | 99.8% | 99.4-99.9% |
| Yersinia pestis | 2139 | 0/0 | - | - | 2139/2139 | 100% | 99.8-100% |

Table 5. BioFire Global Fever Special Pathogens Panel Clinical Performance Summary - Bacteria

ª Evidence of Leptospira spp. was found in 1/1 FN specimens by BioFire Global Fever Special Pathogens Panel retest and by additional PCR, and in 3/4 FP specimens by additional PCR.

b Comparator analysis was not performed for Leptospira on specimens collected after September 2019.

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| BioFire Global Fever Special<br>Pathogens Panel Detected<br>Result | Number<br>Tested | TP/(TP + FN) | PPA | | NPA | |
|--------------------------------------------------------------------|------------------|--------------|-------|------------|-------|------------|
| | | | % | 95% CI | % | 95% CI |
| Leishmania spp. | 2139 | 10/10 | 100% | 72.2-100% | 100% | 99.8-100% |
| Plasmodium spp.a,b | 1875e | 338/343 | 98.5% | 96.6-99.4% | 99.2% | 98.6-99.5% |
| Plasmodium falciparumc | 1875e | 230/248 | 92.7% | 88.8-95.4% | 99.8% | 99.5-99.9% |
| Plasmodium vivax/ovaled | 1875e | 115/124 | 92.7% | 86.8-96.1% | 100% | 99.8-100% |
| TN/(TN +<br>FP) | 2129/2129 | | | | | |
| 1519/1532 | | | | | | |
| 1624/1627 | | | | | | |
| 1751/1751 | | | | | | |

Table 6. BioFire Global Fever Special Pathogens Panel Clinical Performance Summary – Protozoa

® Four (4/5) Plasmodium FN specimens were also P. falciparum FN and one (1/5) was P. vivax/ovale FN. Three (3/13) Plasmodium FP specimens were also P. falciparum FP.

ካ Evidence of Plasmodium spp. was found in 2/5 FN specimen was positive upon BioFire Global Fever Special Pathogens Panel retest and by additional PCR, and one was positive only upon BioFire Global Fever Special Pathogens Panel retest. Evidence of Plasmodium spp. was found in 11/13 FP specimens by additional PCR (10/13) or by species-level comparator assay (1/13).

^ Evidence of P. falciparum was found in 13/18 FN specimens were positive upon BioFire Global Fever Special Pathogens Panel retest and by additional PCR, one was positive only upon BioFire Global Fever Special Pathogens Panel retest, and nine were positive only by additional PCR. Evidence of P. falciparum was found in 2/3 FP specimens by additional PCR. d Evidence of P. vivax/ovale was found in 7/9 FN specimens were positive upon BioFire Global Fever Special Pathogens Panel retest and by additional PCR, two were positive Global Fever Special Pathogens Panel retest, and three were positive only by additional PCR.

e Comparator analysis was not performed for Plasmodium, P. falciparum, or P. vivax/ovale on specimens collected after September 2019.

### Archived Specimen Study

Many of the analytes detectable by the BioFire Global Fever Special Pathogens Panel were not observed or were not encountered in large enough numbers in the prospective clinical evaluation to adequately demonstrate system performance. In this study, archived specimens with known analyte content and/or archived specimens with a high likelihood of containing a given analyte were tested with the BioFire Global Fever Special Pathogens Panel to supplement the prospective clinical evaluation data. Wherever possible, archived whole blood specimens were tested. Where no whole blood specimens could be obtained, blood plasma and blood serum were tested instead. Although plasma and serum are not the intended specimen type for the BioFire Global Fever Special Pathogens Panel, these blood components provide similar results to whole blood specimens in a matrix equivalency study. Archived specimens were collected from a range of ages and sexes (Table 7).

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| | | Overall | Site 01 a | Site 02 | Site 03 a |
|-----|----------|-------------|-------------|------------|------------|
| Sex | Female | 160 (38.5%) | 79 (39.7%) | 49 (59.8%) | 32 (23.7%) |
| | Male | 148 (35.6%) | 97 (48.7%) | 33 (40.2%) | 18 (13.3%) |
| | Unknown | 108 (26.0%) | 23 (11.6%) | 0 (0%) | 85 (63%) |
| Age | <5 | 14 (3.4%) | 14 (7.0%) | 0 (0%) | 0 (0%) |
| | 5 to 21 | 37 (8.9%) | 19 (9.5%) | 14 (17.1%) | 4 (3%) |
| | 22 to 50 | 200 (48.1%) | 121 (60.8%) | 56 (68.3%) | 23 (17%) |
| | >50 | 57 (13.7%) | 22 (11.1%) | 12 (14.6%) | 23 (17%) |
| | Unknown | 108 (26.0%) | 23 (11.6%) | 0 (0%) | 85 (63%) |
| | Total | 416 | 199 | 82 | 135 |

Table 7. Overall and Per Site Archived Demographic Analysis

a Demographic data was not available for 23 specimens from Site 01 and 85 specimens from Site 03.

Specimens were coded and either tested at the clinical site or shipped to BioFire Defense for testing on the BioFire Global Fever Special Pathogens Panel. Nucleic acid was extracted from each specimen and tested using plate-based PCR comparator methods. Samples for which false positive and/or false negative results (i.e., discrepant results) were obtained when comparing the BioFire Global Fever Special Pathogens Panel results to the comparator method results were further investigated. Results are summarized in Table 8.

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| Pathogena | PPA | | | NPA | | |
|------------------------------------------|--------------|-------|------------|--------------|---------|------------|
| | TP/(TP + FN) | % | 95% CI | TN/(TN + FP) | % | 95% CI |
| Viruses | | | | | | |
| Crimean-Congo hemorrhagic fever<br>virus | 0/0 | - | - | 281/281 | 100% | 98.7-100% |
| Ebolavirus spp.b | 0/0 | - | - | 279/279 | 100% | 98.6-100% |
| Lassa virusb,c | 10/12 | 83.3% | 55.2-95.3 | 265/267 | 99.2% | 97.3-99.8% |
| Marburgvirus sp.b | 0/0 | - | - | 279/279 | 100% | 98.6-100% |
| West Nile virusb,d,e,f | 59/65 | 90.8% | 81.3-95.7% | 345/347 | 99.4% | 97.9-99.8% |
| Yellow fever virusb | 0/0 | - | - | 279/279 | 100% | 98.6-100% |
| Bacteria | | | | | | |
| Bacillus anthracis | 0/0 | - | - | 281/281 | 100% | 98.7-100% |
| Francisella tularensis | 0/0 | - | - | 281/281 | 100% | 98.7-100% |
| Yersinia pestis | 0/0 | - | - | 281/281 | 100% | 98.7-100% |
| Protozoa | | | | | | |
| Leishmania spp.g | 0/0 | - | - | 283/283 | 283/283 | 98.7-100% |

### Table 8. BioFire Global Fever Special Pathogens Panel Archived Performance Summary

ª Results are not shown for assays previously granted in DEN20043 (i.e., chikungunya virus, Leptospira spp., and Plasmodium spp.).

b Due to low specimen volume, comparator results for most pathogens were not obtained for two specimens were only tested for a subset of pathogens including Crimean-Congo hemorrhagic fever virus, Bacillus anthracis, Francisella tularensis, Yersinia pestis, and Leishmania spp.

^ Evidence of Lassa virus was found in ½ FN specimens and ½ FP specimens by additional PCR.

ª A set of 133 specimens tested at Site 03 had been previously characterized and were expected to be negative for all panel targets or positive for West Nile virus. These specimens were only evaluated by comparator methods for West Nile virus. e Archived specimens included blood serum and blood plasma.

f Evidence of West Nile virus was found in 6/6 FN specimens were positive upon Global Fever Special Pathogens Panel retesting and three of these were also positive by additional PCR. The other two FN specimens were positive only by

additional PCR testing. Evidence of West Nile virus was detected in 1/2 FP specimens by additional PCR testing.

ß Two specimens tested at Site 03 had been previously characterized and were only tested on comparator assays for Leishmania spp.

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### Contrived Specimen Study

For analytes for which no archived specimens were available, or for which there were an insufficient number of archived specimens, testing was performed using contrived specimens. This study evaluated BioFire Global Fever Special Pathogens Panel sensitivity and specificity when testing whole-blood specimens contrived with Bacillus anthracis. Crimean-Congo hemorrhagic fever virus, Ebolavirus spp., Francisella tularensis, Lassa fever virus, Leishmania spp., Marburgvirus sp., West Nile virus, Yellow fever virus, and Yersinia pestis.

Contrived specimens were prepared using residual human whole blood specimens from patients with signs/symptoms of acute febrile illness. For each analyte, fifty (50) replicates were contrived using quantified isolates at a range of concentrations relative to the limit of detection (LoD). If known, clinically relevant concentrations were used to adjust testing levels. The contrived specimens also served as a negative replicate for all other analytes evaluated. Specimens were prepared and randomized such that the analyte status of each contrived specimen was blinded to the users performing testing. The positive percent agreement (PPA) and negative percent agreement (NPA) were defined as agreement between the BioFire Global Fever Special Pathogens Panel result and the known composition of the contrived specimen. A summary of the results is shown in Table 9.

| | PPA | | | NPA | | |
|------------------------|--------------|-----|-----------|--------------|------|----------|
| Analyte | TP/(TP + FN) | % | 95% CI | TN/(TN + FP) | % | 95% CI |
| Bacillus anthracis | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |
| CCHF virusa | 98/100b | 98 | 93.0-99.5 | 282/282 | 100 | 98.7-100 |
| Ebolavirus spp. | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |
| Francisella tularensis | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |
| Lassa virus | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |
| Leishmania spp. | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |
| Marburgvirus sp.a | 99/100c | 99 | 94.6-99.8 | 282/282 | 100 | 98.7-100 |
| West Nile virus | 50/50 | 100 | 92.9-100 | 331/332d | 99.7 | 98.3-100 |
| Yellow fever virus | 49/50e | 98 | 89.5-99.7 | 332/332 | 100 | 98.9-100 |
| Yersinia pestis | 50/50 | 100 | 92.9-100 | 332/332 | 100 | 98.9-100 |

### Table 9. Summary of BioFire Global Fever Special Pathogens Panel Contrived Specimen Performance Data

ª Tested at additional concentrations to better represent clinically relevant range.

b Two false negative CCHF virus at 2× LoD.

c Single false negative Marburqvirus sp. at 10× LoD.

d Discrepancy testing showed near LoD levels of WNV in a single whole blood sample.

e Single false negative yellow fever virus at 5× LoD.

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# Select Analytical Studies

### Limit of Detection

The Limit of Detection (LoD) for each analyte on the BioFire Global Fever Special Pathogens Panel was determined using quantified stocks within the BioFire Defense BioSafety Level 2 (BSL2) laboratory. For analytes that required BSL3/4 containment, inactivated strains were used. Contrived samples were prepared at known concentrations in a whole blood background. An estimated LoD concentration for each analyte was determined based on results of serial dilutions spanning at least four concentrations bracketing the anticipated LoD. Four replicates were tested at each dilution, with additional dilutions if needed, to reach a concentration at which loss of detection could be observed. The LoD concentration was confirmed by testing 20 replicates at the estimated LoD concentration and another 20 replicates at a ten-fold lower concentration. The required criteria for confirmation of LoD was a detection rate of at least 95% at LoD ((≥19/20) and a detection rate of less than 95% below LoD (<19/20).

The confirmed LoD concentrations for BioFire Global Fever Special Pathogens Panel analytes along with the detection rate at 1× LoD are shown in Table 10. The LoD concentration for each analyte is reported as target copies/mL determined using commercially available quantitative real-time PCR assay kits.

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| Table 10. Limit of Detection for BioFire Global Fever Special Pathogens Panel Test Results | | | | |
|--------------------------------------------------------------------------------------------|-------------------------------------------------------------------------|------------------|-------------------|-------------------|
| Global Fever Special<br>Pathogens Panel<br>Analyte | Isolate Tested | Live/Inactivated | LoD Concentration | |
| | | | Copies/mL¹ | Units/mL |
| BACTERIA | | | | |
| Bacillus anthracis | Ames35 | Live | 4.2E+01 | N/A |
| Francisella tularensis | SCHU S4 | Inactivated | 1.2E+03 | N/A |
| Leptospira spp. | <i>interrogans</i> : serovar icterohaemorrhagiae,<br>Serotype: Budapest | Live | 3.4E+02 | N/A |
| Yersinia pestis | A1122 | Live | 1.3E+02 | N/A |
| VIRUSES | | | | |
| Chikungunya virus | R80422 | Inactivated | 5.5E+02 | 3.6E-01 units/mL |
| Crimean-Congo<br>hemorrhagic fever<br>virus | Strain IbAr10200 | Inactivated | 6.4E+00 | N/A |
| Dengue virus | DENV-1: Hawaii | Live | 2.2E+02 | N/A |
| | DENV-2-1: New Guinea C | Live | 3.4E+02 | N/A |
| | DENV-2-2: Dak AR A1247 | Live | 2.7E+03 | 1.5E+02 TCID50/mL |
| | DENV-3: H87 | Live | 1.3E+02 | 3.7E+00 units/mL |
| | DENV-4: H241 | Live | 6.4E+01 | 1.8E+02 units/mL |
| Ebolavirus spp. | Bundibugyo: 200706291 Uganda | Inactivated | 7.0E+04 | N/A |
| | Taï Forest: Cote d'Ivoire 11/27/94 | Inactivated | 8.3E+03 | N/A |
| | Reston: 119810 RIID<br>(MKY 53) (prototype 1989) | Inactivated | 2.7E+04 | N/A |
| | Sudan: Boniface | Inactivated | 1.1E+04 | N/A |
| | Zaire:<br>Guéckédou/Guinea C07 | Inactivated | 1.0E+02 | 1.5E+02<br>PFU/mL |
| Lassa virus | Josiah | Inactivated | 5.6E+04 | N/A |
| Marburgvirus | Marburg virus: Musoke | Inactivated | 5.0E+02 | N/A |
| | Ravn virus: Kenya Ravn | Inactivated | 2.6E+02 | N/A |
| West Nile virus | NY 2001-6263 (Lineage 1) | Inactivated | 1.1E+03 | 2.7E+01 units/mL |
| | B-956 Uganda (Lineage 2) | Inactivated | 2.3E+04 | 6.2E+00 units/mL |
| Yellow fever virus | Strain 17D | Live attenuated | 1.2E+02 | 1.2E+01 TCID50/mL |
| PROTOZOA | | | | |
| Leishmania spp. | <i>donovani</i> : 9515 (MHOM/IN/95/9515) | Live | 1.0E+01 | 2.2E+01 cells/mL |
| Plasmodium spp. | <i>falciparum</i> , IPC 4884 Pursat<br>Cambodia 2011 | Live | 1.8E+02 | N/A |
| | <i>knowlesi</i> , H strain | gDNA | 2.4E+01 | 20 pg/mL |
| | <i>malariae</i> (Clinical Specimen) | Live | 1.9E+02 | 2.3E-01 cells/mL |
| | <i>ovale, wallikeri</i> (Clinical Specimen) | Live | 2.4E+02 | N/A |
| | <i>vivax</i> , Strain Chesson | Live | 1.5E+02 | N/A |
| Plasmodium falciparum | IPC 4884 Pursat<br>Cambodia 2011 | Live | 1.8E+02 | N/A |
| Plasmodium<br>vivax/ovale | <i>ovale, wallikeri</i> (Clinical Specimen) | Live | 2.4E+02 | N/A |
| | <i>vivax</i> , Strain Chesson | Live | 1.5E+02 | N/A |

### Table 10. Limit of Detection for BioFire Global Fever Special Pathogens Panel Test Results

1 Stock concentrations determined using commercially available quantitative real-time PCR assay kits.

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Since decreased sensitivity may be observed due to nucleic acid damage when evaluating inactivated BSL3/4 pathogens, additional testing was performed using live strains within a subcontracted BSL3/4 laboratory.

The concentrations tested were based on the confirmed LoD for inactivated/attenuated material. In general, four test concentrations were tested for each analyte: 10×, 1×, 0.1×, and 0.01× the LoD for inactivated/attenuated stock. Some analytes required additional concentrations. For each concentration, four unique blood draws were individually spiked and tested on the BioFire Global Fever Special Pathogens Panel. The estimated LoD was identified as the lowest concentration at which 4/4 replicates were positive and fewer than four replicates were positive at the next lower concentration. The estimated LoD values for infectious material are provided in Table 11.

| | Table 11, BioFire Global Fever Special Pathogens Panel Estimated LoD in Whole Blood for Live BSL3/4 | |
|--------|-----------------------------------------------------------------------------------------------------|--|
| Agents | | |
| | | |

| BioFire Global Fever<br>Special Pathogens Panel<br>Analyte | Species/Strain | Estimated LoD Concentration | |
|------------------------------------------------------------|---------------------------------------------|-----------------------------|----------------|
| | | Copies/mL | Units/mL |
| Bacteria | | | |
| Bacillus anthracis | Ames | 6.4E+01 | 3.5E+00 cfu/mL |
| Francisella tularensis | subsp. tularensis Schu | 1.2E+01 | 2.1E+02 cfu/mL |
| Yersinia pestis | CO92 | 1.5E+02 | 3.0E+01 cfu/mL |
| Viruses | | | |
| CCHF Virus | IbAr10200 | 6.4E+02 | 2.9E+03 pfu/mL |
| Chikungunya virus | B8635 | 5.5E+02 | 4.4E+01 pfu/mL |
| Chikungunya virus | Indo23574 | 5.5E+02 | 9.0E+01 pfu/mL |
| Ebolavirus spp. | Bundibugyo virus / Uganda (811250) | 7.0E+02 | 5.6E+00 pfu/mL |
| Ebolavirus spp. | Tai Forest virus / Taï Forest (Ivory Coast) | 1.8E+02 | N/A |
| Ebolavirus spp. | Reston virus / H-28 | 2.7E+03 | 4.7E+01 pfu/mL |
| Ebolavirus spp. | Sudan virus / Boniface | 1.1E+02 | 5.3E+00 pfu/mL |
| Ebolavirus spp. | Zaire ebolavirus / Makona | 1.1E+03 | 1.6E+01 pfu/mL |
| Lassa Virus | Josiah | 5.6E+03 | 2.6E+01 pfu/mL |
| Marburg marburgvirus | Ci67 | 5.0E+02 | 1.5E+03 pfu/mL |
| Marburg marburgvirus | Ravn | 2.6E+02 | 4.7E+01 pfu/mL |
| West Nile virus | Bz NY99 (lin. 1) | 1.6E+02 | 9.5E+00 pfu/mL |
| Yellow fever virus | Asibi | 1.2E+01 | 9.2E+00 pfu/mL |

{20}------------------------------------------------

### Inclusivity (Reactivity)

The analytical reactivity (inclusivity) of the BioFire Global Fever Special Pathogens Panel was evaluated by testing a diverse collection of strains/species/serotypes representing temporal, geographic, and genetic diversity. Available isolates were prepared as contrived whole blood samples with the isolate at a concentration near the LoD of the analyte. If the isolate was detected within 3× the LoD the BioFire Global Fever Special Pathogens Panel was considered inclusive for that isolate and those with similar sequences. Analytes that were detected at ≥10× the established LoD were considered to have reduced sensitivity.

Table 12 shows a summary of the BioFire Global Fever Special Pathogens Panel reactivity based on empirical data. Isolates used to determine LoD are bolded.

| Analyte | # Isolates<br>Detected /<br>Tested | Isolates Tested | Source / ID | Concentration<br>Detected<br>Tested up to<br>100x LoD¹ | Limitations | |
|-------------------------------------------------------------------|------------------------------------|------------------------|---------------------------------------------------------------|--------------------------------------------------------|--------------------------------------------------------|-----------------------------------------------------------------------------|
| BACTERIA | | | | | | |
| <i>Bacillus<br/>anthracis</i> | 4/4 | Ames35 | BEI / NR-10355 | 4.2E+01 copies/mL | None | |
| | | Sterne 34Fs | BEI / NR-1400 | | | |
| | | UM23 | BEI / NR-10351 | 1.3E+02 copies/mL | | |
| | | Weybridge | BEI / NR-10350 | | | |
| <i>Francisella<br/>tularensis</i> | 5/5 | Subspecies | | | | |
| | | <i>tularensis</i> | SCHU S4 | BEI / NR-15753 | 1.2E+03 copies/mL | |
| | | <i>holarctica</i> | LVSR | BEI / NR-597 | | |
| | | | Type B LVS (CDC) | BEI / NR-646 | 3.6E+03 copies/mL | |
| | | <i>novicida</i> | CG62<br>KM14S | BEI / NR-580<br>BEI / NR-573 | | |
| <i>Leptospira spp.</i> | 19/19 | Species | | | | |
| | | <i>interrogans</i> | Serovar (Budapest)<br>HAI0156 (Copenhageni)<br>L495 (Manilae) | ATCC / 23581<br>BEI / NR-19891<br>BEI / NR-19816 | 3.4E+02 copies/mL<br>1.2E+03 copies/mL<br> | |
| | | <i>alexanderi</i> | L60 (Manhao 3) | ATCC / 700520 | 1.0E+03 copies/mL | |
| | | <i>alstonii</i> | Sichuan 79601 | ATCC / BAA-2439 | 1.2E+03 copies/mL | |
| | | <i>borgpetersenii</i> | Castellon 3 (Castellonis)<br>Veldrat Bataviae 46 (Javanica) | ATCC / 23580<br>ATCC / 43292 | 1.2E+03 copies/mL<br>1.0E+03 copies/mL | |
| | | <i>kirschneri</i> | 200701401 (Bogvere) | BEI / NR-19942 | 1.2E+03 copies/mL | |
| | | | 3522 C (Cynopteri) | ATCC / 49945 | 5.3E+02 copies/mL | |
| | | <i>kmetyi</i> | Bejo-Iso9T (Malaysia) | BEI / NR-22254 | | |
| | | <i>mayottensis</i> | 200901116 (undesignated) | KIT / 0254 | 1.2E+03 copies/mL | |
| | | <i>noguchii</i> | CZ 214T (Panama) | BEI / NR-22283 | | |
| | | <i>santarosai</i> | LT 821 (Shermani) | ATCC / 43286 | 8.7E+02 copies/mL | |
| Analyte | # Isolates<br>Detected /<br>Tested | Isolates Tested | Source / ID | Concentration<br>Detected<br>Tested up to<br>100x LoD¹ | Limitations |…