Predicate Devices

Not Found

Reference Devices

The provided text does not contain any K/DEN numbers for Reference Devices. The section titled "Reference Device(s)" explicitly states "Not Found".

AI/ML (Artificial Intelligence / Machine Learning)

No
The description details standard molecular diagnostic techniques (PCR, melt curve analysis) and software interpretation of data, without mentioning AI or ML algorithms.

Therapeutic

No.
The document explicitly states that the device is an "in vitro diagnostic "test intended for use with the FilmArray 2.0 system." Its purpose is to "detects and identifies selected bacterial, viral, and protozoan nucleic acids" to aid in diagnosis, not to provide therapy.

Diagnostic

Yes

The document explicitly states in the "Intended Use / Indications for Use" section that the FilmArray Global Fever Panel is an "in vitro diagnostic test."

SaMD (Software as a Medical Device)

No

The device description clearly outlines a system that includes physical components like the FilmArray 2.0 instrument, pouches containing reagents, inflatable bladders, seal points, pneumatic pistons, Peltier devices, and a digital camera. While software is used for interpretation and guiding the user, it is integral to a larger hardware system that performs the diagnostic test.

IVD (In Vitro Diagnostic)

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

The document explicitly states in the "Intended Use / Indications for Use" section:

"The FilmArray Global Fever Panel is a qualitative, multiplexed, nucleic acid-based in vitro diagnostic test intended for use with the FilmArray 2.0 system."

It also states in the "Intended User / Care Setting" section:

"For in vitro diagnostic use only."

These statements clearly indicate that the device is intended for use in vitro (outside of the body) to diagnose conditions, which is the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The FilmArray Global Fever Panel is a qualitative, multiplexed, nucleic acid-based in vitro diagnostic test intended for use with the FilmArray 2.0 system. The FilmArray Global Fever Panel detects and identifies selected 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 following target pathogens: Leptospira spp., chikungunya virus, dengue virus (serotypes 1, 2, 3 and 4), and Plasmodium spp. (including species differentiation of Plasmodium falciparum and Plasmodium vivax/ovale). 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. 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.

Positive results do not rule out co-infections with pathogens not included on the FilmArray Global Fever Panel. Not all pathogens that cause acute febrile illness are detected by this test, and negative results do not rule out the presence of other infections. Patient travel history and consultation of the CDC Yellow Book should be considered prior to use of the FilmArray Global Fever Panel as some pathogens are more common in certain geographical locations.

Product codes

OMV

Device Description

The FilmArrav Global Fever Panel is a multiplex nucleic acid-based test designed to be used with the FilmArray 2.0 system ("FilmArray system" or "FilmArray instrument"). The FilmArray Global Fever Panel includes a FilmArray Global Fever Panel pouch (pouch) which contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex polymerase chain reaction (PCR) with DNA melt analysis. The FilmArray Global Fever Panel simultaneously conducts six tests for the identification of bacterial, viral, and protozoan organisms from whole blood specimens collected in EDTA tubes. Results from the FilmArray Global Fever Panel are available within about one hour.

A test is initiated by loading Hydration Solution into one port of the pouch and a whole blood or positive blood culture specimen mixed with the provided Sample Buffer and protease into the other port of the pouch and placing it in the FilmArray Instrument. 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 rehvdrates the reagents. After the pouch is prepared, the FilmArray Software on the FilmArray system 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 FilmArray system.

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

Nucleic acid extraction occurs within the 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 multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly 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 2nd stage PCR, or nested PCR, is performed in each well of the array. At the conclusion of the 2nd 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.

The FilmArray software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel. A description of the individual assays and their result interpretation is included below:

Chikungunya Virus: The Global Fever Panel contains two assays for species-level detection of all chikungunya virus strains (CHIKV1 and CHIKV2). The FilmArray software will interpret any single positive chikungunya assay as a Chikungunya Virus Detected result.
Dengue Virus: The Global Fever Panel contains five individual assays for the detection of dengue virus serotypes 1. 2. 3. and 4 with two assays specifically dedicated to detecting dengue virus serotype 2 (DENV1, DENV2 1, DENV2 2, DENV3. and DENV4). The FilmArray software will interpret any single positive dengue virus assay as a Dengue Virus Detected result.
Leptospira: The Global Fever Panel contains a single pan assay for the genus-level detection of all Leptospira Group 1 species (LEPTO1). A positive pan-Leptospira assay will result in a Leptospira spp. Detected call.
Plasmodium: The Global Fever Panel contains three Plasmodium assays, one genus-. level assay and two species-level assays. The genus-level assay (Plasmodium spp.) detects all five Plasmodium species known to infect humans (P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi). One species-level assay detects Plasmodium falciparum and a combined species-level assay detects both Plasmodium vivax and Plasmodium ovale. Each individual assay is reported as a Detected or Not Detected result separately based on the results of the specific Global Fever Panel assay.

Mentions image processing

A digital camera placed in front of the array captures fluorescent images of the PCR2 reactions and software interprets the data.

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

The study enrolled subjects of various ages. Site 7 and Site 14 did not enroll subjects

Regulation Number and Section

§ 866.3966 Device to detect and identify selected microbial agents that cause acute febrile illness.

(a)
Identification. A device to detect and identify selected microbial agents that cause acute febrile illness is identified as an in vitro device intended for the detection and identification of microbial agents in human clinical specimens from patients with signs and symptoms of acute febrile illness who are at risk for exposure or who may have been exposed to these agents. It is intended to aid in the diagnosis of acute febrile illness in conjunction with other clinical, epidemiologic, and laboratory data, including patient travel, pathogen endemicity, or other risk factors.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device.
(2) The labeling required under § 809.10(b) of this chapter must include:
(i) An intended use that includes a detailed description of targets the device detects and measures, the results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended.
(ii) Limiting statements indicating:
(A) Not all pathogens that cause febrile illness are detected by this test and negative results do not rule out the presence of other infections;
(B) Evaluation of more common causes of acute febrile illness should be considered prior to evaluation with this test;
(C) Test results are to be interpreted in conjunction with other clinical, epidemiologic, and laboratory data available to the clinician; and
(D) When using this test, consider patient travel history and exposure risk, as some pathogens are more common in certain geographical locations.
(iii) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens.
(iv) Detailed discussion of the performance characteristics of the device for all claimed specimen types as shown by the analytical and clinical studies required under paragraphs (b)(3)(ii) and (iii) of this section, except specimen stability performance characteristics.
(v) A statement that nationally notifiable results are to be reported to public health authorities in accordance with local, state, and federal law.
(3) Design verification and validation must include:
(i) A detailed device description (
e.g., all device parts, control elements incorporated into the test procedure, reagents required but not provided, the principle of device operation and test methodology), and the computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported result).(ii) Detailed documentation of analytical studies, including those demonstrating Limit of Detection (LoD), inclusivity, cross-reactivity, microbial interference, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, within lab precision, and reproducibility, as appropriate.
(iii) Detailed documentation and performance results from a clinical study that includes prospective (sequentially collected) samples for each claimed specimen type and, when determined to be appropriate by FDA, additional characterized clinical samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from FDA-accepted comparator methods. Documentation from the clinical studies must include the clinical study protocol (including a predefined statistical analysis plan), study report, testing results, and results of all statistical analyses.
(iv) A detailed description of the impact of any software, including software applications and hardware-based devices that incorporate software, on the device's functions.

Summary

0

EVALUATION OF AUTOMATIC CLASS III DESIGNATION FOR FILMARRAY GLOBAL FEVER PANEL DECISION SUMMARY

A. De Novo Number:

DEN200043

B. Purpose for Submission:

De Novo request for evaluation of automatic class III designation for the FilmArray Global Fever Panel.

C. Measurand:

DNA and RNA sequences from the following organisms: Leptospira spp., dengue virus serotypes 1-4, chikungunya virus, and Plasmodium spp.

D. Type of Test:

Multiplex Nucleic Acid Amplification Test

E. Applicant:

BioFire Defense, LLC

F. Proprietary and Established Names:

FilmArray Global Fever Panel

G. Regulatory Information:

1. Regulation section:
  21 CFR 866.3966
1. Classification:
  Class II

1

1. Product code(s):
  OMV
1. Panel:
  Microbiology (83)

H. Intended Use:

1. Intended use(s):
  The FilmArray Global Fever Panel is a qualitative, multiplexed, nucleic acid-based in vitro diagnostic test intended for use with the FilmArray 2.0 system. The FilmArray Global Fever Panel detects and identifies selected 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 following target pathogens: Leptospira spp., chikungunya virus, dengue virus (serotypes 1, 2, 3 and 4), and Plasmodium spp. (including species differentiation of Plasmodium falciparum and Plasmodium vivax/ovale). 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. 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.

Positive results do not rule out co-infections with pathogens not included on the FilmArray Global Fever Panel. Not all pathogens that cause acute febrile illness are detected by this test, and negative results do not rule out the presence of other infections. Patient travel history and consultation of the CDC Yellow Book should be considered prior to use of the FilmArray Global Fever Panel as some pathogens are more common in certain geographical locations.

1. Indication(s) for use:
  Same as intended use.
1. Special conditions for use statement(s):
  For prescription use only.

For in vitro diagnostic use only.

1. Special instrument requirements:
  The FilmArray Global Fever Panel is performed on the FilmArray 2.0 system.

2

I. Device Description:

The FilmArrav Global Fever Panel is a multiplex nucleic acid-based test designed to be used with the FilmArray 2.0 system ("FilmArray system" or "FilmArray instrument"). The FilmArray Global Fever Panel includes a FilmArray Global Fever Panel pouch (pouch) which contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex polymerase chain reaction (PCR) with DNA melt analysis. The FilmArray Global Fever Panel simultaneously conducts six tests for the identification of bacterial, viral, and protozoan organisms from whole blood specimens collected in EDTA tubes. Results from the FilmArray Global Fever Panel are available within about one hour.

A test is initiated by loading Hydration Solution into one port of the pouch and a whole blood or positive blood culture specimen mixed with the provided Sample Buffer and protease into the other port of the pouch and placing it in the FilmArray Instrument. 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 rehvdrates the reagents. After the pouch is prepared, the FilmArray Software on the FilmArray system 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 FilmArray system.

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

Nucleic acid extraction occurs within the 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 multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly 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 2nd stage PCR, or nested PCR, is performed in each well of the array. At the conclusion of the 2nd 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.

The FilmArray software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result

3

for each organism on the panel. A description of the individual assays and their result interpretation is included below:

. Chikungunya Virus: The Global Fever Panel contains two assays for species-level detection of all chikungunya virus strains (CHIKV1 and CHIKV2). The FilmArray software will interpret any single positive chikungunya assay as a Chikungunya Virus Detected result.
. Dengue Virus: The Global Fever Panel contains five individual assays for the detection of dengue virus serotypes 1. 2. 3. and 4 with two assays specifically dedicated to detecting dengue virus serotype 2 (DENV1, DENV2 1, DENV2 2, DENV3. and DENV4). The FilmArray software will interpret any single positive dengue virus assay as a Dengue Virus Detected result.
. Leptospira: The Global Fever Panel contains a single pan assay for the genus-level detection of all Leptospira Group 1 species (LEPTO1). A positive pan-Leptospira assay will result in a Leptospira spp. Detected call.
Plasmodium: The Global Fever Panel contains three Plasmodium assays, one genus-. level assay and two species-level assays. The genus-level assay (Plasmodium spp.) detects all five Plasmodium species known to infect humans (P. falciparum, P. vivax, P. ovale, P. malariae, and P. knowlesi). One species-level assay detects Plasmodium falciparum and a combined species-level assay detects both Plasmodium vivax and Plasmodium ovale. Each individual assay is reported as a Detected or Not Detected result separately based on the results of the specific Global Fever Panel assay.

Materials provided in each FilmArray Global Fever Panel kit:

. Individually packaged FilmArray Global Fever Panel pouches (6)
. Individually-packaged Transfer Pipettes (7)
. Single-use (1.0 mL) Sample Buffer Tubes (7)
Single-use pre-filled (1.5 mL) Hydration Injection Vials (Blue) (7) 0
Single-use Sample Injection Vials (Red) (7) .
Instructions and Documents .
- FilmArray Global Fever Panel Instructions for Use i
- FilmArray Global Fever Panel Quick Guide i

Materials required but not provided:

10% bleach solution .
FilmArray system including:
FilmArray 2.0 instrument, computer, and software .
FilmArray Pouch Loading Station .

4

J. Standard/Guidance Document Referenced (if applicable):

14971:2007/(R)2010 (Corrected 4 October 2007), 'Medical Devices - Applications of Risk Management to Medical Devices'

Guidance for Industry and Food and Drug Administration Staff - De Novo Classification Process (Evaluation of Automatic Class III Designation) (October 30, 2017)

Guidance for Industry and Food and Drug Administration Staff - Highly Multiplexed Microbiological/Medical Countermeasure In Vitro Nucleic Acid Based Diagnostic Devices (August 27, 2014)

Guideline for Industry and Food and Drug Administration Staff - Class II Special Controls Guideline: Dengue Virus Nucleic Acid Amplification Test Reagents

Guideline for Industry and Food and Drug Administration Staff - Class II Special Controls Guidance Document: Plasmodium Species Antigen Detection Assays

Guidance for Industry and FDA Staff - Assayed and Unassayed Quality Control Material

Guidance for Industry and FDA Staff- Statistical Guidance on Reporting Results from Studies Evaluating Diagnostic Tests (March 13, 2007)

MM03-Ed3, Molecular Diagnostic Methods for Infectious Diseases, CLSI Approved Guideline-Third Edition

EP07-Ed3, Interference Testing in Clinical Chemistry; Approved Guideline-Third Edition

Guidance for the Content of Premarket Submissions for Software Contained in Medical Devices, FDA Guidance Document

Guidance for Industry, FDA Reviewers and Compliance on Off-The-Shelf Software Use in Medical Devices

ISO 62304:2006. 'Medical Device Software - Software Life Cycle Processes' - IEC 62304:2006, November 27, 2008

ISO 62304:2006, 'Medical Device Software - Software Life Cycle Processes' - IEC 62304:2006, November 27, 2008

ISO 15223-1:2012. 'Medical Devices - Symbols to be used with medical device labels. labeling and information to be supplied - Part 1: General requirements'

5

Guidance for Industry and FDA on Alternative to Certain Prescription Device Labeling Requirements

K. Test Principle:

The FilmArray Global Fever Panel pouch is a closed system disposable that houses all the chemistry required to isolate, amplify, and detect nucleic acid from multiple biothreat pathogens within whole blood and positive blood culture. The rigid plastic component (fitment) of the pouch contains reagents in freeze-dried form. The flexible plastic portion of the pouch is divided into discrete segments (blisters) where the required chemical processes are carried out. The user of the FilmArray Global Fever Panel loads the sample into the pouch, places the pouch into the FilmArray instrument, and starts the run. All other operations are automated. Operations and processes that occur during a FilmArray run include the following:

(D)(4) . Nucleic Acid Purification- Nucleic acid purification occurs in the of the pouch. The sample is lysed by agitation (bead beating) and the liberated nucleic acid is captured, washed, and eluted using magnetic bead technology. These steps require about ten minutes and the bead-beater apparatus can be heard as a highpitched whine during the first minute of operation.
Reverse Transcription and 1st Stage Multiplex PCR Some pathogens identified . by the FilmArray Global Fever pouch are RNA viruses, and a reverse transcription (RT) step is performed to convert the viral RNA into cDNA prior to amplification. The purified nucleic acid solution is combined with a preheated master mix to initiate the RT step and subsequent thermocycling for multiplex PCR. The effect of 188 stage PCR is to enrich for the target nucleic acids present in the sample.
. 2nd Stage PCR - The products of 1st stage PCR are diluted and mixed with fresh PCR reagents containing an intercalating fluorescent DNA dye (LCGreen Plus, BioFire Defense, LLC). This solution is distributed over the 2nd stage PCR array. The individual wells of the array contain primers for different assays (each present in triplicate) that target specific nucleic acid sequences from each of the pathogens detected, as well as control template material. These primers are "nested" or internal to the specific products of the 185 stage multiplex reaction, which enhances both the sensitivity and specificity of the reactions.
DNA Melting Analysis After 2nd stage PCR, the temperature is slowly increased . and fluorescence in each well of the array is monitored and analyzed to generate a melt curve. The temperature at which a specific PCR product melts (melting temperature or Tm) is consistent and predictable and the FilmArray software automatically evaluates the data from replicate wells for each assay to report results.

The FilmArray software controls the operation of the instrument, collects and analyzes data and automatically generates a test report at the end of the run.

6

L. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

To evaluate the reproducibility of the FilmArray Global Fever Panel, three contrived whole blood samples were prepared with different mixtures of representative panel analytes. For each analyte, one sample was spiked at a Moderate Positive concentration (3 x LoD), another sample at a Low Positive (1 x LoD) and a third sample that was negative (unspiked) for the given analyte. For P. falciparum, a dilution error occurred that resulted in lower than expected analyte concentration being evaluated for all replicates. Six replicates of each sample were tested across 3 different sites on five different days, providing a total of 90 replicate test results per sample. On each test day at each site, two different operators used three FilmArray instruments; GF Panel pouch lot was rotated daily. In total, 270 valid test results were obtained for the reproducibility evaluation of the FilmArray GF Panel.

Organism (Isolate)	Sample 1	Sample 2
Leptospira interrogans
serovar
icterohaemorrhagiae	Low Positive
3.3E+02 copies/mL
1 x LoD	Moderate Positive
1.0E+03 copies/mL
3 x LoD	Negative
Dengue Virus (DENV-2)
New Guinea C	Low Positive
3.3E+02 copies/mL
1 x LoD	Moderate Positive
1.0E+03 copies/mL
3 x LoD	Negative
Plasmodium falciparum
IPC 4884	Moderate Positive
5.4E+02 copies/mL
1.5 x LoD	Negative	Low Positive
9.0E+01
copies/mL
0.5 x LoD

Table 1. Contrived Whole Blood Samples for Reproducibility Testing

Combined reproducibility results are shown below in Table 2.

7

| | | Test Analyte
Isolate | Concentration | Expected
Test
Result | % Agreement with Expected Results | | | | |
|----------|---------------------------------------------------------------------------------------------------------------|-------------------------------------------------------|-------------------------------------------------------|----------------------------|-----------------------------------|----------------|----------------|----------------|-------------------------------|
| | | | | | Site 1 | Site 2 | Site 3 | All
Sites | 95%
Confidence
Interval |
| BACTERIA | | Leptospira interrogans serovar
icterohaemorrhagiae | Moderate Positive
3x LoD
(1.2E+03 copies/mL) | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| | | | Low Positive
1x LoD
(3.9E+02 copies/mL) | Detected | 27/30
90.0% | 28/30
93.4% | 26/30
86.7% | 81/90
90.0% | 81.9-95.3% |
| | | | Negative
(No Analyte) | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| VIRUSES | | Dengue virus
DENV-2
New Guinea C | Moderate Positive
3x LoD
(1.1E+03 copies/mL) | Detected | 29/30
96.7% | 30/30
100% | 30/30
100% | 89/90
98.9% | 94.0-100% |
| | | | Low Positive
1x LoD
(3.6E+02 copies/mL) | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| | | | Negative
(No Analyte) | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| PROTOZOA | | Plasmodium spp.
Detection Results | Moderate Positive
1.5x LoD1
(2.7E+02 copies/mL) | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| | | | Low Positive
0.5× LOD1
(9.0E+01 copies/mL) | Detected | 28/30
93.3% | 30/30
100% | 29/30
96.7% | 87/90
96.7% | 90.7-98.9% |
| | | | Negative
(No Analyte) | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| | IPC 4884 | Plasmodium
falciparum
Detection Results | Moderate Positive
1.5x LoD1
(2.7E+02 copies/mL) | Detected | 29/30
96.7% | 30/30
100% | 28/30
93.4% | 87/90
96.7% | 90.6-99.3% |
| | | | Low Positive
0.5× LoD1
(9.0E+01 copies/mL) | Detected | 18/30
60% | 24/30
80% | 21/30
70% | 63/90
70.0% | 59.4-79.2% |
| | | | Negative
(No Analyte) | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100% | 96.0-100% |
| | Overall Agreement with the Expected Test Result
All Analytes and All Test Levels (95% Confidence Interval) | | | | 1,037/1,080
96.0% (94.7-97.1%) | | | | |

Table 2: Reproducibility of the FilmArray Global Fever Panel Qualitative Results

1 Due to a correction in the stock concentration, P. falciparum was evaluated at 1.5x LoD and 0.5x LoD.

Detected results were as expected for all analytes except for the Low Positive Leptospira interrogans sample which exhibited 90% agreement. The observed negative specimens were distributed across all runs, sites, testing days, and reagent lots and likely reflect underspiking with Leptospira interrogans.

A secondary assessment of reproducibility is based on variability in the melt temperature (Tm) of the amplification products (measured as standard deviation). Melt temperature mean and standard deviations are shown in Table 3. for control and organism assays for the three test sites and overall. Variability in the melt temperatures for the assays evaluated was within the expected range ( LoD of testing either inactivated or live organisms (based on molecular quantification of nucleic acids for each isolate) in whole blood. Several isolates with reduced assay reactivity are described in more detail below.

14

When possible, in silico analysis of sequence data was used to make predictions of assay reactivity for less common strains or serotypes that were not tested.

Table 8 includes a summary of FilmArray Global Fever Panel reactivity based on empirical data.

| FilmArray
Global Fever
Panel Analyte | #
Isolates
Detected
/ Tested | Isolates Tested | Concentration
(Replicates
Detected/Tested) | |--------------------------------------------| | | | | | HAI0156
(Copenhageni)
L495 (Manilae) | | | | | | (Castellonis)
Veldrat Bataviae
46 (Javanica) | | Leptospira spp. | (Bogvere)
3522 C
(Cynopteri) | | (Malaysia) | | | | | | 6712 | | A 102 (Mengrun)
Celledoni
20160426
H 27 (Hekoou)
LT89-68 (Vughia) | | | | | | | Chikungunya
Virus | b/3 DHS4263
St. Martin 2013 | | | | | Strain 12150 | | 276RK1 | | SL-6-6-04 | Dengue virus | | | V1792/2007 | | | | (DENV2 1) | FilmArray
Global Fever
Panel Analyte | # Isolates
Detected / Tested | Detected1 | x LoD Fever Panel Result
(Replicates
Detected/Tested) |
| | (DENV2 2) | | | | | | | | | | | | V1002/2006 | | | | | | | | | | V1329/2006 | | | | | | | | | | | | | | | | | | | | | Plasmodium spp. | | | | | | 02000708 | | | | | | | | | | | | | specimen | | | Plasmodium
falciparum | 4/4 | | | | | | 02000708 | | | | Plasmodium
vivax/ovale | 4/4 | | | | > | x LoD | FilmArray Global
|
---------------------------------------|-----------------|---------------------------------------------------------------------------------------------------|----------------------------|---------------------------------------------------------------------------|---------------------------------------------------------------------------|
| | BACTERIA | | | |
| Species | Strain | | | |
| b/19 | interrogans | Serovar (Budapest)
| (b)(4) | | Detected (b)(41) |
| alexanderi | L60 (Manhao 3) | | | Detected (b) |
| alstonii | Sichuan 79601 | | | Detected (b) |
| borgpetersenii | Castellon 3
| | |
| kirschneri | 200701401
| | | |
| kmetvi | Bejo-Iso9T
| | | Detected (b) |
| mayottensis | 200901116 | | | |
| noguchii | CZ 214T (Panama) | | | |
| santarosai | LT 821 (Shermani)
| | | |
| weilii | 94-79970/3 Topaz
| |
| | | VIRUSES | | |
| | Strain | (b)(4) | | |
| | R80422
| | | Detected (b)(41) |
| | | | | Detected (b) |
| Serotype | Strain | (b)(4) | | |
| b/28 | | Hawaii
| | | Detected (b)(4) |
| | 228690
| | | |
| Serotype 1 | BC89/94
| | | Detected (b) |
| | UIS 1162 | | | |
| | VN/BID-
| | | |
| | | | | |
| Serotype 2 | New Guinea C
| | | Detected (b)(41) |
| Isolates Tested | Concentration
| FilmArray Global
| | DakArA1247
| (b)(4) | | |
| | 1349 | | | |
| | 429557 | | | Detected (b)(4) |
| | ArA6894 | | | |
| | BC102/94 | | | |
| | DKA 811 | | | Detected (b)(4) |
| | VN/BID-
| | | Detected (b) |
| | H87 | | | Detected (b)(4) |
| | 271242 | | | |
| Serotype 3 | BC188/97 | | | |
| | C0360/94 | | | Detected (b) |
| | VN/BID-
| | | |
| | H241 | | | Detected (b)(4) |
| | 703 | | | |
| | BC13/97 | | | |
| Serotype 4 | BC287/97 | | | Detected (b) |
| | BC258/97 | | | |
| | D85-019 | | | |
| | PR 06-65-740 | | | |
| | | PROTOZOA | | |
| Species | Strain | | | |
| | | | | |
| 10/10 | | falciparum | IPC 4884 | (b)(4) | Detected (b)(4) |
| | | SenTh021.09 | | Detected (b) |
| | falciparum | St. Lucia | | |
| | | Tanzania,
| Detected (b) |
| | vivax | Chesson | | Detected (b)(4) |
| | | Panama | | Detected (b) |
| | ovale | Wallikeri | | Detected (b)(4) |
| | | Curtisi | | Detected (b) |
| | knowlesi | Strain H | | |
| | | malariae | | Clinical
|
| | | | | |
| | falciparum | IPC4884 | (b)(4) | |
| | | SenTh021.09 | | Detected (b) |
| | | St. Lucia | | Detected (b) |
| | | Tanzania,
| Detected (b) |
| | | | | |
| | vivax | Chesson | (b)(4) | Detected (b)(4) |
| | | Panama | | Detected (b) |
| | ovale | Wallikeri | | Detected (b) |

Table 8, Summary of FilmArray Global Fever Panel Analytical Reactivity (Inclusivity)

15

Organisms which exhibited reduced or no assay reactivity have limitations included in the labeling and are described specifically in the table below.

16

| Observed

Result	Detection Level	Analyte	Serotype/Strain/Isolate
Detected
(may be
underreported)	10 x LoD	Plasmodium
falciparum1	SenTh021.09
Detected
(may be
underreported)	~100 x LoD	Dengue virus1	Serotype 3 BC188/97
Serotype 4 D85-019
Not Detected	-	Dengue virus2	Serotype 2 DKA 811

The reason for the observed reactivity could not be identified based on in silico sequence analysis. Sequences for these strains were not available in public databases.

21n silico analysis predicted reduced sensitivity or missed detection of this isolate due to sequence variation. Wet testing of this rare sylvatic strain at 10,000 x LoD confirmed detection was significantly impaired.

f. Microbial Interference Studies:

Potentially interfering microorganisms were evaluated for their effect on FilmArray Global Fever Panel performance. To evaluate the potential for interference, FilmArray Global Fever Panel test results from a control blood sample containing representative panel analytes (Leptospira interrogans, P. falciparum, dengue virus type 3) at concentrations near 3×LoD were compared to results from a sample with the same composition plus the potentially interfering microorganism, as well as a negative sample (no analytes) containing only the potentially interfering microorganism. Each potentially competing microorganism was tested at the highest concentration possible (1:10 dilution of the stock). The samples containing the potentially interfering microorganism were evaluated for their effects on the FilmArray Global Fever Panel internal control assays and analyte detection. Reproducible internal control failures or loss of analyte detection associated with the presence of a particular potentially interfering microorganism would be recognized as interference by that microorganism.

Microorganisms	Concentration Tested	Results
Corynebacterium diphtheriae	1:10 dilution of stock	No interference observed
Staphylococcus epidermidis	3.8E+06 CFU/mL	No interference observed
Escherichia coli	1:10 dilution of stock	No interference observed
Klebsiella pneumoniae	5.5E+04 CFU/mL	No interference observed
Haemophilus influenzae	1.0E+08 CFU/mL	No interference observed
Herpes Simplex virus	1.2E+05 TCID50/mL	No interference observed
Epstein-Barr virus	3.3E+07 copies/mL	No interference observed
Cytomegalovirus	1:10 dilution of stock	No interference observed
Human Immunodeficiency virus
(HIV) 1/2	HIV-1: 1.3E+05 U/mL
HIV-2: 2.2E+05 U/mL	No interference observed
Plasmodium vivax	1.5E+06 copies/mL	No interference observed

Table 9, Organisms Evaluated for Potential Microbial Interference

None of the ten microorganisms tested showed interference with the pouch controls or specific Global Fever Panel assay targets.

17

g. Analytical specificity/Cross-reactivity:

The potential for non-specific amplification and detection by the FilmArray Global Fever Panel assays (cross-reactivity) was evaluated by testing high concentrations of on-panel (identified by the FilmArray Global Fever Panel assays) and off-panel (not intended to be identified by the FilmArray Global Fever Panel assays) organisms or purified nucleic acids, as well as by in silico analysis. All on-panel organisms were tested live at high concentration (> 106 copies/mL). As can be seen in Table 10 below, testing of P. knowlesi demonstrated cross-reactivity with the P. vivax/ovale assay at concentrations above 2.2E+03 copies/mL.

Table 10. FilmArray Global Fever Panel Results for On-Panel Organism Testing Assessing Potential Cross-reactivity

| Pathogen | Live or
Inactivated | | Results | |
|---------------------------------------------|-----------------------------------------|-------------|--------------------------------------------------|-----------------------------------------------------------------------|
| | Live | Inactivated | | |
| Leptospira interrogans (Schu S4) | √ | - | As expected (only Leptospira spp. Detected) | |
| Chikungunya virus (R80422 culture
fluid) | √ | - | As expected (only Chikungunya virus
Detected) | |
| Dengue virus | DENV-1 (Hawaii) | √ | - | As expected (only Dengue virus Detected) |
| | DENV-2 ( New Guinea C) | √ | - | As expected (only Dengue virus Detected) |
| | DENV-3 (H87) | √ | - | As expected (only Dengue virus Detected) |
| | DENV-4 (H241) | √ | - | As expected (only Dengue virus Detected) |
| Plasmodium | falciparum
(Pursat Cambodia
2011) | √ | - | As expected (Plasmodium spp. Detected and P.
falciparum Detected) |
| | knowlesi
(Strain H) | √ | - | Plasmodium spp Detected as expected. |
| | malariae
(DLS17-026015) | √ | - | As expected (only Plasmodium spp. Detected) |
| | vivax
(11 Chesson) | √ | - | As expected (Plasmodium spp. Detected and P.
vivax/ovale Detected) |
| | ovale
(Wallikeri) | √ | - | As expected (Plasmodium spp. Detected and P.
vivax/ovale Detected) |

Although no cross-reactivity with P. malariae was observed in this study, in silico analysis shows be mismatches between P. malariae and the P. vivax/ovale assay primers. Wet testing with P. brasilianum (a monkey Plasmodium thought to have descended from P. malariae and which has identical sequence as P. malariae) showed reactivity for the P. vivax/ovale assay at copies/mL.

18

Off-panel organisms were selected for testing based on a combination of several factors including:

Relatedness to the species detected by the Global Fever Panel (near-. neighbors)
. Clinical relevance (causing symptoms similar to the panel pathogens)
. likelihood of being present in blood as a co-infection based on a geographical region or specific population to which a panel pathogen is endemic
genetic similarity to Global Fever Panel assay primers as determined by in . silico analysis

BSL3/4 organisms and viruses that could not be obtained as inactivated stocks or purified nucleic acids were tested live in an appropriate facility | and PMA (b)(4) to assess potential cross-reactivity.

Table 11 below lists the off-panel organisms that were tested at high concentrations (> 106 copies/mL) for potential cross-reactivity with the FilmArray Global Fever Panel.

Bacteria		Viruses
Acinetobacter baumannii	Mycoplasma pneumoniae	Adenovirus 1	Omsk hemorrhagic fever
virus
Bacillus anthracis	Neisseria meningitidis	Adenovirus 3	O'nyong-nyong virus1
Bacillus brevis	Proteus mirabilis	Adenovirus 5	Parvo virus
Bacillus cereus	Pseudomonas aeruginosa	Aura virus	Powassan virus
Bacillus circulans	Rickettsia typhi	Barmah Forest virus	Rabies virus
Bacillus coagulans Hammer	Salmonella enterica subsp.
arizonae	Bunyamwera virus	Rift Valley Fever Virus
Bacillus halodurans	Salmonella enterica subsp.
bongori	Coronavirus NL63	Ross River virus
Bacillus licheniformis	Salmonella enterica subsp.
diarizoniae	Crimean-Congo Hemorrahagic
Fever Virus	Human respiratory syncytial
virus
Bacillus megaterium	Salmonella enterica subsp.
enterica serovar Dublin	Dugbe virus	Rubella virus
Bacillus mycoides	Salmonella enterica subsp.
enterica serovar Enteritidis	Eastern Equine Encephalitis
Virus	Saint Louis encephalitis virus
Bacillus pumilus	Salmonella enterica subsp.
enterica serovar Javiana	Ebolavirus (Zaire, Sudan,
Bundibugyo, Tai Forest,
Reston)	Sindbis virus
Bacillus subtilis	Salmonella enterica subsp.
enterica serovar
Manchester	Enterovirus, HEV-71	Spondweni virus
Bacillus thuringiensis	Salmonella enterica subsp.
enterica serovar
Montevideo	Epstein Barr virus	Tickborne encephalitis virus
Bacteroides fragilis	Salmonella enterica subsp.
enterica serovar
Muenchen	Flexal virus	Una virus
Bordetella bronchiseptica	Salmonella enterica subsp.	Guanarito virus	Usutu virus
	enterica serovar Newport
Borrelia burgdorferi	Salmonella enterica subsp.
enterica serovar Paratyphi	Hantaan virus	Vaccinia virus
Brucella melitensis	Salmonella enterica subsp.
enterica serovar Rubislaw	Hazara virus	Varicella zoster virus
Burkholderia cepacia	Salmonella enterica subsp.
enterica serovar Saintpaul	Hendra virus	Venezuelan Equine
Encephalomyelitis (VEE)
virus
Burkholderia mallei	Salmonella enterica subsp.
enterica serovar Tennessee	Hepatitis A Virus (HAV)	Western Equine Encephalitis
(WEE) virus
Burkholderia pseudomallei	Salmonella enterica subsp.
enterica serovar
Thompson	Hepatitis B Virus (HBV)	West Nile virus Lineage 1
Chlamydophila pneumoniae	Salmonella enterica subsp.
enterica serovar Typhi	Hepatitis C Virus (HCV)	West Nile virus Lineage 2
Clostridium bifermentans	Salmonella enterica subsp.
enterica serovar
Typhimurium	Herpes Simplex Virus 2
(HSV-2)	Yellow Fever virus
Clostridium perfringens	Salmonella enterica subsp.
houtenae	Human Parainfluenza Virus 1	Zika virus
Clostridum sordelli	Salmonella enterica subsp.
indica	Human Parainfluenza Virus 3	Fungi
Clostridium sporogenes	Salmonella enterica subsp.
salamae	Hughes virus	Aspergillus fumigatus
Clostridium perfringens	Serratia marcescens	Human herpesvirus 6B	Cryptococcus neoformans
Coxiella burnetii	Staphylococcus aureus	Human Immunodeficiency
Virus 1	Pathogenic Protozoa
Enterobacter aerogenes	Streptococcus agalactiae	Human Immunodeficiency
Virus 2	Babesia microti
Enterococcus faecalis	Streptococcus pneumoniae	Human T-lymphotropic virus
type 1	Crithidia fasciculata
Enterococcus faecium	Streptococcus pyogenes	Human T-lymphotropic virus
type 2	Cyclospora cayetanensis
Francisella hispaniensis	Vibrio cholerae	Influenza A H1N1-2009	Leishmania donovani
Francisella tularensis subsp.
tularensis	Yersinia aldovae	Influenza A H3N2	Leptomonas seymouri
Francisella persica	Yersinia bercovieri	Influenza B virus	Schistsoma mansoni
Francisella philomiragia	Yersinia entericolitica	Japanese Encephalitis Virus
(JEV)	Toxoplasma gondii
Klebsiella oxytoca	Yersinia fredericksenii	Junin virus	Trypanosoma brucei
Legionella pneumophila	Yersinia intermedia	Lassa virus	Trypanosoma cruzi
Leptospira biflexa	Yersinia kristensenii	Machupo virus	Trypansoma rangeli
Leptospira meyeri	Yersinia mollarettii	Marburg Marburgvirus variant
Musoke	Plasmodium berghei2
Leptospira terpstrae	Yersinia pestis	Marburg Marburgvirus
(RAVN)	Plasmodium brasilianum3
Leptospira vanthielii	Yersinia
pseudotuberculosis	Mayaro virus	Plasmodium cynomolgi4
Leptospira wolbachii	Yersinia rohdei	Measles virus	Plasmodium fieldi5
Leptospira yanagawae		Metapneumovirus	Plasmodium fragile6
Listeria monocytogenes		Mopeia Virus	Plasmodium inui7
Mycobacterium tuberculosis		Mumps Virus	Plasmodium simiovale8

Table 11. Off-Panel Organisms Tested by the FilmArray Global Fever Panel

19

20

Image /page/20/Picture/0 description: The image is a gray rectangle with a red border. There is some text in the middle of the rectangle that is difficult to read. The text appears to be a fraction, with the numerator being (6)(4).

There were two instances of cross-reactivity observed during testing:

Image /page/20/Picture/2 description: The image is a gray rectangle with a red border. The numbers 1 and 2 are in the upper left corner of the rectangle. The text (b)(4) is in the upper center of the rectangle.

No other cross-reactivity was observed in wet testing.

In addition to the experimental testing, in silico analyses directed toward the potential to cross-react with specific pathogens that were unavailable for testing was also performed. This included Avalon virus, Bas-Congo virus, Bacillus luciferensis, Chalmydophila psittaci, Francisella mediasiatica, Lymphocytic choriomeningitis virus, Middleburg virus, Murray Valley encephalitis virus, Orientia chuto (tsutsugamushi), Pirital virus, Rickettsia prowazekii, Rickettsia ricketsii, Sabia virus,

21

Semliki Forest virus, Tonate virus, and Variola major. No expected cross-reactivity with off-panel pathogens were predicted by in silico analysis.

h. Interfering Substances:

Potentially interfering substances that could be present in whole blood or introduced during specimen collection and testing were evaluated for their effect on the FilmArray Global Fever Panel performance. Each substance was added to contrived samples containing representative FilmArray Global Fever Panel organisms (Leptospira interrogans, dengue virus Type 3, and P. falciparum) at concentrations equivalent to approximately 3×LoD. The concentrations of endogenous and exogenous substances tested were based on a reference concentration of "normal" to "high" levels expected to be present in clinical specimens.

The following two tables show the results for tested endogenous and exogenous substances, and for technique specific substances and anticoagulants, respectively.

| Endogenous Substances | Concentration
Tested a | Results |
|---------------------------------------------|---------------------------|--------------------------|
| Albumin | 60.0 mg/mL | No interference observed |
| Bilirubin (Conjugated) | 0.41 mg/mL | No interference observed |
| Bilirubin (Unconjugated) | 0.41 mg/mL | No interference observed |
| Cholesterol (total) | 4.2 mg/mL | No interference observed |
| Glucose | 10.1 mg/mL | No interference observed |
| Hemoglobin | 137.0 mg/mL | No interference observed |
| Immunoglobulins | 60 mg/mL | No interference observed |
| Triglycerides | 15.1 mg/mL | No interference observed |
| Human cells
(K-562 Human Leukemia Cells) | 6.1E+06 cells/mL | No interference observed |
| Exogenous Substances | Concentration Tested a | Results |
| Artemether-Lumefantrine | 0.0004 mg/mL | No interference observed |
| Atovaquone | 0.005 mg/mL | No interference observed |
| Proguanil | 0.001 mg/mL | No interference observed |
| Mefloquine | 0.0017 mg/mL | No interference observed |
| Amphotericin B | 0.002 mg/mL | No interference observed |
| Pentamidine | 0.0015 mg/mL | No interference observed |
| Fluconazole | 0.026 mg/mL | No interference observed |
| Amoxicillin | 0.062 mg/mL | No interference observed |
| Azithromycin | 0.011mg/mL | No interference observed |
| Cetriazone | 1.0 mg/mL | No interference observed |
| Ciprofloxacin | 0.012 mg/mL | No interference observed |
| Clindamycin | 0.055 mg/mL | No interference observed |
| Doxycycline | 0.02 mg/mL | No interference observed |

Table 12. Results for Potentially Interfering Substances Tested on the FilmArray
Global Fever Panel (Endogenous and Exogenous Substances)

22

Gentamicin	0.036 mg/mL	No interference observed
Meropenem	0.39 mg/mL	No interference observed
Sulfamethoxazole	0.38 mg/mL	No interference observed
Vancomycin	0.12 mg/mL	No interference observed
Cycloserine	75.0 mg/mL	No interference observed
Isoniazid	0.06 mg/mL	No interference observed
Oseltamivir	0.0005 mg/mL	No interference observed
Ribavirin	0.011 mg/mL	No interference observed
Tenofovir	0.001 mg/mL	No interference observed
Acetaminophen	0.16 mg/mL	No interference observed
Aspirin	0.03 mg/mL	No interference observed
Ibuprofen	0.22 mg/mL	No interference observed
Prednisone	0.0001 mg/mL	No interference observed
Prednisolone	1.2 mg/mL	No interference observed
Cortisone	0.001 mg/mL	No interference observed
Artesunate	0.1 mg/mL	No interference observed

4Concentrations of interfering substances were based on guidelines contained in CLSI guidelines for interference testing (EP07 and EP37) when available.


	Global Fever Panel (Disinfectants, Solvents, and Anticoagulants)
Technique Specific Substances	Concentration Tested	Results
Bleach	1% (v/v) or 525 ppm NaOCl	No interference observed
Povidone-iodine	1% (v/v)	No interference observed
Ethanol	2% (v/v)	No interference observed
TRIzol	2-3% (v/v)	Potentially Interfering
DMSO1	2% (v/v)	No interference observed
Methanol1	2% (v/v)	No interference observed
Saline1	2% (v/v)	No interference observed
Chloroform1	2% (v/v)	No interference observed
Acetone1	2% (v/v)	No interference observed
Hydrochloric Acid1	0.0005 N	No interference observed
Anticoagulant	Concentration Tested	Results
Citrate (sodium)	~0.32 % (10.9 mmol/L)	No interference observed
K · EDTA in excess (5x) 2	~9 mg/mL	No interference observed
Na · Heparin	~19 USP/mL	Potentially Interfering
Acid-citrate-dextrose (ACD)	~2.2 g/L citrate
0.8 g/L citric acid
2.5 g/L dextrose	No interference observed
Sodium polyanethenole
sulfonate (SPS)	0.72 mg/mL	No interference observed
Serum separation tubes	N/A	No interference observed

1 Testing of these specific substances was performed to account for their use as solvents for resuspension of other potential interferants.

2 Testing was performed to evaluate potential interference caused by a short blood draw and a collection tube that thus contained higher anticoagulant concentration

23

dengue virus, Leptospira spp., and Plasmodium spp. vaccines were not evaluated in this study but are predicted to be reactive with the corresponding Global Fever Panel assay targets.

All evaluated exogenous substances exhibited no interference effect. Among technique specific substances, initial testing of TRIzol exhibited an interference effect wherein P. falciparum replicates were incorrectly reported as a Not Detected result. Subsequent retesting at the same interferent level and additionally with samples containing 1% TRIzol (v/v) did not produce unexpected assay results but may result in delaved Cp values. Similarly, initial testing with Heparin resulted in two unexpected Not Detected results for a single analyte with only Preplicates positive. However, repeat testing at the same heparin concentration successfully met acceptance criteria with replicates positive. These data demonstrate that Heparin levels near the evaluated concentration may be inhibitory for analytes near the LoD.

i. Assay cut-off:

The FilmArray Global Fever Panel Melt Detector software determines whether a FilmArray Global Fever Panel assay result is positive or negative using a predefined algorithm that includes Tm values, fluorescence values, and analysis of melting curves.

Initial melt ranges for each analyte-specific FilmArray Global Fever Panel assay were determined based on a combination of mathematical modeling using known sequence variations of different strains/isolates/variants of targeted organisms as well as data from testing of clinical specimens and known isolates.

After completion of the analytical and clinical studies on the FilmArray Global Fever Panel, a final validation of the melt ranges was performed and included review of data from the Inclusivity study and clinical studies. The observed sensitivity and specificity rates for the individual melt curves and assay calls as compared to expert annotation was greater than 99.8% and 99.9% respectively. The sensitivity, specificity, and accuracy for the validation data were determined to be well above the acceptance criteria.

j. Specimen Transport and Storage (Specimen Stability)

Table 22. Global Fever Panel Clinical Study Specimen Co-Detections

"Chikungunya virus was detected by Global Fever Panel but not by comparator testing in one of the two co-detections 6 Comparator testing did not identify Leptospira spp or Plasmodium spp. in this specimen.

b. Clinical specificity:
See section L.3a above.
c. Other clinical supportive data (when a. and b. are not applicable):
See section L.3a above.
1. Clinical cut-off:
  Not applicable.
1. Expected values/Reference range:
  The prevalence of individual analytes detected by the Global Fever Panel as observed during the clinical study is described in Table 23 below.

33

| | Overall
(N=1875) | | USA
Site 07
(N=179) | | USA
Site 14
(N=9) | | Africa
Site 01
(N=134) | | Africa
Site 02
(N=108) | | Africa
Site 05
(N=199) | | Africa
Site 11
(N=158) | | Southeast Asia
Site 08
(N=249) | | Southeast Asia
Site 09
(N=404) | | Central & S. America
Site 12
(N=297) | | Central & S. America
Site 13
(N=136) | |
|-----------------------------------------------|---------------------|-------|---------------------------|------|-------------------------|------|------------------------------|-------|------------------------------|-------|------------------------------|-------|------------------------------|-------|--------------------------------------|-------|--------------------------------------|-------|--------------------------------------------|-------|--------------------------------------------|-------|
| Analyte | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV | # | EV |
| Chikungunya virus | 27 | 1.4% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 27 | 6.7% | 0 | 0.0% | 0 | 0.0% |
| Dengue virus
(Serotypes 1, 2,
3, and 4) | 266 | 14.2% | 0 | 0.0% | 0 | 0.0% | 1 | 0.7% | 0 | 0.0% | 0 | 0.0% | 0 | 0.0% | 90 | 36.1% | 54 | 13.4% | 20 | 6.7% | 101 | 74.3% |
| Leptospira spp. | 19 | 1.0% | 1 | 0.6% | 0 | 0.0% | 0 | 0.0% | 1 | 0.9% | 0 | 0.0% | 0 | 0.0% | 4 | 1.6% | 4 | 1.0% | 9 | 3.0% | 0 | 0.0% |
| Plasmodium spp. | 351 | 18.7% | 0 | 0.0% | 0 | 0.0% | 16 | 11.9% | 50 | 46.3% | 141 | 70.9% | 49 | 31.0% | 7 | 2.8% | 4 | 1.0% | 84 | 28.3% | 0 | 0.0% |
| P. falciparum | 233 | 12.4% | 0 | 0.0% | 0 | 0.0% | 14 | 10.4% | 44 | 40.7% | 125 | 62.8% | 42 | 26.6% | 3 | 1.2% | 0 | 0.0% | 5 | 1.7% | 0 | 0.0% |
| P. vivax/ovale | 115 | 6.1% | 0 | 0.0% | 0 | 0.0% | 3 | 2.2% | 0 | 0.0% | 12 | 6.0% | 12 | 7.6% | 4 | 1.6% | 4 | 1.0% | 80 | 26.9% | 0 | 0.0% |

Table 23. Prevalence of Detected Analytes Stratified by Region and Clinical Study Site

34

The Global Fever Panel detected at least one analyte in 657 of the 1875 included specimens (35.1% positivity rate).

| FilmArray Result | Number Detected | % of Total
(% of Positives) |
|--------------------------------|-----------------|--------------------------------|
| Detected (at least one result) | 657 | 35.0%
(100%) |
| One analyte result | 629 | 33.5%
(95.7%) |
| Two analyte resultsa | 28 | 1.5%
(4.3%) |

Table 24. Expected Values Summary (Detections) as Determined in the Global Fever Panel Clinical Study

"Data for discernible co-detections only. The Plasmodium spp. assay is not considered a unique Global Fever Panel detection when co-detected with a Plasmodium species-level assay. Specimens containing multiple species within a genus are not always discemble (e.g., a specimen containing P. malariae and P. falciparum would not produce a discernible co-detection.)

	Table 25. Expected Values Summary (Analytes) as Determined in the Global Fever
Panel Clinical Study
FilmArray Result	Number Detected	% of Positives
Chikungunya virus	27	4.1%
Dengue virus (serotypes 1,
2, 3, and 4)	266	40.5%
Leptospira spp.	19	2.9%
Plasmodium spp.	351	53.4%
Plasmodium falciparum	233	35.5%
Plasmodium vivax/ovale	115	17.5%

The most prevalent analyte result was Plasmodium spp. (351/657; 53.4%), of which, 233/351 (66.4%) also had Plasmodium falciparum identified, 115/351 (32.8%) also had Plasmodium vivax/ovale identified, 25/351 (7.1%) had no species-level identification, and 22/351 (6.3%) had a combination of P. falciparum and P. vivax/ovale. The second most prevalent analyte was dengue virus (266/657, 40.5%). Chikungunya virus was detected in 4.1% (27/657) of specimens and Leptospira spp. was detected in 2.9% (19/657).

35

M. Instrument Name:

Not applicable. The device does not utilize an instrument for result generation.

N. System Descriptions:

1. Modes of Operation:

Does the applicant's device contain the ability to transmit data to a computer, webserver, or mobile device?

Yes __________________________________________________________________________________________________________________________________________________________________________

Does the applicant's device transmit data to a computer, webserver, or mobile device using wireless transmission?

Yes X - or No - - - -

1. Software:
  FDA has reviewed applicant's Hazard Analysis and software development processes for this line of product types:

Yes X or No or No

The device does not contain any software or instrument components.

3. Specimen Identification:

The Sample ID can be entered manually or scanned in by using the FilmArray barcode scanner

1. Specimen Sampling and Handling:
  Not applicable.
1. Calibration:
  Not applicable.
1. Quality Control:
  See Quality Control Section above (L.1.c "Traceability, Stability, Expected Values (controls, calibrators, or methods)")

36

O. Other Supportive Instrument Performance Characteristics Data Not Covered in the "Performance Characteristics" Section above:

None.

P. Proposed Labeling:

The labeling supports the decision to grant the De Novo request for this device.

Q. Identified Risks to Health and Identified Mitigations

Identified Risks to Health	Mitigation Measures
Risk of an inaccurate test result (false
positive or false negative result) leading
to improper patient management	Certain labeling information, including certain
limiting statements and performance
information.
Certain design verification and validation,
including certain analytical studies and clinical
studies.
Use of certain specimen collection devices.
Misinterpretation of test results leading to
misdiagnosis and associated risk of false
test results	Certain labeling information, including certain
limiting statements and performance
information.
Certain design verification and validation,
including certain analytical studies and clinical
studies.
Failure to correctly operate the device
leading to inaccurate test results	Certain labeling information, including certain
limiting statements and performance
information.
Certain design verification and validation,
including certain analytical studies and clinical
studies.
Use of certain specimen collection devices.

R. Benefit/Risk Analysis:

Patient Perspectives

This submission did not include specific information on patient perspectives for this device.

Summary of the Assessment of Benefit

Fever is a host response to infection that is not pathognomonic of a specific disease. The benefit of the assay is aiding the accurate diagnosis of infections that cause acute febrile illness in the specific population of patients who have been potentially exposed to pathogens detected by the Global Fever Panel. When guided by appropriate risk factors and epidemiological information, the Global Fever Panel can be helpful to identify

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the specific cause of fever and initiate appropriate treatment, including, but not limited to. antimicrobial therapy. Earlier identification of fever causing pathogens and an appropriate course of treatment may improve patient outcomes. Additionally, the Global Fever Panel fills an unmet need for in vitro diagnostics as no currently approved or cleared tests exist for many of the pathogens on the Global Fever Panel.

While the performance of the device in the clinical and analytical studies suggests that patients will benefit from the assay, expected and acceptable sources of uncertainty are the wide confidence intervals around point estimates of performance. The special controls, including the interpretation of results and the limiting statements in device labeling will help to ensure that errors will be uncommon and will facilitate accurate assay implementation and interpretation of results.

Summary of the Assessment of Risk

The risks associated with the device, when used as intended, are those related to the risk of inaccurate test results, the failure to correctly interpret test results, and failure to correctly operate the device.

The risk of a false positive test result is improper patient management, including potentially inappropriate administration of unnecessary antibiotics or anti-malarial medications. Inappropriate administration of prolonged courses of antibiotics is associated with toxicity, allergic reactions, and other adverse outcomes, including secondary infections such as C. difficile colitis. Inappropriate administration of antimalarial medications may also have adverse effects that depend upon the drug administered. No specific treatments exist for dengue and chikungunya virus infections, although a vaccine was recently developed for dengue virus. This vaccine is only recommended in individuals who had previously had confirmed dengue infection and individuals who have not been infected are at increased risk of severe dengue if they are infected after being vaccinated. In the broader population, individual false positive results could lead to increased burden on the CDC to perform confirmatory testing since all targets detected by the Global Fever Panel are nationally notifiable.

During the analytical evaluation of exclusivity, some pathogen cross-reactivity was identified that could pose risks of patient harm. Specifically, P. knowlesi and P. malariae may cross-react with the P. vivax/ovale assay on the Global Fever Panel. P. vivax/ovale infections require a specific course of treatment to clear hypnozoite liver stage of infection and prevent recrudescence. A false positive P. vivax/ovale result could therefore lead to patients receiving unnecessary and potentially toxic treatment.

The risk of a false negative test result is delayed identification of the cause of the disease in the patient, which could lead to improper patient management, including administration of unnecessary treatment and/or discontinuation of appropriate treatment. An undiagnosed infection or delaved diagnosis, particularly with P, falciparum or Leptospira can result in increased morbidity and mortality.

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Failure to correctly operate the device can lead to false test results. Failure to correctly interpret test results can lead to treatment of a clinically positive patient in the same manner as a false negative test result and a clinically negative patient in the same manner as a false positive test result, with the corresponding implications discussed above.

Summary of the Assessment of Benefit-Risk

General controls are insufficient to mitigate the risks associated with the device. However, the probable clinical benefits outweigh the probable risks for the proposed assay, considering the mitigations of the risks provided for in the listed special controls established for this device, as well as general controls. The required special controls will help ensure that errors will be uncommon and will facilitate accurate assay implementation and interpretation of results.

The risk of inaccurate test results (both positive and negative) is mitigated by the intended use clearly stating that the assay results are intended to be used with other clinical, epidemiologic, and laboratory data. The risk of false results is also mitigated by the inclusion of performance characteristics from analytical and clinical studies in the labeling. Risks from cross-reactivity are mitigated by appropriate limitations in the labeling that indicate potential cross-reactivity of these malaria strains and further state that a P. vivax/ovale result should be further confirmed. Additionally, a separate limitation states that all Plasmodium spp. detected results from patients exposed in Southeast Asia should be further investigated for possible P. knowlesi infection.

Risks of failure to correctly interpret the test results are mitigated through the inclusion in the labeling of a detailed description of what the device detects, the specimen type for which testing is indicated, the type of results provided to the user in the intended use statement, as well as a detailed explanation of the interpretation of results. Finally, the risk of failure to correctly operate the device is mitigated by the inclusion of detailed directions for use in the package insert, such that the operator can successfully use the instrument.

The clinical performance observed in the clinical trial suggests that errors will be uncommon and that the assay will provide substantial benefits to patients in the diagnosis of acute febrile illness and when used in conjunction with other clinical and diagnostic findings.

Given the combination of the device's indications for use, labeling, the required general controls, and the special controls established for this device, the probable benefits would outweigh the probable risks.

S. Conclusion

The De Novo request is granted and the device is classified under the following and subject to the special controls identified in the letter granting the De Novo request:

Product Code: OMV

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Device Type: Device to detect and identify selected microbial agents that cause acute febrile illness
Class: II Regulation: 21 CFR 866.3966