Who is the manufacturer of FilmArray Meningitis/Encephalitis(ME) Panel?

Biofire Diagnostics, LLC filed the De Novo classification request for FilmArray Meningitis/Encephalitis(ME) Panel (DEN150013).

What is the FDA product code for FilmArray Meningitis/Encephalitis(ME) Panel?

PLO. It is classified under 21 CFR 866.3970 as a Class 2 device in the Microbiology panel.

What is the regulatory pathway for FilmArray Meningitis/Encephalitis(ME) Panel?

De Novo classification (submission type: Direct).

Who can help prepare an FDA De Novo submission like FilmArray Meningitis/Encephalitis(ME) Panel?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA De Novo submissions. See https://innolitics.com/services/regulatory/ or contact https://innolitics.com/contact.

FilmArray Meningitis/Encephalitis(ME) Panel

DEN150013 · Biofire Diagnostics, LLC · PLO · Oct 8, 2015 · Microbiology

Device Facts

Record ID	DEN150013
Device Name	FilmArray Meningitis/Encephalitis(ME) Panel
Applicant	Biofire Diagnostics, LLC
Product Code	PLO · Microbiology
Decision Date	Oct 8, 2015
Decision	DENG
Submission Type	Direct
Regulation	21 CFR 866.3970
Device Class	Class 2

Intended Use

The FilmArray Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray and FilmArray 2.0 systems. The FilmArray ME Panel is capable of simultaneous detection and identification of multiple bacterial, viral, and yeast nucleic acids directly from cerebrospinal fluid (CSF) specimens obtained via lumbar puncture from individuals with signs and/or symptoms of meningitis and/or encephalitis. The following organisms are identified using the FilmArray ME Panel: Bacteria: Escherichia coli K1, Haemophilus influenzae, Listeria monocytogenes, Neisseria meningitidis (encapsulated), Streptococcus agalactiae, Streptococcus pneumoniae. Viruses: Cytomegalovirus, Enterovirus, Herpes simplex virus 1, Herpes simplex virus 2, Human herpesvirus 6, Human parechovirus, Varicella zoster virus. Yeast: Cryptococcus neoformans/gattii. The FilmArray ME Panel is indicated as an aid in the diagnosis of specific agents of meningitis and/or encephalitis and results are meant to be used in conjunction with other clinical, epidemiological, and laboratory data. Results from the FilmArray ME Panel are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out coinfection with organisms not included in the FilmArray ME Panel. The agent detected may not be the definite cause of the disease. Negative results do not preclude central nervous system (CNS) infection. Not all agents of CNS infection are detected by this test and sensitivity in clinical use may differ from that described in the package insert. The FilmArray ME Panel is not intended for testing of specimens collected from indwelling CNS medical devices. The FilmArray ME Panel is intended to be used in conjunction with standard of care culture for organism recovery, serotyping, and antimicrobial susceptibility testing.

Device Story

Multiplexed nucleic acid-based in vitro diagnostic test; uses FilmArray and FilmArray 2.0 systems. Input: CSF specimens via lumbar puncture. Process: automated nucleic acid extraction, amplification, and detection of specific bacterial, viral, and yeast pathogens. Output: qualitative identification of target organisms. Used in clinical laboratory settings; operated by trained laboratory personnel. Results aid clinicians in diagnosis alongside standard-of-care culture, clinical, and epidemiological data. Benefits: rapid identification of CNS pathogens to support timely clinical decision-making.

Clinical Evidence

Prospective multi-center clinical study (n=1560) compared FilmArray ME Panel to culture (bacteria) and PCR/sequencing (viruses/yeast). Sensitivity/PPA and specificity/NPA varied by analyte. Overall, the device demonstrated high specificity (>99%) across most analytes. False positives for S. pneumoniae were noted. Performance was supplemented by archived specimen testing (n=235) and contrived specimen testing to address low-prevalence analytes.

Technological Characteristics

Multiplexed nucleic acid-based assay; utilizes FilmArray/FilmArray 2.0 systems. Methodology involves nucleic acid extraction, amplification, and detection. Requires standard-of-care culture for confirmation. Designed for use with CSF specimens. Subject to 21 CFR 820 design controls.

Indications for Use

Indicated for patients with signs/symptoms of meningitis or encephalitis. Used for qualitative detection/identification of 14 bacterial, viral, and yeast pathogens in CSF specimens obtained via lumbar puncture. Prescription use only.

Regulatory Classification

Identification

A device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid is a qualitative in vitro device intended for the detection and identification of microbial-associated nucleic acid sequences from patients suspected of meningitis or encephalitis. A device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid is intended to aid in the diagnosis of meningitis or encephalitis when used in conjunction with clinical signs and symptoms and other clinical and laboratory findings.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed device description documentation, including the device components, ancillary reagents required but not provided, and a detailed explanation of the methodology, including primer/probe sequence, design, and rationale for sequence selection. (2) Premarket notification submissions must include detailed documentation from the following analytical studies: Analytical sensitivity (limit of detection), inclusivity, reproducibility, interference, cross reactivity, and specimen stability. (3) Premarket notification submissions must include detailed documentation from a clinical study. The study, performed on a study population consistent with the intended use population, must compare the device performance to results obtained from well-accepted comparator methods. (4) Premarket notification submissions must include detailed documentation for device software, including, but not limited to, software applications and hardware-based devices that incorporate software. (5) The Intended Use statement in the device labeling must include a statement that the device is intended to be used in conjunction with standard of care culture. (6) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling. (7) The device labeling must include a limitation stating that the negative results do not preclude the possibility of central nervous system infection. (8) The device labeling must include a limitation stating that device results are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. (9) The device labeling must include a limitation stating that positive results do not mean that the organism detected is infectious or is the causative agent for clinical symptoms. (10) As part of the risk management activities performed under 21 CFR 820.10(c) design and development, you must document an appropriate end user device training program that will be offered as part of your efforts to mitigate the risk of failure to correctly operate the instrument.

Related Devices

K160462 — FilmArray Meningitis/Encephalitis (ME) Panel for use with FilmArray Torch · Biofire Diagnostics, LLC · Mar 17, 2016
K242256 — QIAstat-Dx Meningitis/Encephalitis (ME) Panel · QIAGEN GmbH · Oct 29, 2024
K143080 — FilmArray Respiratory Panel (RP) for use with Multi-instrument FilmArray System · Biofire Diagnostics, LLC · Feb 17, 2015

Submission Summary (Full Text)

{0}------------------------------------------------ ## EVALUATION OF AUTOMATIC CLASS III DESIGNATION FOR FilmArray® Meningitis/Encephalitis (ME) Panel ## DECISION SUMMARY ## A. DEN Number: DEN150013 ## B. Purpose for Submission: De Novo request for evaluation of automatic class III designation for the FilmArray Meningitis/Encephalitis (ME) Panel ## C. Measurands: The assay detects and identifies nucleic acids of the following organisms: Escherichia coli K1, Haemophilus influenzae, Listeria monocytogenes, Neisseria meningitidis (encapsulated), Streptococcus agalactiae, Streptococcus pneumoniae, Cytomegalovirus, Enterovirus, Herpes simplex virus 1, Herpes simplex virus 2, Human herpesvirus 6, Human parechovirus, Varicella zoster virus, Cryptococcus neoformans/gattii ## D. Type of Test: The FilmArray Meningitis/Encephalitis (ME) Panel ("FilmArray ME Panel"), performed with FilmArray and FilmArray 2.0 systems, is a nucleic acid-based test for the detection of the above listed bacteria, viruses, and yeast from cerebrospinal fluid (CSF) specimens obtained from patients with signs and symptoms of meningitis or encephalitis. ## E. Applicant: BioFire Diagnostics, LLC ## F. Proprietary and Established Names: FilmArray Meningitis/Encephalitis (ME) Panel ## G. Regulatory Information: ## 1. Regulation section: 21 CFR 866.3970, Device to detect and identify microbial pathogen nucleic acids in cerebrospinal fluid ## 2. Classification: Class II (Special Controls) {1}------------------------------------------------ - 3. Product code(s): PLO, OOI, NSU - 4. Panel: 83- Microbiology ## H. Intended Use: - 1. Intended use(s): The FilmArray Meningitis/Encephalitis (ME) Panel is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with FilmArray and FilmArray 2.0 systems. The FilmArray ME Panel is capable of simultaneous detection and identification of multiple bacterial, viral, and yeast nucleic acids directly from cerebrospinal fluid (CSF) specimens obtained via lumbar puncture from individuals with signs and/or symptoms of meningitis and/or encephalitis. The following organisms are identified using the FilmArray ME Panel: ### Bacteria: Escherichia coli K1 Haemophilus influenzae Listeria monocytogenes Neisseria meningitidis (encapsulated) Streptococcus agalactiae Streptococcus pneumoniae - Viruses: Cytomegalovirus Enterovirus Herpes simplex virus 1 Herpes simplex virus 2 Human herpesvirus 6 Human parechovirus Varicella zoster virus #### Yeast: #### Cryptococcus neoformans/gattii The FilmArray ME Panel is indicated as an aid in the diagnosis of specific agents of meningitis and/or encephalitis and results are meant to be used in conjunction with other clinical, epidemiological, and laboratory data. Results from the FilmArray ME Panel are not intended to be used as the sole basis for diagnosis, treatment, or other patient management decisions. Positive results do not rule out co-infection with organisms not included in the FilmArray ME Panel. The agent {2}------------------------------------------------ detected may not be the definite cause of the disease. Negative results do not preclude central nervous system (CNS) infection. Not all agents of CNS infection are detected by this test and sensitivity in clinical use may differ from that described in the package insert. The FilmArray ME Panel is not intended for testing of specimens collected from indwelling CNS medical devices. The FilmArray ME Panel is intended to be used in conjunction with standard of care culture for organism recovery, serotyping, and antimicrobial susceptibility testing. - 2. Indication(s) for use: Same as Intended Use - 3. Special conditions for use statement(s): For prescription use only. - 4. Special instrument requirements: The FilmArray ME Panel is performed on the FilmArray and FilmArray 2.0 systems. # I. Device Description: The FilmArray ME Panel is a multiplex nucleic acid-based test designed to be used with FilmArray or FilmArray 2.0 system ("FilmArray systems" or "FilmArray instruments"). The FilmArray ME panel includes a FilmArray ME Panel pouch which contains freeze-dried reagents to perform nucleic acid purification and nested, multiplex polymerase chain reaction (PCR) with DNA melt analysis. The FilmArray ME Panel simultaneously conducts 14 tests for the identification of potential CNS pathogens from CSF specimens obtained via lumbar puncture. Results from the FilmArray ME Panel are available within about one hour. A test is initiated by loading Hydration Solution into one port of the pouch and a CSF sample mixed with the provided Sample Buffer ampoules into the other port of the pouch and placing it in the FilmArray Instrument. The pouch contains all of 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 FilmArray Software on the FilmArray systems guides the user though the steps of placing the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run on the FilmArray systems. The 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 channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. b(4) {3}------------------------------------------------ Nucleic acid extraction occurs within the 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. b(4) . 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 21d 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. ## Materials provided in each FilmArray ME Panel kit: - Individually packaged FilmArray ME Panel pouches - Single-use (1.0 mL) Sample Buffer ampoules - Single-use pre-filled (1.5 mL) Hydration Injection Vials ● - Single-use Sample Injection Vials ● - Individually packaged Transfer Pipettes Materials required but not provided: FilmArray system including: - FilmArray or FilmArray 2.0 instrument and software - FilmArray Pouch Loading Station # J. Standard/Guidance Document Referenced (if applicable): - CLSI EP12-A2, User Protocol for Evaluation of Qualitative Test Performance, 2008 - CLSI EP07-A2, Interference Testing in Clinical Chemistry, 2005 ● - CLSI MM03-A2, Molecular Diagnostic Methods for Infectious Diseases, 2006 ● - EN ISO 14971:2012, 'Medical devices – Application of risk management to medical devices' - Guidance for Sponsors, Institutional Review Boards, Clinical Investigators and FDA ● Staff - Guidance on Informed Consent for In Vitro Diagnostic Device Studies Using Leftover Human Specimens that are Not Individually Identifiable. April 25, 2005 - Guidance for Industry and Food and Drug Administration Staff Assay Migration ● Studies for In Vitro Diagnostic Devices, April 25, 2013 - Guidance for Industry and Food and Drug Administration Staff Highly Multiplexed ● {4}------------------------------------------------ Microbiological/Medical Countermeasure In Vitro Nucleic Acid Based Diagnostic Devices, August 27, 2014 - Statistical Guidance on Reporting Results from Studies Evaluating Diagnostic Tests, ● FDA Guidance Document, March 13, 2007 - Class II Special Controls Guidance Document: Nucleic Acid Amplification Assay for the ● Detection of Enterovirus RNA, January 2, 2009 # K. Test Principle: The FilmArray ME Panel pouch) is a closed system disposable that houses all the chemistry required to isolate, amplify and detect nucleic acid from multiple meningitis and encephalitis pathogens within a single CSF specimen obtained from a lumbar puncture. 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 ME Panel loads the sample into the pouch, places the pouch into the FilmArray systems, and starts the run. All other operations are automated. Operations and processes that occur during a FilmArray run include the following: - . Nucleic Acid Purification - b(4) 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 approximately ten minutes and the bead-beater apparatus can be heard as a high-pitched whine during the first minute of operation. - Reverse Transcription and 1st Stage Multiplex PCR Some pathogens identified by . the pouch are RNA viruses, and a reverse transcription (RT) step is performed to convert the viral RNA into cDNA prior to amplification. b(4) for multiplex PCR. The effect of 1st 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 "" 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 1st 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. {5}------------------------------------------------ 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. ## L. Performance Characteristics: ## 1. Analytical performance: ## a. Reproducibility Reproducibility studies were performed with the FilmArray ME Panel on both the FilmArray and FilmArray 2.0 systems. Testing for the FilmArray system was performed using multiple instruments at three different testing sites, Biofire Diagnostics and two external laboratories. Testing for the FilmArray 2.0 system was performed internally at BioFire Diagnostics using multiple instruments at three different locations within BioFire Diagnostics. Assay reproducibility was evaluated for both the FilmArray and FilmArray 2.0 system using a panel of contrived CSF samples prepared in artificial CSF matrix (aCSF) and spiked with combinations of nine different ME Panel analytes, including at least one representative gram-negative bacterium, gram-positive bacterium, yeast, DNA virus and RNA virus. Each spiked analyte was evaluated at three different concentrations: Negative (no analyte), Low Positive (1× the limit of detection (LoD)) and Moderate Positive (3× LoD). Testing on both FilmArray systems incorporated a range of potential testing variables including different operators, three different pouch lots, and different FilmArray Instruments. Samples were tested on five different days with a total of 90 replicates tested per panel member. For the FilmArray system. 366 runs were initiated and 360 runs were completed (98.4% initially valid results). Of the six initially invalid runs, one invalid run was due to a control failure and five invalid tests were due to instrument or software errors. Retesting of initially invalid specimens gave valid results for all six specimens. There were seven unexpected false positive results observed in the study: six for Streptococcus pneumoniae (6/360 = 1.7%) and one for Human herpesvirus 6 (HHV-6) (1/360 = 0.3%). For the FilmArray 2.0 system. 365 runs were initiated and 360 runs were completed (98.6% initially valid results). Of the five initially invalid runs, three runs were invalid because they were aborted by the operator due to being run on an incorrect instrument. The two other invalid results were due to a software error (1/365 = 0.3%) and due to an incomplete result caused by a data transfer error (1/365 = 0.3%). A summary of qualitative results from both reproducibility studies (percent agreement with the expected result) for each analyte and organism concentration is provided in the following tables. {6}------------------------------------------------ ## Reproducibility of the FilmArray ME Panel on FilmArray | Organism/Isolate Tested | Concentration | Expected Test Result | FilmArray Agreement with Expected Results | | | All Sites (95% Confidence Interval) | | |----------------------------|---------------------------------------------------------------------|-----------------------------------------------------|-------------------------------------------|-------------------------------------------|------------------|----------------------------------------------|----------------------------------| | | | | Site A | Site B | Site C | | | | | BACTERIA | | | | | | | | E. coli K1 b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | E. coli K1 b(4) | Low Positive 1× LoD 1×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 96.7% (96%-100%) | | | E. coli K1 b(4) | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | | H. influenzae b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | H. influenzae b(4) | Low Positive 1× LoD 1×103 CFU/mL | Detected | 29/30 96.7% | 30/30 100% | 30/30 100% | 89/90 98.9% (94.0%-100%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | | L. monocytogenes b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 29/30 96.7% | 30/30 100% | 30/30 100% | 89/90 98.9% (94.0%-100%) | | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | | N. meningitidis | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | | S. agalactiae b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 29/30 96.7% | 30/30 100% | 27/30 90.0% | 86/90 95.6% (89.0%-98.8%) | | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 26/30 86.7% | 30/30 100% | 27/30 90.0% | 83/90 92.2% (84.6%-96.8%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | | S. pneumoniae | Negative (No analyte) | Not Detected | 118/120 98.3% | 118/120 98.3% | 118/120 98.3% | 354/360a 98.3% (96.4%-99.4%) | | | VIRUSES/YEAST | | | | | | | | | CMV | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | | | Concentration | Expected Test Result | FilmArray Agreement with Expected Results | | | All Sites (95% Confidence Interval) | | | Organism/Isolate Tested | | | Site A | Site B | Site C | | | | EV | Moderate Positive $3\times LoD$ 15 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Coxsackievirus A9 b(4) | Low Positive $1\times LoD$ 5 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 28/30 93.3% | 88/90 97.8% (92.2%-99.7%) | | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | | HSV-1 | | | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | | | Moderate Positive $3\times LoD$ 150 TCID50/mL | | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | HSV-2 b(4) | Low Positive $1\times LoD$ 50 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | HHV-6 | | Negative (No analyte) | Not Detected | 120/120 100% | 119/120 99.2% | 120/120 100% | 359/360 99.7% (98.5%-100%) | | | Moderate Positive $3\times LoD$ $1.5\times 10^3$ TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | HPeV b(4) | Low Positive $1\times LoD$ 500 TCID50/mL | Detected | 29/30 96.7% | 30/30 100% | 30/30 100% | 89/90 98.9% (94.0%-100%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | VZV b(4) | | Moderate Positive $3\times LoD$ 0.3 TCID50/mL | Detected | 29/30 96.7% | 30/30 100% | 30/30 100% | 89/90 98.9% (94.0%-100%) | | | Low Positive $1\times LoD$ 0.1 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | | C. gattii b(4) | Moderate Positive $3\times 10^3$ CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Low Positive $1\times 10^3$ CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | | Organism/Isolate Tested | Concentration | Expected Test Result | FilmArray Agreement with Expected Results | | | | | Site A | | | | Site B | Site C | All Sites (95% Confidence Interval) | | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | {7}------------------------------------------------ {8}------------------------------------------------ 4Six false positive S. pneumoniae results were reported. The unexpected results were observed at all three test sites, in different samples, on different days, and with different pouch lots. The overall incidence of false S. pneumoniae results observed in all reproducibility testing was <1%. * C. gattii was tested at concentrations equivalent to 10× and 30× LoD for the combined Cryptococcus neoformans/gattii test result on the FilmArray. #### Reproducibility of the FilmArray ME Panel on FilmArray 2.0 | Organism/Isolate Tested | Concentration | Expected Test Result | FilmArray 2.0 Agreement with Expected Results | | | All Systems (95% Confidence Interval) | |----------------------------|-----------------------------------------------------|-------------------------|-----------------------------------------------|-----------------|-----------------|---------------------------------------------| | | | | System A | System B | System C | | | BACTERIA | | | | | | | | E. coli K1 b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 29/30 96.7% | 29/30 96.7% | 29/30 96.7% | 87/90 96.7% (90.6%-99.3%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | H. influenzae b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | L. monocytogenes b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | N. meningitidis | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | S. agalactiae b(4) | Moderate Positive 3× LoD 3×103 CFU/mL | Detected | 29/30 96.7% | 30/30 100% | 29/30 96.7% | 88/90 97.8% (92.2%-99.7%) | | Organism/Isolate | Concentration | Expected Test Result | FilmArray 2.0 Agreement with Expected Results | | | | | Tested | | | System A | System B | System C | All Systems (95% Confidence Interval) | | | Low Positive 1× LoD 1×103 CFU/mL | Detected | 29/30 96.7% | 29/30 96.7% | 30/30 100% | 88/90 97.8% (92.2%-99.7%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | S. pneumoniae | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | | | | VIRUSES/YEAST | | | | | CMV | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | EV | Moderate Positive 3× LoD 15 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | Coxsackievirus A9 b(4) | Low Positive 1× LoD 5 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | HSV-I | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | | Moderate Positive 3× LoD 150 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | HSV-2 b(4) | Low Positive 1× LoD 50 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 29/30 96.7% | 89/90 98.9% (94.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | HHV-6 | Negative (No analyte) | Not Detected | 120/120 100% | 120/120 100% | 120/120 100% | 360/360 100% (99.0%-100%) | | | Moderate Positive 3× LoD 1.5×103 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | HPeV b(4) | Low Positive 1× LoD 500 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | VZV b(4) | Moderate Positive 3× LoD 0.3 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 29/30 96.7% | 89/90 98.9% (94.0%-100%) | | Organism/Isolate Tested | Concentration | Expected Test Result | FilmArray 2.0 Agreement with Expected Results | | | | | | | | System A | System B | System C | All Systems (95% Confidence Interval) | | | Low Positive 1× LoD 0.1 TCID50/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100% | 180/180 100% (98.0%-100%) | | C. gattii b(4) | Moderate Positive 3× LoD 300 CFU/mL | Detected | 30/30 100% | 30/30 100% | 30/30 100% | 90/90 100% (96.0%-100%) | | | Low Positive 1× LoD 100 CFU/mL | Detected | 30/30 100% | 30/30 100% | 28/30 93.3% | 88/90 97.8% (92.2%-99.7%) | | | Negative (No analyte) | Not Detected | 60/60 100% | 60/60 100% | 60/60 100 % | 180/180 100% (98.0%-100%) | {9}------------------------------------------------ {10}------------------------------------------------ The following table includes analyses of assay reproducibility by analyte and panel member based on the Tm for each positive result observed during both the FilmArray system and FilmArray 2.0 system Reproducibility studies. Results demonstrated similar performance for both instruments with Tm standard deviations of ≤ 0.5℃ for all analytes and concentrations evaluated. Reproducibility of Tm values for Positive FilmArray ME Assays on FilmArray and FilmArray 2.0 systems | | Assay | Concentration Tested (× LoD) | Mean Tm (°C) (±StDev) | | | | | | | | |------------------------------------------|----------------|---------------------------------|-----------------------|--------|--------|---------------|----------|----------|----------|-------------| | Test Result (Organism/Isolate Tested) | | | FilmArray | | | FilmArray 2.0 | | | | | | | | | Site A | Site B | Site C | All Sites | System A | System B | System C | All Systems | | | yeastRNA | | 82.4 | 82.5 | 82.0 | 82.3 | 81.7 | 81.9 | 81.7 | 81.8 | | RNA Process Control | | | ±0.2 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | PCR2 | | 76.5 | 76.6 | 76.1 | 76.4 | 75.5 | 75.7 | 75.5 | 75.6 | | PCR2 Control | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | BACTERIA | | | | | | | | | | | | | Ecoli 3 | 3× LoD | 82.9 | 82.8 | 82.6 | 82.8 | 82.1 | 82.3 | 82.1 | 82.2 | | E. coli K1 b(4) | | | ±0.1 | ±0.3 | ±0.1 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | | 1× LoD | 83.0 | 83.1 | 82.6 | 82.9 | 82.2 | 82.4 | 82.3 | 82.3 | | | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | Hinfluenzae 1 | 3× LoD | 78.6 | 78.6 | 78.2 | 78.5 | 77.9 | 77.9 | 77.7 | 77.8 | | | | | ±0.1 | ±0.4 | ±0.2 | ±0.4 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | | 1× LoD | 78.6 | 78.7 | 78.3 | 78.5 | 78.0 | 78.0 | 77.9 | 78.0 | | H. influenzae b(4) b(4) | | | ±0.2 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | Hinfluenzae 2 | 3× LoD | 82.0 | 81.9 | 81.6 | 81.8 | 81.0 | 81.1 | 81.0 | 81.0 | | | | | ±0.1 | ±0.4 | ±0.2 | ±0.3 | ±0.2 | ±0.2 | ±0.2 | ±0.2 | | | | 1× LoD | 82.0 | 82.0 | 81.7 | 81.9 | 81.2 | 81.3 | 81.2 | 81.2 | | | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.2 | ±0.2 | ±0.3 | ±0.2 | | | Lmonocytogenes | 3× LoD | 80.4 | 80.6 | 80.2 | 80.4 | 80.0 | 80.1 | 80.0 | 80.0 | | L. monocytogenes (b(4) b(4) | | | ±0.2 | ±0.3 | ±0.2 | ±0.3 | ±0.4 | ±0.3 | ±0.3 | ±0.3 | | | | 1× LoD | 80.5 | 80.6 | 80.1 | 80.4 | 80.0 | 80.1 | 79.9 | 80.0 | | | | | ±0.2 | ±0.4 | ±0.2 | ±0.4 | ±0.4 | ±0.3 | ±0.2 | ±0.3 | | | Sagalactiae | 3× LoD | 82.0 | 82.0 | 81.6 | 81.9 | 81.1 | 81.3 | 81.2 | 81.2 | | S. agalactiae b(4) b(4) | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | | 1× LoD | 82.1 | 82.2 | 81.7 | 82.0 | 81.2 | 81.4 | 81.2 | 81.3 | {11}------------------------------------------------ | | Assay | Concentration Tested (× LoD) | Mean Tm (°C) (±StDev) | | | | | | | | |----------------------------------------------|--------------|---------------------------------|-----------------------|--------|--------|---------------|----------|----------|----------|-------------| | Test Result (Organism/Isolate Tested) | | | FilmArray | | | FilmArray 2.0 | | | | | | | | | Site A | Site B | Site C | All Sites | System A | System B | System C | All Systems | | | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.2 | ±0.2 | ±0.3 | | VIRUSES | | | | | | | | | | | | EV (Coxsackievirus A9) b(4) | EV2 | 3× LoD | 89.7 | 89.7 | 89.3 | 89.6 | 89.1 | 89.3 | 89.2 | 89.2 | | | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | | 1× LoD | 89.7 | 89.7 | 89.3 | 89.6 | 89.1 | 89.3 | 89.2 | 89.2 | | b(4) | | | ±0.1 | ±0.4 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | | 3× LoD | 75.6 | 75.5 | 75.1 | 75.4 | 74.6 | 74.8 | 74.6 | 74.7 | | | | | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | HSV2 1 | 1× LoD | 75.9 | 76.0 | 75.4 | 75.7 | 74.9 | 75.1 | 74.9 | 75.0 | | HSV-2 | | | ±0.2 | ±0.2 | ±0.3 | ±0.4 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | b(4) | HSV2 2 | 3× LoD | 88.8 | 88.9 | 88.4 | 88.7 | 88.1 | 88.3 | 88.2 | 88.2 | | | | | ±0.2 | ±0.4 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | | 1× LoD | 88.9 | 89.0 | 88.5 | 88.8 | 88.2 | 88.5 | 88.3 | 88.3 | | | | | ±0.2 | ±0.3 | ±0.2 | ±0.4 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | HPeV b(4) | HPeV | 3× LoD | 82.8 | 82.8 | 82.5 | 82.7 | 82.2 | 82.3 | 82.1 | 82.2 | | | | | ±0.2 | ±0.4 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | | 1× LoD | 82.8 | 82.9 | 82.5 | 82.7 | 82.3 | 82.3 | 82.2 | 82.3 | | | | | ±0.2 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | | | VZV 1 | 3× LoD | 88.9 | 89.0 | 88.5 | 88.8 | 88.5 | 88.5 | 88.4 | 88.5 | | | | | ±0.2 | ±0.4 | ±0.2 | ±0.4 | ±0.4 | ±0.2 | ±0.2 | ±0.3 | | | | 1× LoD | 89.0 | 89.0 | 88.5 | 88.8 | 88.5 | 88.5 | 88.3 | 88.4 | | VZV b(4) | | | ±0.2 | ±0.5 | ±0.2 | ±0.4 | ±0.3 | ±0.3 | ±0.2 | ±0.3 | | | VZV 2 | 3× LoD | 82.0 | 82.1 | 81.7 | 81.9 | 81.5 | 81.5 | 81.4 | 81.5 | | | | | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.2 | ±0.2 | ±0.2 | | | | 1× LoD | 82.1 | 82.1 | 81.7 | 82.0 | 81.5 | 81.5 | 81.4 | 81.5 | | | | | ±0.1 | ±0.4 | ±0.2 | ±0.3 | ±0.3 | ±0.2 | ±0.2 | ±0.2 | | YEAST | | | | | | | | | | | | C. neoformans/gattii (C. gattii) (b(4) | Cryptococcus | 30× LoDa (FilmArray) | 82.0 | 82.0 | 81.6 | 81.8 | 81.2 | 81.4 | 81.3 | 81.3 | | | | 3× LoD (FilmArray 2.0) | ±0.1 | ±0.3 | ±0.2 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | ±0.3 | | | | 10× LoDa (FilmArray) | 82.0 | 82.1 | 81.6 | 81.9 | 81.3 | 81.5 | 81.3 | 81.4 | | | | 1× LoD (FilmArray 2.0) | ±0.2 | ±0.2 | ±0.2 | ±0.3 | ±0.3 | ±0.4 | ±0.2 | ±0.3 | 9 C. gattii was tested at concentrations equivalent to 10× and 30× LoD for the combined Cryptococcus neoformans/gattii test result on the FilmArray system, while C. gattii was tested at the intended 1× and 3× LoD concentrations on the FilmArray 2.0 system. - b. Linearity/assay Reportable Range: Not Applicable - c. Traceability, Stability, Expected Values (controls, calibrators, or methods): ## Internal Controls: Two internal controls are included in each FilmArray ME Panel pouch: - RNA Process Control: The RNA Process Control assay targets an RNA transcript ● from the yeast Schizosaccharomyces pombe. b(4) . The control material is carried through all stages of the test process, including lysis, nucleic acid purification, reverse transcription, 1st stage PCR, dilution, 2nd stage PCR and DNA melting. A positive control result indicates that all steps carried out in the FilmArray ME Panel pouch were successful. {12}------------------------------------------------ - . PCR2 Control: The PCR2 Control assay detects a DNA target b(4) . A positive result indicates that the 2nd stage PCR was successful. Both internal control assays must be positive for the test run to pass. When either control fails, the Controls field of the test report will display "Failed" and all results will be listed as Invalid. If the controls fail, the user is instructed to repeat the test using a new pouch. Of the six pouch control failures observed in the prospective clinical study, five were attributed to both RNA Process Control and PCR 2 Control failures and one was attributed to RNA Process Control failure only. ## Recommended External Controls: External controls are not provided with the FilmArray ME Panel, but are recommended in the package insert. Molecular grade water or artificial CSF can be used as an external negative control. Previously characterized CSF specimens or negative matrix spiked with well-characterized organisms can be used as external positive controls. External controls should be used in accordance with the appropriate accrediting organization requirements, as applicable. ## d. Detection Limit: ## Evaluation of the Limit of Detection: The limit of detection (LoD) for FilmArray ME Panel analytes was estimated by testing dilutions of contrived samples containing known concentrations of bacteria, viruses or yeast detected by the FilmArray ME Panel. Representative strains were chosen in order to obtain positive results for every assay on the panel and multiple strains were evaluated to cover clinically important species or variants for some analytes. Samples for LoD testing were prepared in aCSF (artificial CSF obtained from a commercial provider) matrix and contained one or up to five targeted organisms. | b(4) | | | |------|--|-------------------------------| | | | . b(4) | | | | | | | | | | | | | | | | The LoD was confirmed for all | | | | 1 | analytes on both FilmArray and FilmArray 2.0 systems. Data presented in the following table are from LoD confirmation testing on the FilmArray system. {13}------------------------------------------------ | ME Panel Test Result | Species/Isolate Tested | LoD Concentration | Detection at LoD Concentration | | | |-------------------------|-------------------------------------------------------|----------------------------------------|--------------------------------------|--|--| | | BACTERIA | | | | | | E. coli K1 | E. coli K1, strain C5 b(4) b(4) | 1×103 CFU/mL | 20/20 100% | | | | H. influenzae | H. influenzae, strain AMC 36-A-1 b(4) | 1×103 CFU/mL | 20/20 100% | | | | L. monocytogenes | L. monocytogenes, strain 1071/53, b(4) | 1×103 CFU/mL | 20/20 100% | | | | N. meningitidis | N. meningitidis, strain M-1574 b(4) | 100 CFU/mL | 19/20 95% | | | | S. agalactiae | S. agalactiae, type strain, G19, group B b(4) | 1×103 CFU/mL | 20/20 100% | | | | S. pneumoniae | S. pneumoniae, strain SV 1, serotype 1 b(4) | 100 cells/mL | 19/20 95% | | | | | VIRUSES | | | | | | CMV | CMV, strain AD-169 b(4) | 100 TCID50/mL (4.30×103 copies/mL) | 20/20 100% | | | | EV (Species A-D) | Coxsackievirus A6, species A, b(4) | 50 TCID50/mL | 20/20 100% | | | | | Coxsackievirus A9, species B b(4) | 5 TCID50/mL | 20/20 100% | | | | | Coxsackievirus A17, species C, strain G-12 b(4) | 5 TCID50/mL | 20/20 100% | | | | | EV 70, species D, b(4) b(4) | 50 TCID50/mL | 20/20 100% | | | | HSV-1 | HSV-1, strain MacIntyre b(4) | 250 TCID50/mL (1.51×103 copies/mL) | 20/20 100% | | | | HSV-2 | HSV-2, strain MS b(4) | 50 TCID50/mL (1.29×103 copies/mL) | 20/20 100% | | | | HHV-6 | HHV-6A, strain U1102 b(4) | 1×104 copies/mL | 19/20 95% | | | | | HHV-6B, strain HST b(4) | 1×104 copies/mL | 19/20 95% | | | | HPeV | HPeV, type 3 b(4) | 500 TCID50/mL | 19/20 95% | | | | VZV | VZV, strain Ellen b(4) | 0.10 TCID50/mL (1.66×103 copies/mL) | 20/20 100% | | | | | YEAST | | | | | ## Limit of Detection Results for FilmArray ME Panel Analytes {14}------------------------------------------------ | ME Panel Test Result | Species/Isolate Tested | LoD Concentration | Detection at LoD Concentration | |-------------------------|-----------------------------------------|-------------------|-----------------------------------| | C. neoformans/gattii | C. neoformans var. grubii, type b(4) | 100 CFU/mL | 20/20 100% | | | C. gattii, strain A6MR38, b(4) | 100 CFU/mL | 20/20 100% | ## Evaluation of Effect on Analyte LoDs in Multi-Spiked Samples: A study was performed to determine the applicability of evaluating multiple analytes in a single CSF sample during the analytical studies and evaluated for potential loss of detection when multiple targets are present in clinical specimens (co-infections). Samples consisting of three different organism mixes with up to five targeted organisms each were compared to contrived single-spike samples for the following representative panel analytes: bacterium (E. coli K1), yeast (C. neoformans), DNA virus (HSV-1), and RNA virus (HPeV). All samples were prepared in aCSF matrix. Serial dilutions were prepared for both single-spike samples and multiple spiked samples containing the corresponding analyte for comparison. For E. coli K1 and HSV-1 samples prepared with analyte concentrations at LoD, 4/4 replicates were positive for both single and multi-spiked samples. Dilutions with concentrations below LoD showed a loss of detection between the single-spiked and multi-spiked specimens at the same organism concentration, thus demonstrating no difference in assay detection for E. coli K1 and HSV-1 for single versus multi-spiked samples. For C. neoformans, 4/4 samples were positive at both 1x LoD and 0.1× LoD, At 0.01× LoD, 4/4 were positive for single-spiked samples and 1/4 replicates were positive for multi-spiked samples. To evaluate whether the observed difference in detection was due to multi-spiking versus single-spiking, mean PCR crossing point (Cp) values were evaluated for each concentration evaluated. The analysis showed that mean Cp values for each concentration tested showed no trend toward lower or higher Cp values between single and multi-spiked samples. For HPeV at 1× LoD. 3/4 replicates were detected for multi-spiked samples and 4/4 replicates were detected for single-spiked samples. However, at two different dilutions below the LoD, there was no trend in detection differences between single and multispiked samples. In summary, testing of single versus multi-spiked samples did not show a significant difference in assay performance for the four representative analytes evaluated. The study results supported the use of multi-spiked samples in various analytical studies. e. Analytical Reactivity (Inclusivity): Inclusivity testing was performed using contrived samples consisting of 96 isolates {15}------------------------------------------------ spiked into artificial CSF (aCSF) sample matrix at a concentration near the LoD for each analyte (1× to 3× LoD). Strains were selected to represent relevant species, subspecies, or serotypes. For any isolate that was not detected at the initial test concentration, results were reviewed and if reactivity was expected, the isolate was retested at the same concentration up to 5 times. If needed, retesting of the isolate was also performed at a higher concentration (typically 10× LoD) and a detected result was a demonstration of reactivity with the isolate at the elevated concentration. The FilmArray ME panel gave positive results for the majority of isolates evaluated when tested at sample concentrations near the LoD (1× to 3× LoD). One strain of HpeV (Serotype 5) was detected at 10× LoD as compared to the HpeV strain evaluated in the LoD study. 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 evaluated with empirical testing. The following tables includes a summary of ME Panel reactivity based on empirical data with footnotes describing predictions of assay reactivity based on in silico analysis. | FilmArray ME Panel Test Result | # of Isolates Tested and Detected | Concentration Detected | Isolates Tested and Detected | |--------------------------------------|--------------------------------------|------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------| | Bacteria | | | | | E. coli K1 | 5 | 1,000 - 3,000 CFU/mL | E. coli strains of the K1 serotype only | | H. influenzae | 9 | 1,000 - 3,000 CFU/mL | Non-typeable and typeable (types a-f) strains of H. influenzae | | L. monocytogenes | 6 | 1,000 - 3,000 CFU/mL | Types 1/2a, 1/2b, and 4b of L. monocytogenes a | | N. meningitidis | 7 | 100 - 300 CFU/mL | Encapsulated N. meningitidis (serotypes W135, A, B, C, D, Y and DNA from a strains with a variant ctrA gene) | | S. agalactiae | 5 | 1,000 - 3,000 CFU/mL | Multiple serotypes or isolates of S. agalactiae (Group B Streptococcus ) | | S. pneumoniae | 6 | 100 - 300 cells/mL | Multiple serotypes of S. pneumoniae | | Viruses | | | | | CMV | 5 | 100 - 300 TCID50/mL (4.3×103 - 1.3×104 copies/mL) | Multiple strains of Cytomegalovirus (CMV). | | EV | 18 | 5 - 50 TCID50/mL | Representative isolates from all species (A-D) and several serotypes of human Enterovirus, Coxsackievirus, and Echovirusb | | HSV-1 | 5 | 250 - 750 TCID50/mL (1.5×103 - 4.5×103 copies/mL) | Multiple strains of Herpes simplex virus 1 (HSV-1) | | HSV-2 | 5 | 50 - 150 TCID50/mL (1.3×103 - 3.9×103 copies/mL) | Multiple strains of Herpes simplex virus 2 (HSV-2) | | HHV-6 | 4 | 1×104 - 3×104 copies/mL | A and B variants of Human herpesvirus 6 (HHV-6) | Summary of FilmArray ME Panel Analytical Reactivity (Inclusivity) {16}------------------------------------------------ | FilmArray ME Panel Test Result | # of Isolates Tested and Detected | Concentration Detected | Isolates Tested and Detected | |--------------------------------------|--------------------------------------|--------------------------------------------------|-----------------------------------------------------------------------------------------------------| | HPeV | 6 | 500 - 5,000 TCID50/mL | Serotypes 1-6 of Human parechovirus (HPeV)c | | VZV | 5 | 0.1 - 0.3 TCID50/mL (1.7×103-5×103 copies/mL) | Multiple strains of Varicella zoster virus (VZV) | | Yeast | | | | | C. neoformans/gattii | 10 (5 per species) | 100 - 300 CFU/mL | Multiple strains, serotypes, and genotypes of Cryptococcus neoformans and Cryptococcus gattii | ª In silco analysis of available sequences predicts that the FilmArray ME Panel with all currently characterized strains and serotypes of L. monocytogenes. 1 1200 condysis of available sequences predicts that the FilmArray ME Panel will react with all currently characterized serotypes (>100) of human enteroviruses (including enteroviruses, coxsackieviruses, and echoviruses). 6 Based on sequence analysis, the FilmArray ME Panel is also predicted to react with HPeV serotypes 7 and 8. No sequence data were available for predicting reactivity with other serotypes. ## The following tables include specific strains evaluated with the FilmArray ME Panel. #### Results for Escherichia coli K1 Inclusivity Testing | Organism | Isolate ID | Serotype [Strain-Year Isolated] | Concentration Tested | Test Result | |---------------------|------------|-------------------------------------------------|----------------------------------|----------------| | Escherichia coli K1 | b(4) | Serotype O18ac:K1:H7 [Strain C5 [Bort]-1975] | $1\times10^3$ CFU/mL (1x LoD) | Detected | | | | Serotype O2:K1:H4 [Strain U9-41] | $3\times10^3$ CFU/mL | Detected | | | | Serotype O16:K1:H- [Strain F11119-41-1952] | $3\times10^3$ CFU/mL | Detected | | | | Serotype O9:K1:H- [Strain Bi 7509/41-1952] | $3\times10^3$ CFU/mL | Detected | | | | Serotype O45:K1:H10 [Strain H61-1952] | $3\times10^3$ CFU/mL | Detected | | Results for Haemophilus influenzae Inclusivity Testing | | | | |--------------------------------------------------------|--|--|--| |--------------------------------------------------------|--|--|--| | | | Type | Concentration | Test | |-------------|------------|------------------------------------------------------------|-------------------------------|----------| | Organism | Isolate ID | [Strain] | Tested | Result | | | b(4) | Non-typeable [strain Rd [KW20]] | $3 × 10^3$ CFU/mL | Detected | | | | Non-typeable biogroup aegyptius [type strain, 180-a] | $3 × 10^3$ CFU/mL | Detected | | Haemophilus | | Type a [strain AMC 36-A-3] | $3 × 10^3$ CFU/mL | Detected | | influenzae | | Type b [strain Rab] | $3 × 10^3$ CFU/mL | Detected | | | | Type b [biotype 1] | $1 × 10^3$ CFU/mL (1× LoD) | Detected | | | | Type c [strain C 9007] | $3 × 10^3$ CFU/mL | Detected | {17}------------------------------------------------ | Organism | Isolate ID | Type [Strain] | Concentration Tested | Test Result | |----------|------------|-------------------------------|-------------------------|----------------| | | b(4) | Type d [strain AMC 36-A-6] | $3 × 10^3$ CFU/mL | Detected | | | | Type e [strain AMC 36-A-7] | $3 × 10^3$ CFU/mL | Detected | | | | Type f [strain GA-1264] | $3 × 10^3$ CFU/mL | Detected | | | Results for Listeria monocytogenes Inclusivity Testing | | |--|--------------------------------------------------------|--| | | | | | Organism | Source | Isolate ID | Type | Concentration Tested | Test Result | |-----------------------------------|--------|------------|-----------|-------------------------------------|----------------| | Listeria monocytogenes | b(4) | | Type 1/2a | $3 \times 10^3$ CFU/mL | Detected | | | | | Type 1/2a | $3 \times 10^3$ CFU/mL | Detected | | | | | Type 1/2b | $3 \times 10^3$ CFU/mL | Detected | | | | | Type 1/2b | $3 \times 10^3$ CFU/mL | Detected | | | | | Type 4b | $3 \times 10^3$ CFU/mL | Detected | | | | | Type 4b | $1 \times 10^3$ CFU/mL (1 × LoD) | Detected | Note: At least 12 serotypes of L. monocytogenes have been recognized (i.e., 1/2a, 1/2c, 3a, 3b, 3c, 4a, 4b, 4c, 4d, 4e and 7) based on somatic (O) and flagellar (H) antigens, but more than 90% of human isolates belong to only three serotypes: 1/2a, 1/2b, and 4b. In silico analysis based on available sequence data suggest that all serotypes are expected to be amplified by the assay and detected by the FilmArray ME Panel. | Organism | Isolate ID | Serotype | Concentration Tested | Test Result | |------------------------------------------|------------|-------------------------------|---------------------------------------------------|----------------| | | | Serotype W135 | 100 CFU/mL (1× LoD) [~1.9×103 copies/mL] | Detected | | | | Serotype A | 300 CFU/mL | Detected | | Neisseria meningitidis (Encapsulated) | | Serotype B | 300 CFU/mL | Detected | | | | Serotype C | 300 CFU/mL | Detected | | | | Serotype D | 300 CFU/mL | Detected | | | | Serotype Y | 300 CFU/mL | Detected | | | | DNA with variant ctrA gene | 5.6×103 copies/mL (~3× LoD) | Detected | Results for Neisseria meningitidis Inclusivity Testing {18}------------------------------------------------ | Organism | Isolate ID | Serotype | Concentration Tested | Test Result | |-----------------------------|------------|-------------------------------|------------------------------------|----------------| | Streptococcus agalactiae | b(4) | Serotype I a/c Type strain | $1 \times 10^3$ CFU/mL (1× LoD) | Detected | | | | Serotype III | $3 \times 10^3$ CFU/mL | Detected | | | | Serotype V | $3 \times 10^3$ CFU/mL | Detected | | | | Unknown | $3 \times 10^3$ CFU/mL | Detected | | | | Unknown | $3 \times 10^3$ CFU/mL | Detected | Results for Streptococcus agalactiae Inclusivity Testing Note: In silico analysis predicts that the FilmArray ME panel should detect S. agalactiae serotypes Ia, Ia/c, Ib, II, III, V and VIII strains. | Organism | Isolate ID | Serotype | Concentration Tested | Test Result | |--------------------------|------------|--------------|--------------------------|-------------| | | b(4) | Serotype 1 | 100 cells/mL (1× LoD) | Detected | | | b(4) | Serotype 4 | 300 cells/mL | Detected | | Streptococcus pneumoniae | b(4) | Serotype 5 | 300 cells/mL | Detected | | | b(4) | Serotype 11A | 300 cells/mL | Detected | | | b(4) | Serotype 14 | 300 cells/mL | Detected | | | b(4) | Serotype 19A | 300 cells/mL | Detected | Results for Streptococcus pneumoniae Inclusivity Testing Note: Based on the serotype information associated with sequences available in public databases, in silico analysis indicates the assay will react with all serotypes of S. pneumoniae, including those covered by the PPSV23 pneumococcal vaccine (1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F, and 33F). | Organism | Isolate ID | Strain | Location/ Year Isolated | Concentration Tested | Test Result | |------------------------------------------------|------------|--------|----------------------------|---------------------------------------------------------------------|----------------| | Cytomegalovirus (CMV) [Human Herpesvirus 5] | b(4) | AD-169 | unknown | 100 TCID50/mL [ $4.3 \times 10^3$ copies/mL] (1 $\times$ LoD) | Detected | | | | Towne | Unknown | $1.3 \times 10^4$ copies/mL | Detected | | | | Merlin | Wales, 2003 | $1.3 \times 10^4$ copies/mL | Detected | ## Results for Cytomegalovirus (CMV) Inclusivity Testing {19}------------------------------------------------ | Organism | Isolate ID | Strain | Location/ Year Isolated | Concentration Tested | Test Result | |----------|------------|--------|----------------------------|-----------------------------|----------------| | | b(4) | Davis | 1957 | $1.3 \times 10^4$ copies/mL | Detected | | | | Toledo | Virginia, USA 2011 | $1.3 \times 10^4$ copies/mL | Detected | ## Results for Enterovirus (EV) Inclusivity Testing | Species | Virus/Serotype | Strain Location/Year | Concentration Tested | Test Result | |---------|---------------------------------------|------------------------|---------------------------|--------------| | A | Coxsackievirus A6 | Gdula b(4) | 50 TCID50/mL (1 × LoD) | Detected | | | Coxsackievirus A10 | M.K. (Kowalik) b(4) | 150 TCID50/mL | Detected | | | Coxsackievirus A16 | H07314 334 | 50 TCID50/mL | Detected | | | Enterovirus 71 | BrCr | 150 TCID50/mL | Detected | | |…