ePlex Respiratory Pathogen Panel
Device Facts
| Record ID | K163636 |
|---|---|
| Device Name | ePlex Respiratory Pathogen Panel |
| Applicant | Genmark Diagnostics, Incorporated |
| Product Code | OCC · Microbiology |
| Decision Date | Jun 9, 2017 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3980 |
| Device Class | Class 2 |
Intended Use
The ePlex® Respiratory Pathogen (RP) Panel is a multiplexed nucleic acid in vitro diagnostic test intended for use on the ePlex® Instrument for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals exhibiting signs and symptoms of respiratory tract infection. The following virus types, subtypes, and bacteria are identified using the ePlex RP Panel: adenovirus, coronavirus, human metapneumovirus, human rhinovirus/enterovirus, influenza A, influenza A H1, influenza A H1-2009, influenza A H3, influenza B, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, respiratory syncytial virus (RSV) A, respiratory syncytial virus (RSV) B, Chlamydia pneumoniae, and Mycoplasma pneumoniae. The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of respiratory tract infection aids in the diagnosis of respiratory infection when used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms; the organism(s) detected by the ePlex RP Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection. Due to the genetic similarity between human rhinovirus and enterovirus, the ePlex RP Panel cannot reliably differentiate them. If differentiation is required, an ePlex RP Panel positive human rhinovirus/enterovirus result should be followed-up using an alternative method (e.g., cell culture or sequence analysis). Performance characteristics for influenza A were established when influenza A H1-2009 and A H3 were the predominant influenza A viruses in circulation. Performance of detecting influenza A may vary if other influenza A strains are circulating or a novel influenza A virus emerges. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL-3+ facility is available to receive and culture specimens.
Device Story
The ePlex RP Panel is an automated, multiplexed, qualitative in vitro diagnostic test for respiratory pathogens. It processes nasopharyngeal swab samples on the ePlex Instrument using a single-use cartridge. The device automates nucleic acid extraction, amplification (RT-PCR), and detection. It utilizes electrowetting (digital microfluidics) to manipulate droplets on a PCB and eSensor technology for electrochemical detection via voltammetry, avoiding optical/fluorescent methods. The system identifies 15 viral and 2 bacterial targets. It is intended for use in clinical laboratories by trained personnel. The instrument provides automated interpretation and reporting; raw data is inaccessible to users. Results aid clinicians in diagnosing respiratory infections, though they must be interpreted alongside clinical and epidemiological data. The device benefits patients by providing rapid, simultaneous identification of multiple pathogens, potentially guiding appropriate treatment and infection control decisions.
Clinical Evidence
Clinical performance evaluated in 2,462 prospective nasopharyngeal swab samples across 8 sites. Comparator was an FDA-cleared multiplexed molecular panel and bi-directional sequencing for RSV subtypes. PPA/NPA varied by target; for example, Influenza A H1-2009 showed 98.6% PPA and 99.7% NPA. Co-detection rate was 5.5%. Reproducibility study across 3 sites showed 100% agreement for most targets. Bench testing confirmed LoD, inclusivity, and exclusivity (no cross-reactivity with 41 off-panel organisms).
Technological Characteristics
Multiplex NAAT using electrowetting (digital microfluidics) and eSensor electrochemical detection. Reagents contained in single-use cartridges stored at 2-8°C. Automated nucleic acid extraction, RT-PCR, and voltammetric detection. No optical/fluorescent components. Software provides automated interpretation and report generation. Moderate complexity.
Indications for Use
Indicated for individuals of all ages exhibiting signs/symptoms of respiratory tract infection. Used for qualitative detection/identification of 17 respiratory viral/bacterial targets in nasopharyngeal swabs. Not for sole diagnostic use; results must be used with clinical/epidemiological data. Contraindicated for sole diagnosis; requires follow-up for rhinovirus/enterovirus differentiation and novel influenza A screening.
Regulatory Classification
Identification
A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B; (2) Influenza A subtype H1 and Influenza A subtype H3; (3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B; (4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus; (5) Human Metapneumovirus; (6) Rhinovirus; and (7) Adenovirus.
Special Controls
*Classification.* Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;” (2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and (3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.
Predicate Devices
- FilmArray Respiratory Panel (K160068)
Related Devices
- K183597 — QIAstat-Dx Respiratory Panel · QIAGEN GmbH · May 18, 2019
- K250080 — QIAstat-Dx Respiratory Panel Plus; QIAstat-Dx Respiratory Panel Mini · QIAGEN GmbH · Aug 27, 2025
- K233100 — QIAstat-Dx® Respiratory Panel Plus · QIAGEN GmbH · May 10, 2024
- K113731 — ESENSOR RESPIRATORY VIRAL PANEL · Clinical Micro Sensors, Inc. · Sep 10, 2012
Submission Summary (Full Text)
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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is an abstract symbol that resembles a stylized caduceus or a series of human profiles.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
June 9, 2017
GenMark Diagnostics, Incorporated Alan Maderazo, Ph.D., RAC Vice President, Quality, Regulatory and Clinical Affairs 5964 La Place Court Carlsbad, CA 92008
Re: K163636
Trade/Device Name: ePlex® Respiratory Pathogen (RP) Panel Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: II Product Code: OCC, OEM, OEP, OOU, OTG, OZE, OZX, OZY, OQW, NSU Dated: May 3, 2017 Received: May 5, 2017
Dear Dr. Maderazo:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours.
# Tamara V. Feldblyum -S for
Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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# Indications for Use
#### 510(k) Number (if known) K163636
#### Device Name
ePlex® Respiratory Pathogen (RP) Panel
#### Indications for Use (Describe)
The ePlex® Respiratory Pathogen (RP) Panel is a multiplexed nucleic acid in vitro diagnostic test intended for use on the ePlex® Instrument for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals exhibiting signs and symptoms of respiratory tract infection.
The following virus types, subtypes, and bacteria are identified using the ePlex RP Panel: adenovirus, coronavirus, human metapneumovirus, human rhinovirus/enterovirus, influenza A, influenza A H1, influenza A H1-2009, influenza A H3, influenza B, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, respiratory syncytial virus (RSV) A, respiratory syncytial virus (RSV) B, Chlamydia pneumoniae, and Mycoplasma pneumoniae.
The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of respiratory tract infection aids in the diagnosis of respiratory infection when used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms; the organism(s) detected by the ePlex RP Panel may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.
Due to the genetic similarity between human rhinovirus and enterovirus, the ePlex RP Panel cannot reliably differentiate them. If differentiation is required, an ePlex RP Panel positive human rhinovirus/enterovirus result should be followed-up using an alternative method (e.g., cell culture or sequence analysis).
Performance characteristics for influenza A were established when influenza A H1-2009 and A H3 were the predominant influenza A viruses in circulation. Performance of detecting influenza A may vary if other influenza A strains are circulating or a novel influenza A virus emerges. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL-3+ facility is available to receive and culture specimens.
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Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)
| | Over-The-Counter Use (21 CFR 801 Subpart C)
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# 510(k) Summary
# Summary of Safety and Effectiveness
| Submitter Information | |
|------------------------------------|----------------------------------------------------------|
| Submitter: | GenMark Diagnostics, Incorporated |
| | 5964 La Place Court |
| | Carlsbad, CA 92008 |
| Manufacturer: | GenMark Diagnostics, Incorporated |
| | 5964 La Place Court |
| | Carlsbad, CA 92008 |
| Establishment Registration Number: | 3008632402 |
| Contact: | Alan Maderazo, Ph.D., RAC |
| | Vice President, Quality, Regulatory and Clinical Affairs |
| Phone: | 760-448-4308 |
| Fax: | 760-683-6961 |
| E-mail: | Al.Maderazo@genmarkdx.com |
| Alternate Contact: | Joseph McMullen |
| | Consultant, Regulatory Affairs |
| Phone: | 760-410-5052 |
| Fax: | 760-683-6961 |
| E-mail: | Joseph.McMullen@genmarkdx.com |
| Date Prepared: | December 21, 2016 |
# Name of Device and Classification
| Product Name: | ePlex® Respiratory Pathogen (RP) Panel |
|------------------------|-------------------------------------------------------------------------|
| Device Classification: | 866.3980, Respiratory viral panel multiplex nucleic acid assay, Class I |
| Product Code(s): | OCC, OEM, OOU, OEP, OTG, OQW, OZE, OZY, OZX, NSU |
# Predicate Device
- Predicate: FilmArray Respiratory Panel; BioFire Diagnostics; K160068
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# Device Description
The ePlex RP Panel is an automated qualitative nucleic acid multiplex in vitro diagnostic test for simultaneous detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS). The test is able to detect 15 respiratory viral targets and 2 bacterial targets as summarized in the table below. This test is performed on the ePlex Instrument.
| Target | Classification (Genome Type) |
|----------------------------------|------------------------------|
| Adenovirus (A-F) | Adenovirus (DNA) |
| Coronavirus | Coronavirus (RNA) |
| (229E, HKU1, NL63, OC43) | |
| Human Metapneumovirus | Paramyxovirus (RNA) |
| Human Rhinovirus/<br>Enterovirus | Picornavirus (RNA) |
| Influenza A | |
| Influenza A H1 | |
| Influenza A H1-2009 | Orthomyxovirus (RNA) |
| Influenza A H3 | |
| Influenza B | |
| Parainfluenza Virus 1 | |
| Parainfluenza Virus 2 | Paramyxovirus (RNA) |
| Parainfluenza Virus 3 | |
| Parainfluenza Virus 4 | |
| Respiratory Syncytial Virus A | Paramyxovirus (RNA) |
| Respiratory Syncytial Virus B | |
| Chlamydia pneumoniae | Bacterium (DNA) |
| Mycoplasma pneumoniae | Bacterium (DNA) |
# Targets Detected by the ePlex RP Panel
The ePlex Instrument automates all aspects of nucleic acid testing including extraction, amplification, and detection, combining electrowetting and GenMark's eSensor® technology in a single-use cartridge. eSensor technology is based on the principles of competitive DNA hybridization and electrochemical detection. which is highly specific and is not based on fluorescent or optical detection.
Electrowetting, or digital microfluidics, uses electrical fields to directly manipulate discrete droplets on the surface of a hydrophobically coated printed circuit board (PCB). Sample and
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reagents are moved in a programmable fashion in the ePlex cartridge to complete all portions of the sample processing from nucleic acid extraction to detection.
A sample is loaded onto the ePlex cartridge and nucleic acids are extracted and purified from the specimen via magnetic solid phase extraction. For RNA targets, a reverse transcription step is performed to generate complementary DNA from the RNA. followed by PCR to amplify the targets. Exonuclease digestion creates single-stranded DNA in preparation for eSensor detection.
The target DNA is mixed with ferrocene-labeled signal probes that are complementary to the specific targets on the panel. Target DNA hybridizes to its complementary signal probe and capture probes, which are bound to gold-plated electrodes. The presence of each target is determined by voltammetry which generates specific electrical signals from the ferrocenelabeled signal probe.
# Intended Use/Indications for Use
The ePlex® Respiratory Pathogen (RP) Panel is a multiplexed nucleic acid in vitro diagnostic test intended for use on the ePlex Instrument for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals exhibiting signs and symptoms of respiratory tract infection.
The following virus types, subtypes, and bacteria are identified using the ePlex RP Panel: adenovirus, coronavirus, human metapneumovirus, human rhinovirus/enterovirus, influenza A, influenza A H1, influenza A H1-2009, influenza A H3, influenza B, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 3, parainfluenza virus 4, respiratory syncytial virus (RSV) A, respiratory syncytial virus (RSV) B, Chlamydia pneumoniae, and Mycoplasma pneumoniae.
The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of respiratory tract infection aids in the diagnosis of respiratory infection when used in conjunction with other clinical and epidemiological
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information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other patient management decisions.
Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test, or lower respiratory tract infection that may not be detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the organism(s) detected by the ePlex RP Panel may not be the definite cause of disease. Additional laboratory testing (e.g., bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.
Due to the genetic similarity between human rhinovirus and enterovirus, the ePlex RP Panel cannot reliably differentiate them. If differentiation is required, an ePlex RP Panel positive human rhinovirus/enterovirus result should be followed-up using an alternative method (e.g., cell culture or sequence analysis).
Performance characteristics for influenza A were established when influenza A H1-2009 and A H3 were the predominant influenza A viruses in circulation. Performance of detecting influenza A may vary if other influenza A strains are circulating or a novel influenza A virus emerges. If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL-3+ facility is available to receive and culture specimens.
# Summary of Technological Characteristics of the Device Compared to the Predicate Device
The GenMark ePlex Respiratory Pathogen (RP) Panel ("Proposed Device") and the legally marketed device, FilmArray Respiratory Panel ("Predicate Device") are described below:
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| Characteristic | Proposed Device | Predicate Device |
|---------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Product Name | ePlex Respiratory Pathogen (RP) Panel | FilmArray Respiratory Panel |
| Manufacturer | GenMark Diagnostics | BioFire Diagnostics |
| Regulation | 866.3980<br>Respiratory viral panel multiplex<br>nucleic acid assay | 866.3980<br>Respiratory viral panel multiplex<br>nucleic acid assay |
| Product Code(s) | OCC, OEM, OOU, OEP, OTG, OZE,<br>OQW, OZY, OZX, NSU | OCC, OEM, OOU, OEP, OTG,<br>OQW, OOI, OZZ, OZY, OZX |
| Device Class | Class II | Class II |
| Organisms<br>Detected | Influenza A<br>Influenza A subtype H1<br>Influenza A subtype H3<br>Influenza A subtype H1-2009<br>Influenza B<br>Respiratory Syncytial Virus A and B<br>Parainfluenza Virus 1, 2, 3, 4<br>Human Metapneumovirus<br>Adenovirus<br>Human Rhinovirus/Enterovirus<br>Coronavirus<br>Chlamydia pneumoniae<br>Mycoplasma pneumoniae | Influenza A<br>Influenza A subtype H1<br>Influenza A subtype H3<br>Influenza A subtype H1-2009<br>Influenza B<br>Respiratory Syncytial Virus<br>Parainfluenza Virus 1, 2, 3, 4<br>Human Metapneumovirus<br>Adenovirus<br>Human Rhinovirus/Enterovirus<br>Coronavirus HKU1, NL63, 229E,<br>OC43<br>Bordetella pertussis<br>Chlamydia pneumoniae<br>Mycoplasma pneumoniae |
| Analyte | RNA/DNA | RNA/DNA |
| Technological<br>Principles | Multiplex nucleic acid amplification test<br>(NAAT) | Multiplex nucleic acid amplification<br>test (NAAT) |
| Specimen Type | Nasopharyngeal swabs (NPS) in VTM | Nasopharyngeal swabs (NPS) in<br>VTM |
| Chemistry | Reagents contained within cartridge to<br>allow: sample lysis and nucleic acid<br>extraction, RT-PCR amplification, and<br>hybridization-based electrochemical<br>detection reagents. | Nested multiplex RT-PCR followed<br>by high resolution melting analysis to<br>confirm identity of amplified<br>product. |
| Instrumentation | ePlex Instrument | FilmArray Instrument |
| Test<br>Interpretation | Automated test interpretation and report<br>generation. User cannot access raw data. | Automated test interpretation and<br>report generation. User cannot<br>access raw data. |
| Sample<br>Preparation<br>Method | Sample processing is automated in the<br>ePlex RP cartridge. | Sample processing is automated in<br>the FilmArray RP pouch. |
| Controls | Eight internal controls are used to<br>demonstrate that sample extraction,<br>amplification and detection processes<br>functioned as intended. | Two controls are included in each<br>reagent pouch to control for sample<br>processing and both stages of PCR<br>and melt analysis. |
| Characteristic | Proposed Device | Predicate Device |
| User Complexity | Moderate | Moderate |
| Reagent Storage | Cartridge (which contains reagents) is<br>stored at refrigerated (2-8°C) temperature. | Reagents are stored at room<br>temperature. |
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Analysis of the similarities and differences indicate that the devices are substantially equivalent in their intended uses/indications for use, and are generally the same regarding user process, ease of use and general operator protocol. Comparison of technological similarities and differences between the proposed device and the predicate do not raise new or different questions of safety and effectiveness, and therefore render the proposed device as substantially equivalent to the predicate device.
# Expected Values
A prospective, multicenter clinical study was conducted to evaluate the clinical performance of the ePlex RP Panel in nasopharyngeal swab samples. 2462 nasopharyngeal swab samples were prospectively-collected at 8 collection sites in 2 phases from patients of all ages and genders presenting with signs and/or symptoms of respiratory infection. In the first phase from March 2013 through August 2014, 1951 samples were prospectively-collected and frozen; from September 2016 through October 2016, 511 samples were prospectively-collected and tested fresh (never frozen). The expected values of individual analytes based on ePlex RP Panel results in prospective samples for each phase are summarized in Tables 1-4.
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| By Age Group in the Prospective Clinical Evaluation (Phase 1: March 2013 – August 2014) | | | | | | |
|-----------------------------------------------------------------------------------------|----------------------|--------------------|---------------------|----------------------|-----------------------|--------------------|
| | All Ages<br>(N=1951) | Age 0-1<br>(N=315) | Age >1-5<br>(N=250) | Age >5-21<br>(N=246) | Age >21-65<br>(N=745) | Age >65<br>(N=395) |
| Organism | n (%) | n (%) | n (%) | n (%) | n (%) | n (%) |
| Adenovirus | 72 (3.7) | 31 (9.8) | 24 (9.6) | 7 (2.8) | 7 (0.9) | 3 (0.8) |
| Coronavirus | 102 (5.2) | 19 (6.0) | 18 (7.2) | 16 (6.5) | 32 (4.3) | 17 (4.3) |
| Human Metapneumovirus | 113 (5.8) | 22 (7.0) | 28 (11.2) | 6 (2.4) | 31 (4.2) | 26 (6.6) |
| Human<br>Rhinovirus/Enterovirus | 388 (19.9) | 113 (35.9) | 94 (37.6) | 58 (23.6) | 87 (11.7) | 36 (9.1) |
| Influenza A | 110 (5.6) | 6 (1.9) | 18 (7.2) | 20 (8.1) | 49 (6.6) | 17 (4.3) |
| Influenza A H1 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza A H1-2009 | 76 (3.9) | 4 (1.3) | 13 (5.2) | 14 (5.7) | 37 (5.0) | 8 (2.0) |
| Influenza A H3 | 34 (1.7) | 1 (0.3) | 5 (2.0) | 6 (2.4) | 12 (1.6) | 10 (2.5) |
| Influenza B | 62 (3.2) | 4 (1.3) | 9 (3.6) | 10 (4.1) | 24 (3.2) | 15 (3.8) |
| Parainfluenza Virus 1 | 24 (1.2) | 4 (1.3) | 12 (4.8) | 4 (1.6) | 3 (0.4) | 1 (0.3) |
| Parainfluenza Virus 2 | 10 (0.5) | 4 (1.3) | 4 (1.6) | 0 (0.0) | 2 (0.3) | 0 (0.0) |
| Parainfluenza Virus 3 | 99 (5.1) | 31 (9.8) | 20 (8.0) | 3 (1.2) | 27 (3.6) | 18 (4.6) |
| Parainfluenza Virus 4 | 7 (0.4) | 3 (1.0) | 2 (0.8) | 1 (0.4) | 1 (0.1) | 0 (0.0) |
| RSV A | 28 (1.4) | 13 (4.1) | 6 (2.4) | 3 (1.2) | 2 (0.3) | 4 (1.0) |
| RSV B | 83 (4.3) | 33 (10.5) | 19 (7.6) | 6 (2.4) | 15 (2.0) | 10 (2.5) |
| Chlamydia pneumoniae | 3 (0.2) | 0 (0.0) | 0 (0.0) | 1 (0.4) | 1 (0.1) | 1 (0.3) |
| Mycoplasma pneumoniae | 5 (0.3) | 1 (0.3) | 1 (0.4) | 2 (0.8) | 1 (0.1) | 0 (0.0) |
Table 1: Expected Value (As Determined by ePlex RP Panel) Summary
Eroup in the Prospective Clinical Fraluation (Phase 1: March 2013 – Aug Ry Age August 2014)
Table 2: Expected Value (As Determined by ePlex RP Panel) Summary By Age Group in the Prospective Clinical Evaluation (Phase 2: September 2016 – October 2016)
| By Age Group in the Prospective Clinical Evaluation (Phase 2: September 2016 – October 2016) | | | | | | |
|----------------------------------------------------------------------------------------------|------------------------------|----------------------------|-----------------------------|------------------------------|--------------------------------|-----------------------------|
| Organism | All Ages<br>(N=511)<br>n (%) | Age 0-1<br>(N=73)<br>n (%) | Age >1-5<br>(N=75)<br>n (%) | Age >5-21<br>(N=75)<br>n (%) | Age >21-65<br>(N=181)<br>n (%) | Age >65<br>(N=107)<br>n (%) |
| Adenovirus | 10 (2.0) | 3 (4.1) | 4 (5.3) | 1 (1.3) | 1 (0.6) | 1 (0.9) |
| Coronavirus | 8 (1.6) | 2 (2.7) | 0 (0.0) | 1 (1.3) | 4 (2.2) | 1 (0.9) |
| Human Metapneumovirus | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Human Rhinovirus/Enterovirus | 188 (36.8) | 37 (50.7) | 40 (53.3) | 33 (44.0) | 58 (32.0) | 20 (18.7) |
| Influenza A | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza A H1 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza A H1-2009 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza A H3 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza B | 2 (0.4) | 0 (0.0) | 0 (0.0) | 1 (1.3) | 1 (0.6) | 0 (0.0) |
| Parainfluenza Virus 1 | 1 (0.2) | 0 (0.0) | 1 (1.3) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Parainfluenza Virus 2 | 13 (2.5) | 3 (4.1) | 4 (5.3) | 3 (4.0) | 2 (1.1) | 1 (0.9) |
| Parainfluenza Virus 3 | 5 (1.0) | 2 (2.7) | 1 (1.3) | 1 (1.3) | 1 (0.6) | 0 (0.0) |
| Parainfluenza Virus 4 | 8 (1.6) | 1 (1.4) | 4 (5.3) | 2 (2.7) | 1 (0.6) | 0 (0.0) |
| RSV A | 8 (1.6) | 5 (6.8) | 3 (4.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| RSV B | 9 (1.8) | 3 (4.1) | 4 (5.3) | 0 (0.0) | 2 (1.1) | 0 (0.0) |
| Chlamydia pneumoniae | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Mycoplasma pneumoniae | 4 (0.8) | 0 (0.0) | 1 (1.3) | 2 (2.7) | 1 (0.6) | 0 (0.0) |
{11}------------------------------------------------
| 2014) | | | | | | |
|------------------------------|--------------------------------|----------------------------|----------------------------|----------------------------|----------------------------|----------------------------|
| Organism | All Sites<br>(N=1951)<br>n (%) | Site 1<br>(N=165)<br>n (%) | Site 2<br>(N=248)<br>n (%) | Site 3<br>(N=350)<br>n (%) | Site 4<br>(N=892)<br>n (%) | Site 5<br>(N=296)<br>n (%) |
| Adenovirus | 72 (3.7) | 4 (2.4) | 8 (3.2) | 28 (8.0) | 23 (2.6) | 9 (3.0) |
| Coronavirus | 102 (5.2) | 8 (4.8) | 11 (4.4) | 32 (9.1) | 29 (3.3) | 22 (7.4) |
| Human Metapneumovirus | 113 (5.8) | 10 (6.1) | 23 (9.3) | 27 (7.7) | 30 (3.4) | 23 (7.8) |
| Human Rhinovirus/Enterovirus | 388 (19.9) | 27 (16.4) | 33 (13.3) | 61 (17.4) | 185 (20.7) | 82 (27.7) |
| Influenza A | 110 (5.6) | 5 (3.0) | 21 (8.5) | 48 (13.7) | 19 (2.1) | 17 (5.7) |
| Influenza A H1 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) |
| Influenza A H1-2009 | 76 (3.9) | 3 (1.8) | 22 (8.9) | 31 (8.9) | 5 (0.6) | 15 (5.1) |
| Influenza A H3 | 34 (1.7) | 2 (1.2) | 0 (0.0) | 18 (5.1) | 12 (1.3) | 2 (0.7) |
| Influenza B | 62 (3.2) | 9 (5.5) | 9 (3.6) | 9 (2.6) | 19 (2.1) | 16 (5.4) |
| Parainfluenza Virus 1 | 24 (1.2) | 0 (0.0) | 0 (0.0) | 5 (1.4) | 2 (0.2) | 17 (5.7) |
| Parainfluenza Virus 2 | 10 (0.5) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 10 (1.1) | 0 (0.0) |
| Parainfluenza Virus 3 | 99 (5.1) | 13 (7.9) | 3 (1.2) | 28 (8.0) | 41 (4.6) | 14 (4.7) |
| Parainfluenza Virus 4 | 7 (0.4) | 0 (0.0) | 0 (0.0) | 1 (0.3) | 4 (0.4) | 2 (0.7) |
| RSV A | 28 (1.4) | 4 (2.4) | 6 (2.4) | 7 (2.0) | 4 (0.4) | 7 (2.4) |
| RSV B | 83 (4.3) | 6 (3.6) | 15 (6.0) | 24 (6.9) | 15 (1.7) | 23 (7.8) |
| Chlamydia pneumoniae | 3 (0.2) | 0 (0.0) | 0 (0.0) | 1 (0.3) | 2 (0.2) | 0 (0.0) |
| Mycoplasma pneumoniae | 5 (0.3) | 1 (0.6) | 0 (0.0) | 3 (0.9) | 0 (0.0) | 1 (0.3) |
Table 3: Expected Value (As Determined by ePlex RP Panel) Summary By Sample Collection Site in the Prospective Clinical Evaluation (Phase 1: March 2013 – August
#### Table 4: Expected Value (As Determined by ePlex RP Panel) Summary By Sample Collection Site in the Prospective Clinical Evaluation (Phase 2: September 2016 – Octobe 2016)
| October 2010) | | | | | | | | |
|------------------------------|------------|-----------|-----------|-----------|-----------|--|--|--|
| | All Sites | Site 5 | Site 6 | Site 7 | Site 8 | | | |
| Organism | (N=511) | (N=49) | (N=101) | (N=161) | (N=200) | | | |
| | n (%) | n (%) | n (%) | n (%) | n (%) | | | |
| Adenovirus | 10 (2.0) | 2 (4.1) | 3 (3.0) | 3 (1.9) | 2 (1.0) | | | |
| Coronavirus | 8 (1.6) | 0 (0.0) | 2 (2.0) | 4 (2.5) | 2 (1.0) | | | |
| Human Metapneumovirus | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Human Rhinovirus/Enterovirus | 188 (36.8) | 24 (49.0) | 49 (48.5) | 62 (38.5) | 53 (26.5) | | | |
| Influenza A | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Influenza A H1 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Influenza A H1-2009 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Influenza A H3 | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Influenza B | 2 (0.4) | 1 (2.0) | 0 (0.0) | 0 (0.0) | 1 (0.5) | | | |
| Parainfluenza Virus 1 | 1 (0.2) | 0 (0.0) | 0 (0.0) | 1 (0.6) | 0 (0.0) | | | |
| Parainfluenza Virus 2 | 13 (2.5) | 2 (4.1) | 4 (4.0) | 3 (1.9) | 4 (2.0) | | | |
| Parainfluenza Virus 3 | 5 (1.0) | 2 (4.1) | 2 (2.0) | 0 (0.0) | 1 (0.5) | | | |
| Parainfluenza Virus 4 | 8 (1.6) | 1 (2.0) | 1 (1.0) | 4 (2.5) | 2 (1.0) | | | |
| RSV A | 8 (1.6) | 0 (0.0) | 8 (7.9) | 0 (0.0) | 0 (0.0) | | | |
| RSV B | 9 (1.8) | 1 (2.0) | 4 (4.0) | 0 (0.0) | 4 (2.0) | | | |
| Chlamydia pneumoniae | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | 0 (0.0) | | | |
| Mycoplasma pneumoniae | 4 (0.8) | 0 (0.0) | 3 (3.0) | 0 (0.0) | 1 (0.5) | | | |
{12}------------------------------------------------
# Summary of Performance Data
# Clinical performance
# Comparator Method
The performance of the ePlex RP Panel was compared to an FDA-cleared multiplexed molecular respiratory pathogen panel and analytically validated PCR tests with bi-directional sequencing for confirmation of RSV subtypes.. Details of the comparator method are described in Table 5.
| Target | Comparator Method |
|-------------------------------|---------------------------------------------------------------------------------------------------------------------------------------|
| Adenovirus | FDA-cleared multiplexed molecular respiratory pathogen<br>panel |
| Coronavirus | |
| Human Metapneumovirus | |
| Human Rhinovirus/Enterovirus | |
| Influenza A | |
| Influenza A H1 | |
| Influenza A H1-2009 | |
| Influenza A H3 | |
| Influenza B | |
| Parainfluenza Virus 1 | |
| Parainfluenza Virus 2 | |
| Parainfluenza Virus 3 | |
| Parainfluenza Virus 4 | |
| Respiratory Syncytial Virus A | FDA-cleared multiplexed molecular respiratory pathogen |
| Respiratory Syncytial Virus B | FDA-cleared multiplexed molecular respiratory pathogen<br>panel followed by a PCR test with bi-directional sequencing<br>confirmation |
| Chlamydia pneumoniae | FDA-cleared multiplexed molecular respiratory pathogen<br>panel |
| Mycoplasma pneumoniae | |
Table 5: Comparator Methods Used to Assess ePlex RP Panel Clinical Performance
# Prospective Clinical Samples
Clinical performance was evaluated in clinical nasopharyngeal swab samples in VTM prospectively-collected at 8 clinical sites in 2 phases. From March 2013 through August 2014, 2218 samples were prospectively-collected and frozen; from September 2016 through October
{13}------------------------------------------------
2016, 514 samples were prospectively-collected and tested fresh (never frozen). A total of 2732 samples were collected across the 2 phases. Prior to the start of investigational testing, 263 samples were withdrawn (251 had sample handling deviations, 9 were tested outside of protocol timelines, 2 had insufficient volume, and 1 had incomplete documentation). Of the 2469 prospectively-collected samples eligible for testing, 2462 were evaluable. Samples with final, valid results and a valid comparator result were considered evaluable. Seven prospectivelycollected samples were not evaluable because they did not have final, valid ePlex RP Panel results and were excluded from performance evaluations. Demographic information for prospectively-collected samples is described in Table 6. Subjects enrolled in this study were from a diverse demographic distribution and represent the intended patient population.
Table 6: Subject Demographic Data for Prospectively-Collected Samples by Collection Site (N=2462)
| | All Sites<br>N=2462<br>n (%) | Site 1<br>N=165<br>n (%) | Site 2<br>N=248<br>n (%) | Site 3<br>N=350<br>n (%) | Site 4<br>N=892<br>n (%) | Site 5<br>N=345<br>n (%) | Site 6<br>N=101<br>n (%) | Site 7<br>N=161<br>n (%) | Site 8<br>N=200<br>n (%) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
|-------------|------------------------------|--------------------------|--------------------------|--------------------------|--------------------------|--------------------------|--------------------------|--------------------------|--------------------------|-----|------------|-----------|----------|-----------|------------|-----------|-----------|---------|-----------|-------|------------|----------|----------|-----------|----------|------------|-----------|----------|----------|--------|------------|----------|---------|-----------|----------|------------|-----------|-----------|----------|---------|------------|-----------|------------|-----------|------------|-----------|---------|-----------|-----------|------|------------|-----------|-----------|-----------|------------|----------|---------|-----------|-----------|
| Sex | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| Male | 1247 (50.6) | 96 (58.2) | 118 (47.6) | 186 (53.1) | 450 (50.4) | 188 (54.5) | 43 (42.6) | 84 (52.2) | 82 (41.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| Female | 1215 (49.4) | 69 (41.8) | 130 (52.4) | 164 (46.9) | 442 (49.6) | 157 (45.5) | 58 (57.4) | 77 (47.8) | 118 (59.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| Age (years) | | | | | | | | | | 0-1 | 388 (15.8) | 17 (10.3) | 21 (8.5) | 74 (21.1) | 164 (18.4) | 45 (13.0) | 28 (27.7) | 3 (1.9) | 36 (18.0) | > 1-5 | 325 (13.2) | 12 (7.3) | 22 (8.9) | 62 (17.7) | 64 (7.2) | 100 (29.0) | 39 (38.6) | 16 (9.9) | 10 (5.0) | > 5-21 | 321 (13.0) | 15 (9.1) | 6 (2.4) | 38 (10.9) | 82 (9.2) | 116 (33.6) | 34 (33.7) | 18 (11.2) | 12 (6.0) | > 21-65 | 926 (37.6) | 87 (52.7) | 131 (52.8) | 98 (28.0) | 385 (43.2) | 55 (15.9) | 0 (0.0) | 92 (57.1) | 78 (39.0) | > 65 | 502 (20.4) | 34 (20.6) | 68 (27.4) | 78 (22.3) | 197 (22.1) | 29 (8.4) | 0 (0.0) | 32 (19.9) | 64 (32.0) |
| Age (years) | | | | | | | | | | 0-1 | 388 (15.8) | 17 (10.3) | 21 (8.5) | 74 (21.1) | 164 (18.4) | 45 (13.0) | 28 (27.7) | 3 (1.9) | 36 (18.0) | > 1-5 | 325 (13.2) | 12 (7.3) | 22 (8.9) | 62 (17.7) | 64 (7.2) | 100 (29.0) | 39 (38.6) | 16 (9.9) | 10 (5.0) | > 5-21 | 321 (13.0) | 15 (9.1) | 6 (2.4) | 38 (10.9) | 82 (9.2) | 116 (33.6) | 34 (33.7) | 18 (11.2) | 12 (6.0) | > 21-65 | 926 (37.6) | 87 (52.7) | 131 (52.8) | 98 (28.0) | 385 (43.2) | 55 (15.9) | 0 (0.0) | 92 (57.1) | 78 (39.0) | > 65 | 502 (20.4) | 34 (20.6) | 68 (27.4) | 78 (22.3) | 197 (22.1) | 29 (8.4) | 0 (0.0) | 32 (19.9) | 64 (32.0) |
| Age (years) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| 0-1 | 388 (15.8) | 17 (10.3) | 21 (8.5) | 74 (21.1) | 164 (18.4) | 45 (13.0) | 28 (27.7) | 3 (1.9) | 36 (18.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| > 1-5 | 325 (13.2) | 12 (7.3) | 22 (8.9) | 62 (17.7) | 64 (7.2) | 100 (29.0) | 39 (38.6) | 16 (9.9) | 10 (5.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| > 5-21 | 321 (13.0) | 15 (9.1) | 6 (2.4) | 38 (10.9) | 82 (9.2) | 116 (33.6) | 34 (33.7) | 18 (11.2) | 12 (6.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| > 21-65 | 926 (37.6) | 87 (52.7) | 131 (52.8) | 98 (28.0) | 385 (43.2) | 55 (15.9) | 0 (0.0) | 92 (57.1) | 78 (39.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
| > 65 | 502 (20.4) | 34 (20.6) | 68 (27.4) | 78 (22.3) | 197 (22.1) | 29 (8.4) | 0 (0.0) | 32 (19.9) | 64 (32.0) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | |
# Prospective Clinical Performance
Positive percent agreement (PPA) was calculated by dividing the number of true positive (TP) results by the sum of TP and false negative (FN) results, while negative percent agreement (NPA) was calculated by dividing the number of true negative (TN) results by the sum of TN and false positive (FP) results. A TP result was one where the detected ePlex RP Panel result matched the detected comparator method result, while a TN result was one where a negative ePlex RP Panel result matched a negative comparator method result. The two-sided 95% confidence interval was also calculated.
{14}------------------------------------------------
A total of 2462 prospectively-collected samples (511 tested fresh and 1951 tested after previously frozen) were evaluated for 17 ePlex RP Panel organisms. PPA and NPA results are summarized by target in Tables 7 and 8 below.
| Organism | Prevalence | TP/TP+FN | PPA (95% CI) | TN/TN+FP | NPA (95% CI) |
|------------------------------|------------|----------|------------------|----------|------------------|
| Adenovirus | 1.6% | 6/8a | 75.0 (40.9-92.9) | 499/503a | 99.2 (98.0-99.7) |
| Coronavirus | 1.4% | 7/7 | 100 (64.6-100) | 503/504 | 99.8 (98.9-100) |
| Human Metapneumovirus | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Human Rhinovirus/Enterovirus | 35.8% | 176/183b | 96.2 (92.3-98.1) | 316/328b | 96.3 (93.7-97.9) |
| Influenza A | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Influenza A H1 | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Influenza A H1-2009 | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Influenza A H3 | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Influenza B | 0.2% | 1/1 | 100 (20.7-100) | 509/510 | 99.8 (98.9-100) |
| Parainfluenza Virus 1 | 0.2% | 1/1 | 100 (20.7-100) | 510/510 | 100 (99.3-100) |
| Parainfluenza Virus 2 | 2.5% | 12/13 | 92.3 (66.7-98.6) | 497/498 | 99.8 (98.9-100) |
| Parainfluenza Virus 3 | 1.0% | 5/5 | 100 (56.6-100) | 506/506 | 100 (99.2-100) |
| Parainfluenza Virus 4 | 0.6% | 3/3 | 100 (43.9-100) | 503/508c | 99.0 (97.7-99.6) |
| RSV A | 1.8% | 8/9 | 88.9 (56.5-98.0) | 501/501 | 100 (99.2-100) |
| RSV B | 2.0% | 9/10 | 90.0 (59.6-98.2) | 500/500 | 100 (99.2-100) |
| Chlamydia pneumoniae | 0.0% | 0/0 | --- | 511/511 | 100 (99.3-100) |
| Mycoplasma pneumoniae | 0.6% | 3/3 | 100 (43.9-100) | 507/508d | 99.8 (98.9-100) |
Table 7: Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) in the ePlex RP Panel Clinical Study (Fresh)
ª Adenovirus was not detected in 2 of 2 FN samples and detected in 4 of 4 FP samples using PCR/sequencing.
b Human rhinovirus/enterovirus was not detected in 1 of 7 FN samples using of 12 FP samples using PCR/sequencing.
& Parainfluenza virus 4 was detected in 3 of 5 FP samples using PCR/sequencing.
d M. pneumoniae was detected in the 1 FP sample using PCR/sequencing.
{15}------------------------------------------------
| Organism | Prevalence | TP/TP+FN | Positive % Agreement<br>PPA (95% CI) | TN/TN+FP | Negative % Agreement<br>NPA (95% CI) |
|------------------------------|------------|----------|--------------------------------------|------------|--------------------------------------|
| Adenovirus | 2.7% | 48/53a | 90.6 (79.7-95.9) | 1874/1898a | 98.7 (98.1-99.1) |
| Coronavirus | 5.6% | 89/110b | 80.9 (72.6-87.2) | 1828/1841b | 99.3 (98.8-99.6) |
| Human Metapneumovirus | 5.8% | 107/113c | 94.7 (88.9-97.5) | 1832/1838c | 99.7 (99.3-99.9) |
| Human Rhinovirus/Enterovirus | 17.2% | 317/336d | 94.3 (91.3-96.4) | 1544/1615d | 95.6 (94.5-96.5) |
| Influenza Ae | 5.7% | 106/111f | 95.5 (89.9-98.1) | 1836/1840f | 99.8 (99.4-99.9) |
| Influenza A H1 | 0.0% | 0/0 | --- | 1951/1951 | 100 (99.8-100) |
| Influenza A H1-2009 | 3.6% | 70/71 | 98.6 (92.4-99.8) | 1874/1880g | 99.7 (99.3-99.9) |
| Influenza A H3 | 1.9% | 34/37h | 91.9 (78.7-97.2) | 1914/1914 | 100 (99.8-100) |
| Influenza B | 3.3% | 58/65i | 89.2 (79.4-94.7) | 1882/1886i | 99.8 (99.5-99.9) |
| Parainfluenza Virus 1 | 1.2% | 23/24 | 95.8 (79.8-99.3) | 1926/1927 | 99.9 (99.7-100) |
| Parainfluenza Virus 2 | 0.5% | 9/9 | 100 (70.1-100) | 1941/1942 | 99.9 (99.7-100) |
| Parainfluenza Virus 3 | 5.3% | 94/104j | 90.4 (83.2-94.7) | 1842/1847j | 99.7 (99.4-99.9) |
| Parainfluenza Virus 4 | 0.3% | 5/5 | 100 (56.6-100) | 1944/1946 | 99.9 (99.6-100) |
| RSV A | 1.6% | 27/31 | 87.1 (71.1-94.9) | 1917/1918 | 99.9 (99.7-100) |
| RSV B | 4.4% | 81/86 | 94.2 (87.1-97.5) | 1861/1863k | 99.9 (99.6-100) |
| Chlamydia pneumoniae | 0.3% | 2/5l | 40.0 (11.8-76.9) | 1945/1946l | 99.9 (99.7-100) |
| Mycoplasma pneumoniae | 0.3% | 4/5m | 80.0 (37.6-96.4) | 1945/1946 | 99.9 (99.7-100) |
#### Table 8: Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) in the ePlex RP Panel Clinical Study (After Previously Frozen)
ే Adenovirus was not detected in 1 of 5 FN samples and detected in 9 of 24 FP samples using PCR/sequencing.
b Coronavirus was not detected in 2 of 21 FN samples and detected in 3 of 13 FP samples using PCR/sequencing.
° Human Metapneumovirus was not detected in 1 of 6 FN samples and detected in 4 of 6 FP samples using PCR/sequencing.
9 Human rhinovirus/enterovirus was not detected in 33 of 71 FP samples using PCR/sequencing.
ື Influenza A comparator results contain 71 samples with A H3, and 3 samples with A H3, and 3 samples with no subtype detected.
် Influenza A was not detected in 1 of 3 FN samples were not tested by PCR/sequencing) and detected in 1 of 4 FP samples using PCR/sequencing.
8 Influenza A H1-2009 was detected in 4 of 6 FP samples using PCR/sequencing.
" Influenza A H3 was not detected in 1 of 3 FN samples using PCR/sequencing.
් Influenza B was not detected in 3 of 7 FN samples and detected in 2 of 4 FP samples using PCR/sequencing.
1 Parainfluenza virus 3 was not detected in 3 of 10 FN samples and detected in 4 of 5 FP samples using PCR/sequencing.
* RSV B was detected in 1 of 2 FP samples using PCR/sequencing.
¹ C. pneumoniae was not detected in 1 of 3 FN samples and detected in the 1 FP sample using PCR/sequencing.
10 M. pneumoniae was not detected in the 1 FN sample using PCR/sequencing.
{16}------------------------------------------------
### Retrospective Clinical Samples
To supplement the number of positives for targets that were not sufficiently represented in the prospective collection, additional nasopharyngeal swab in VTM samples were retrospectively collected from 6 sites. A total of 535 nasopharyngeal swab samples that had previously tested positive for one or more of the target organisms during standard-of-care (SOC) testing were collected and stored frozen. Prior to the start of investigational testing, 11 samples were withdrawn due to noncompliance with the study protocol, and 52 samples were withdrawn because the organisms present had sufficient representation in other samples. In addition, the composition and integrity of the retrospective samples were confirmed with the same comparator method employed in the prospective clinical study (i.e., an FDA-cleared multiplexed respiratory pathogen panel). As the result of this confirmation testing using the comparator method, 26 additional samples were withdrawn because the original SOC testing positive results for the intended organisms were not confirmed when tested with the comparator method. Of the remaining 446 retrospectively-collected samples eligible for testing, all 446 were evaluable. Demographic information for retrospectively-collected samples is described in Table 9. Subjects enrolled in this study were from a diverse demographic distribution and represent the intended patient population.
{17}------------------------------------------------
| | All Sites<br>N=446 | Site 1<br>N=1 | Site 2<br>N=1 | Site 3<br>N=129 | Site 4<br>N=18 | Site 5<br>N=131 | Site 6<br>N=166 |
|-------------|--------------------|---------------|---------------|-----------------|----------------|-----------------|-----------------|
| | n (%) | n (%) | n (%) | n (%) | n (%) | n (%) | n (%) |
| Sex | | | | | | | |
| Male | 232 (52.0) | 0 (0.0) | 1 (100) | 76 (58.9) | 11 (61.1) | 68 (51.9) | 76 (45.8) |
| Female | 214 (48.0) | 1 (100) | 0 (0.0) | 53 (41.1) | 7 (38.9) | 63 (48.1) | 90 (54.2) |
| Age (years) | | | | | | | |
| 0 - 1 | 122 (27.4) | 0 (0.0) | 0 (0.0) | 24 (18.6) | 5 (27.8) | 56 (42.7) | 37 (22.3) |
| > 1 - 5 | 107 (24.0) | 0 (0.0) | 1 (100) | 51 (39.5) | 3 (16.7) | 16 (12.2) | 36 (21.7) |
| > 5 - 21 | 59 (13.2) | 0 (0.0) | 0 (0.0) | 9 (7.0) | 2 (11.1) | 19 (14.5) | 29 (17.5) |
| > 21 - 65 | 99 (22.2) | 1 (100) | 0 (0.0) | 11 (8.5) | 8 (44.4) | 31 (23.7) | 48 (28.9) |
| > 65 | 59 (13.2) | 0 (0.0) | 0 (0.0) | 34 (26.4) | 0 (0.0) | 9 (6.9) | 16 (9.6) |
Table 9: Subject Demographic Data for Retrospectively-Collected Samples by Collection Site (N=446)
### Retrospective Clinical Performance
A total of 446 retrospectively-collected samples were evaluated for 17 ePlex RP Panel organisms. The following specimens with the original positive SOC results for the unintended organisms that were not confirmed by the comparator method were excluded from the performance calculation for the respective organism: 1 coronavirus positive specimen, 3 human rhinovirus/enterovirus positive specimens, 1 influenza A positive specimen, 1 influenza A H3 positive specimen. 1 parainfluenza virus positive specimen. In addition, 5 unintended RSV positive specimens by the comparator method were not confirmed by PCR/sequencing with regard to determining RSV subtypes and therefore were excluded from the performance calculations for RSV A and RSV B. PPA and NPA results are summarized by target in Table 10 below.
| | | Positive % Agreement | Negative % Agreement | |
|------------------------------|----------|----------------------|----------------------|------------------|
| Organism | TP/TP+FN | PPA (95% CI) | TN/TN+FP | NPA (95% CI) |
| Adenovirus | 55/56a | 98.2 (90.6-99.7) | 386/390a | 99.0 (97.4-99.6) |
| Coronavirus | 121/138b | 87.7 (81.2-92.2) | 307/307 | 100 (98.8-100) |
| Human Metapneumovirus | 5/7 | 71.4 (35.9-91.8) | 439/439 | 100 (99.1-100) |
| Human Rhinovirus/Enterovirus | 37/41 | 90.2 (77.5-96.1) | 384/402 | 95.5 (93.0-97.1) |
| Influenza Ac | 75/82d | 91.5 (83.4-95.8) | 363/363 | 100 (99.0-100) |
| Influenza A H1 | 0/0 | --- | 446/446 | 100 (99.1-100) |
| Influenza A H1-2009 | 27/31e | 87.1 (71.1-94.9) | 415/415 | 100 (99.1-100) |
| Influenza A H3 | 45/51f | 88.2 (76.6-94.5) | 394/394 | 100 (99.0-100) |
| Influenza B | 1/1 | 100 (20.7-100) | 445/445 | 100 (99.1-100) |
| Parainfluenza Virus 1 | 43/48g | 89.6 (77.8-95.5) | 396/397 | 99.7 (98.6-100) |
Table 10: Positive Percent Agreement (PPA) and Negative Percent Agreement (NPA) of…
