Who is the manufacturer of BioCode Respiratory Pathogen Panel (RPP)?

Applied BioCode, Inc. filed the 510(k) submission for BioCode Respiratory Pathogen Panel (RPP) (K192485).

What is the FDA product code for BioCode Respiratory Pathogen Panel (RPP)?

OCC. It is classified under 21 CFR 866.3980 as a Class 2 device in the Microbiology panel.

What is the regulatory pathway for BioCode Respiratory Pathogen Panel (RPP)?

510(k) clearance (submission type: Traditional).

What are the predicate devices for BioCode Respiratory Pathogen Panel (RPP)?

BioFire FilmArray Respiratory Pathogen Panel 2 (RP2) (K170604).

Who can help prepare an FDA 510(k) submission like BioCode Respiratory Pathogen Panel (RPP)?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

BioCode Respiratory Pathogen Panel (RPP)

K192485 · Applied BioCode, Inc. · OCC · Dec 23, 2019 · Microbiology

Device Facts

Record ID	K192485
Device Name	BioCode Respiratory Pathogen Panel (RPP)
Applicant	Applied BioCode, Inc.
Product Code	OCC · Microbiology
Decision Date	Dec 23, 2019
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 866.3980
Device Class	Class 2

Intended Use

The BioCode® Respiratory Pathogen Panel (RPP) is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with BioCode MDx-3000 Instrument. The BioCode RPP is capable of the simultaneous detection and identification of nucleic acids from multiple viruses and bacteria extracted from nasopharyngeal swab (NPS) samples obtained from individuals with signs and/or symptoms of respiratory tract infection. The following pathogens and subtypes are identified using the BioCode RPP: Adenovirus, Coronavirus (229E, OC43, HKU1, and NL63), Human Metapneumovirus A/B, Influenza A, including subtypes H1, H1 2009 Pandemic, and H3, Influenza B, Parainfluenza 1, Parainfluenza 2, Parainfluenza 3, Parainfluenza 4, Respiratory Syncytial Virus A/B, Rhinovirus/Enterovirus, Bordetella pertussis, Chlamydia pneumoniae, Mycoplasma pneumoniae. The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of respiratory infection if 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 agent(s) detected by the BioCode RPP 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.

Device Story

BioCode RPP is a multiplexed nucleic acid-based in vitro diagnostic test for respiratory pathogens; used with BioCode MDx-3000 system. Input: nasopharyngeal swab (NPS) samples in transport media. Process: automated nucleic acid extraction (NucliSens easyMAG or Roche MagNA Pure 96); PCR amplification; target capture; signal generation; optical detection using barcoded magnetic beads (BMB) capturing biotinylated products with streptavidin conjugate. Output: qualitative detection/identification of 17 viral/bacterial targets. Used in clinical laboratories by trained personnel. Results aid diagnosis of respiratory infections; must be used with other clinical/epidemiological data. Benefits: simultaneous detection of multiple pathogens from single specimen; aids clinical decision-making for patient management.

Clinical Evidence

Multi-center prospective study (2649 specimens) and retrospective study (165 specimens) compared BioCode RPP to FDA-cleared molecular multiplexed respiratory pathogen panel. Prospective study showed high positive/negative agreement across targets (e.g., Influenza A PA 96.4%, NA 99.0%). Bench testing included LoD, inclusivity, specificity, and interference studies. No clinical data for rare pathogens; supplemented by contrived samples.

Technological Characteristics

Multiplex RT-PCR; probe hybridization; fluorescence detection; barcoded magnetic beads (BMB) for target capture; streptavidin conjugate. Automated system (BioCode MDx-3000). Compatible with NucliSens easyMAG or Roche MagNA Pure 96 extraction. Software-based interpretation algorithm. Class II device.

Indications for Use

Indicated for individuals with signs/symptoms of respiratory tract infection. Used for qualitative detection/identification of 17 viral and bacterial pathogens/subtypes from nasopharyngeal swab (NPS) samples. Not for sole diagnostic use; results must be used with clinical/epidemiological data. Cannot differentiate Rhinovirus/Enterovirus.

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

BioFire FilmArray Respiratory Pathogen Panel 2 (RP2) (K170604)

Related Devices

K233410 — LIAISON PLEX Respiratory Flex Assay · Luminex Corporation · Mar 1, 2024
K143653 — Verigene Respiratory Pathogens Flex Nucleic Acid Test (RP Flex) · Nanosphere, Inc. · Sep 4, 2015
K193167 — NxTAG Respiratory Pathogen Panel, NxTAG Respiratory Pathogen Panel Files, SYNCT Software · Luminex Molecular Diagnostics, Inc. · Dec 11, 2019
K242613 — NxTAG® Respiratory Pathogen Panel · Luminex Molecular Diagnostics, Inc. · Oct 2, 2024

Submission Summary (Full Text)

{0}------------------------------------------------ December 23, 2019 Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue. Applied BioCode, Inc. Robert Tullio Regulatory Consultant 10020 Pioneer Blvd Suite 102 Santa Fe Springs, California 90670 Re: K192485 Trade/Device Name: BioCode Respiratory Pathogen Panel (RPP) Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: Class II Product Code: OCC, OZE, OEP, OEM, OOU, OTG, OZX, OZY, OZZ, NSU Dated: December 5, 2019 Received: December 6, 2019 Dear Robert Tullio: We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmp/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading. If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see {1}------------------------------------------------ https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems. For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100). Sincerely, Tamara Feldblyum, Ph.D. Chief Viral Respiratory and STI Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health Enclosure {2}------------------------------------------------ # Indications for Use 510(k) Number (if known) K192485 Device Name BioCode Respiratory Pathogen Panel (RPP) ### Indications for Use (Describe) The BioCode Respiratory Pathogen Panel (RPP) is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with BioCode MDx 3000 Instrument. The BioCode of the simultaneous detection and identification of nucleic acids from multiple viruses and bacteria extracted from nasopharyngeal swab (NPS) samples obtained from individuals with signs and/or symptoms of respiratory tract infection. The following pathogens and subtypes are identified using the BioCode RPP: - Adenovirus - · Coronavirus (229E, OC43, HKU1, and NL63) - Human Metapneumovirus A/B - · Influenza A, including subtypes H1, H1 2009 Pandemic, and H3 - Influenza B - Parainfluenza Virus 1 - Parainfluenza Virus 2 - · Parainfluenza Virus 3 - · Parainfluenza Virus 4 - · Respiratory Syncytial Virus A/B - Rhinovirus/Enterovirus - · Bordetella pertussis - Chlamydia pneumoniae - Mycoplasma pneumoniae The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of respiratory infection if 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 agent(s) detected by the BioCode RPP 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, the BioCode RPP cannot differentiate them. A positive BioCode RPP Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. The BioCode RPP detects Human Rhinovirus with reduced sensitivity. If a more accurate Rhinovirus result is required, it is recommended that specimens found to be negative for Human Rhinovirus/Enterovirus after examination using BioCode RPP be confirmed by an alternate method (e.g. FDA cleared molecular tests). Performance characteristics for Influenza A were established when Influenza A H1 2009 Pandemic 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. {3}------------------------------------------------ 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. | Type of Use (Select one or both, as applicable) | | |-------------------------------------------------|--| |-------------------------------------------------|--| X Prescription Use (Part 21 CFR 801 Subpart D) | | Over-The-Counter Use (21 CFR 801 Subpart C) ### CONTINUE ON A SEPARATE PAGE IF NEEDED. This section applies only to requirements of the Paperwork Reduction Act of 1995. ### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: > Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {4}------------------------------------------------ # 1.0 510(k) SUMMARY Introduction: According to the requirements of 21 CFR 807.92, the following provides information to understand the basis for a determination of substantial equivalence. ## Submitted by: Applied BioCode®, Inc. 10020 Pioneer Blvd. Suite 102 Santa Fe Springs, CA 90670 ### Contact: Robert Di Tullio Regulatory Consultant rditullio@apbiocode.com Telephone: 310 801 1235 Fax: 323 372 3816 Date Submitted: September 9, 2019 ### Trade Name: BioCode® Respiratory Pathogen Panel (RPP) ### Classification Name and Regulation Number: Respiratory Viral Panel Multiplex Nucleic Acid Assay (21 CFR 866.3980) ### Predicate Device: K170604 – BioFire FilmArray Respiratory Pathogen Panel 2 (RP2) ### Intended Use: # BioCode® Respiratory Pathogen Panel (RPP) The BioCode® Respiratory Pathogen Panel (RPP) is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with BioCode MDx-3000 Instrument. The BioCode RPP is capable of the simultaneous detection and identification of nucleic acids from multiple viruses and bacteria extracted from nasopharyngeal swab (NPS) samples obtained from individuals with signs and/or symptoms of respiratory tract infection. The following pathogens and subtypes are identified using the BioCode RPP: - Adenovirus - Coronavirus (229E, OC43, HKU1, and NL63) - Human Metapneumovirus A/B - Influenza A, including subtypes H1, H1 2009 Pandemic, and H3 - Influenza B - Parainfluenza 1 - Parainfluenza 2 - Parainfluenza 3 - Parainfluenza 4 {5}------------------------------------------------ - Respiratory Syncytial Virus A/B - Rhinovirus/Enterovirus - Bordetella pertussis - Chlamydia pneumoniae - . Mycoplasma pneumoniae The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of respiratory infection if 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 agent(s) detected by the BioCode RPP 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 BioCode RPP cannot differentiate them. A positive BioCode RPP Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. The BioCode RPP detects Human Rhinovirus/Enterovirus with reduced sensitivity. If a more accurate HRV/EV result is required, it is recommended that specimens found to be negative for Human Rhinovirus/Enterovirus after examination using BioCode RPP be confirmed by an alternate method (e.g. FDA cleared molecular tests). Performance characteristics for Influenza A were established when Influenza A H1 2009 Pandemic 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 virulenza 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. {6}------------------------------------------------ # Device Description: The BioCode® Respiratory Pathogen Panel is a respiratory pathogen multiplex nucleic acid test designed for use with the BioCode® MDx-3000 system. The BioCode® MDx-3000 is an automated system that integrates PCR amplification, target capture, signal generation and optical detection for multiple viral and bacterial pathogens from a single nasopharyngeal swab specimen collected in transport media. Specimens are processed and nucleic acids extracted with the NucliSens easyMAG or Roche MagNA Pure 96 automated systems. Once the PCR plate is set up and sealed, all other operations are automated on MDx-3000. The BioCode® RPP simultaneously tests for 17 pathogens and/or subtypes (see table below) from nasopharyngeal swab specimens collected in UTM or VTM. Results from the BioCode RPP test are available within about 5 hours, including off-board nucleic acids extraction. | Viruses | Viruses | |----------------------|------------------------------------------| | Influenza A | Respiratory Syncytial Virus A and B | | • Subtype H1 | Human Metapneumovirus A and B | | • Subtype H1 2009pdm | Rhinovirus/Enterovirus | | • Subtype H3 | Coronavirus (229E, OC43, HKU1, and NL63) | | Influenza B | Adenovirus | | Parainfluenza 1 | Bacteria | | Parainfluenza 2 | Mycoplasma pneumoniae | | Parainfluenza 3 | Chlamydia pneumoniae | | Parainfluenza 4 | Bordetella pertussis | | | Internal Control (MS2) | ### Device Comparison: ### Comparison of the BioCode RPP with the Predicate Device | Characteristic | Proposed Device | Predicate | |----------------|------------------------------------------------------------|---------------------------------------------------------| | Name | BioCode® Respiratory Pathogen Panel (RPP) | BioFire FilmArray Respiratory Pathogen Panel 2 (RP2) | | Common Name | Respiratory Pathogen Panel Multiplex Nucleic acid assay | Respiratory Viral Panel Multiplex Nucleic acid assay | | 510(k) No. | K192485 | K170604 | | Regulation | 21CFR 866.3980 | 21CFR 866.3980 | | Product Code | OCC, OZE, OEM, OOU, OEP, OTG, OZX, OZY, OZZ, NSU | OCC, OEM, OOU, OEP, OTG, OZX, OZY, OZZ, OOI | | Device Class | II | II | ### Similarities {7}------------------------------------------------ Premarket Notification 510(k) | Intended Use | | | |--------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | | The BioCode® Respiratory Pathogen Panel (RPP) is a qualitative multiplexed nucleic acid-based in vitro diagnostic test intended for use with BioCode MDx-3000 Instrument. The BioCode RPP is capable of the simultaneous detection and identification of nucleic acids from multiple viruses and bacteria extracted from nasopharyngeal swab (NPS) samples obtained from individuals with signs and/or symptoms of respiratory tract infection. The following pathogens and subtypes are identified using the BioCode RPP: • Adenovirus • Coronavirus (229E, OC43, HKU1, and NL63) • Human Metapneumovirus A/B • Influenza A, including subtypes H1, H1 2009 Pandemic, and H3 • Influenza B • Parainfluenza 1 • Parainfluenza 2 • Parainfluenza 3 • Parainfluenza 4 • Respiratory Syncytial Virus A/B • Rhinovirus/Enterovirus • Bordetella pertussis • Chlamydia pneumoniae • Mycoplasma pneumoniae | The FilmArray Respiratory Panel 2 (RP2) is a multiplexed nucleic acid test intended for use with FilmArray® 2.0 or FilmArray® Torch systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray RP2: • Adenovirus • Coronavirus 229E • Coronavirus HKU1 • Coronavirus NL63 • Coronavirus OC43 • Human Metapneumovirus • Human Rhinovirus/Enterovirus • Influenza A, including subtypes H1, H1-2009, and H3 • Influenza B • Parainfluenza Virus 1 • Parainfluenza Virus 2 • Parainfluenza Virus 3 • Parainfluenza Virus 4 • Respiratory Syncytial Virus • Bordetella parapertussis (IS1001) • Bordetella pertussis (ptxP) • Chlamydia pneumoniae • Mycoplasma pneumoniae | | | The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of respiratory infection if 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 agent(s) detected by the BioCode RPP may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence and radiography) | The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and/or symptoms of a respiratory infection aids in the diagnosis of respiratory infection if 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 agent(s) detected by the FilmArray RP2 may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and) | | | may be necessary when evaluating a patient with possible respiratory tract infection. Due to the genetic similarity between Human Rhinovirus and Enterovirus, the BioCode RPP cannot differentiate them. A positive BioCode RPP Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. The BioCode RPP detects Human Rhinovirus/Enterovirus with reduced sensitivity. If a more accurate HRV/EV result is required, it is recommended that specimens found to be negative for Human Rhinovirus/Enterovirus after examination using BioCode RPP be confirmed by an alternate method (e.g. FDA cleared molecular tests). Performance characteristics for Influenza A were established when Influenza A H1 2009 Pandemic 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. | radiography) may be necessary when evaluating a patient with possible respiratory tract infection. Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray RP2 cannot reliably differentiate them. A positive FilmArray RP2 Rhinovirus/Enterovirus result should be followed up using an alternate method (e.g., cell culture or sequence analysis) if differentiation is required. Performance characteristics for Influenza A were established when Influenza A H1-2009, A H1, 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. | | Sample Type | Nasopharyngeal swab in Viral or Universal Transport Media | Same | | Controls | Externally Sourced | Same | | Differences | | | | Methodology | Multiplex RT-PCR in a single reaction and probe hybridization followed by fluorescence detection and decoding of barcoded magnetic beads (BMB) that capture biotinylated products with streptavidin conjugate | Nested multiplex PCR executed in two stages. First, a single, large volume, highly multiplexed reverse transcription PCR (RT- PCR) reaction. Second, nested PCR, is performed in singleplex fashion in each well of the array. Followed by fluorescent detection of images of the PCR reactions. | {8}------------------------------------------------ {9}------------------------------------------------ ## Summary of Performance Characteristics of the BioCode RPP® ### Clinical Performance ### Study Overview The clinical performance of the BioCode RPP was established in a multi-center study conducted during periods of the 2017-2019 respiratory illness seasons. Residual (leftover) and de-identified nasopharyngeal swab (NPS) specimens in VTM or UTM that were prospectively collected from patients suspected of respiratory tract infections at five geographically diverse clinical sites in the U.S. were enrolled and tested with the BioCode RPP at five testing sites during the prospective clinical study. The enrolled prospective specimens were tested freshly with an FDA-cleared molecular multiplexed respiratory pathogen panel as part of the Stand of Care (SOC), and were either tested freshly with the BioCode RPP (i.e., specimens that were stored in a 2-8°C refrigerator for no more than 7 days), or stored frozen and then thawed and tested with the BioCode RPP at a testing site at a later date (i.e., specimens that were initially stored in a 2-8°C refrigerator but were not able to be tested by the BioCode RPP within 7 days from specimen collection). A waiver of the informed consent requirement was obtained from the Institutional Review Boards (IRBs) at each specimen enrollment site for the use of residual NPS in VTM or UTM specimens. The following information was recorded on the Case Report Form (CRF) for each subject from whom a specimen was enrolled: - · Age and sex - Date and time of specimen collection - Standard of care (SOC) comparator test result - Specimen storage status, i.e., fresh or frozen A total of 2654 residual NPS specimens in VTM that were prospectively collected at the five clinical sites from August 2017 to May 2019 were enrolled initially for the clinical study. Five specimens were withdrawn from the clinical study due to incomplete data collection and testing, resulted in a total of 2649 prospective specimens (1401 fresh and 1248 frozen specimens) that were included in the prospective clinical study. The prospective specimens enrolled for evaluation were tested at the five testing sites by trained laboratory personnel. DNA/RNA was extracted using either the BioMerieux NucliSENS easyMAG system or Roche MagNA Pure 96 system. After extraction, the samples were tested using the BioCode RPP on the BioCode MDx-3000 System according to the instructions for use. | Characteristic | Prospective Study Specimens | |------------------|-----------------------------| | Total Specimens | 2649 | | Gender (n/N (%)) | | | Male | 1346/2649 (50.8%) | ### Demographics - Prospective Samples {10}------------------------------------------------ | Characteristic | Prospective Study Specimens | |------------------------|-----------------------------| | Female | 1303/2649 (49.2%) | | Age Category (n/N (%)) | | | 0-5 yrs | 1004/2649 (37.9%) | | 6-21 yrs | 609/2649 (23.0%) | | 22-59 yrs | 531/2649 (20.0%) | | 60+ yrs | 505/2649 (19.1%) | | Status (n/N (%))a | | | Inpatient | 1555/2646 (58.8%) | | Outpatient | 1091/2646 (41.2%) | a - Inpatient/outpatient status was unavailable for 3 specimens. | | | Storage Method | | Extraction Method | | |---------|----------------|----------------|--------|-------------------|---------------| | Site | Samples Tested | Fresh | Frozen | easyMAG | MagNA Pure 96 | | Site 01 | 530 | 250 | 280 | 530 | 0 | | Site 02 | 419 | 182 | 237 | 0 | 419 | | Site 03 | 600 | 366 | 234 | 600 | 0 | | Site 04 | 550 | 300 | 250 | 550 | 0 | | Site 05 | 550 | 303 | 247 | 0 | 550 | | Total: | 2649 | 1401 | 1248 | 1680 | 969 | #### Prospective Sample Type and Test Method Breakdown Performance of the BioCode RPP was evaluated by comparing the BioCode RPP test results with those from an FDA-cleared molecular multiplexed respiratory pathogen panel. Positive agreement was calculated as TP/(TP + FN). TP = true positive by both the comparator test and BioCode RPP; FN = false negative or negative by BioCode RPP only. Negative agreement was calculated as TN/(TN + FP). TN = true negative or negative by both the comparator test and BioCode RPP; FP = false positive or positive by BioCode RPP only. The two-sided 95% confidence interval was calculated with Score method (per CLSI EP12-A2). Samples for which false positive and/or false negative results (i.e., discrepant results) were obtained when comparing the BioCode RPP results to the comparator test results were further investigated. The discrepancy investigation was mainly conducted by performing independent molecular tests, including analytically validated PCR followed by bi-directional sequencing assays and alternate NAATs. Of the 2649 specimens included in the prospective clinical study, two specimens, one fresh and one frozen specimen, obtained a final "invalid" result from the BioCode RPP, and were excluded from the performance analyses for all analytes. In addition, three specimens, one fresh and two frozen specimens, obtained a final influenza A "indeterminate" result by the BioCode RPP, and two specimens, one fresh and one frozen, obtained an influenza A "equivocal" result from the comparator test. They were included in the performance analyses for all analytes but excluded from the performance {11}------------------------------------------------ calculations for Flu A and Flu A subtypes. Furthermore, two frozen specimens obtained a valid influenza A result from the comparator test without the accompanying Flu A subtyping results. They were included in the performance analyses for all analytes but excluded from the performance calculations for Flu A subtypes. The Prospective study results stratified by storage condition are presented in the table below. | | | | Positive Agreement | | Negative Agreement | | |------------------------------------|---------|------|--------------------|----------------|--------------------|----------------| | Target | Storage | (n) | PA (%) | 95% Cl | NA (%) | 95% Cl | | Adenovirusa | Fresh | 1400 | 31/40 (77.5%) | (62.5%, 87.7%) | 1340/1360 (98.5%) | (97.7%, 99.0%) | | | Frozen | 1247 | 37/38 (97.4%) | (86.5%, 99.5%) | 1188/1209 (98.3%) | (97.4%, 98.9%) | | | Total | 2647 | 68/78 (87.2%) | (78.0%, 92.9%) | 2528/2569 (98.4%) | (97.8%, 98.8%) | | Bordetella pertussisb | Fresh | 1400 | 1/1 (100%) | (20.7%, 100%) | 1387/1399 (99.1%) | (98.5%, 99.5%) | | | Frozen | 1247 | 1/1 (100%) | (20.7%, 100%) | 1239/1246 (99.4%) | (98.8%, 99.7%) | | | Total | 2647 | 2/2 (100%) | (34.2%, 100%) | 2626/2645 (99.3%) | (98.9%, 99.5%) | | Chlamydia pneumoniaec | Fresh | 1400 | 2/2 (100%) | (34.2%, 100%) | 1397/1398 (99.9%) | (99.6%, 100%) | | | Frozen | 1247 | 2/2 (100%) | (34.2%, 100%) | 1245/1245 (100%) | (99.7%, 100%) | | | Total | 2647 | 4/4 (100%) | (51.0%, 100%) | 2642/2643 (100%) | (99.8%, 100%) | | Coronavirusd | Fresh | 1400 | 35/50 (70%) | (56.2%, 80.9%) | 1338/1350 (99.1%) | (98.5%, 99.5%) | | | Frozen | 1247 | 76/83 (91.6%) | (83.6%, 95.9%) | 1154/1164 (99.1%) | (98.4%, 99.5%) | | | Total | 2647 | 111/133 (83.5%) | (76.2%, 88.8%) | 2492/2514 (99.1%) | (98.7%, 99.4%) | | Human Metapneumoviruse | Fresh | 1400 | 89/93 (95.7%) | (89.5%, 98.3%) | 1299/1307 (99.4%) | (98.8%, 99.7%) | | | Frozen | 1247 | 46/49 (93.9%) | (83.5%, 97.9%) | 1189/1198 (99.2%) | (98.6%, 99.6%) | | | Total | 2647 | 135/142 (95.1%) | (90.2%, 97.6%) | 2488/2505 (99.3%) | (98.9%, 99.6%) | | Human Rhinovirus/Enterovirusf | Fresh | 1400 | 221/261 (84.7%) | (79.8%, 88.5%) | 1119/1139 (98.2%) | (97.3%, 98.9%) | | | Frozen | 1247 | 162/213 (76.1%) | (69.9%, 81.3%) | 1020/1034 (98.6%) | (97.7%, 99.2%) | | | Total | 2647 | 383/474 (80.8%) | (77.0%, 84.1%) | 2139/2173 (98.4%) | (97.8%, 98.9%) | | Influenza Ag | Fresh | 1398 | 115/120 (95.8%) | (90.6%, 98.2%) | 1265/1278 (99.0%) | (98.3%, 99.4%) | | | Frozen | 1244 | 98/101 (97.0%) | (91.6%, 99.0%) | 1131/1143 (99.0%) | (98.2%, 99.4%) | | | Total | 2642 | 213/221 (96.4%) | (93.0%, 98.2%) | 2396/2421 (99.0%) | (98.5%, 99.3%) | | Influenza A H1 | Fresh | 1398 | N/A+ | N/A+ | 1398/1398 (100%) | (99.7%, 100%) | | | Frozen | 1242 | N/A+ | N/A+ | 1242/1242 (100%) | (99.7%, 100%) | | | Total | 2640 | N/A+ | N/A+ | 2640/2640 (100%) | (99.9%, 100%) | | Influenza A H1 2009pdmh | Fresh | 1398 | 29/30 (96.7%) | (83.3%, 99.4%) | 1365/1368 (99.8%) | (99.4%, 99.9%) | | | Frozen | 1242 | 23/23 (100%) | (85.7%, 100%) | 1213/1219 (99.5%) | (98.9%, 99.8%) | | | Total | 2640 | 52/53 (98.1%) | (90.1%, 99.7%) | 2578/2587 (99.7%) | (99.3%, 99.8%) | | | | | Positive Agreement | | Negative Agreement | | | Target | Storage | (n) | PA (%) | 95% CI | NA (%) | 95% CI | | Influenza A H3 | Fresh | 1398 | 82/88 (93.2%) | (85.9%, 96.8%) | 1306/1310 (99.7%) | (99.2%, 99.9%) | | | Frozen | 1242 | 65/69 (94.2%) | (86.0%, 97.7%) | 1168/1173 (99.6%) | (99.0%, 99.8%) | | | Total | 2640 | 147/157 (93.6%) | (88.7%, 96.5%) | 2474/2483 (99.6%) | (99.3%, 99.8%) | | Influenza B | Fresh | 1400 | 7/7 (100%) | (64.6%, 100%) | 1388/1393 (99.6%) | (99.2%, 99.8%) | | | Frozen | 1247 | 44/47 (93.6%) | (82.8%, 97.8%) | 1191/1200 (99.2%) | (98.6%, 99.6%) | | | Total | 2647 | 51/54 (94.4%) | (84.9%, 98.1%) | 2579/2593 (99.5%) | (99.1%, 99.7%) | | Mycoplasma pneumoniae | Fresh | 1400 | 8/8 (100%) | (67.6%, 100%) | 1381/1392 (99.2%) | (98.6%, 99.6%) | | | Frozen | 1247 | 10/10 (100%) | (72.2%, 100%) | 1228/1237 (99.3%) | (98.6%, 99.6%) | | | Total | 2647 | 18/18 (100%) | (82.4%, 100%) | 2609/2629 (99.2%) | (98.8%, 99.5%) | | Parainfluenza Virus 1 | Fresh | 1400 | 4/4 (100%) | (51.0%, 100%) | 1396/1396 (100%) | (99.7%, 100%) | | | Frozen | 1247 | 11/13 (84.6%) | (57.8%, 95.7%) | 1234/1234 (100%) | (99.7%, 100%) | | | Total | 2647 | 15/17 (88.2%) | (65.7%, 96.7%) | 2630/2630 (100%) | (99.9%, 100%) | | Parainfluenza Virus 2 | Fresh | 1400 | 2/3 (66.7%) | (20.8%, 93.9%) | 1396/1397 (99.9%) | (99.6%, 100%) | | | Frozen | 1247 | 8/9 (88.9%) | (56.5%, 98.0%) | 1236/1238 (99.8%) | (99.4%, 100%) | | | Total | 2647 | 10/12 (83.3%) | (55.2%, 95.3%) | 2632/2635 (99.9%) | (99.7%, 100%) | | Parainfluenza Virus 3 | Fresh | 1400 | 77/79 (97.5%) | (91.2%, 99.3%) | 1312/1321 (99.3%) | (98.7%, 99.6%) | | | Frozen | 1247 | 41/43 (95.3%) | (84.5%, 98.7%) | 1196/1204 (99.3%) | (98.7%, 99.7%) | | | Total | 2647 | 118/122 (96.7%) | (91.9%, 98.7%) | 2508/2525 (99.3%) | (98.9%, 99.6%) | | Parainfluenza Virus 4 | Fresh | 1400 | 1/1 (100%) | (20.7%, 100%) | 1399/1399 (100%) | (99.7%, 100%) | | | Frozen | 1247 | 15/17 (88.2%) | (65.7%, 96.7%) | 1228/1230 (99.8%) | (99.4%, 100%) | | | Total | 2647 | 16/18 (88.9%) | (67.2%, 96.9%) | 2627/2629 (99.9%) | (99.7%, 100%) | | Respiratory Syncytial Virus | Fresh | 1400 | 91/93 (97.8%) | (92.5%, 99.4%) | 1293/1307 (98.9%) | (98.2%, 99.4%) | | | Frozen | 1247 | 109/111 (98.2%) | (93.7%, 99.5%) | 1129/1136 (99.4%) | (98.7%, 99.7%) | | | Total | 2647 | 200/204 (98.0%) | (95.1%, 99.2%) | 2422/2443 (99.1%) | (98.7%, 99.4%) | Table. Summary of Clinical Study results: Prospective specimens stratified by storage condition. {12}------------------------------------------------ * No positive reference results recorded a – Adenovirus: The 10 FNs were not detected by PCR/bi-directional sequencing or alternative NAAT. Of 41 FPs, 37 were not detected and 4 were indeterminate by PCR/bi-directional sequencing. b – Bordetella pertussis: Of 19 FPs, 4 were detectional sequencing, 3 were indeterminate and 12 were not detected by PCR/bi-directional sequencing. c – Chlamydia pneumoniae: The 1 FP was detected by PCR/bi-directional sequencing. d – Coronavirus: Of 22 FNs, 5 were detectional sequencing. 2 were not detected by PCR/bi-directional sequencing but detected by alternative NAAT. 12 were not detected by either alternative NAAT or PCR/bi-directional sequencing. 3 were not detected by PCR/bi-directional sequencing and were not tested by alternative NAAT. The 22 FPs were not detectional sequencing. e – Human Metapneumovirus: Of 7 FNs, 3 were detected by PCR/bi-directional sequencing. Of 17 FPs, 7 were detected while 10 were not detected by PCR/bi-directional sequencing. f – Human Rhinovirus/Enterovirus: Of 91 FNs, 26 were indeterminate by PCR/bi-directional sequencing. 25 were not detected by PCR/bi-directional sequencing but were detected by alternative NAAT. 11 were not detectional {13}------------------------------------------------ sequencing or by alternative NAAT. 27 were not detectional sequencing and had insufficient volume for alternative NAAT. Of 34 FPs, 8 were detected and 26 were not detected by PCR/bi-directional sequencing. g – Influenza A: Of the 8 FNs, 7 were not detectional sequencing. 1 had insufficient volume for follow-up testing. Of 25 FPS, 5 were detected by PCR/bi-directional sequencing, 19 were not detectional sequencing, and 1 had insufficient volume for follow-up testing. h – Influenza A H1 2009pdm: The 1 FN had insufficient volume for follow-up testing. Of 9 FPs, 4 were detectional sequencing, and 5 were not detected by PCR/bi-directional sequencing. i – Influenza A H3: Of 10 FNs, 7 were not detected by PCR/bi-directional sequencing. 2 had insufficient volume for followup testing. Of 9 FPs, 3 were detected, 1 was indeterminate, and 4 were not detectional sequencing. 1 had insufficient volume for follow-up testing. j – Influenza B: The 3 FNs were not detected by PCR/bi-directional sequencing. Of 14 FPs, 1 was detected by PCR/bidirectional sequencing, and 1 had insufficient volume for follow-up testing. k – Mycoplasma pneumonioe: Of 20 FPs, 7 were detected, and 1 was invalid by PCR/bi-directional sequencing. 2 had insufficient volume for follow-up testing. I – Parainfluenza Virus 1: The 2 FNs were not detected by PCR/bi-directional sequencing. m – Parainfluenza Virus 2: Of 2 FNs, 1 was detected by PCR/bi-directional sequencing. The 3 FPs were detected by PCR/bi-directional sequencing. n – Parainfluenza Virus 3: Of 4 FNs, 2 were detected, and 1 was indeterminate, by PCR/bi-directional sequencing. Of 17 FPs, 8 were detected, 8 were not detected, and 1 was indeterminate, by PCR/bi-directional sequencing. o – Parainfluenza Virus 4: The 2 FNs were detected by PCR/bi-directional sequencing. The 2 FPs were detectional sequencing. p – Respiratory Syncytial Virus: The 4 FNs were not detectional sequencing. Of 21 FPs, 18 were not detected, 2 were detected, and 1 was indeterminate, by PCR/bi-directional sequencing. Prospective prevalence as detected by BioCode® RPP, stratified by age or study site, are presented in tables below. #### Table. Prospective prevalence detected by BioCode® RPP stratified by Age | Analyte | Overall (N=2647) | ≤5 yrs (N=1004) | 6-21 yrs (N=609) | 22-59 yrs (N=531) | 60+ yrs (N=503) | |------------------------------|---------------------|--------------------|---------------------|----------------------|--------------------| | Adenovirus | 109 (4.1%) | 80 (8.0%) | 20 (3.3%) | 5 (0.9%) | 4 (0.8%) | | Bordetella pertussis | 21 (0.8%) | 8 (0.8%) | 12 (2.0%) | 0 (0.0%) | 1 (0.2%) | | Chlamydia pneumoniae | 5 (0.2%) | 1 (0.1%) | 3 (0.5%) | 1 (0.2%) | 0 (0.0%) | | Coronavirus | 133 (5.0%) | 63 (6.3%) | 27 (4.4%) | 19 (3.6%) | 24 (4.8%) | | Human Metapneumovirus | 152 (5.7%) | 94 (9.4%) | 26 (4.3%) | 15 (2.8%) | 17 (3.4%) | | Human Rhinovirus/Enterovirus | 417 (15.8%) | 234 (23.3%) | 101 (16.6%) | 51 (9.6%) | 31 (6.2%) | | Influenza A | 238 (9.0%) | 71 (7.1%) | 84 (13.8%) | 47 (8.9%) | 36 (7.2%) | | Influenza A H1 | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | | Influenza A H1 2009pdm | 62 (2.3%) | 24 (2.4%) | 15 (2.5%) | 15 (2.8%) | 8 (1.6%) | | Influenza A H3 | 157 (5.9%) | 45 (4.5%) | 61 (10.0%) | 26 (4.9%) | 25 (5.0%) | | Influenza B | 65 (2.5%) | 13 (1.3%) | 26 (4.3%) | 15 (2.8%) | 11 (2.2%) | | Mycoplasma pneumoniae | 38 (1.4%) | 9 (0.9%) | 21 (3.4%) | 6 (1.1%) | 2 (0.4%) | | Parainfluenza Virus 1 | 15 (0.6%) | 4 (0.4%) | 1 (0.2%) | 4 (0.8%) | 6 (1.2%) | | Parainfluenza Virus 2 | 13 (0.5%) | 6 (0.6%) | 4 (0.7%) | 3 (0.6%) | 0 (0.0%) | | Parainfluenza Virus 3 | 135 (5.1%) | 74 (7.4%) | 21 (3.4%) | 21 (4.0%) | 19 (3.8%) | {14}------------------------------------------------ | Analyte | Overall (N=2647) | ≤5 yrs (N=1004) | 6-21 yrs (N=609) | 22-59 yrs (N=531) | 60+ yrs (N=503) | |-----------------------------|---------------------|--------------------|---------------------|----------------------|--------------------| | Parainfluenza Virus 4 | 18 (0.7%) | 12 (1.2%) | 1 (0.2%) | 2 (0.4%) | 3 (0.6%) | | Respiratory Syncytial Virus | 221 (8.3%) | 156 (15.5%) | 30 (4.9%) | 19 (3.6%) | 16 (3.2%) | ### Table. Prospective prevalence detected by BioCode® RPP stratified by site. | Analyte | Overall (N=2647) | Site 1 (N=529) | Site 2 (N=419) | Site 3 (N=599) | Site 4 (N=550) | Site 5 (N=550) | |------------------------------|---------------------|-------------------|-------------------|-------------------|-------------------|-------------------| | Adenovirus | 109 (4.1%) | 8 (1.5%) | 27 (6.4%) | 21 (3.5%) | 17 (3.1%) | 36 (6.5%) | | Bordetella pertussis | 21 (0.8%) | 0 (0.0%) | 19 (4.5%) | 1 (0.2%) | 0 (0.0%) | 1 (0.2%) | | Chlamydia pneumoniae | 5 (0.2%) | 0 (0.0%) | 3 (0.7%) | 0 (0.0%) | 1 (0.2%) | 1 (0.2%) | | Coronavirus | 133 (5.0%) | 36 (6.8%) | 21 (5.0%) | 14 (2.3%) | 26 (4.7%) | 36 (6.5%) | | Human Metapneumovirus | 152 (5.7%) | 18 (3.4%) | 22 (5.3%) | 26 (4.3%) | 38 (6.9%) | 48 (8.7%) | | Human Rhinovirus/Enterovirus | 417 (15.8%) | 48 (9.1%) | 83 (19.8%) | 65 (10.9%) | 125 (22.7%) | 96 (17.5%) | | Influenza A | 238 (9.0%) | 49 (9.3%) | 50 (11.9%) | 35 (5.8%) | 41 (7.5%) | 63 (11.5%) | | Influenza A H1 | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | 0 (0.0%) | | Influenza A H1 2009pdm | 62 (2.3%) | 13 (2.5%) | 7 (1.7%) | 11 (1.8%) | 22 (4.0%) | 9 (1.6%) | | Influenza A H3 | 157 (5.9%) | 32 (6.0%) | 40 (9.5%) | 19 (3.2%) | 17 (3.1%) | 49 (8.9%) | | Influenza B | 65 (2.5%) | 12 (2.3%) | 32 (7.6%) | 15 (2.5%) | 2 (0.4%) | 4 (0.7%) | | Mycoplasma pneumoniae | 38 (1.4%) | 4 (0.8%) | 15 (3.6%) | 5 (0.8%) | 3 (0.5%) | 11 (2.0%) | | Parainfluenza Virus 1 | 15 (0.6%) | 4 (0.8%) | 0 (0.0%) | 11 (1.8%) | 0 (0.0%) | 0 (0.0%) | | Parainfluenza Virus 2 | 13 (0.5%) | 0 (0.0%) | 0 (0.0%) | 3 (0.5%) | 8 (1.5%) | 2 (0.4%) | | Parainfluenza Virus 3 | 135 (5.1%) | 12 (2.3%) | 28 (6.7%) | 41 (6.8%) | 9 (1.6%) | 45 (8.2%) | | Parainfluenza Virus 4 | 18 (0.7%) | 0 (0.0%) | 1 (0.2%) | 9 (1.5%) | 5 (0.9%) | 3 (0.5%) | | Respiratory Syncytial Virus | 221 (8.3%) | 21 (4.0%) | 35 (8.4%) | 25 (4.2%) | 49 (8.9%) | 91 (16.5%) | The overall success rate for initial specimen testing in the prospective study was 98.8% (2618/2649) (95% Cl: 98.3% - 99.2%); 31 tests were unsuccessful (26 tests with an invalid result and 5 tests due to low BMB count/instrument error). Upon a single retest per the instructions for use, 29 of the 31 initially unsuccessful specimens generated a valid result. The final validity rate was 99.9% (2647/2649) (95% Cl: 99.7%-100%). {15}------------------------------------------------ There were 193 samples with mixed infections by the BioCode® RPP in the prospective clinical study (193/2649 or 7.3%). The distribution, prevalence and most common co-infections combinations detected by BioCode RPP in the prospective clinical study are summarized in the tables below. Table. Distribution of co-infection combinations detected by BioCode® RPP from prospective clinical study | Analytes Detected Simultaneously | Number of Specimen | |-------------------------------------|--------------------| | 2 | 168 (87.0%) | | 3 | 24 (12.4%) | | 5 | 1 (0.5%) | | Total Co-Infections | 193 | Table. Prevalence of targets in co-infection combinations detected by BioCode® RPP from prospective clinical study | Analyte | Prevalence in Co-Infections (N=193) | |------------------------------|-------------------------------------| | Adenovirus | 51 (26.4%) | | Bordetella pertussis | 14 (7.3%) | | Chlamydia pneumoniae | 1 (0.5%) | | Coronavirus | 47 (24.4%) | | Human Metapneumovirus | 35 (18.1%) | | Human Rhinovirus/Enterovirus | 101 (52.3%) | | Influenza A | 37 (19.2%) | | Influenza B | 8 (4.1%) | | Mycoplasma pneumoniae | 7 (3.6%) | | Parainfluenza Virus 1 | 2 (1.0%) | | Parainfluenza Virus 2 | 4 (2.1%) | | Parainfluenza Virus 3 | 39 (20.2%) | | Parainfluenza Virus 4 | 8 (4.1%) | | Respiratory Syncytial Virus | 59 (30.1%) | Table. Most prevalent multiple detection combinations (5 or more instances) detected by BioCode® RPP from prospective clinical study. | Co-Infection Combination | Number of Specimen | |------------------------------------------------------------|--------------------| | Human Rhinovirus/Enterovirus + Respiratory Syncytial Virus | 17 | | Adenovirus + Human Rhinovirus/Enterovirus | 13 | | Human Metapneumovirus + Human Rhinovirus/Enterovirus | 13 | | Human Rhinovirus/Enterovirus + Influenza A | 11 | | Adenovirus + Respiratory Syncytial Virus | 10 | | Human Rhinovirus/Enterovirus + Parainfluenza Virus 3 | 10 | | Coronavirus + Human Rhinovirus/Enterovirus | 8 | | Coronavirus + Respiratory Syncytial Virus | 7 | | Bordetella pertussis + Human Rhinovirus/Enterovirus | 6 | {16}------------------------------------------------ | Co-Infection Combination | Number of Specimen | |-------------------------------------|--------------------| | Adenovirus + Parainfluenza Virus 3 | 5 | | Coronavirus + Human Metapneumovirus | 5 | | Coronavirus + Influenza A | 5 | ### Testing of Preselected Archived Specimens Some of the pathogens on the BioCode RPP were of low prevalence and were not encountered in sufficiently large numbers during the prospective study to adequately demonstrate system performance. To supplement the results of the prospective clinical study, an evaluation of preselected archived retrospective specimens was performed. These specimens were archived NPS in VTM or UTM specimens that were selected because they had previously tested positive for one of the following pathogens at the source laboratory: coronavirus 229E, coronavirus HKU1, parainfluenza virus 1, parainfluenza virus 2, parainfluenza virus 4, Bordetella pertussis, Chlamydia pneumoniae, and Mycoplasma pneumoniae, or had been negative in previous laboratory testing. A total of 165 clinical specimens were enrolled for testing in this retrospective study. The specimens were randomized such that the users performing the BioCode RPP assay were blinded to the expected test result and shipped to one of two of the testing sites participated in the prospective clinical study for testing. A summary of the demographic information of the tested samples is provided in the following table: | Characteristic | Archived Study Specimens | |------------------------|--------------------------| | Total Specimens | 165 | | Gender (n/N (%)) | | | Male | 96/165 (58.2%) | | Female | 69/165 (41.8%) | | Age Category (n/N (%)) | | | 0-5 yrs | 77/165 (46.7%) | | 6-21 yrs | 58/165 (35.2%) | | 22-59 yrs | 15/165 (9.1%) | | 60+ yrs | 15/165 (9.1%) | # Demographics of Archived Specimens The performance of the BioCode RPP was evaluated by comparing the BioCode RPP test results with those from an FDA-cleared molecular multiplexed respiratory pathogen panel, the same panel test as the one used as the comparator in the prospective clinical study. The BioCode RPP retrospective performance data expressed as positive percent and negative percent agreements against the comparator method are presented by pathogen in the table below. {17}------------------------------------------------ | Target | (n) | PA (%) | 95% CI | NA (%) | 95% CI | |-------------------------------|-----|---------------|----------------|-----------------|----------------| | Adenovirusa | 165 | 7/7 (100%) | (64.6%, 100%) | 155/158 (98.1%) | (94.6%, 99.4%) | | Bordetella pertussisb | 165 | 10/10 (100%) | (72.2%, 100%) | 144/155 (92.9%) | (87.7%, 96.0%) | | Chlamydia pneumoniae | 165 | 10/10 (100%) | (72.2%, 100%) | 155/155 (100%) | (97.6%, 100%) | | Coronavirusc | 165 | 52/59 (88.1%) | (77.5%, 94.1%) | 99/106 (93.4%) | (87.0%, 96.8%) | | Human Metapneumovirus | 165 | 4/4 (100%) | (51.0%, 100%) | 161/161 (100%) | (97.7%, 100%) | | Human Rhinovirus/Enterovirusd | 165 | 16/23 (69.6%) | (49.1%, 84.4%) | 141/142 (99.3%) | (96.1%, 99.9%) | | Influenza A | 165 | N/A+ | N/A+ | 165/165 (100%) | (97.7%, 100%) | | Influenza A H1 | 165 | N/A+ | N/A+ | 165/165 (100%) | (97.7%, 100%) | | Influenza A H1 2009pdm | 165 | N/A+ | N/A+ | 165/165 (100%) | (97.7%, 100%) | | Influenza A H3 | 165 | N/A+ | N/A+ | 165/165 (100%) | (97.7%, 100%) | | Influenza Be | 165 | 2/3 (66.7%) | (20.8%, 93.9%) | 162/162 (100%) | (97.7%, 100%) | | Mycoplasma pneumoniaef | 165 | 7/7 (100%) | (64.6%, 100%) | 153/158 (96.8%) | (92.8%, 98.6%) | | Parainfluenza Virus 1g | 165 | 12/13 (92.3%) | (66.7%, 98.6%) | 152/152 (100%) | (97.5%, 100%) | | Parainfluenza Virus 2h | 165 | 19/20 (95%) | (76.4%, 99.1%) | 144/145 (99.3%) | (96.2%, 99.9%) | | Parainfluenza Virus 3 | 165 | 1/1 (100%) | (20.7%, 100%) | 164/164 (100%) | (97.7%, 100%) | | Parainfluenza Virus 4i | 165 | 14/15 (93.3%) | (70.2%, 98.8%) | 150/150 (100%) | (97.5%, 100%) | | Respiratory Syncytial Virusj | 165 | 11/12 (91.7%) | (64.6%, 98.5%) | 152/153 (99.3%) | (96.4%, 99.9%) | #### Table. Results from Archived Specimens tested by the BioCode® RPP with easyMAG extraction system. * No positive reference results recorded a - Adenovirus: Of 3 FPs, 1 was detected by PCR/bi-directional sequencing and 2 were not detected by PCR/bi-directional sequencing. b - Bordetella pertussis: Of 11 FPs, 7 were detected by PCR/bi-directional sequencing and 4 were not detected by PCR/bidirectional sequencing. c - Coronavirus: Of 7 FNs, 4 were detected by PCR/bi-directional sequencing while 3 were not detectional sequencing. The 7 FPs were not detected by PCR/bi-directional sequencing. All had low MFls (<620) near the MFL cut off on initial BioCode RPP results. d - Human Rhinovirus/Enterovirus: The 7 FNs were not detected by PCR/bi-directional sequencing. The 1 FP was not detected by PCR/bi-directional sequencing. e - Influenza B: 1 FN was not detected by PCR/bi-directional sequencing. f - Mycoplasma pneumoniae: Of 5 FPs, 3 were detected by PCR/bi-directional sequencing and 2 were not detected by PCR/bi-directional sequencing. g - Parainfluenza Virus 1: 1 FN was not detected by PCR/bi-directional sequencing. h - Parainfluenza Virus 2: 1 FN was not detected by PCR/bi-directional sequencing. 1 FP was not detected by PCR/bidirectional sequencing. i - Parainfluenza Virus 4: 1 FN was not detected by PCR/bi-directional sequencing. j - Respiratory Syncytial Virus: 1 FN was not detected by PCR/bi-directional sequencing. 1 FP was not detected by PCR/bidirectional sequencing. {18}------------------------------------------------ # Testing of Contrived Specimens Some analytes are so rare that both prospective and archived specimen collection efforts were insufficient to demonstrate the clinical performance. To supplement the prospective and archived data, an evaluation of contrived specimens was performed for two pathogens: Chlamydia pneumoniae and Influenza A H1. These contrived clinical specimens were prepared using 50 unique natural NPS in VTM or UTM specimens that were previously tested negative for all BioCode RPP analytes. Contrived specimens were spiked at concentrations of 2X LOD or greater using different strains for each pathogen. The 50 positive samples of each pathogen were prepared, interspersed with negative samples and randomized before testing at one of the five testing sites participated in the prospective clinical study. A total of 110 samples, including 100 positives, were tested. The results of the BioCode RPP testing are presented in the following table: | Organism | Source | Strain/Isolate | Fold LoD | Concentration | PA (%) | 95% CI | NA (%) | 95% CI | |-------------------------|--------------------------|--------------------------|---------------|-----------------|-----------------|-------------|-----------------|----------------| | | ATCC 53592 | AR-39 | 2 | 33.4 CFU/mL | 9/9 (100%) | 70.1%, 100% | | | | | | | 10 | 167 CFU/mL | 5/5 (100%) | 56.6%, 100% | | | | | | | 100 | 1670 CFU/mL | 4/4 (100%) | 51.0%, 100% | | | | Chlamydia pneumoniae | ATCC VR- 1360 | CM-1 | 2 | 33.4 CFU/mL | 8/8 (100%) | 67.6%, 100% | 60/60 (100%) | 94.0%, 100% | | | | | 10 | 167 CFU/mL | 5/5 (100%) | 56.6%, 100% | | | | | | | 100 | 1670 CFU/mL | 3/3 (100%) | 43.9%, 100% | | | | | ATCC VR- 1310 | CWL-029 | 2 | 33.4 CFU/mL | 8/8 (100%) | 67.6%, 100% | | | | | | | 10 | 167 CFU/mL | 5/5 (100%) | 56.6%, 100% | | | | | | | 100 | 1670 CFU/mL | 3/3 (100%) | 43.9%, 100% | | | | | | | | Combined | 50/50 (100%) | 92.9%, 100% | | | | Influenza A H1N1 | Zeptometrix 0810036CF | A/New Caledonia/20/99 | 2 | 30 TCID50/ mL | 5/5 (100%) | 56.6%, 100% | | 94.0%, 100% | | | | | | 10 | 150 TCID50/ mL | 3/3 (100%) | 43.9%, 100% | | | | | | | 100 | 1500 TCID50/ mL | 3/3 (100%) | 43.9%, 100% | | | | Zeptometrix 0810247CF | A/Taiwan/42/06 | 2 | 30 TCID50/ mL | 5/5 (100%) | 56.6%, 100% | | | | | | | | 10 | 150 TCID50/ mL | 3/3 (100%) | 43.9%, 100% | | | | | | | 100 | 1500 TCID50/ mL | 2/2 (100%) | 34.2%, 100% | | |…