BioCode® Respiratory Pathogen Panel (RPP)
Device Facts
| Record ID | K254139 |
|---|---|
| Device Name | BioCode® Respiratory Pathogen Panel (RPP) |
| Applicant | Applied BioCode, Inc. |
| Product Code | OCC · Microbiology |
| Decision Date | May 1, 2026 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.3980 |
| Device Class | Class 2 |
Indications for 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 a 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.
Device Story
BioCode RPP is a multiplexed nucleic acid test for respiratory pathogens; uses nasopharyngeal swab (NPS) samples in VTM/UTM. Nucleic acids extracted via KingFisher Flex (subject of this submission), NucliSENS easyMAG, or MagNA Pure 96. MDx-3000 instrument performs automated RT-PCR, target capture, signal generation, and optical detection. PCR products labeled with biotin; captured by target-specific probes on barcoded magnetic beads (BMBs). Streptavidin-phycoerythrin conjugate added for fluorescence labeling. Instrument images wells to decode BMB patterns and measure fluorescence intensity (MFI). Software calculates MFI to determine pathogen presence relative to validated cutoffs. Used in clinical laboratories by trained personnel. Results aid diagnosis alongside clinical/epidemiological data. Benefits include simultaneous identification of 17 respiratory pathogens/subtypes from a single specimen.
Clinical Evidence
No clinical prospective/retrospective studies performed. Evidence consists of analytical performance (reproducibility, competitive inhibition, LoD) and method comparison. Method comparison used 735 archived clinical NPS samples and 300 contrived samples to compare KingFisher Flex extraction against the predicate MagNA Pure 96 system. PPA and NPA were calculated; results showed high agreement (e.g., 100% PPA for Influenza A, 95.9% PPA for B. pertussis).
Technological Characteristics
Multiplex RT-PCR; target capture via biotinylated PCR products and barcoded magnetic beads (BMBs); fluorescence detection (streptavidin-phycoerythrin). Automated processing on BioCode MDx-3000. Extraction via KingFisher Flex, NucliSENS easyMAG, or MagNA Pure 96. High-complexity CLIA status. Internal control: bacteriophage MS2 (13,000 MFI cutoff).
Indications for Use
Indicated for qualitative detection/identification of respiratory viral and bacterial nucleic acids in nasopharyngeal swab (NPS) samples from individuals with signs/symptoms of respiratory tract infection. Pathogens: Adenovirus, Coronavirus (229E, OC43, HKU1, NL63), hMPV A/B, Flu A (H1, H1pdm09, H3), Flu B, PIV 1-4, RSV A/B, HRV/HEV, B. pertussis, C. pneumoniae, M. pneumoniae. Prescription use only.
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
- BioCode Respiratory Pathogen Panel (RPP) (K192485)
Submission Summary (Full Text)
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FDA
U.S. FOOD & DRUG
ADMINISTRATION
# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
ASSAY AND INSTRUMENT
## I Background Information:
A 510(k) Number
K254139
B Applicant
Applied BioCode, Inc
C Proprietary and Established Names
BioCode Respiratory Pathogen Panel (RPP)
D Regulatory Information
| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| OCC, OZE, OEP, OEM, OOU, OTG, OZX, OZY, OZZ, NSU | II | 21 CFR 866.3980 - Respiratory Viral Panel
Multiplex Nucleic Acid Assay | MI - Microbiology |
## II Submission/Device Overview:
### A Purpose for Submission:
To obtain a substantial equivalence determination for the BioCode RPP for the detection of microbial and viral nucleic acids extracted from nasopharyngeal swab (NPS) samples for use on the Applied BioCode MDx 3000 instrument, using an alternate sample extraction system, the KingFisher Flex.
### B Measurand:
Adenovirus, Human Metapneumovirus (hMPV) A/B, Influenza A (Flu A), Influenza A subtype H1 (Flu A/H1), Influenza A subtype H1 2009 Pandemic (Flu A/H1pdm09), Influenza A subtype H3 (Flu A/H3), Influenza B (Flu B), Coronavirus (229E, HKU1, OC43, and NL63), Parainfluenza virus 1 (PIV 1), Parainfluenza virus 2 (PIV 2), Parainfluenza virus 3 (PIV 3), Parainfluenza virus 4 (PIV 4), Human Rhinovirus/Enterovirus (HRV/HEV), Respiratory Syncytial Virus (RSV) A/B, Bordetella pertussis, Chlamydia pneumoniae, and Mycoplasma pneumoniae nucleic acid target sequences.
### C Type of Test:
A multiplexed nucleic acid test intended for use with the BioCode MDx-3000 Instrument for the simultaneous qualitative in vitro detection and identification of multiple respiratory viral and
Food and Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993-0002
www.fda.gov
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bacterial nucleic acids in nasopharyngeal swabs (NPS) collected in viral transport media (VTM) or universal transport media (UTM) and obtained from individuals suspected of respiratory tract infections.
## Intended Use/Indications for Use:
### A Intended Use(s):
See Indications for Use below.
### B Indication(s) for 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 a 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
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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.
## C Special Conditions for Use Statement(s):
Rx – For Prescription Use Only
## D Special Instrument Requirements:
BioCode MDx-3000 Instrument
bioMérieux NucliSENS easyMAG system
Roche MagNA Pure 96 system
Thermo Fisher KingFisher Flex automated systems
## IV Device/System Characteristics:
## A Device Description:
The BioCode Respiratory Pathogen Panel (RPP) 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 VTM or UTM. Liquid specimens are processed, and nucleic acids extracted with the bioMérieux NucliSENS easyMAG, Roche MagNA Pure 96 or Thermo Fisher KingFisher Flex automated systems. Once the PCR plate is manually set up and sealed, all other operations are automated on the MDx-3000. The BioCode RPP simultaneously tests for 17 pathogens and/or subtypes from nasopharyngeal swab specimens collected in VTM or UTM and are listed below.
## Viruses
- 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
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- Rhinovirus/Enterovirus
Bacteria
- Bordetella pertussis
- Chlamydia pneumoniae
- Mycoplasma pneumoniae
Results from the BioCode RPP test are available within about 5 hours.
## B Principle of Operation:
The BioCode MDx-3000 is an automated system that integrates PCR amplification, target capture, signal generation and optical detection for multiple respiratory viruses and bacteria from a single nasopharyngeal swab (NPS) specimen, in either VTM or UTM. Nucleic acids are extracted with the bioMérieux NucliSENS easyMAG, the Roche MagNA Pure 96 or the Thermo Fisher KingFisher Flex automated systems. Once the PCR plate is set up and sealed, all other operations are automated on the MDx-3000.
## Nucleic Acid Extraction
Nucleic acids (both RNA and DNA) are captured by silica coated magnetic beads and eluted on the bioMérieux NucliSENS easyMAG, Roche MagNA Pure 96 or Thermo Fisher KingFisher Flex automated systems according to the manufacturer provided protocol.
## Overview of a BioCode MDx-3000 Run
1. Reverse Transcription and Multiplex PCR – Since targets of the BioCode RPP include RNA viruses, a reverse transcription (RT) step is performed to convert the viral RNA into cDNA prior to amplification. The purified nucleic acid solution is combined with a freshly prepared reaction mixture for the RT step and subsequent thermal cycling for multiplex PCR to enrich the target nucleic acids present in the sample. One of the target-specific primers for each pathogen is biotinylated at the 5'-end to generate labeled PCR product for subsequent detection.
2. Dispensing Barcoded Magnetic Bead (BMB) – Probe Mix – Toward the end of PCR amplification, the robotic head dispenses BMB – Probe mix into the designated reaction wells of the capture plate using disposable pipette tips.
3. PCR Product Transfer – After PCR amplification is completed, the robotic head pierces the foil seal with disposable pipette tips and transfers PCR products into corresponding wells of the capture plate.
4. Target Capture – Amplified PCR products labeled with biotin are captured at a defined temperature by target-specific probes that are covalently coupled to designated BMBs. During this step, BMBs are kept in suspension by gentle agitation. Differentiation of captured targets is achieved by assigning a unique barcode pattern BMB for each pathogen and the internal control.
5. Signal Generation – After washing off unbound PCR products and unused primers, a
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streptavidin-phycoerythrin (SA-PE) conjugate is automatically added to the reaction by the robot. High affinity binding between biotin and streptavidin ensures that captured PCR products with the biotin moiety are labeled with phycoerythrin in close proximity to the BMBs.
6. Optical Detection – Optical detection is performed for each reaction well of the capture plate, an optically clear, flat-bottom microtiter plate. After washing off unbound SA-PE, excitation of the fluorophore at the designated wavelength emits fluorescence signal from BMBs tagged with SA-PE conjugates. Each reaction well is imaged at a specific emission wavelength for fluorescent signal and under bright field for identifying the barcode patterns (decoding).
The BioCode MDx-3000 Software controls the operation of the instrument, collects and analyzes data, and automatically generates interpretation for test reports at the end of the run. Fluorescent signals from BMBs with the same barcode are sorted and calculated to generate a median fluorescence intensity (MFI) for each analyte. The presence or absence of a pathogen is determined relative to the validated assay cutoff by MFI. The software also analyzes the results of external and internal controls to validate the run and individual specimen results for reporting.
C Instrument Description Information:
1. Instrument Name: BioCode MDx-3000 Instrument
2. Specimen Identification: Specimen identity is provided by barcoded magnetic beads (BMB).
3. Specimen Sampling and Handling: After nucleic acid extraction with the bioMérieux NucliSENS easyMAG, the Roche MagNA Pure 96 or the Thermo Fisher KingFisher Flex automated systems and manually loading samples into a 96-well formatted plate, the BioCode MDx 3000 processes all RT-PCR, target capture, signal generation, and optical detection steps automatically.
4. Calibration: Optical calibration of the BioCode MDx 3000 is performed twice a year by Applied BioCode. No calibration kit is available.
5. Quality Control: Each laboratory is expected to establish its own Quality Control ranges and frequency of QC testing based on applicable local laws, regulations, and good laboratory practices. The BioCode RPP uses an internal control (bacteriophage MS2) which is added to each sample prior to extraction. The internal control monitors the efficiency of the extraction, reverse transcription, amplification, and detection stages of the assay. Positive results may be reported in the absence of RNA IC detection. The BioCode RPP software will suppress negative results for any wells with invalid RNA IC results.
V Substantial Equivalence Information:
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A Predicate Device Name(s):
BioCode Respiratory Pathogen Panel (RPP)
B Predicate 510(k) Number(s):
K192485
C Comparison with Predicate(s):
| Device & Predicate Device(s): | K254139 | K192485 (Predicate) |
| --- | --- | --- |
| Device Trade Name | BioCode Respiratory Pathogen Panel (RPP) | BioCode Respiratory Pathogen Panel (RPP) |
| Common Name | Respiratory Virus Panel Nucleic Acid Assay System | Respiratory Virus Panel Nucleic Acid Assay System |
| Regulation | 21CFR 866.3980 | 21CFR 866.3980 |
| Product Code | OCC | OCC |
| Device Class | II | II |
| General Device Characteristic Similarities | | |
| 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 | Same |
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| | • 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 coinfection 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 a 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. |
| --- | --- |
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| | 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. | |
| --- | --- | --- |
| Specimen Types | NPS (in VTM or UTM) | Same |
| Pathogens Detected | Adenovirus, Coronavirus (229E, HKU1, NL63, and OC43), Human Metapneumovirus A/B, Influenza A, Influenza A subtype H1, Influenza A subtype H3, Influenza A subtype 2009 Pandemic, Influenza B, Parainfluenza Virus 1, Parainfluenza Virus 2, | Same |
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| | Parainfluenza Virus 3, Parainfluenza Virus 4, Rhinovirus/Enterovirus, Respiratory Syncytial Virus A/B, Bordetella pertussis, Chlamydia pneumoniae, and Mycoplasma pneumoniae | |
| --- | --- | --- |
| Analyte | RNA/DNA | Same |
| Instrumentation | Nucleic Acid Purification System BioCode MDx-3000 | Same |
| Time to Result | About 5.0 hours | Same |
| Test Interpretation | Automated test interpretation and report generation | Same |
| Controls | An RNA Internal Control is added to each sample during extraction. External Positive and Negative Controls are externally sourced. | Same |
| Methodology | Multiplex RT-PCR and probe hybridization to biotinylated PCR product(s) followed by fluorescence detection and decoding of barcoded magnetic beads (BMB) that are coupled to target-specific probes | Same |
| CLIA Complexity | High | Same |
| Sample Extraction | easyMAG, Roche MagNA Pure 96, KingFisher Flex | easyMAG, Roche MagNA Pure 96 |
| Internal Control Cut-off Values | 13,000 MFI | 8,000 MFI |
VI Standards/Guidance Documents Referenced:
- FDA guidance document issued on August 27, 2014, titled “Highly Multiplexed Microbiological/Medical Countermeasure In Vitro Nucleic Acid Based Diagnostic Devices”
- FDA guidance document issued on October 9, 2009, titled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay”
- FDA guidance document issued on October 9, 2009, titled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Assays”
- FDA guidance document issued on October 9, 2009, titled “Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays”
- FDA guidance document issued on July 15, 2011, titled “Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection or Detection and Differentiation of Influenza Viruses”
- FDA guidance document issued on April 25, 2005, titled “Guidance on Informed Consent for In Vitro Diagnostic Device Studies Using Leftover Human Specimens that are Not Individually Identifiable”
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VII Performance Characteristics (if/when applicable):
# A Analytical Performance:
# 1. Precision/Reproducibility:
A reproducibility study was conducted to assess the reproducibility of BioCode RPP with nucleic acid extraction performed using the KingFisher Flex system.
The reproducibility panel includes eight contrived samples composed of combinations of twelve representative targets (nine viral and three bacterial) at 1.5x Limit of Detection (LoD) (Low) and 3x LoD (Medium) spiked into pooled negative synthetic nasopharyngeal swab (sNPS) matrix, and one negative control sample (Table 1). Synthetic NPS matrix (sNPS) was previously determined to be equivalent to NPS matrix and was used for the LoD and reproducibility studies for the original 510(k) submission of BioCode RPP (K192485).
One lot of BioCode RPP was tested by three operators on five non-consecutive days. Each day the samples were extracted in triplicate on KingFisher Flex extraction instruments by each operator and assayed using BioCode RPP for a total of 90 extraction/PCR replicates per panel member.
Table 1. Reproducibility Panel - Sample Composition
| Medium (3x LoD) | Medium (3x LoD) | Low (1.5x LoD) | Low (1.5x LoD) | Sample Name |
| --- | --- | --- | --- | --- |
| Human Rhinovirus/Enterovirus | Parainfluenza Virus 2 | N/A | N/A | RP1a |
| N/A | N/A | Human Rhinovirus/Enterovirus | Parainfluenza Virus 2 | RP1b |
| Human Metapneumovirus (hMPV) | Bordetella pertussis | N/A | N/A | RP2a |
| N/A | N/A | Human Metapneumovirus (hMPV) | Bordetella pertussis | RP2b |
| Influenza B | Coronavirus NL63 | Chlamydia pneumoniae | Parainfluenza Virus 3 | RP3 |
| Chlamydia pneumoniae | Parainfluenza Virus 3 | Influenza B | Coronavirus NL63 | RP4 |
| Influenza A H3N2 | Mycoplasma pneumoniae | Respiratory Syncytial Virus (RSV) | Adenovirus C (ADV) | RP5 |
| Respiratory Syncytial Virus (RSV) | Adenovirus C (ADV) | Influenza A H3N2 | Mycoplasma pneumoniae | RP6 |
| Negative matrix (synthetic NPS) | | | | RP7 |
All targets tested at 3x LoD had 90/90 (100%) agreement with expected results except for Human Metapneumovirus and Bordetella pertussis (RP2a) which had 89/90 (98.9%) agreement with expected results. Reproducibility results are summarized in Table 2.
Table 2. Reproducibility Results
| Analyte | Concentration Tested | Expected Result | Agreement with Expected Results |
| --- | --- | --- | --- |
| | | | KingFisher Flex Extraction (95% CI) |
| Viruses | | | |
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| Adenovirus | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| --- | --- | --- | --- |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Coronavirus NL63 | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Coronavirus HKU | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Coronavirus OC43 | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Coronavirus 229E | None (no analyte) | Not Detected | 809/810 / 99.9% / (99.3% - 100%) |
| Human Metapneumovirus | 3x LoD | Detected | 89/90* / 98.9% / (94.0%-99.8%) |
| | 1.5x LoD | Detected | 88/90 / 97.8% / (92.3%-99.4%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Human Rhinovirus/Enterovirus | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (89.1% - 98.3%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Influenza A/H3 | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Influenza A/H1pdm09 | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Influenza A/H1 | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Influenza B | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Parainfluenza Virus 1 | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Parainfluenza Virus 2 | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 88/90 / 97.8% / (92.3% - 99.4%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Parainfluenza Virus 3 | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Parainfluenza Virus 4 | None (no analyte) | Not Detected | 810/810 / 100% / (99.6% - 100%) |
| Respiratory Syncytial Virus | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 629/630 / 99.8% / (99.1% - 100%) |
| Bacteria | | | |
| Mycoplasma pneumoniae | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
| Bordetella pertussis | 3x LoD | Detected | 89/90* / 98.9% / (94.0% - 100%) |
| | 1.5x LoD | Detected | 88/90 / 97.8% / (92.3% - 99.4%) |
| | None (no analyte) | Not Detected | 629/630 / 99.8% / (99.1% - 100%) |
| Chlamydia pneumoniae | 3x LoD | Detected | 90/90 / 100% / (95.9% - 100%) |
| | 1.5x LoD | Detected | 89/90 / 98.9% / (94.0% - 99.8%) |
| | None (no analyte) | Not Detected | 630/630 / 100% / (99.4% - 100%) |
* Missed replicate due to PCR set up error detected in (3/3) repeat replicates on the same instrument. It was not due to KFF extraction.
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Descriptive statistical calculations of Median Fluorescence Intensity (MFI) (average, standard deviation and percent coefficient of variation) between runs and between days, are presented in Table 3.
Table 3. Median Fluorescence Intensity (MFI) Summary Statistics Between Runs and Days
| Target | Concentration | Average ± Standard Deviation (%CV) | |
| --- | --- | --- | --- |
| | | Between Runs | Between Days |
| Adenovirus 1 | 3x LoD | 9884 ± 2049 (21%) | 9878 ± 892 (9%) |
| | 1.5x LoD | 9362 ± 2204 (24%) | 9261 ± 659 (7%) |
| Adenovirus 2 | 3x LoD | 14595 ± 2668 (18%) | 14633 ± 937 (6%) |
| | 1.5x LoD | 13813 ± 2887 (21%) | 13694 ± 770 (6%) |
| Coronavirus NL63 | 3x LoD | 31076 ± 2403 (8%) | 30814 ± 830 (3%) |
| | 1.5x LoD | 27451 ± 1648 (6%) | 27369 ± 629 (2%) |
| Human Metapneumovirus 1 | 3x LoD | 34769 ± 2343 (7%) | 34552 ± 632 (2%) |
| | 1.5x LoD | 32704 ± 2984 (9%) | 32761 ± 1189 (4%) |
| Human Metapneumovirus 2 | 3x LoD | 27057 ± 3351 (12%) | 27047 ± 1483 (5%) |
| | 1.5x LoD | 24061 ± 2560 (11%) | 24435 ± 1320 (5%) |
| Human Rhinovirus | 3x LoD | 26049 ± 3607 (14%) | 26356 ± 678 (3%) |
| | 1.5x LoD | 20419 ± 3665 (18%) | 20713 ± 933 (5%) |
| Influenza A/H3 | 3x LoD | 37536 ± 3920 (10%) | 37233 ± 1124 (3%) |
| | 1.5x LoD | 33004 ± 3120 (9%) | 33045 ± 303 (1%) |
| Influenza B1 | 3x LoD | 7590 ± 1495 (20%) | 7685 ± 675 (9%) |
| | 1.5x LoD | 8453 ± 1273 (15%) | 8352 ± 1059 (13%) |
| Influenza B2 | 3x LoD | 28916 ± 4178 (14%) | 28880 ± 1641 (6%) |
| | 1.5x LoD | 24176 ± 4530 (19%) | 24800 ± 1468 (6%) |
| Parainfluenza Virus 2 | 3x LoD | 22276 ± 2278 (10%) | 22081 ± 860 (4%) |
| | 1.5x LoD | 17004 ± 2547 (15%) | 16910 ± 1476 (9%) |
| Parainfluenza Virus 3 | 3x LoD | 25157 ± 2082 (8%) | 25087 ± 287 (1%) |
| | 1.5x LoD | 23631 ± 2247 (10%) | 23624 ± 780 (3%) |
| Respiratory Syncytial Virus | 3x LoD | 36077 ± 4711 (13%) | 36023 ± 1561 (4%) |
| | 1.5x LoD | 38821 ± 3397 (9%) | 38874 ± 1477 (4%) |
| Mycoplasma pneumoniae | 3x LoD | 44239 ± 4679 (11%) | 44153 ± 2598 (6%) |
| | 1.5x LoD | 39045 ± 7167 (18%) | 38573 ± 3577 (9%) |
| Bordetella pertussis | 3x LoD | 14422 ± 4935 (34%) | 14666 ± 1934 (13%) |
| | 1.5x LoD | 11114 ± 3865 (35%) | 11519 ± 1510 (13%) |
| Chlamydia pneumoniae | 3x LoD | 26886 ± 4646 (17%) | 26443 ± 2692 (10%) |
| | 1.5x LoD | 22558 ± 4124 (18%) | 22400 ± 1425 (6%) |
2. Linearity:
Not applicable, BioCode RPP is a qualitative assay
3. Analytical Specificity/Interference:
Inclusivity, microbial interference, cross-reactivity and potential interfering substance of BioCode RPP was evaluated in the original 510(k) Premarket Notification (K192485). No additional testing was conducted.
Competitive Inhibition
An analytical study was performed to evaluate potential competitive inhibition for Adenovirus species C Serotype 2 (AdVC2) and Bordetella pertussis (Bp) on the BioCode RPP when using the KFF extraction method.
K254139 - Page 12 of 18
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Competitive inhibition of AdVC2 and Bp were evaluated by spiking potential inhibitory analytes in competitive inhibition samples at high concentration ( $\geq 10^{5}$ for viruses) and the two targets at low concentration (3x LoD).
Summary results of this study are presented in Table 4 below. No inhibition was observed at the concentrations tested in this study.
Table 4. Competitive Inhibition Study Results
| Panel Designation | Viral/Bacterial Strain | Source | Level | Titer Tested | Detected (n of 3) | Pass/Fail |
| --- | --- | --- | --- | --- | --- | --- |
| Competitive Inhibition Sample 1 | Respiratory Syncytial Virus Type A | Zeptometrix 0810040ACF | High | 1x105TCID50/mL | 3/3 | Pass |
| | Influenza A H3N2 A/Alice | ATCC VR-776 | Low | 81 CEID50/mL | 3/3 | |
| | Adenovirus species C Serotype 2 | ATCC VR-846 | Low | 54 TCID50/mL | 3/3 | |
| Competitive Inhibition Sample 2 | Influenza A H3N2 A/Alice | ATCC VR-776 | High | 1x105CEID50/mL | 3/3 | Pass |
| | Adenovirus species C Serotype 2 | ATCC VR-846 | Low | 54 TCID50/mL | 3/3 | |
| | Respiratory Syncytial Virus Type A | Zeptometrix 0810040ACF | Low | 0.99 TCID50/mL | 3/3 | |
| Competitive Inhibition Sample 3 | Coronavirus OC43 | Zeptometrix 0810024CF | High | 1x105TCID50/mL | 3/3 | Pass |
| | Human Metapneumovirus | Zeptometrix 0810161CF | Low | 5 TCID50/mL | 3/3 | |
| | Bordetella pertussis | Zeptometrix 081459 | Low | 810 CFU/mL | 3/3 | |
| Competitive Inhibition Sample 4 | Human Metapneumovirus | Zeptometrix 0810161CF | High | 1x105TCID50/mL | 3/3 | Pass |
| | Bordetella pertussis | Zeptometrix 081459 | Low | 810 CFU/mL | 3/3 | |
| | Coronavirus OC43 | Zeptometrix 0810024CF | Low | 0.27 TCID50/mL | 3/3 | |
# 4. Detection Limit and Assay Reportable Range:
The LoD of BioCode RPP was evaluated with both KFF and MagNA Pure 96 (MP96) extraction systems using simulated NPS matrix samples spiked with target analytes. An estimated LoD for each analyte was determined by testing serial dilutions of contrived samples. Four extraction replicates per dilution on each extraction system, KFF and MP96, were tested using the BioCode RPP.
The LoD for each analyte was confirmed by testing 20 replicates extracted with each extraction system. LoD for each isolate was defined as the lowest concentration with $\geq 95\%$ detection of 20 replicates (at least 19 out of 20 replicates). Summary results of the LoD confirmation study are presented in Table 5.
Table 5. Limit of Detection for the KingFisher Extraction System - Comparison with MagNA Pure 96
| Organism | Strain | Source | MagNA Pure 96 | KingFisher Flex |
| --- | --- | --- | --- | --- |
| KFF | 20 | KFF | KFF | KFF |
| MagNA Pure 96 | 20 | KFF | KFF | KFF |
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| | | | LoD | Detection (n/20) | LoD | Detection (n/20) |
| --- | --- | --- | --- | --- | --- | --- |
| Influenza A H1N1 | A/New Caledonia/20/99 | Zeptometrix 0810036CF | 1.67x10^{0} TCID50/mL | 20/20 | 5.00x10^{0} TCID50/mL | 20/20 |
| Influenza A H1N1 | A/NWS/33 | ATCC VR-219 | 2.70x10^{1} TCID50/mL | 20/20 | 2.70x10^{1} TCID50/mL | 20/20 |
| Influenza A H1N1 pdm09 | H1N1 California/07/09 | Zeptometrix 0810165CF | 4.00x10^{-1} TCID50/mL | 20/20 | 4.00x10^{-1} TCID50/mL | 20/20 |
| Influenza A H3N2 | A/Wisconsin/67/05 | Zeptometrix 0810252CF | 4.33x10^{-1} TCID50/mL | 19/20 | 1.30x10^{0} TCID50/mL | 20/20 |
| Influenza A H3N2 | A/Alice | ATCC VR-776 | 9.00x10^{0} CEID50/.2mL | 19/20 | 2.70x10^{1} CEID50/mL | 20/20 |
| Influenza B | Flu B/Florida/4/2006 (Yamagata) | Zeptometrix 0810255CF | 1.00x10^{-2} TCID50/mL | 20/20 | 6.00x10^{-2} TCID50/mL | 20/20 |
| Influenza B | B/Hong Kong/5/1972 (Victoria) | ATCC VR-823 | 1.80x10^{0} TCID50/mL | 20/20 | 5.40x10^{0} TCID50/mL | 19/20 |
| Respiratory Syncytial Virus | Type A | Zeptometrix 0810040ACF | 1.10x10^{-1} TCID50/mL | 20/20 | 3.30x10^{-1} TCID50/mL | 20/20 |
| Human Metapneumovirus | 16; Type A1 IA10-2003 | Zeptometrix 0810161CF | 5.56x10^{-1} TCID50/mL | 20/20 | 1.67x10^{0} TCID50/mL | 20/20 |
| Parainfluenza Virus 1 | [C-35/Washington DC/1957] | Zeptometrix 0810014CF | 9.00x10^{0} TCID50/mL | 20/20 | 9.00x10^{0} TCID50/mL | 20/20 |
| Parainfluenza Virus 2 | [Greer/Ohio/1955] | ATCC VR-92 | 1.80x10^{0} TCID50/mL | 20/20 | 5.40x10^{0} TCID50/mL | 19/20 |
| Parainfluenza Virus 3 | N/A | Zeptometrix 0810016CF | 5.00x10^{0} TCID50/mL | 19/20 | 1.50x10^{1} TCID50/mL | 20/20 |
| Parainfluenza Virus 4 | type 4a | Zeptometrix 0810060CF | 2.70x10^{1} TCID50/mL | 20/20 | 8.10x10^{1} TCID50/mL | 20/20 |
| Adenovirus | Species B Serotype 7A | Zeptometrix 0810021CF | 1.20x10^{0} TCID50/mL | 20/20 | 3.60x10^{0} TCID50/mL | 20/20 |
| Adenovirus | Species C Serotype 2 | ATCC VR-846 | 2.00x10^{0} TCID50/mL | 19/20 | 1.80x10^{1} TCID50/mL | 20/20 |
| Adenovirus | Species E Serotype 4 | Zeptometrix 0810070CF | 4.00x10^{-2} TCID50/mL | 20/20 | 1.33x10^{-1} TCID50/mL | 19/20 |
| Coronavirus 229E | 229E | Zeptometrix 0810229CF | 6.00x10^{-1} TCID50/mL | 20/20 | 1.80x10^{0} TCID50/mL | 20/20 |
| Coronavirus HKU1 | HKU1^{a} | Clinical Sample | 1.50x10^{3} copies/mL | 20/20 | 1.50x10^{3} copies/mL | 19/20 |
| Coronavirus NL63 | NL63 | Zeptometrix 0810228CF | 1.33x10^{-2} TCID50/mL | 20/20 | 4.00x10^{-2} TCID50/mL | 20/20 |
| Coronavirus OC43 | OC43 | Zeptometrix 0810024CF | 3.00x10^{-2} TCID50/mL | 20/20 | 9.00x10^{-2} TCID50/mL | 20/20 |
| Human Rhinovirus/Enterovirus | Rhinovirus Type A1 | Zeptometrix 0810012CFN | 4.00x10^{-1} TCID50/mL | 20/20 | 4.00x10^{-1} TCID50/mL | 20/20 |
| Enterovirus | Enterovirus D68 | Zeptometrix 0810300CF | 9.00x10^{0} CFU/mL | 19/20 | 9.00x10^{0} CFU/mL | 19/20 |
| Bordetella pertussis | A639 | Zeptometrix 0801459 | 1.50x10^{1} CFU/mL | 20/20 | 2.70x10^{2} CFU/mL | 20/20 |
| Chlamydia pneumoniae | TW-183 (AR39) | ATCC 53592 | 3.30x10^{1} IFU/mL | 20/20 | 3.30x10^{1} IFU/mL | 20/20 |
| Mycoplasma pneumoniae | M129 | Zeptometrix 0801579 | 1.50x10^{1} CCU/mL | 20/20 | 4.50x10^{1} CCU/mL | 20/20 |
a Coronavirus HKU1 is not available as a titered stock. A positive patient sample, quantified in copies/mL using a PCR standard curve created using quantified in vitro RNA, was used.
5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):
No modifications were made to BioCode RPP or MDx 3000 instrument system. Please refer to the published decision summary for the original 510(k) submission for additional information (K192485).
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# 6. Assay Cut-Off:
MFI cutoff values were not modified for analyte detection, please refer to the published decision summary for the original $510(\mathrm{k})$ submission for additional information (K192485). The MFI cutoff value for the Internal Control (IC) RNA (bacteriophage MS2) was increased from 8,000 to 13,000 MFI.
# B Comparison Studies:
# 1. Method Comparison with Predicate Device:
To support the performance of the BioCode RPP when used with the KFF extraction system, a total of 735 (629 positive and 106 negative) archived NPS clinical samples collected from patients across five geographically diverse investigational sites were evaluated with both the KFF extraction system and previously cleared methods employing the MP96 extraction system. These remnant samples were used to support the original $510(\mathrm{k})$ clearance of the BioCode RPP device.
Ten (10) deidentified, frozen remnant Bp positive samples were used for Bp method comparison testing. In addition, a total of 300 samples were contrived at $2\mathrm{x}$ LoD and $4\mathrm{x}$ LoD (25 each) and tested to determine the performance characteristics for Bordetella pertussis, Chlamydia pneumonia, Mycoplasma pneumonia, Influenza A H1N1, Parainfluenza 1 and Parainfluenza 4. The demographics of the enrolled subjects in the clinical study are shown in Table 6 and the demographics of the Bp positive samples are shown in Table 9.
Table 6. Demographic data for Archived Samples
| Archived Samples | |
| --- | --- |
| Total Specimen Count | 735 |
| Sex | |
| Male | 374/735 (50.9%) |
| Female | 361/735 (49.1%) |
| Age | |
| ≤ 5 yrs | 368/735 (50.1%) |
| 6-21 yrs | 197/735 (26.8%) |
| 22-59 yrs | 89/735 (12.1%) |
| 60+ yrs | 81/735 (11.0%) |
Positive percent agreement (PPA) was calculated as $\mathrm{TP / (TP + FN)(TP =}$ true positive or positive by both the MP96 and KFF; $\mathrm{FN} =$ false negative or negative by the KFF only). Negative percent agreement was calculated as $\mathrm{TN / (TN + FP)(TN =}$ true negative or negative by the MP96 and KFF; $\mathrm{FP} =$ false positive or positive by KFF only). Performance of BioCode RPP using KFF extraction method is shown in Tables 7, 8 and 10.
Table 7. Results from archived samples tested with BioCode RPP using KFF extraction compared to MP96 extraction
| Target | Positive Agreement | | Negative Agreement | |
| --- | --- | --- | --- | --- |
| | PPA % | 95% CI | NPA (%) | 95% CI |
| Influenza A | 110/110 (100%) | 96.6-100% | 624/625 (99.8%) | 99.1-100% |
| Influenza A (H1N1 seasonal) | N/A | N/A | 735/735 (100%) | 99.5-100% |
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Table 8. Results from contrived samples tested by BioCode RPP using the KFF extraction system compared to MP96
| Target | Strain/ Isolate | Fold LoD | Concentration | PPA (%) | 95% CI | NPA (%) | 95 CI |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Bordetella pertussis | A639 | 2x | 5.40x102CFU/mL | 22/23R(95.7%) | 79.0%-99.2% | 249/251b(99.2%) | 97.1%-99.8% |
| | | 4x | 1.08x103CFU/mL | 25/25 (100%) | 86.7%-100% | | |
| | Combined | | | 47/49 (95.9%) | 86.3%-98.9% | | |
| Chlamydia pneumoniae | TW-183 (AR39) | 2x | 6.67x101IFU/mL | 25/25 (100%) | 86.7%-100% | 250/250(100%) | 98.5%-100% |
| | | 4x | 1.33x102IFU/mL | 25/25 (100%) | | | |
| | Combined | | | 50/50 (100%) | 92.9%-100% | | |
| | A/New | 2x | 1.00x101TCID50/mL | 25/25 (100%) | 86.7%-100% | | 98.5%- |
| | | | | | | | |
| | | | | | | | |
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| Influenza A H1 | Caledonia/20/99 | 4x | 2.00x10¹ TCID50/mL | 25/25 (100%) | | 250/250 (100%) | 100% |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | Combined | | | 50/50 (100%) | 92.9%-100% | | |
| Mycoplasma pneumoniae | M129 | 2x | 9.00x10¹ CCU/mL | 25/25 (100%) | 86.7%-100% | 250/250 (100%) | 98.5%-100% |
| | | 4x | 1.80x10² CCU/mL | 25/25 (100%) | | | |
| | Combined | | | 50/50 (100%) | 92.9%-100% | | |
| Parainfluenza Virus 1 | C-35/ Washington DC/ 1957 | 2x | 1.80x10¹ TCID50/mL | 25/25 (100%) | 86.7%-100% | 250/250 (100%) | 98.5%-100% |
| | | 4x | 3.60x10¹ TCID50/mL | 25/25 (100%) | | | |
| | Combined | | | 50/50 (100%) | 92.9%-100% | | |
| Parainfluenza Virus 4 | Type 4a | 2x | 1.62x10² TCID50/mL | 25/25 (100%) | 86.7%-100% | 250/250 (100%) | 98.5%-100% |
| | | 4x | 3.24x10² TCID50/mL | 25/25 (100%) | | | |
| | Combined | | | 50/50 (100%) | 92.9%-100% | | |
a25 samples contrived with BP at 2x LoD were tested. BP was detected in 23/25 from the MP96 extracts and 24/25 from the KFF extracts.
b Bordetella pertussis was detected in one of the 4x LoD PIV4 MP96 extracted contrived samples
Table 9. Demographic data for Bordetella pertussis samples
| Remnant Samples | |
| --- | --- |
| Total Specimen Count | 10 |
| Sex | |
| Male | 3/10 (30%) |
| Female | 7/10 (70%) |
| Age Category | |
| ≤ 5 yrs | 3/10 (30%) |
| 6-21 yrs | 6/10 (60%) |
| 22-59 yrs | 1/10 (10%) |
| 60 + yrs | 0/10 (0%) |
Table 10. Method Comparison Results: RPP Bp (MP96 Extraction vs KFF Extraction)
| Target | Positive Agreement (MP96 extraction vs KFF extraction) | |
| --- | --- | --- |
| | PPA % a | 95% CI |
| Bordetella pertussis | 10/10 (100%) | 74.1-100% |
a Positive percent agreement (PPA; [true positive/true positive + false negative] x 100)
2. Matrix Comparison: Not applicable
C Clinical Studies:
1. Clinical Sensitivity: Not applicable.
2. Clinical Specificity: Not applicable.
{17}
3. Clinical Cut-Off
Not applicable.
4. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
None
D Expected Values/Reference Range:
Refer to the published decision summary for the original clearance in K192485.
E Other Supportive Instrument Performance Characteristics Data:
Refer to the published decision summary for the original clearance in K192485.
VIII Proposed Labeling:
The labeling supports the finding of substantial equivalence for this device.
IX Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
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