(227 days)
The Quidel Molecular RSV + hMPV assay is a multiplex Real Time RT-PCR assay for the in vitro qualitative detection and identification of respiratory syncytial virus and human metapneumovirus viral RNA extracted from nasal and nasopharyngeal swabs specimens from patients with signs and symptoms of respiratory infection. This in vitro diagnostic test is intended to aid in the differential diagnosis of respiratory syncytial virus and human metapneumovirus infections. This test is not intended to differentiate the four genetic sub-lineages of hMPV.
Negative results do not preclude RSV or hMPV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
The Quidel Molecular RSV + hMPV Assay detects viral nucleic acids that have been extracted from a patient sample using the NucliSENS® easyMAG® automated extraction platform. A multiplex RT-PCR reaction is carried out under optimized conditions in a single tube generating amplicons for each of the target viruses present in the sample. This reaction is performed utilizing either the Cepheid SmartCycler® II or the Applied Biosystems 7500 Fast DX. Identification of RSV and hMPV and the PRC occurs by the use of target-specific primers and fluorescent-labeled probes that hybridize to conserved regions in the genomes of RSV and hMPV and the PRC.
Here's a summary of the acceptance criteria and study details for the Quidel Molecular RSV + hMPV Assay, based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The acceptance criteria provided in the document are primarily for analytical performance (LoD, Reproducibility, Inclusivity, Specificity) and clinical performance (Sensitivity and Specificity/Positive and Negative Percent Agreement). The clinical performance is reported compared to predicate devices or established methods.
Device: Quidel Molecular RSV + hMPV Assay
| Performance Measure | Acceptance Criteria (Implicit from study results) | Reported Device Performance (Cepheid SmartCycler II) | Reported Device Performance (Applied Biosystems 7500 Fast DX) |
|---|---|---|---|
| Analytical Performance | |||
| Limit of Detection (LoD) | Defined as the lowest concentration at which 95% of replicates tested positive. | Ranges from 1.89E+00 TCID50/mL (RSV A) to 2.645E+01 TCID50/mL (hMPV-A1) | Ranges from 6.29E-01 TCID50/mL (RSV A) to 1.7E+01 TCID50/mL (hMPV-A1) |
| Reproducibility | High concordance for positive and negative controls/high and medium positives; acceptable %CV for Ct values. | RSV Low Positive 2x LoD: 89/89 (99-100%)RSV Med Positive 5x LoD: 90/90 (100%)hMPV Low Positive 2x LoD: 90/90 (100%)hMPV Med Positive 5x LoD: 90/90 (100%)Negative Controls: 0/90 (0%) positive | RSV Low Positive 2x LoD: 90/90 (100%)RSV Med Positive 5x LoD: 90/90 (100%)hMPV Low Positive 2x LoD: 87/90 (96.7%)hMPV Med Positive 5x LoD: 89/90 (98.9%)Negative Controls: 0/90 (0%) positive |
| Analytical Reactivity (Inclusivity) | All tested strains of RSV and hMPV should be detected as positive. | All 13 RSV strains and 12 hMPV strains tested were Positive. | All 13 RSV strains and 12 hMPV strains tested were Positive. |
| Analytical Specificity (Cross-Reactivity) | No false positives with common respiratory pathogens or flora. | 100% specificity against 27 viruses, 24 bacteria, and 1 yeast strain. | 100% specificity against 27 viruses, 24 bacteria, and 1 yeast strain. |
| Clinical Performance (RSV - vs. DSFA & Cell Culture w/DFA) | Good sensitivity and specificity (implicitly high values) | Sensitivity: 97.9% (95% CI: 93.9% - 99.3%)Specificity: 97.6% (95% CI: 96.3% - 98.4%) | Sensitivity: 98.6% (95% CI: 94.9% - 99.6%)Specificity: 96.8% (95% CI: 95.4% - 97.8%) |
| Clinical Performance (hMPV - vs. Pro hMPV+) | Good positive and negative percent agreement (implicitly high values) | Positive percent agreement: 96.7% (95% CI: 92.4% - 98.6%)Negative percent agreement: 99.6% (95% CI: 98.9% - 99.9%) | Positive percent agreement: 98.0% (95% CI: 94.3% - 99.3%)Negative percent agreement: 99.3% (95% CI: 98.4% - 99.7%) |
2. Sample Size Used for the Test Set and Data Provenance
- Clinical Performance Test Set Samples:
- Total samples collected: 1014 specimens (414 fresh, 600 frozen) for RSV comparison.
- RSV testing (SmartCycler II): 1009 specimens after excluding contaminated cell cultures.
- hMPV testing (SmartCycler II): 951 specimens after excluding invalid comparative device results.
- RSV testing (7500 Fast Dx): 1007 specimens after excluding contaminated cell cultures and invalid subject method results.
- hMPV testing (7500 Fast Dx): 946 specimens after excluding invalid comparative and subject method results.
- Data Provenance: The samples were collected prospectively during the 2012 respiratory virus season (January to March 2012) from symptomatic patients at four sites across the United States. The study specifically used "fresh (414) and frozen (600) swab specimens."
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts
The document does not explicitly state the number or qualifications of experts used. However, the ground truth for RSV was established using "DSFA & Cell Culture w/DFA" (Direct Specimen Fluorescent Antibody and Cell Culture with DFA), which implies interpretation by trained laboratory personnel or specialists, although their specific qualifications or number are not detailed. For hMPV, the ground truth was established by comparing it to the "FDA Cleared hMPV molecular test" (Gen-Probe Prodesse Pro hMPV+, K082688), which is a molecular diagnostic method rather than expert interpretation of raw data.
4. Adjudication Method for the Test Set
- RSV Discrepant Results:
- For the SmartCycler II, all 21 originally discordant specimens (QM RSV + hMPV positive, reference method negative) were positive for RSV by an FDA-cleared RT-PCR assay and by bi-directional sequence analysis.
- For the 7500 Fast Dx, 25 of 28 originally discordant specimens were positive by an FDA-cleared RT-PCR assay, and 27 of 28 were positive by bi-directional sequence analysis.
- hMPV Discrepant Results:
- For both the SmartCycler II and the 7500 Fast Dx, all originally discordant specimens (QM RSV + hMPV positive, reference method negative) were positive for hMPV by bi-directional sequence analysis.
This indicates a form of post-hoc adjudication or discrepancy resolution using additional, more definitive molecular methods (FDA-cleared RT-PCR and bi-directional sequencing) for cases where the subject device and the initial reference method disagreed.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, If So, What Was the Effect Size of How Much Human Readers Improve with AI vs. Without AI Assistance
No, this was not an MRMC study. The device is an in vitro diagnostic (molecular assay) for direct detection of viral RNA, not an imaging device requiring human reader interpretation or AI assistance in interpretation. Therefore, a multi-reader multi-case comparative effectiveness study on human reader improvement with or without AI assistance is not applicable here.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done
Yes, the clinical performance study evaluates the standalone performance of the Quidel Molecular RSV + hMPV Assay. The results are presented as the device's agreement (sensitivity, specificity, positive/negative percent agreement) compared directly to the reference methods, without human interpretation of the assay results impacting the reported performance metrics. The assay itself provides a qualitative (positive/negative) result based on its internal thresholding (e.g., fluorescence achieved by 50 cycles on SmartCycler II or 35 cycles on ABI 7500 Fast Dx).
7. The Type of Ground Truth Used
- For RSV: The ground truth for clinical performance was established using Direct Specimen Fluorescent Antibody (DSFA) and Cell Culture with DFA.
- For hMPV: The ground truth for clinical performance was established using an FDA Cleared hMPV molecular test (Gen-Probe Prodesse Pro hMPV+).
- For discrepant results: Bi-directional sequence analysis and/or another FDA-cleared RT-PCR assay were used as definitive ground truth.
8. The Sample Size for the Training Set
The document does not explicitly state a sample size for a "training set" in the context of machine learning or algorithm development. For this type of molecular diagnostic assay, analytical studies (LoD, inclusivity, specificity) and clinical validation are performed. The LoD study involved replicates of serially diluted viral cultures, and inclusivity/specificity studies used panels of various strains/organisms. These analytical studies are analogous to "training" or "development" data in that they inform and validate the assay's operational parameters, but they are not framed as a classic machine learning training set.
9. How the Ground Truth for the Training Set Was Established
Given that this is a molecular diagnostic assay, the "ground truth" for establishing analytical parameters (like LoD, inclusivity, and specificity) is based on:
- Quantified viral cultures (TCID50/mL): Used for LoD studies, where the exact concentration of virus is known.
- Known viral strains or bacterial/yeast cultures: Used for inclusivity (known to contain the target virus) and specificity (known to contain other organisms to test for cross-reactivity) studies. The identity and concentration of these cultures are established through standard microbiological and virological techniques.
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K 122189
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Quidel Corporation
Quidel Molecular RSV + hMPV Assay 7/20/2012 Page 1 of 15
Section 05, 510(k) Summary
Applicant:
Quidel Corporation 10165 McKellar Court San Diego, California 92121 Telephone: 858-552-7910 Fax: 858-646-8045
Contact Information:
Ronald H. Lollar, Senior Director Clinical and Quality Affairs 1055 East State Street Suite 100 Athens, Ohio 45701 740-589-3300 - Corporate number 740-589-3373 - Desk phone 740-593-8437 - Fax lollar@dhiusa.com
Date of preparation of 510(k) summary:
July 20, 2012
Device Name:
Trade name - Quidel Molecular RSV + hMPV Assay Classification name - Respiratory viral panel multiplex nucleic acid assay Product Code - OEM. OCC Regulation - 21 CFR 866.3980
Legally marketed devices to which equivalence is claimed:
Gen-Probe Prodesse ProFlu+ (K092500)
The ProFluTM+ Assay is a multiplex Real-Time PCR (RT-PCR) in vitro diagnostic test for the rapid and qualitative detection and discrimination of Influenza A Virus, Influenza B Virus, and Respiratory Syncytial Virus (RSV) nucleic acids isolated and purified from nasopharyngeal (NP) swab specimens obtained from symptomatic patients. This test is intended for use to aid in the differential diagnosis of Influenza A, Influenza B and RSV viral infections in humans and is not intended to detect Influenza C.
Negative results do not preclude influenza or RSV virus infection and should not be used as the sole basis for treatment or other management decisions. It is recommended that negative RSV results be confirmed by culture.
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Performance characteristics for Influenza A Virus were established when Influenza A/H3 and A/H1 were the predominant Influenza A viruses in circulation. When other Influenza A viruses are emerging, performance characteristics may vary. 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 department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.
Gen-Probe Prodesse Pro hMPV+ (K082688)
The Pro hMPV+ Assay is a Real Time RT-PCR in vitro diagnostic test for the qualitative detection of human Metapneumovirus (hMPV) nucleic acid isolated and purified from nasopharyngeal swab (NP) specimens obtained from individuals exhibiting signs and symptoms of acute respiratory infection. This assay targets a highly conserved region of the Nucleocapsid gene of hMPV. The detection of hMPV nucleic acid from symptomatic patients aids in the diagnosis of human respiratory hMPV infection if used in conjunction with other clinical and laboratory findings. This test is not intended to differentiate the four genetic sub-lineages of hMPV.
Negative results do not preclude hMPV infection and should not be used as the sole basis for diagnosis, treatment or other management decisions.
Intended Use:
The Ouidel Molecular RSV + hMPV assay is a multiplex Real Time RT-PCR assay for the in vitro qualitative detection and identification of respiratory syncytial virus and human metapneumovirus viral RNA extracted from nasal and nasopharyngeal swabs specimens from patients with signs and symptoms of respiratory infection. This in vitro diagnostic test is intended to aid in the differential diagnosis of respiratory syncytial virus and human metapneumovirus infections. This test is not intended to differentiate the four genetic sub-lineages of hMPV.
Negative results do not preclude RSV or hMPV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Device Description:
The Quidel Molecular RSV + hMPV Assay detects viral nucleic acids that have been extracted from a patient sample using the NucliSENS® easyMAG® automated extraction platform. A multiplex RT-PCR reaction is carried out under optimized conditions in a single tube generating amplicons for each of the target
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viruses present in the sample. This reaction is performed utilizing either the Cepheid SmartCycler® II or the Applied Biosystems 7500 Fast DX. Identification of RSV and hMPV and the PRC occurs by the use of target-specific primers and fluorescent-labeled probes that hybridize to conserved regions in the genomes of RSV and hMPV and the PRC.
The following is a summary of the procedure:
- I. Sample Collection: Obtain nasal or nasopharyngeal swabs specimens using standard techniques from symptomatic patients. Transport, store, and process these specimens according to established laboratory procedures.'
-
- Nucleic Acid Extraction: Extract Nucleic Acids from the specimens with the NucliSENS® easyMAG® System following the manufacturer's instructions and using the appropriate reagents (See Materials Required but Not Provided).
Prior to the extraction procedure, add 20 uL of the Process Control (PRC) to each 180 µL aliquot of specimen. The PRC serves to monitor inhibitors in the extracted specimen, assures that adequate amplification has taken place, and confirms that the nucleic acid extraction was sufficient.
-
- Rehydration of Master Mix: Rehydrate the lyophilized Master Mix using the Rehydration Solution. The Master Mix contains oligonucleotide primers, fluorophore and quencher-labeled probes targeting conserved regions of RSV and hMPV, as well as the process control sequence.
-
- Nucleic Acid Amplification and Detection: Add 15 µL of the rehydrated Master Mix to each reaction tube or plate well. Then add 5 uL of extracted nucleic acids (specimen with PRC) to the plate well or appropriately labeled reaction tube. Place the tube or plate into either the SmartCycler® II or 7500 Fast Dx instruments, respectively.
Once the reaction tube or plate is added to the instrument, initiate the assay protocol. This protocol initiates reverse transcription of the RNA targets generating complementary DNA, and the subsequent amplification of the target amplicons occurs. The Quidel Molecular RSV + hMPV assay is based on TagMan® chemistry and uses an enzyme with reverse transcriptase, DNA polymerase, and 5'-3' exonuclease activities. During DNA amplification, this enzyme cleaves the probe bound to the complementary DNA sequence, separating the quencher dye from the reporter dye. This step generates an increase in fluorescent signal upon excitation by a light source of the appropriate wavelength. With each cycle, additional dye molecules are separated from their quenchers resulting in additional signal. If sufficient
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.
fluorescence is achieved by 50 cycles on the SmartCycler® II or 35 cycles on the ABI 7500 Fast Dx, the sample is reported as positive for the detected nucleic acid.
Device Comparison
.
The Quidel Molecular RSV + hMPV assay will be compared to legally marketed RT-PCR assays. The characteristics of Quidel Molecular RSV + hMPV assay ("Subject Device") and the Prodesse ProFlu + and Pro hMPV+ ("Predicate Devices") are described in the table below:
| Subject Device and Comparator Device Comparison | |||
|---|---|---|---|
| Item | Subject DeviceQuidel MolecularRSV + hMPV Assay | Predicate DeviceProdesse ProFlu+ | Predicate DeviceProdesse ProhMPV+ |
| Intended Use | The Quidel MolecularRSV + hMPV assay isa multiplex Real TimeRT-PCR assay for thein vitro qualitativedetection andidentification ofrespiratory syncytialvirus and humanmetapneumovirusviral RNA extractedfrom nasal andnasopharyngeal swabsspecimens with signsand symptoms ofrespiratory infection.This in vitrodiagnostic test isintended to aid in thedifferential diagnosisof respiratorysyncytial virus andhumanmetapneumovirusinfections. This test isnot intended to | The ProFlu™+ Assayis a multiplex Real-Time PCR (RT-PCR)in vitro diagnostic testfor the rapid andqualitative detectionand discrimination ofInfluenza A Virus,Influenza B Virus, andRespiratory SyncytialVirus (RSV) nucleicacids isolated andpurified fromnasopharyngeal (NP)swab specimensobtained fromsymptomatic patients.This test is intendedfor use to aid in thedifferential diagnosisof Influenza A,Influenza B and RSVviral infections inhumans and is notintended to detectInfluenza C | The Pro hMPV+Assay is a Real TimeRT-PCR in vitrodiagnostic test for thequalitative detection ofhumanMetapneumovirus(hMPV) nucleic acidisolated and purifiedfrom nasopharyngealswab (NP) specimensobtained fromindividuals exhibitingsigns and symptoms ofacute respiratoryinfection. This assaytargets a highlyconserved region ofthe Nucleocapsid geneof hMPV. Thedetection of hMPVnucleic acid fromsymptomatic patientsaids in the diagnosis ofhuman respiratoryhMPV infection if |
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| Item | Subject DeviceQuidel Molecular | Predicate DeviceProdesse ProFlu+ | Predicate DeviceProdesse ProhMPV+ | ||
|---|---|---|---|---|---|
| RSV + hMPV Assay | |||||
| differentiate the fourgenetic sub-lineagesof hMPV.Negative results donot preclude RSV orhMPV infection andshould not be used asthe sole basis fordiagnosis, treatmentor other patientmanagementdecisions. | Negative results do notpreclude influenza orRSV virus infectionand should not be usedas the sole basis fortreatment or othermanagement decisions.It is recommended thatnegative RSV resultsbe confirmed byculture.Performancecharacteristics forInfluenza A Viruswere established whenInfluenza A/H3 andA/H1 were thepredominant InfluenzaA viruses incirculation. Whenother Influenza Aviruses are emerging,performancecharacteristics mayvary.If infection with anovel Influenza Avirus is suspectedbased on currentclinical andepidemiologicalscreening criteriarecommended by | used in conjunctionwith other clinical andlaboratory findings.This test is notintended todifferentiate the fourgenetic sub-lineages ofhMPV.Negative results do notpreclude hMPVinfection and shouldnot be used as the solebasis for diagnosis,treatment or othermanagement decisions. |
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| Subject Device and Comparator Device Comparison | |||
|---|---|---|---|
| Item | Subject DeviceQuidel MolecularRSV + hMPV Assay | Predicate DeviceProdesse ProFlu+ | Predicate DeviceProdesse ProhMPV+ |
| with appropriateinfection controlprecautions for novelvirulent Influenzaviruses and sent tostate or local healthdepartment for testing.Viral culture shouldnot be attempted inthese cases unless aBSL 3+ facility isavailable to receiveand culture specimens. | |||
| Assay Target | respiratory syncytialvirus, hMPV | Influenza A virus,influenza B virus,respiratory syncytialvirus | hMPV |
| SampleTypes | nasal swab,nasopharyngeal swab | nasopharyngeal swab | nasopharyngeal swab |
| ExtractionMethods | bioMérieuxeasyMAG AutomatedMagnetic ExtractionReagents | Roche MagNA PureLC Total Nucleic AcidIsolation Kit or thebioMérieux easyMAGAutomated MagneticExtraction Reagents | Roche MagNA PureLC Total Nucleic AcidIsolation Kit or thebioMérieux easyMAGAutomated MagneticExtraction Reagents |
| AssayMethodology | PCR-based system fordetecting the presenceor absence of viralRNA in clinicalspecimens | PCR-based system fordetecting the presenceor absence of viralRNA in clinicalspecimens | PCR-based system fordetecting the presenceor absence of viralRNA in clinicalspecimens |
| DetectionTechniques | Multiplex assay usingdifferent reporter dyesfor each target | Multiplex assay usingdifferent reporter dyesfor each target | Multiplex assay usingdifferent reporter dyesfor each target |
| Viral Targets | RSV: L viralpolymerase and NS2genes | Influenza A: MatrixGene;Influenza B: Non- | Nucleocapsid |
| Subject Device and Comparator Device Comparison | |||
| Item | Subject DeviceQuidel MolecularRSV + hMPV Assay | Predicate DeviceProdesse ProFlu+ | Predicate DeviceProdesse ProhMPV+ |
| hMPV: RNApolymerase gene | structural NS1 andNS2 | ||
| LoD | The analyticalsensitivity (limit ofdetection or LoD) ofthe Quidel MolecularhMPV assay wasdetermined usingquantified(TCID50/mL) culturesof 2 RSV strains and4 hMPV strains (A1,A2, B1, B2) seriallydiluted in negativenasopharyngealmatrix. Each dilutionwas extracted usingthe NucliSENSeasyMAG System andtested in replicates of20 per concentrationof virus on both theApplied Biosystems®7500 Fast Dxplatform. Analyticalsensitivity (LoD) isdefined as the lowestconcentration atwhich 95% of allreplicates testedpositive, ranged from10 -1 to 101TCID50/mL. | The analyticalsensitivity (limit ofdetection or LoD) ofthe ProFlu+ Assay wasdetermined usingquantified(TCID50/mL) culturesof 4 Influenza A (2H1N1 and 2 H3N2), 2Influenza B, 2Respiratory SyncytialVirus Type A, and 2Respiratory SyncytialVirus Type B strainsserially diluted innasopharyngealclinical matrix. Eachviral strain wasextracted using theRoche MagNA PureLC instrument andtested in replicates of20 per concentration ofvirus.Analytical sensitivity(LoD), as defined asthe lowestconcentration at which95% of all replicatestested positive, rangedfrom 102 to 101TCID50/mL. | The analyticalsensitivity (limit ofdetection or LoD) ofthe Pro hMPV+ Assaywas determined usingquantified(TCID50/mL) culturesof 2 hMPV (subtypeA2 and subtype B2)strains serially dilutedin nasopharyngealclinical matrix. Eachviral strain wasextracted using theRoche MagNA PureLC instrument andtested in replicates of20 per concentration ofvirus.Analytical sensitivity(LoD) as defined asthe lowestconcentration at which≥ 95% of all replicatestested positive, rangedfrom 102 - 101TCID50/mL. |
·
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Quidel Molecular RSV + hMPV Assay 7/20/2012
Section 05, Page 7 of 15
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Analytical Performance:
Reproducibility:
The reproducibility of the Quidel Molecular Influenza RSV + hMPV assay was evaluated at 3 laboratory sites. Reproducibility was assessed using a panel of 4 simulated samples that include medium (5x LoD) and low (2x LoD), high negative (0.3x LoD) RSV, hMPV and negative samples. Panels and controls were tested at each site by 2 operators for 5 days (triplicate testing x 2 operators x 5 days x 3 sites = 90 results per level for each virus). The LoD values are based on the values obtained in the LoD study. The panels and controls were extracted using the bioMérieux easyMAG system and tested on the Cepheid SmartCycler® II and the Applied Biosystems® 7500 Fast DX.
| Reproducibility Results - Cepheid SmartCycler | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Panel MemberID | Site 1 | Site 2 | Site 3 | TotalResults | ||||||
| Results | AVECt | %CV | Results | AVECt | %CV | Results | AVECt | %CV | ||
| RSV HighNegative 0.06xLoD | 18/30 | 43.15 | 9.84 | 18/30 | 41.9 | 11.0 | 7/30 | 45.3 | 11.8 | 43/90 |
| RSV LowPositive2x LoD | 29/29 | 31.79 | 6.32 | 30/30 | 31.2 | 4.1 | 30/30 | 31.9 | 4.4 | 89/89 |
| RSV MedPositive 5xLoD | 30/30 | 30.35 | 9.57 | 30/30 | 29.7 | 2.5 | 30/30 | 29.2 | 9.1 | 90/90 |
| RSV Negative | 1/30 | 48.50 | N/A | 0/30 | N/A | N/A | 1/30 | 43.0 | N/A | 2/90 |
| RSV NegativeControl | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| RSV PositiveControl | 30/30 | 28.25 | 5.31 | 30/30 | 27.5 | 2.9 | 30/30 | 26.8 | 9.7 | 90/90 |
| hMPV HighNegative 0.06xLoD | 25/30 | 37.27 | 10.75 | 20/30 | 40.43 | 7.75 | 11/30 | 42.3 | 5.4 | 56/90 |
| hMPV LowPositive2x LoD | 30/30 | 32.35 | 7.68 | 30/30 | 31.55 | 3.19 | 30/30 | 32.2 | 3.5 | 90/90 |
| hMPV MedPositive 5xLoD | 30/30 | 30.19 | 4.01 | 30/30 | 29.95 | 2.02 | 30/30 | 30.4 | 1.7 | 90/90 |
| hMPVNegative | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| hMPVNegativeControl | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| hMPV PositiveControl | 30/30 | 27.51 | 2.53 | 30/30 | 26.83 | 2.37 | 30/30 | 27.0 | 1.1 | 90/90 |
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| Reproducibility Results - Applied Biosystems 7500 Fast Dx | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Panel MemberID | Site 1 | Site 2 | Site 3 | TotalResults | ||||||
| Results | AVE Ct | %CV | Results | AVE Ct | %CV | Results | AVE Ct | %CV | ||
| RSV HighNegative 0.06xLoD | 4/30 | 26.4 | 18.2 | 5/30 | 34.6 | 1.0 | 7/30 | 31.7 | 9.1 | 16/90 |
| RSV LowPositive2x LoD | 30/30 | 24.5 | 6.64 | 30/30 | 22.8 | 7.3 | 30/30 | 24.5 | 8.8 | 90/90 |
| RSV MedPositive 5xLoD | 30/30 | 21.02 | 5.57 | 30/30 | 20.8 | 6.2 | 30/30 | 21.5 | 8.4 | 90/90 |
| RSV Negative | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| RSV NegativeControl | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| RSV PositiveControl | 30/30 | 19.2 | 3.75 | 30/30 | 7.21 | 2.8 | 30/30 | 7.8 | 8.0 | 90/90 |
| hMPV HighNegative 0.06xLoD | 6/30 | 28.5 | 20.9 | 2/30 | 31.6 | 0.67 | 10/30 | 27.3 | 13.2 | 18/90 |
| hMPV LowPositive2x LoD | 27/30 | 24.5 | 11.07 | 30/30 | 24.6 | 12.4 | 30/30 | 24.6 | 12.8 | 87/90 |
| hMPV MedPositive 5xLoD | 29/30 | 23.1 | 7.58 | 30/30 | 23.3 | 4.9 | 30/30 | 21.7 | 6.4 | 89/90 |
| hMPVNegative | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| hMPVNegativeControl | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 |
| hMPV PositiveControl | 30/30 | 18.89 | 2.49 | 30/30 | 11.8 | 2.5 | 30/30 | 11.4 | 4.2 | 90/90 |
The data from the combined sites indicates that the Quidel Molecular RSV + hMPV assay generates reproducible results for RSV and hMPV when tested with the Cepheid SmartCycler® Il and the Applied Biosystems® 7500 Fast Dx.
Limit of Detection
The analytical sensitivity (limit of detection or LoD) of the Quidel Molecular RSV + hMPV assay was determined using quantified (TCID50/mL) cultures of 2 RSV strains and 4 hMPV strains (1 A1, 1 A2, 1 B1, 1B2) serially diluted in negative nasopharyngeal matrix. Each dilution was extracted in replicates of 20 per concentration of virus using the NucliSENS easyMAG System and tested on both the 7500 Fast Dx or Cepheid SmartCycler® II platforms. Analytical sensitivity (LoD) is defined as the lowest concentration at which 95% of all replicates tested positive.
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| LoD Value Summary for the 7500 Fast Dx & SmartCycler | ||
|---|---|---|
| Virus | LoD TCID50/mL 7500 Fast Dx | LoD TCID50/mLSmartCycler |
| RSV A | 6.29E-01 | 1.89E+00 |
| RSV B | 7.5E-01 | 7.5E-01 |
| hMPV-A1 | 1.7E+01 | 2.645E+01 |
| hMPV-A2 | 2.91E+00 | 2.91E+00 |
| hMPV-B1 | 1.05E+00 | 7.88E+00 |
| hMPV-B2 | 2.25E+00 | 2.25E+00 |
Analytical reactivity (inclusivity)
The reactivity of the Quidel Molecular RSV + hMPV assay was evaluated against multiple strains of RSV and hMPV. The clinical panel consisted of 13 RSV strains and 12 hMPV strains (3 A-1, 2 A-2, 3 B-1, and 4 B-2). Each panel member was extracted using the NucliSens easyMAG instrument and tested in triplicate.
| RSV Inclusivity Panel | ||||
|---|---|---|---|---|
| Subtype | Strain | TCID 50/mL | (7500 Dx) | (SmartCycler) |
| A | Long | $1.89E+0$ | Positive | Positive |
| A | A-2 | $9.38E-1$ | Positive | Positive |
| B | Washington | $1.58E0$ | Positive | Positive |
| B | 9320 | $1.95E+0$ | Positive | Positive |
| NA | WC-026 | NA | Positive | Positive |
| NA | WC-189 | NA | Positive | Positive |
| NA | MHIA - 0022 | NA | Positive | Positive |
| NA | MHIA - 0020 | NA | Positive | Positive |
| NA | MHIA - 0016 | NA | Positive | Positive |
| NA | Chile - 424 | NA | Positive | Positive |
| NA | Chile - 457 | NA | Positive | Positive |
| NA | WC - 229 | NA | Positive | Positive |
| NA | WC - 192 | NA | Positive | Positive |
Sec05 Summary_QM_RSV + hMPV
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| hMPV Inclusivity Panel | ||||
|---|---|---|---|---|
| Subtype | Strain | TCID50/mL | (7500 Dx) | (SmartCycler) |
| A1 | Italy | 1.11E+2 | Positive | Positive |
| B1 | Italy | 2.20E+0 | Positive | Positive |
| B2 | Italy | 4.50E+1 | Positive | Positive |
| B1 | Peru2-2002 G Gene | 4.17E+2 | Positive | Positive |
| B2 | Peru1-2002 G Gene | 1.26E+2 | Positive | Positive |
| B1 | Peru3-2003 G Gene | 1.26E+2 | Positive | Positive |
| B2 | Peru6-2003 | 5.01E+2 | Positive | Positive |
| A1 | IA3-2002 G Gene | 1.51E+2 | Positive | Positive |
| A1 | IA10-2003 | 3.80E+2 | Positive | Positive |
| B2 | IA18-2003 G Gene | 6.61E+2 | Positive | Positive |
| A2 | IA14-2003 G Gene | 1.95E+2 | Positive | Positive |
| A2 | Clinical Isolate | 1.05E+2 | Positive | Positive |
Analytical specificity (cross-reactivity)
The analytical specificity of the Quidel Molecular RSV + hMPV assay was evaluated by testing a panel consisting of 27 viruses, 24 bacteria and 1 yeast strain representing common respiratory pathogens or flora commonly present in nasopharynx. Samples were extracted using the NucliSENS easyMAG instrument and tested in triplicate. Analytical specificity of the Quidel Molecular RSV + hMPV assay was 100%.
Organisms used for the study were the following:
Viruses
Influenza A/Mexico/4108/2009, Influenza B/Florida/04/2006, Adenovirus 1/Adenoid 71, Adenovirus 2, Adenovirus 3, Adenovirus 4, Adenovirus 5, Adenovirus 7, Adenovirus 11, Adenovirus 14, Adenovirus 31, Coronavirus NL63, Coronavirus 229E. Coronavirus OC43, Coxsackievirus B4, Coxsackievirus B5/10/2006, Cytomegalovirus, Echovirus 7, Echovirus 9, Echovirus 6, Echovirus
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- Enterovirus 71. Enterovirus 70, Epstein Barr Virus, HSV Type 1 Maclnytre strain. Human Rhinovirus. HSV Type 2 G strain. Measles virus. Mumps virus. Parainfluenza Type 1, Parainfluenza Type 2, Parainfluenza Type 3, Parainfluenza Type 4. Varicella Zoster Virus
Bacteria
Bordetella pertussis. . Bordetella bronchiseptica, Chlamydophila pneumonia. Chlamydia trachomatis, Legionella pneumophila, Mycobacterium intracellulare, Mvcobacterium tuberculosis. Mycobacterium avium. Mycoplasma pneumoniae, Haemophilus influenza. Pseudomonas aeruginosa, Proteus vulgaris, Proteus mirabilis. Neisseria gonorrhoeae, Neisseria menigitidis, Neisseria mucosa, Klebsiella pneumonia, Escherichia coli, Moraxella catarrhalis, Corynebacterium diptheriae, Lactobacillus plantarum, Streptococcus pneumonia, Streptococcus pyogenes, Streptococcus salivarius, Staphylococcus epidermidis, Staphylococcus aureus
Yeast
Candida albicans
Clinical Performance:
Performance characteristics of the Ouidel Molecular RSV + hMPV assay using the Cepheid SmartCycler® II instrument and the Applied Biosystems® 7500 Fast Dx platform were established during a prospective study during the 2012 respiratory virus season (January to March 2012). Samples used for this study were fresh (414) and frozen (600) swab specimens that were collected for routine respiratory virus testing four sites across the United States. A single specimen was collected per patient and tested (direct specimen DFA and culture with DFA) for RSV immediately after collection. The specimens were extracted with the bioMériuex easyMAG and tested with Ouidel Molecular RSV + hMPV assay. Aliquots of each specimen were sent to a central location for testing with a FDA Cleared hMPV molecular test.
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Cepheid SmartCycler® II
One thousand and fourteen (1014) specimens were tested by both the subject and comparative methods for RSV. Five specimens were contaminated in cell culture (0.5%). Results for the remaining 1009 specimens are detailed in the table below.
| Combined Site - Respiratory syncytial virus | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| DSFA & CellCulturew/DFA | 95% CI | |||||||||
| POS | NEG | Total | Sensitivity | 97.9% | 93.9% | 99.3% | ||||
| QM RSV +hMPV | POS | 137 | 21* | 158 | Specificity | 97.6% | 96.3% | 98.4% | ||
| NEG | 3 | 848 | 851 | |||||||
| Total | 140 | 869 | 1009 | Prevalence | 13.9% |
- All originally discordant specimens were positive for RSV by an FDA-cleared RT-PCR assay. All specimens were also positive for RSV by bi-directional sequence analysis.
Nine hundred and sixty (960) specimens were tested by both the subject and comparative devices for hMPV (the comparator device was unavailable to complete comparison testing). Nine specimens were invalid in the comparative device (0.9%). Results for the remaining 951 specimens are detailed in the table below.
| Combined Site - Human metapneumovirus | ||||||||
|---|---|---|---|---|---|---|---|---|
| Pro hMPV+ | 95% CI | |||||||
| POS | NEG | Total | Positive percent agreement | 96.7% | 92.4% | 98.6% | ||
| QM RSV + hMPV | POS | 145 | 3* | 148 | Negative percent agreement | 99.6% | 98.9% | 99.9% |
| NEG | 5 | 798 | 803 | |||||
| Total | 150 | 801 | 951 | Prevalence | 15.8% |
- All originally discordant specimens were positive for hMPV by bi-directional sequence analysis.
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Applied Biosystems 7500 Fast Dx
One thousand and fourteen (1014) specimens were tested by both the subject and comparative methods for RSV. Five specimens were contaminated in cell culture (0.5%). Two specimens were invalid in the subject methods (0.2%). Results for the remaining 1,007 specimens are detailed in the table below.
| Combined Site - Respiratory syncytial virus | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| DSFA & CellCulture w/DFA | 95% CI | ||||||||
| POS | NEG | Total | Sensitivity | 98.6% | 94.9% | 99.6% | |||
| QM RSV + hMPV | POS | 138 | 28* | 166 | Specificity | 96.8% | 95.4% | 97.8% | |
| NEG | 2 | 839 | 841 | ||||||
| Total | 140 | 867 | 1007 | Prevalence | 13.9% |
*25 of 28 originally discordant specimens were positive for RSV by a FDA-cleared RT-PCR assay. 27 of 28 specimens were positive for RSV by bi-directional sequence analysis.
Nine hundred and fifty seven (957) specimens were tested by both the subject and comparative devices for hMPV (the comparator device was unavailable to complete comparison testing). Nine (9) specimens were invalid in the comparative device (0.9%). Two (2) specimens were invalid in the subject methods (0.2%). Results for the remaining 946 specimens are detailed in the table below.
| Combined Site - Human metapneumovirus | ||||||||
|---|---|---|---|---|---|---|---|---|
| Pro hMPV+ | 95% CI | |||||||
| POS | NEG | Total | Positivepercentagreement | 98.0% | 94.3% | 99.3% | ||
| QM RSV +hMPV | POS | 147 | 6* | 153 | Negativepercentagreement | 99.3% | 98.4% | 99.7% |
| NEG | 3 | 790 | 793 | |||||
| Total | 150 | 796 | 946 | Prevalence | 15.9% |
- All originally discordant specimens were positive for hMPV by bi-directional sequence analysis.
Conclusions
Quidel Molecular RSV + hMPV Assay yielded good sensitivity and specificity for RSV. with nasal and nasopharyngeal swabs when compared to a direct specimen DFA and cell culture with DFA.
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Quidel Molecular RSV + hMPV Assay 7/20/2012 . Section 05, Page 15 of 15
Quidel Molecular RSV + hMPV Assay yielded good positive and negative percent agreement with nasal and nasopharyngeal swabs compared to a 510(k) cleared molecular device.
.
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DEPARTMENT OF HEALTH & HUMAN SERVICES
Image /page/15/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of an eagle or bird-like figure with three curved lines representing its body and wings.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-002
Quidel Corporation C/O Ronald H. Lollar 1055 East State Street, Suite 100 Athens, Ohio 45701
March 8, 2013
Re: K122189
Trade/Device Name: Quidel® Molecular RSV + hMPV Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: Class II Product Code: OEM, OCC Dated: January 28, 2013 Received: January 29, 2013
Dear Mr. Lollar:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.
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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostics and Radiological Health at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.
Sincerely yours,
Uwesschierf -S for
Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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510(k) Number (if known): K122189
Device Name: Quidel® Molecular RSV + hMPV Assay
Indication for Use:
The Quidel Molecular RSV + hMPV assay is a multiplex Real Time RT-PCR assay for the qualitative detection and identification of respiratory syncytial virus and human metapneumovirus RNA extracted from nasal and nasopharyngeal swabs specimens from patients with signs and symptoms of respiratory infection. This in vitro diagnostic test is intended to aid in the differential diagnosis of respiratory syncytial virus and human metapneumovirus infections in humans in conjunction with clinical and epidemiological risk factors. This test is not intended to differentiate the two subtypes of RSV or the four genetic sub-lineages of hMPV.
Negative results do not preclude RSV infection and/or hMPV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Prescription Use Over-The-Counter Use AND/OR X (21 CFR 801 Subpart C) (Part 21 CFR 801 Subpart D)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED
Concurrence of CDRH, Office of Device Evaluation (ODE)
Tamara V. Feldblyum -S 2013.03.08 15:11:17 -05'00'
Division Sign-Off Office of In Vitro Diagnostics and Radiological Health 510(k) K122189
§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.
(a)
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.
(b)
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.