(125 days)
The Focus Diagnostics Simplexa™ Influenza A H1N1 (2009) assay is intended for use on the 3M Integrated Cycler as part of the Microfluidic Molecular System for the in vitro qualitative detection and differentiation of influenza A and 2009 H1N1 influenza viral RNA in nasopharyngeal swabs (NPS), nasal swabs (NS), and nasopharyngeal aspirates (NPA) from human patients with signs and symptoms of respiratory infection in conjunction with clinical and epidemiological risk factors.
Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2009-2010 influenza season when 2009 H1N1 influenza was the predominant influenza A virus 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.
Simplexa™ Influenza A H1N1 (2009) test kit is a nucleic acid amplification test that uses real-time reverse transcriptase polymerase chain reaction (RT-PCR) amplification to enable simultaneous and distinct detection of influenza A and 2009 H1N1 influenza in a single reaction from nasophayngeal swabs (NPS), nasal swabs (NS), and nasopharyngeal aspirates (NPA). The assay combines real-time PCR amplification with fluorescent signal detection technology. A bi-functional fluorescent probe-primer is used together with a reverse primer to amplify a specific target (for each analyte and internal control). A fluorescent signal is generated after the separation of the fluorophore from the quencher as a result of the binding of a probe element to the extended RNA fragment synthesized during amplification.
The 3M Integrated Cycler is a rapid real-time Polymerase Chain Reaction thermocycler used for the identification of nucleic acid from prepared biological samples. The instrument utilizes disk media to contain and to process samples. The instrument uses real time flourometric detection to identify targets within the sample wells. The instrument is controlled by an external computer running the Integrated Cycler Studio Software. Together, the instrument, software and test kit are referred to as the "Microfluidic Molecular System."
Here's a breakdown of the acceptance criteria and study details for the Simplexa™ Influenza A H1N1 (2009) assay based on the provided document:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly state "acceptance criteria" in a separate table with target values for clinical performance as commonly seen for medical devices. However, the reported performance metrics (Positive Percent Agreement - PPA, Negative Percent Agreement - NPA) from the clinical agreement studies serve as the de-facto acceptance criteria demonstrated by the device.
| Performance Metric | Acceptance Criteria (Implied by Study Results) | Reported Device Performance (Simplexa™ Influenza A H1N1 (2009)) |
|---|---|---|
| 2009 H1N1 Influenza - Swabs (Prospective) | ||
| Positive Agreement | High, ideally 95% or higher | 100% (101/101); 95% CI: 96.3-100% |
| Negative Agreement | High, ideally 95% or higher | 95.5% (171/179); 95% CI: 91.4-97.7% |
| 2009 H1N1 Influenza - Aspirates (Prospective) | ||
| Positive Agreement | High, ideally 95% or higher | 100% (24/24); 95% CI: 86.2-100% |
| Negative Agreement | High, ideally 95% or higher | 92.5% (74/80); 95% CI: 84.6-96.5% |
| Influenza A - Swabs (Prospective) | ||
| Positive Agreement | High, ideally 95% or higher | 100% (116/116); 95% CI: 96.8-100% |
| Negative Agreement | High, ideally 95% or higher | 92.5% (160/173); 95% CI: 87.6-95.6% |
| Influenza A - Aspirates (Prospective) | ||
| Positive Agreement | High, ideally 95% or higher | 100% (31/31); 95% CI: 89-100% |
| Negative Agreement | High, ideally 95% or higher | 96.1% (73/76); 95% CI: 89-98.6% |
| 2009 H1N1 Influenza - Swabs (Retrospective) | ||
| Positive Agreement | High, ideally 95% or higher | 100% (57/57); 95% CI: 93.7-100% |
| Negative Agreement | High, ideally 95% or higher | 90.8% (139/153); 95% CI: 85.2-94.5% |
| Influenza A - Swabs (Retrospective) | ||
| Positive Agreement | High, ideally 95% or higher | 99.2% (131/132); 95% CI: 95.8-99.9% |
| Negative Agreement | High, ideally 95% or higher | 83.5% (66/79); 95% CI: 73.9-90.1% |
2. Sample Size Used for the Test Set and Data Provenance
-
Prospective Data:
- Nasal/Nasopharyngeal Swabs: 299 specimens initially collected. After exclusions due to lack of consensus among reference assays (10 for Influenza A, 9 additional for H1N1 subtype sequencing), the sample sizes used for agreement calculations are:
- H1N1 Detected: 101
- H1N1 Not Detected: 179
- Influenza A Detected: 116
- Influenza A Not Detected: 173
- Nasopharyngeal Aspirates: 112 specimens initially collected. After exclusions due to lack of consensus among reference assays (5 for Influenza A, 3 additional for H1N1 subtype sequencing), the sample sizes used for agreement calculations are:
- H1N1 Detected: 24
- H1N1 Not Detected: 80
- Influenza A Detected: 31
- Influenza A Not Detected: 76
- Provenance: Collected from patients with signs and symptoms of influenza-like illness at three sites: Austin, TX (September 2009) and the New South Wales region of Australia (July - September 2009). The collected specimens were blinded and randomly distributed to 3 U.S. clinical laboratories for testing.
- Nasal/Nasopharyngeal Swabs: 299 specimens initially collected. After exclusions due to lack of consensus among reference assays (10 for Influenza A, 9 additional for H1N1 subtype sequencing), the sample sizes used for agreement calculations are:
-
Retrospective Data:
- Nasal/Nasopharyngeal Swabs: 214 specimens initially collected from the Focus Sample Bank. After exclusions due to lack of consensus among reference assays (3 for Influenza A, 1 additional for H1N1 subtype sequencing), the sample sizes used for agreement calculations are:
- H1N1 Detected: 57
- H1N1 Not Detected: 153 (including 1 indeterminate)
- Influenza A Detected: 132 (including 1 indeterminate)
- Influenza A Not Detected: 79
- Nasal Washes: 2 specimens (both positive for 2009 H1N1 influenza and Influenza A by both methods).
- Provenance: Focus Sample Bank (implying archival samples). No specific country of origin is mentioned beyond "Focus Sample Bank."
- Nasal/Nasopharyngeal Swabs: 214 specimens initially collected from the Focus Sample Bank. After exclusions due to lack of consensus among reference assays (3 for Influenza A, 1 additional for H1N1 subtype sequencing), the sample sizes used for agreement calculations are:
3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications
The document does not specify the number of experts or their qualifications for establishing the ground truth. It refers to a "composite reference method" which includes multiple validated assays, but not human expert review for establishing ground truth.
4. Adjudication Method for the Test Set
The ground truth was established using a composite reference method which included:
- Luminex xTAG RVP Flu A target
- A validated PCR assay using primer and probe sequences published by the CDC
- A well-characterized PCR followed by sequencing.
Specimens were excluded from analysis if there was "no consensus among the reference assays" for the influenza A result or if sequencing data was not available for subtyping. This implies a form of consensus/adjudication among the reference methods, but not by human experts in the traditional sense as a separate step.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done
No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not performed. This study evaluates the Simplexa™ Influenza A H1N1 (2009) assay in a standalone capacity against a composite reference (ground truth), not in comparison to or in assistance with human readers. Therefore, there is no effect size reported for human readers improving with AI vs without AI assistance.
6. If a Standalone (i.e. algorithm only without human-in-the-loop performance) was done
Yes, this was a standalone performance study. The Simplexa™ Influenza A H1N1 (2009) assay's performance was evaluated by directly comparing its results to the established composite reference ground truth. There is no mention of human-in-the-loop involvement in the performance evaluation.
7. The Type of Ground Truth Used
The ground truth used was a composite reference method comprised of:
- Luminex xTAG RVP Flu A target
- A validated PCR assay using primer and probe sequences published by the CDC
- A well-characterized PCR followed by sequencing.
8. The Sample Size for the Training Set
The document does not explicitly mention a separate "training set" or its sample size for the development of the Simplexa™ Influenza A H1N1 (2009) assay. The provided studies focus solely on the performance of the developed assay, typically referring to validation or test sets.
9. How the Ground Truth for the Training Set was Established
Since a "training set" is not explicitly discussed, the method for establishing its ground truth is also not specified. It can be inferred that similar validated molecular methods would have been used during the assay's development phase.
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Image /page/0/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in large, bold, sans-serif font, with the word "Diagnostics" in a smaller font size underneath. A curved, black shape is positioned to the left of the word "FOCUS", adding a visual element to the logo.
510(k) Summary of Safety and Effectiveness
Simplexa™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 1 of 9
| Applicant | Focus Diagnostics, Inc.11331 Valley View StreetCypress, California 90630USA | |
|---|---|---|
| Establishment Registration No. | 2023365 | |
| Contact Person | Tara Vivianitel 714.822.2115fax 714.822.3898tviviani@focusdx.com | MAY 24 2010 |
| Summary Date | May 18, 2010 | |
| Proprietary Name | Simplexa™ Influenza A H1N1 (2009) | |
| Generic Name | Influenza A H1N1 2009 Real Time RT-PCR | |
| Classification | Class II, Special ControlsLuminex Diagnostics XTAG RESPIRATORY VIRAL PANEL(K091667, K081483, K063765)CDC Human Influenza Virus Real-Time RT- PCR Detection andCharacterization Panel (K080570) |
Intended Use
The Focus Diagnostics Simplexa™ Influenza A H1N1 (2009) assay is intended for use on the 3M Integrated Cycler as part of the Microfluidic Molecular System for the in vitro qualitative detection and differentiation of influenza A and 2009 H1N1 influenza viral RNA in nasopharyngeal swabs (NS), and nasopharyngeal aspirates (NPA) from human patients with signs and symptoms of respiratory in conjunction with clinical and epidemiological risk factors.
Neqative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2009-2010 influenza season when 2009 H1N1 influenza was the predominant influenza A virus 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.
Device Description
Simplexa™ Influenza A H1N1 (2009) test kit is a nucleic acid amplification test that uses real-time reverse transcriptase polymerase chain reaction (RT-PCR) amplification to enable simultaneous and distinct detection of influenza A and 2009 H1N1 influenza in a single reaction from nasophayngeal swabs (NPS), nasal swabs (NS), and nasopharyngeal aspirates (NPA). The assay combines real-time PCR amplification with fluorescent signal detection technology. A bi-functional fluorescent probe-primer is used together with a reverse primer to amplify a specific target (for each analyte and internal control). A fluorescent signal is generated after the separation of the fluorophore from the quencher as a result of the binding of a probe element to the extended RNA fragment synthesized during amplification.
The 3M Integrated Cycler is a rapid real-time Polymerase Chain Reaction thermocycler used for the identification of nucleic acid from prepared biological samples. The instrument utilizes disk media to contain and to process samples. The instrument uses real time flourometric detection to identify targets within the sample wells. The instrument is controlled by an external computer running the Integrated Cycler Studio Software. Together, the instrument, software and test kit are referred to as the "Microfluidic Molecular System."
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Image /page/1/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features a stylized, curved shape resembling a crescent or a checkmark above the word "FOCUS" in bold, sans-serif font. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.
510(k) Summary of Safety and Effectiveness Simplexa™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 2 of 9
Predicate Device Information
| Trade Name / Method | 510(k)submitter | 510(k)number | Decision Date | Panel | ProductCode(s) |
|---|---|---|---|---|---|
| XTAG RespiratoryViral Panel - FluA | Luminex | K091667K081483K063765 | 06/25/200906/25/200811/30/2007 | Microbiology(83) | OCC, OEM,OEP |
| CDC Human InfluenzaVirus Real-Time RT-PCR Detection andCharacterization Panel | Centers forDiseaseControl andPrevention | K080570 | 09/30/2008 | Microbiology(83) | NXD, OEP,OCC, NSU |
| ItemName | Device | Predicate | |
|---|---|---|---|
| Simplexa™ Influenza AH1N1 (2009) | xTAG Respiratory ViralPanel - FLUA | CDC Human InfluenzaVirus Real-Time RT-PCRDetection andCharacterization Panel | |
| Intended Use | The Focus DiagnosticsSimplexa™ Influenza A H1N1(2009) assay is intended foruse on the 3M IntegratedCycler as part of theMicrofluidic Molecular Systemfor the in vitro qualitativedetection and differentiation ofinfluenza A and 2009 H1N1influenza viral RNA innasopharyngeal swabs (NPS),nasal swabs (NS), andnasopharyngeal aspirates(NPA) from human patientswith signs and symptoms ofrespiratory infection inconjunction with clinical andepidemiological risk factors.Negative results do notpreclude influenza virusinfection and should not beused as the sole basis fortreatment or other patientmanagement decisions.Performance characteristicsfor influenza A wereestablished during the 2009-2010 influenza season when2009 H1N1 influenza was thepredominant influenza Aviruses in circulation. Whenother Influenza A viruses areemerging, performancecharacteristics may vary.If infection with a novelInfluenza A virus is suspected | The xTAG® RespiratoryViral Panel (RVP) is aqualitative nucleic acidmultiplex test intended forthe simultaneous detectionand identification ofmultiple respiratory virusnucleic acids innasopharyngeal swabsfrom individuals suspectedof respiratory tractinfections. The followingvirus types and subtypesare identified using RVP:Influenza A, Influenza Asubtype H1, Influenza Asubtype H3, Influenza B,Respiratory Syncytial Virussubtype A, RespiratorySyncytial Virus subtype B,Parainfluenza 1,Parainfluenza 2, andParainfluenza 3 virus,Human Metapneumovirus,Rhinovirus, andAdenovirus. The detectionand identification of specificviral nucleic acids fromindividuals exhibiting signsand symptoms ofrespiratory infection aids inthe diagnosis of respiratoryviral infection if used inconjunction with other | The Human Influenza VirusReal-time RT-PCRDetection andCharacterization Panel(rRT-PCR Flu Panel) isintended for use in Real-time RT-PCR assays on anABI 7500 Fast Dx Real-time PCR instrument inconjunction with clinicaland epidemiologicalinformation:* for qualitative detectionof influenza virus type A orB in symptomatic patientsfrom viral RNA innasopharyngeal and/ornasal swab specimens, for determination of thesubtype of seasonal humaninfluenza A virus, asseasonal A/HI or A/H3, ifpresent, from viral RNA innasopharyngeal and/ornasal swab specimens, for presumptiveidentification of virus inpatients who may beinfected with influenza Asubtype A/H5 (Asianlineage) from viral RNA inhuman respiratoryspecimens and viral culturein conjunction with clinical |
| ItemName | Device | Predicate | |
| Simplexa™ Influenza AH1N1 (2009) | xTAG Respiratory ViralPanel - FLUA | CDC Human InfluenzaVirus Real-Time RT- PCRDetection andCharacterization Panel | |
| based on current clinical andepidemiological screeningcriteria recommended bypublic health authorities,specimens should be collectedwith appropriate infectioncontrol precautions for novelvirulent Influenza viruses andsent to state or local healthdepartment for testing. Viralculture should not beattempted in these casesunless a BSL 3+ facility isavailable to receive andculture specimens. | clinical and laboratoryfindings. It is recommendedthat specimens found to benegative for Influenza B,Respiratory Syncytial Virussubtype A and B,Parainfluenza 1,Parainfluenza 2,Parainfluenza 3 andAdenovirus, afterexamination using RVP beconfirmed by cell culture.Negative results do notpreclude respiratory virusinfection and should not beused as the sole basis fordiagnosis, treatment orother managementdecisions. Positive resultsdo not rule out bacterialinfection, or co-infectionwith other viruses. Theagent detected may not bethedefinite cause of disease.The use of additionallaboratory testing (e.g.bacterial culture,immunofluorescence,radiography) and clinicalpresentation must be takeninto consideration in orderto obtain the final diagnosisof respiratory viral infection.Due to seasonalprevalence, performancecharacteristics for InfluenzaA/H1 were establishedprimarily with retrospectivespecimens.The RVP assay cannotadequately detectAdenovirus species C, orserotypes 7a and 41. TheRVP primers for detectionof rhinovirus cross-reactwith enterovirus. Arhinovirus reactive result | and epidemiological riskfactors.* to provide epidemiologicinformation for surveillancefor influenza viruses.Performancecharacteristics for influenzaA were established wheninfluenza A/H3 and A/H1were the predominantinfluenza A viruses incirculation. When otherinfluenza A viruses areemerging, performancecharacteristics may vary.Testing with the influenzaH5a and H5b primer andprobe sets should not beperformed unless thepatient meets the mostcurrent U.S. Department ofHealth and HumanServices (DHHS) clinicaland epidemiologic criteriafor testing suspect A/H5specimens. The definitiveidentification of influenzaA/H5 (Asian lineage) eitherdirectly from patientspecimens or from viruscultures requires additionallaboratory testing, alongwith clinical andepidemiologicalassessment in consultationwith nationalinfluenza surveillanceexperts.Negative results do notpreclude influenza virusinfection and should not beused as the sole basis fortreatment or other patientmanagement decisions. Allusers, analysts, and anyperson reporting diagnosticresults from use of thisdevice should be trained to | |
| ItemName | Device | Predicate | |
| Simplexa™ Influenza AH1N1 (2009) | xTAG Respiratory ViralPanel - FLUA | CDC Human InfluenzaVirus Real-Time RT- PCRDetection andCharacterization Panel | |
| should be confirmed by analternate method (e.g. cellculture). Performancecharacteristics for InfluenzaA Virus were establishedwhen Influenza A/H3 andA/H1 were the predominantInfluenza A viruses incirculation. When otherInfluenza A viruses areemerging, performancecharacteristics may vary. Ifinfections with a 2009H1N1 Influenza A virus issuspected based oncurrent clinical andepidemiological screeningcriteria recommended bypublic health authorities,specimens should becollected with appropriateinfection controlprecautions for 2009 H1N1virulent Influenza virusesand sent to a state or localhealth department fortesting. Viral culture shouldnot be attempted in thesecases unless a BSL 3+facility is available toreceive and culturespecimens. | perform and interpret theresults from this procedureby a CDC instructor ordesignee prior to use. CDCInfluenza Division will limitthe distribution of thisdevice to only those userswho have successfullycompleted training providedby CDC instructors ordesignees. | ||
| Assay Targets | Influenza A2009 H1N1 Influenza | Influenza A, Influenza Asubtype H1, Influenza Asubtype H3, Influenza B,Respiratory Syncytial Virussubtype A, RespiratorySyncytial Virus subtype B,Parainfluenza 1,Parainfluenza 2, andParainfluenza 3 virus,Human Metapneumovirus,Rhinovirus, andAdenovirus. | Influenza A/H1Influenza A/H3Influenza A/H5 (asianlineage)Influenza B |
| ItemName | Device | Predicate | |
| Simplexa™ Influenza AH1N1 (2009) | xTAG Respiratory ViralPanel - FLUA | CDC Human InfluenzaVirus Real-Time RT- PCRDetection andCharacterization Panel | |
| ExtractionMethods | Roche MagNA Pure LCTotal Nucleic Acid IsolationKit,QIAGEN QIAamp ViralRNA Mini Kit | QIAGEN QIAamp MiniEluteBiomérieux EasyMag,Biomérieux MiniMag | QIAamp® Viral RNA MiniKit.Qiagen RNeasy® Mini Kit,MagNA Pure LC RNAIsolation Kit IIRoche MagNA Pure LCTotal Nucleic Acid IsolationKit, |
| AssayMethodology | PCR-based system fordetecting the presence /absence of viral RNA inclinical specimens | PCR-based system fordetecting the presence /absence of viral DNA/RNAin clinical specimens | PCR-based system fordetecting the presence /absence of viral RNA inclinical specimens |
| DetectionTechniques | Multiplex assay usingdifferent reporter dyes foreach target. | Multiplex assay using acombination of color codedbeads and differentreporter dyes. | A panel of oligonucleotideprimers and dual-labeledhydrolysis (TaqMan®)probes for the qualitativedetection and differentiationof influenza virus type andsubtype target sequences. |
| Influenza AViral Target | Well conserved region ofthe matrix gene | Well conserved region ofthe matrix gene | Well conserved region ofthe matrix gene |
| H1N1 (2009)Viral Target | Well conserved region ofthe hemagglutinin genespecific for H1N1 (2009) | n/a | n/a |
| LoD | Influenza A StrainsTCID50/mL in a range of$1x10^{-1}$ to $2.7x10^{1}$2009 H1N1TCID50/mL in a range of$1x10^{-1}$ to $2.7x10^{1}$Please refer to detailed tablebelow. | Influenza AA/PR/8/34 (TCID50/mL =$8x10^{-1}$ )A/Victoria/3/75 (TCID50/mL= $1x10^{2}$ ) | Influenza AA/New Caledonia/20/1999(TCID50/mL = $10^{1.2}$ )A/Hawaii/15/2001(TCID50/mL = $10^{1.5}$ )A/New York/55/2004(TCID50/mL = $10^{2.2}$ )A/Wisconsin/67/2005(TCID50/mL = $10^{1.2}$ ) |
| Reproducibility | FLUA Inter-AssayTotal %CV range 0.0 to 4.8FLUA Intra-AssayTotal %CV range 0.0 to 6.6H1N1 Inter-AssayTotal %CV range 0.0 to 1.8H1N1 Intra-AssayTotal %CV range 0.0 to 4.7 | Influenza A - low positive%CV range 29.41 to 60.22Influenza A - medium titer%CV range 5.6 to 35.6 | Influenza A - 1:10 of lowpositive %CV range 1.94 to7.09Influenza A - low positive%CV range 2.12 to 7.89 |
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Image /page/2/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with the word "Diagnostics" in a smaller, non-bold font underneath. To the left of the word "FOCUS" is a curved, black shape that resembles a crescent or a stylized checkmark.
510(k) Summary of Safety and Effectiveness Simplexa ™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010
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Page 3 of 9
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510(k) Summary of Safety and Effectiveness Simplexa™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 4 of 9
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Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font. Above and to the left of the word is a black, curved shape that resembles a stylized checkmark or swoosh. Below the word "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin black line.
510(k) Summary of Safety and Effectiveness Simplexa™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 5 of 9
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K1001 510(k) Summary of Safety and Effectiveness Simplexa ™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 6 of 9
Reproducibility:
Three investigative sites assessed the device's inter-laboratory reproducibility and inter/intra-assay reproducibility. Each of the three laboratories tested eighteen samples, the Positive Control and the No Template Control, in triplicate on five different days. Each site had two operators who each ran the assay once per day, for a total of two runs per Two sites performed the extraction using the MagNA Pure LC Total Nucleic Acid Isolation Kit; one site day. performed the extraction step using the Qiagen QlAamp Viral RNA Mini Kit. Combined results for all sites are summarized below.
| FLUA Inter-Assay | Total %CV range (0.0 to 4.8) |
|---|---|
| FLUA Intra-Assay | Total %CV range (0.0 to 6.6) |
| H1N1 Inter-Assay | Total %CV range (0.0 to 1.8) |
| H1N1 Intra-Assay | Total %CV range (0.0 to 4.7) |
Limit of Detection
Simplexa™ Influenza A H1N1 (2009) Limit of Detection - FLUA
| Influenza A Strain | LoD MagNA Pure extraction(TCID50/mL) | LoD QIAgen extraction(TCID50/mL) | ||
|---|---|---|---|---|
| Swab | Aspirate | Swab | Aspirate | |
| A/California/7/2009 NYMC x-179-A | 1.3x101 | 1.3x101 | 1.3x101 | 2.7x101 |
| A/Swine NY/02/2009 H1N1 | 1x10-1 | 1x10-1 | 1x10-1 | 1x10-1 |
| A/Solomon Island/03/06 H1 | 5x100 | 5x100 | 1x100 | 1x100 |
| A/Brisbane/59/07 H1 | 1x100 | 1x100 | 1x100 | 1x100 |
| A/Brisbane/10/07 H3 | 1x10-1 | 1x10-1 | 5x10-1 | 5x10-1 |
| A/Wisconsin/67/05 H3 | 1x10-1 | 5x10-1 | 1x10-1 | 1x10-1 |
Simplexa™ Influenza A H1N1 (2009) Limit of Detection - H1N1
| 2009 Influenza A Strain | LoD MagNA Pure extraction(TCID50/mL) | LoD QIAgen extraction(TCID50/mL) | ||
|---|---|---|---|---|
| Swab | Aspirate | Swab | Aspirate | |
| A/California/7/2009 NYMC x-179-A | $1.3\times10^1$ | $2.7\times10^1$ | $2.7\times10^1$ | $2.7\times10^1$ |
| A/Swine NY/02/2009 H1N1 | $1\times10^1$ | $1\times10^1$ | $1\times10^1$ | $1\times10^1$ |
Analytical Reactivity
| Influenza A Strain | Estimated LoD |
|---|---|
| A/PR/8/34 H1N1 | $1x10^0$ |
| A/New Caledonia/20/99 H1N1 | $1x10^0$ |
| A/Taiwan/42/06 H1N1 | $1x10^0$ |
| A/WS/33 H1N1 | $1x10^0$ |
| A/Hong Kong/8/68 H3N2 | $1x10^0$ |
Cross-Reactivity
No cross reactivity was detected for either Influenza A or 2009 H1N1 to organisms that are closely related to influenza A or 2009 H1N1, or cause similar clinical symptoms as influenza A or 2009 H1N1, or present as normal flora in the specimen types of interest.
Interference
Potentially interfering substances were not tested with this methodology; common medications taken by study participants do not appear to inhibit the PCR process.
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510(k) Summary of Safety and Effectiveness Simplexa™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 7 of 9
Clinical Agreement
Specimens were prospectively collected at three sites from patients with signs and symptoms of influenza like illness (Austin, TX (September 2009) and the New South Wales region of Australia (July - September 2009). Specimens were blinded and randomly distributed to 3 U.S. clinical laboratories for testing. Specimens were determined to be positive for 2009 H1N1 influenza by a composite reference method including the Luminex xTAG RVP Flu A target, a validated PCR assay using primer and probe sequences published by the CDC and a well characterized PCR followed by sequencing. Two results were generated for each specimen, an influenza A result and a 2009 H1N1 influenza sub-typing result. Both results must be positive to determine that a specimen is 2009 H1N1 influenza positive.
299 prospectively collected nasal/nasopharyngeal swabs and 112 nasopharyngeal aspirates were analyzed using the Simplexa™ Influenza A H1N1 (2009) assay. The data presented below are stratified by both result and specimen type. Fifteen (15) specimens (10 swabs and 5 aspirates) were excluded from the analysis because there was no consensus among the reference assays for the influenza A result. Twelve (12) specimens (9 swabs and 3 aspirates) were excluded from the 2009 H1N1 Influenza Clinical Acreement Summary tables as sequencing data used to determine subtype was not available. These 12 specimens are included in the Influenza A Clinical Agreement Summary tables.
2009 H1N1 Influenza Clinical Agreement Summary Simplexa™ Influenza A H1N1 (2009) vs. Composite Reference Result - Prospectively Collected Swabs'
| H1N1 Result - Simplexa™ Influenza A H1N1 (2009) | ||||
|---|---|---|---|---|
| CompositeReference Result | n | H1N1Detected | H1N1Not Detected | % Agreement |
| H1N1Detected | 101 | 101 | 0 | % Positive Agreement100%(101/101)95% CI:96.3-100% |
| H1N1Not Detected | 179 | 8 | 171 | % Negative Agreement95.5%(171/179)95% CI:91.4-97.7% |
- Ten (10) samples were excluded from the analysis because there was no consensus among the influenza A reference assays. Nine (9) samples were excluded because sequencing results to determine sub-types were not available.
| H1N1 Result - Simplexa™ Influenza A H1N1 (2009) | |||||
|---|---|---|---|---|---|
| n | H1N1Detected | H1N1Not Detected | % Agreement | ||
| CompositeReference Result | H1N1Detected | 24 | 24 | 0 | % Positive Agreement100%(24/24)95% CI:86.2-100% |
| H1N1Not Detected | 80 | 6 | 74 | % Negative Agreement92.5%(74/80)95% CI:84.6-96.5% |
2009 H1N1 Influenza Clinical Agreement Summary
(1N1 (2009) vs. Composite Reference Result - Prospectively Colle
- Five (5) samples were excluded from the analysis because there was no consensus among the influenta A reference assays. Three (3) samples were excluded because sequencing results to determine sub-types were not available.
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Image /page/7/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font. A curved, black shape resembling a stylized checkmark or swoosh is positioned to the left of the word, partially encompassing the "F". Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin black line.
510(k) Summary of Safety and Effecti Simplexa ™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010 Page 8 of 9
Influenza A Clinical Agreement Summary
Simplexa™ Influenza A H1N1 (2009) vs. Composite Reference Result - Prospectively Collected Swabs'
| Influenza A Result - Simplexa™ Influenza A H1N1 (2009) | ||||
|---|---|---|---|---|
| n | Influenza A Detected | Influenza A Not Detected | % Agreement | |
| CompositeReference Result | ||||
| Influenza ADetected | 116 | 116 | 0 | % Positive Agreement100%(116/116)95% CI:96.8-100% |
| Influenza ANot Detected | 173 | 13 | 160 | % Negative Agreement92.5%(160/173)95% CI:87.6-95.6% |
- Due to the low prevalence of other strains of intrasting period, all FLU A responses from prospectively collected sware combined to demonstrate the performance of the FLU A bi-functional fluorescent primer-probe. Of the 116 speciment to be positive for FLU A: 101 were 2009 H N1 influenza positive, zero (0) were H3N2, two (2) were not detected by the atternate PCR and could not be sequenced, and nine (9) were not sub-typed. Ten (10) samples were excluded from the analysis because there was no consensus arnong the influenza A reference assays.
Influenza A Clinical Agreement Summary
Simplexa™ Influenza A H1N1 (2009) vs. Composite Reference Result – Prospectively Collected Aspirates
| Influenza A Result - Simplexa™ Influenza A H1N1 (2009) | ||||
|---|---|---|---|---|
| n | Influenza A Detected | Influenza A Not Detected | % Agreement | |
| Influenza ADetected | 31 | 31 | 0 | % Positive Agreement100%(31/31)95% CI:89-100% |
| Influenza ANot Detected | 76 | 3 | 73 | % Negative Agreement96.1%(73/76)95% CI:89-98.6% |
- Due to the low prevalence of other strains of intributional FLU A responses from prospecively collected aspirates were combined to demonstrate the performance of the FLU A bi-functional fluorescent primer-probe. Of the 31 specimens delermined to be positive for FLU A, 24 were 2009 H1N1 influenza positive, one (1) was sequenced but the sub-ype could not be determined, three (3) were not detected by the allernate PCR and could not be sequenced three sufficient volume to sequence to determine sub-type. Five (5) samples were excluded from the analysis because there was no consensus of the influenza A reference assays.
An additional 214 retrospectively collected nasal/nasopharyngeal swabs and 2 nasal washes from the Focus Sample Bank were also tested at 3 sites. Three (3) swab specimens were excluded from the analysis because there was no consensus among the reference assay results. One (1) swab specimen was excluded from the 2009 H1N1 Influenza Clinical Agreement Summary tables as sequencing data used to determine subtype was not available. This specimen is included in the Influenza A Clinical Agreement Summary tables.
2009 H1N1 Influenza Clinical Agreement Summary
Simplexa™ Influenza A H1N1 (2009) vs. Composite Reference Result - Retrospectively Collected Swabs
| H1N1 Result - Simplexa™ Influenza A H1N1 (2009) | ||||||
|---|---|---|---|---|---|---|
| CompositeReference Result | n | H1N1Detected | H1N1Not Detected | Indeterminate | % Agreement | |
| H1N1Detected | 57 | 57 | 0 | 0 | % Positive Agreement100%(57/57)95% CI:93.7-100% | |
| H1N1Not Detected | 153 | 13 | 139 | 1 | % Negative Agreement90.8%(139/153)95% CI:85.2-94.5% |
- Three (3) samples were excluded from the analysis because there was no consensus of the influenza A reference assays. One (1) sample was excluded from the analysis because sequencing results there was insufficient sample to perform sequencing.
Two retrospectively collected washes were found to be positive for 2009 H1N1 influenza by the composite reference method and by the Simplexa assay.
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Image /page/8/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with the word "Diagnostics" in a smaller font size underneath. A curved, crescent-shaped graphic is positioned to the left of the word "FOCUS", adding a visual element to the logo.
510(k) Summary of Safety and Effectiveness
Simplexa ™ Influenza A H1N1 (2009) Catalog No. MOL2500 Prepared Date: May 18, 2010
Page 9 of 9
Influenza A Clinical Agreement Summary
Simplexa™ Influenza A H1N1 (2009) vs. Composite Reference Result - Retrospectively Collected Swabs
| Influenza A Result - Simplexa™ Influenza A H1N1 (2009) | ||||||
|---|---|---|---|---|---|---|
| n | Influenza A Detected | Influenza A Not Detected | Indeterminate | % Agreement | ||
| CompositeReference Result | Influenza A Detected | 132 | 131 | 0 | 1 | % Positive Agreement99.2%(131/132)95% CI:95.8-99.9% |
| Influenza A Not Detected | 79 | 13 | 66 | 0 | % Negative Agreement83.5%(66/79)95% CI:73.9-90.1% |
- Due to the low prevalence of other strains of influenza A during the testing period; all FLU A responses from retrospectively collected samples were combined to demonstrate the performan fluorescent primer-probe. Of the 132 specimens determined to be positive for FLU A, 57 were 2009 H1N1 influenza positive, two (2) were H1N1, 59 were H3N2, one (1) was sequenced but the sub-ype could not be determined, one (1) was indeterminate by Simplexa, 11 were not detected by the alternate PCR and one (1) did not have sufficient volume to sequence to delemine sub-lype. Three (3) samples were excluded from the analysis because of the influenza A reference assays.
Two retrospectively collected washes were found to be positive for influenza A by the composite reference method and by the Simplexa assay.
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Image /page/9/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized caduceus symbol, which is a staff with two snakes coiled around it, and the words "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" arranged in a circular pattern around the symbol. The logo is black and white.
Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center - WO66-0609 Silver Spring, MD 20993-0002
Focus Diagnostics, Inc. c/o Tara Viviani Regulatory Affairs Project Manager 11331 Valley View St. Cypress, California 90630
MAY 2 4 2010
K100148 Regulation Number: Regulation Name: Regulatory Class: Class II Product Code: OOW Dated: Received: April 29, 2010
Trade/Device Name: Simplexa ™ Influenza A H1N1 (2009) 21CFR 8866.3332 Influenza A HIN1 2009 Real Time RT-PCR April 27, 2010
Dear Ms. Viviani:
Re:
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. Iisting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.
If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a
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legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety 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,
ValgaArs
Sally A. Hojvat, M.Sc., Ph.D Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known): K100148
Device Name:
Simplexa™ Influenza A H1N1 (2009)
:
Indications for Use:
The Focus Diagnostics Simplexa™ Influenza A H1N1 (2009) assay is intended for use on the 3M Integrated Cycler as part of the Microfluidic Molecular System for the in vitro qualitative detection and differentiation of influenza A and 2009 H1N1 influenza viral RNA in nasopharyngeal swabs (NPS), nasal swabs (NS), and nasopharyngeal aspirates (NPA) from human patients with signs and symptoms of respiratory infection in conjunction with clinical and epidemiological risk factors.
Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other patient management decisions.
Performance characteristics for influenza A were established during the 2009-2010 influenza season when 2009 H1N1 influenza was the predominant influenza A virus 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 focal 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.
Prescription Use X (Part 21 CFR 801 Subpart D)
AND/OR AND/OR AND/OR
Over-The-Counter Use (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)
Concurrence of CDRH, Office of In Vitro Diagnostics (OIVD)
Une Schif
Division Sign-Off
Office of In Vitro Diagnostic Device Evaluation and Safety
510(k) k 100148
§ 866.3332 Reagents for detection of specific novel influenza A viruses.
(a)
Identification. Reagents for detection of specific novel influenza A viruses are devices that are intended for use in a nucleic acid amplification test to directly detect specific virus RNA in human respiratory specimens or viral cultures. Detection of specific virus RNA aids in the diagnosis of influenza caused by specific novel influenza A viruses in patients with clinical risk of infection with these viruses, and also aids in the presumptive laboratory identification of specific novel influenza A viruses to provide epidemiological information on influenza. These reagents include primers, probes, and specific influenza A virus controls.(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Reagents for Detection of Specific Novel Influenza A Viruses.” See § 866.1(e) for information on obtaining this document.
(2) The distribution of these devices is limited to laboratories with experienced personnel who have training in standardized molecular testing procedures and expertise in viral diagnosis, and appropriate biosafety equipment and containment.