(328 days)
The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay is a qualitative in vitro diagnostic system used to simultaneously detect 139 clinically relevant cystic fibrosis disease-causing mutations and variants of the cystic fibrosis transmembrance regulator (CFTR) gene in genomic DNA isolated from human peripheral whole blood specimens. The variants include those recommended in 2004 by the American College of Medical Genetics (ACMG) and in 2011 by the American College of Obstetricians and Gynecologists (ACOG). The test is intended for carrier screening in adults of reproductive age, in confirmatory diagnostic testing of newborns and children, and as an initial test to aid in the diagnosis of individuals with suspected cystic fibrosis. The results of this test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available laboratory and clinical information. This test is not indicated for use for newborn screening, fetal diagnostic testing, pre-implantation testing, or for stand-alone diagnostic purposes.
The test is intended to be used on the Illumina MiSeqDxTM Instrument.
The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay consists of library preparation and sample indexing reagents, sequencing reagents and consumables, MiSeqDx instrument and data analysis software. Testing begins with genomic DNA from a peripheral whole blood sample. The genomic DNA is processed through the library preparation steps, which specifically amplifies the intended genomic regions of each sample while also adding the indexes for sample identification. Flow cell capture sequences are also added to the amplified products. The resulting sample libraries are then transferred into a MiSeqDx reagent cartridge which contains all of the reagents required for cluster generation and sequencing (Sequencing By Synthesis). The MiSeqDx Cartridge, MiSeqDx Flow Cell, and MiSeqDx SBS Solution (PR2) are then inserted into the MiSeqDx instrument, which performs cluster generation, sequencing and data analysis.
Here's a breakdown of the acceptance criteria and the study proving the device meets them, based on the provided text:
Acceptance Criteria and Device Performance
| Measure | Acceptance Criteria (Not explicitly stated as criteria, but implied by 100% and >99.99% results in accuracy study and high metrics in reproducibility) | Reported Device Performance (Accuracy Study) | Reported Device Performance (Reproducibility Study) |
|---|---|---|---|
| Genotype-level PA | High (implied 100% or near 100%) | 100% for all variants | 99.77% |
| Negative Agreement (NA) | High (implied >99%) | >99.99% for all wild types | 99.88% |
| Overall Agreement (OA) | High (implied >99%) | >99.99% for all reported positions | 99.88% |
| Sample Pass Rate | Not explicitly stated for accuracy study, but for reproducibility study, samples passing QC on first attempt | N/A for accuracy study (all tested samples were successfully genotyped) | 99.9% (number of samples passing QC metrics on the first attempt) |
| Accuracy (PolyTG/PolyT) | High (implied 100% in most cases, with explanations for deviations) | Varies by genotype, generally 100%, with exceptions explained (e.g., 50.0% for one less common genotype, 90.9% for another) | N/A for this specific sub-analysis in the reproducibility section, but overall reproducibility metrics apply. |
| Accuracy (Interfering Substances) | 100% call rate and 100% reproducibility | 100% call rate and 100% reproducibility in genotype calls | N/A (tested in interference study) |
| Call Rate (DNA Extraction) | 100% | 100% | N/A (tested in extraction study) |
| Accuracy (DNA Extraction) | 100% | 100% | N/A (tested in extraction study) |
| Sample First Pass Rate (DNA Extraction) | 100% | 100% | N/A (tested in extraction study) |
| Accuracy (DNA Input) | 100% | 100% | N/A (tested in DNA input study) |
| Sample First Pass Rate (DNA Input) | >95% for upper and lower bounds; 100% for specific tested input levels | >95% for 1250 ng and 25 ng; 100% for 1250 ng, 250 ng, 100 ng, and 25 ng samples | N/A (tested in DNA input study) |
| Reproducibility & Accuracy (Sample Indexing) | 100% | 100% | 100% (for sample/index primer combinations) |
Note regarding "Acceptance Criteria": The document K124006 primarily presents the results of validation studies rather than explicitly stating pre-defined "acceptance criteria" in a separate section. The "acceptance criteria" implied above are derived from the consistently high performance reported across various studies and the overall conclusion of the device being substantially equivalent.
Study Details
2. Sample Sizes Used for the Test Set and Data Provenance
- Accuracy Study (Primary Source):
- Test Set Size: 500 samples in total, comprised of:
- 366 samples from a clinical accuracy study (primary source).
- 68 cell line samples from the reproducibility study (supplementary).
- 14 clinical samples from the extraction method evaluation analytical study (supplementary).
- 52 synthetic plasmid samples (supplementary).
- Data Provenance:
- Clinical Samples: Majority (n=355) were archived, anonymized clinical gDNA specimens isolated from human blood. (No specific country of origin mentioned, but implied to be human clinical samples).
- Cell Line Samples: 11 samples were commercially available cell line specimens.
- Synthetic Plasmid Samples: Designed to include genomic context of rare variants, linearized, diluted, and blended with human genomic DNA (lab-generated, not from patients).
- Retrospective/Prospective: The 355 clinical gDNA specimens were "archived, anonymized clinical gDNA specimens," suggesting a retrospective approach for these samples. The cell line and synthetic samples were laboratory-generated.
- Test Set Size: 500 samples in total, comprised of:
- Reproducibility Study:
- Test Set Size: 46 samples (in each of two panels, total 92 unique samples) were tested. Each sample was tested for a total of 810 calls per site.
- Data Provenance: Genomic DNA from lymphoblastoid cell lines with known mutations and leukocyte-depleted blood spiked with lymphoblastoid cell lines with known mutations. This suggests laboratory-controlled samples, not directly from clinical patients in a prospective manner.
- DNA Extraction Study:
- Test Set Size: 14 unique blood samples per extraction method, with 2 operators performing 3 runs each, and each run having 2 replicates, totaling 168 (14 x 2 x 3 x 2) for each of the three extraction methods.
- Data Provenance: K2EDTA anti-coagulated whole blood samples, including wild type and three mutant genotypes (samples with F508del, I506V, and D110H).
- DNA Input Study:
- Test Set Size: 14 representative DNA samples (with 16 unique CF variants) tested in duplicate at 9 DNA input levels. Additional testing involved 4 representative samples with 20 replicates each (n=80), and 14 samples with 20 replicates each (n=280).
- Data Provenance: Representative DNA samples containing CF variants.
- Interfering Substances Study:
- Test Set Size: 8 whole blood specimens (including 3 CF positive samples with unique genotypes). 16 replicates each for specific interferents.
- Data Provenance: Whole blood specimens.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
The document specifies the reference methods used to establish ground truth, but does not explicitly state the number or qualifications of experts involved in reviewing these reference results or establishing consensus.
- For 137 SNA/small InDel sites and PolyTG/Poly T region: Sanger bi-directional sequence analysis was the reference method.
- For two large deletions: A PCR-based assay was the reference method, confirmed for accuracy using Sanger Sequencing.
While Sanger sequencing and PCR assays are considered robust reference methods, the process of interpreting and confirming these results as "ground truth" and addressing any discrepancies would typically involve qualified molecular geneticists or lab personnel. However, this level of detail is not provided.
4. Adjudication Method for the Test Set
The document does not describe a formal adjudication method (e.g., 2+1, 3+1, none) for the test set. Ground truth was established by reference methods (Sanger bi-directional sequencing and a PCR-based assay). Any "miscalls" (one in the accuracy study and a few in the reproducibility study) or "no calls" are noted and sometimes explained (e.g., insufficient coverage, sample handling issues), implying internal review rather than a multi-expert adjudication process for primary ground truth establishment.
5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study
No Multi-Reader Multi-Case (MRMC) comparative effectiveness study was done. This device is an in vitro diagnostic system for detecting genetic mutations, not an imaging device or a diagnostic aid meant to directly assist human readers in interpreting complex visual data. Therefore, the concept of "human readers improve with AI vs. without AI assistance" is not applicable in this context.
6. Standalone (Algorithm Only Without Human-in-the-Loop Performance)
Yes, a standalone performance assessment was conducted. The accuracy and reproducibility studies directly evaluated the device's ability (including its data analysis software) to detect mutations and variants in genomic DNA. The "results of this test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available laboratory and clinical information," indicating that while human interpretation is part of the overall clinical process, the performance metrics (PA, NA, OA) are for the device (algorithm + instrument) alone.
7. Type of Ground Truth Used
The ground truth used was primarily based on:
- Sanger bi-directional sequence analysis: For 137 SNA/small InDel sites and the PolyTG/Poly T region. This is considered a gold standard for sequence verification.
- PCR-based assay: For two large deletions, confirmed by Sanger Sequencing.
This represents established molecular diagnostic laboratory methods, widely accepted as reliable for genetic variant detection.
8. Sample Size for the Training Set
The document does not report the sample size for a training set. This is typical for in vitro diagnostic devices based on established sequencing technology and a defined set of targets, where the "training" may involve internal development and optimization using design verification samples rather than a formally described, distinct "training set" in the context of machine learning model development. The focus is on validation against known samples to determine performance characteristics.
9. How the Ground Truth for the Training Set Was Established
As no explicit "training set" is described, how its ground truth might have been established is not mentioned in the document. Development and optimization would have relied on samples with known CFTR genotypes, likely confirmed by methods similar to those used for the test set (Sanger sequencing, PCR).
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MiSeqDx Cystic Fibrosis System Premarket Notification
510(k) Summary
GENERAL INFORMATION
-
Submitted by: Illumina Inc. 5200 Illumina Way San Diego, CA 92122 858-202-4500 (phone)
NOV 1 9 2013 -
Leanne M. Kiviharju Company Contact: Sr. Director, Regulatory Affairs 858-246-8811 (phone) lkiviharju@illumina.com
Date Prepared: November 18, 2013
DEVICE IDENTIFICATION
Assay:
| Trade or Proprietary Name: Illumina MiSeqDx™ Cystic Fibrosis 139-Variant Assay | |
|---|---|
| Assay Common Name: | Next generation sequencing cystic fibrosis test |
| Classification Name: | CFTR (cystic fibrosis transmembrane conductanceregulator) gene mutation detection (21 CFR 866.5900,Product Code PFR) |
| Predicate Device: | Luminex xTAG® Cystic Fibrosis 60 Kit v2 (K083845) |
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DEVICE DESCRIPTION
The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay consists of library preparation and sample indexing reagents, sequencing reagents and consumables, MiSeqDx instrument and data analysis software. Testing begins with genomic DNA from a peripheral whole blood sample. The genomic DNA is processed through the library preparation steps, which specifically amplifies the intended genomic regions of each sample while also adding the indexes for sample identification. Flow cell capture sequences are also added to the amplified products. The resulting sample libraries are then transferred into a MiSeqDx reagent cartridge which contains all of the reagents required for cluster generation and sequencing (Sequencing By Synthesis). The MiSeqDx Cartridge, MiSeqDx Flow Cell, and MiSeqDx SBS Solution (PR2) are then inserted into the MiSeqDx instrument, which performs cluster generation, sequencing and data analysis.
INTENDED USE
Illumina MiSeqDx™ Cystic Fibrosis 139-Variant Assay
The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay is a qualitative in vitro diagnostic system used to simultaneously detect 139 clinically relevant cystic fibrosis disease-causing mutations and variants of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in genomic DNA isolated from human peripheral whole blood specimens. The variants include those recommended in 2004 by the American College of Medical Genetics (ACMG) and in 2011 by the American College of Obstetricians and Gynecologists (ACOG). The test is intended for carrier screening in adults of reproductive age, in confirmatory diagnostic testing of newborns and children, and as an initial test to aid in the diagnosis of individuals with suspected cystic fibrosis. The results of this test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available laboratory and clinical information. This test is not indicated for use for newborn screening, fetal diagnostic testing, pre-implantation testing, or for stand-alone diagnostic purposes.
The test is intended to be used on the Illumina MiSeqDxTM Instrument.
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Image /page/2/Picture/0 description: The image shows the word "illumina" in a simple, sans-serif font. The letters are black and appear to be bolded. There is a superscript symbol after the last letter.
SUBSTANTIAL EQUIVALENCE
MiSeqDx Cystic Fibrosis 139-Variant Assay
| Characteristic | Illumina | Luminex (K083845) |
|---|---|---|
| Assay Name | Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay | Luminex xTAG®CysticFibrosis 60 Kit v2 |
| Intended Use | The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay is a qualitative in vitrodiagnostic system used to simultaneouslydetect 139 clinically relevant cystic fibrosisdisease-causing mutations and variants ofthe cystic fibrosis transmembraneconductance regulator (CFTR) gene ingenomic DNA isolated from humanperipheral whole blood specimens. Thevariants include those recommended in 2004by the American College of Medical Genetics(ACMG) and in 2011 by the AmericanCollege of Obstetricians and Gynecologists(ACOG). The test is intended for carrierscreening in adults of reproductive age, inconfirmatory diagnostic testing of newbornsand children, and as an initial test to aid inthe diagnosis of individuals with suspectedcystic fibrosis. The results of this test areintended to be interpreted by a board-certified clinical molecular geneticist orequivalent and should be used inconjunction with other available laboratoryand clinical information. This test is notindicated for use for newborn screening,fetal diagnostic testing, pre-implantationtesting, or for stand-alone diagnosticpurposes.The test is intended to be used on theIllumina MiSeqDx™ instrument. | The xTAG® Cystic Fibrosis 60kit v2 is a device used tosimultaneously detect andidentify a panel of mutationsand variants in the cysticfibrosis transmembraneconductance regulator (CFTR)gene in human bloodspecimens. The panel includesmutations and variantscurrently recommended by theAmerican College of MedicalGenetics and AmericanCollege of Obstetricians andGynecologists (ACMG/ACOG)plus some of the world's mostcommon and North Americanprevalent mutations. ThexTAG Cystic Fibrosis 60 kit v2is a qualitative genotyping testwhich provides informationintended to be used for carriertesting in adults ofreproductive age, as an aid innewborn screening, and inconfirmatory diagnostic testingin newborns and children.The kit is not indicated for usein fetal diagnostic or pre-implantation testing. The kit isalso not indicated for stand-alone diagnostic purposes. |
| Assay type | Next generation sequencing test | Qualitative nucleic acidmultiplex test |
| Variants | 139 clinically relevant variants | 60 CFTR mutations and 4 |
| Characteristic | Illumina | Luminex (K083845) |
| Detected | variants (benignpolymorphisms) | |
| Technology | Sequencing by Synthesis (SBS) | Multiplex PCR followed bymultiplex allele specific primerextension for genotyping,hybridized to multiplexfluorescent microparticles,detected by flow cytometry. |
| Sample Type | Nucleic acid from EDTA anticoagulatedblood | Nucleic acid from whole bloodanticoagulated with eitherEDTA or citrate. |
| SamplePreparation | DNA extraction using validated laboratorymethod | Same |
| Contra-indications | Not indicated for newborn screening, fetaldiagnostic testing, pre-implantation testing,or for stand-alone diagnostic purposes. | The kit is not indicated for usein fetal diagnostic or pre-implantation testing. This kit isalso not indicated for stand-alone diagnostic purposes |
| Assay Controls | Positive and negative controls required, notsupplied | Negative controls required, notsupplied. Positive controlsrecommended, not supplied. |
| InstrumentSystem | MiSeqDx instrument | Luminex 100 or 200 IS |
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PERFORMANCE CHARACTERISTICS
Accuracy
Accuracy of the Illumina MiSeqDx Cystic Fibrosis 139-Variant assay was assessed by evaluating 500 samples representing a wide variety of CFTR variants from four separate sources. The primary source of accuracy data was a clinical accuracy study conducted using a panel of 366 samples. The majority (n=355) of samples consisted of archived, anonymized clinical gDNA specimens isolated from human blood, the remaining 11 samples were obtained from commercially available cell line specimens.
Data from this study was supplemented with accuracy data from 68 cell line samples evaluated in the reproducibility study, 14 clinical samples from the extraction method evaluation analytical study, and 52 synthetic plasmid samples. The synthetic plasmids
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were designed to include the genomic context of the rare variants, and contained anywhere from 1 to 9 variants within the same construct. They were linearized, diluted to genomic DNA equivalent copy numbers, and blended with human genomic DNA samples of wild type genotype at equivalent copy numbers to mimic a heterozygous sample.
The genotyping results for 137 SNA/small InDel sites, including the PolyTG/Poly T region were compared to Sanger bi-directional sequence analysis. A PCR based assay was used as the reference method for the two large deletions in the panel. Each duplex PCR assay made use of 2 primer sets to discriminate between wild type, heterozygous, and homozygous genotypes. One of the primer sets was designed to flank the deletion breakpoints, whereas the other amplified a region internal to the deletion. The two products were detected by size separation on an agarose gel.
The assays were validated using a panel of 28 samples in all (22 samples for each deletion) consisting of cell line and blood derived genomic DNA samples, and synthetic plasmids which encompassed the WT, HET and HOM genotypes for each large deletion. The PCR assays were confirmed to have 100% specificity and reproducibility for all samples tested, by evaluation of PCR products on an agarose gel. The accuracy of the PCR assays was confirmed using Sanger Sequencing and found to be 100% for all sample
Accuracy was determined for each genotype through three statistical measures. Positive Agreement (PA) was calculated for each variant genotype by dividing the number of samples with agreeing variant calls by the total number of samples with that variant as identified by the reference methods. Negative Agreement (NA) was calculated across all wild type (WT) positions by dividing the number of concordant WT positions by the total number of WT positions as defined by the reference methods. Overall Agreement (OA) was calculated across all reported positions by dividing the number of concordant WT and variant positions by the total number of reported positions as determined by the reference methods.
The MiSegDx Cystic Fibrosis 139-Variant Assay had a genotype-level PA of 100%. The NA for all wild types was >99.99% and the OA for all reported positions was >99.99%. All test results were based on initial testing.
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able 1: Overall Accuracy for the MiSeqDx Cystic Fibrosis 139-Variant Assa
| Mutation(Common Name) | Total callspermutation | Positive calls (Variants) | Negative calls(Wild Type) | # Miscalls | # NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | ||
|---|---|---|---|---|---|---|---|---|---|---|
| ClinicalSamples | Cell LineSamples | SyntheticSamples | ||||||||
| CFTR dele2, 3 | 500 | 4 | 1 | 0 | 495 | 0 | 0 | 100 | 100 | 100 |
| E60X | 500 | 6 | 1 | 0 | 493 | 0 | 0 | 100 | 100 | 100 |
| P67L | 500 | 1 | 0 | 1 | 498 | 0 | 0 | 100 | 100 | 100 |
| R75X | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| G85E | 500 | 6 | 2 | 0 | 492 | 0 | 0 | 100 | 100 | 100 |
| 394delTT | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| 406-1G>A | 500 | 4 | 0 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| E92X | 500 | 0 | 1 | 1 | 498 | 0 | 0 | 100 | 100 | 100 |
| D110H | 500 | 1 | 0 | 1 | 498 | 0 | 0 | 100 | 100 | 100 |
| R117C | 500 | 4 | 0 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| R117H | 500 | 17 | 2 | 0 | 481 | 0 | 0 | 100 | 100 | 100 |
| Y122X | 500 | 0 | 1 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| 621+1G>T | 500 | 7 | 5 | 0 | 488 | 0 | 0 | 100 | 100 | 100 |
| 663delT | 500 | 1 | 1 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| G178R | 500 | 1 | 1 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| 711+1G>T | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| P205S | 500 | 1 | 0 | 1 | 498 | 0* | 0 | 100 | 100 | 100 |
| L206W | 500 | 8 | 1 | 0 | 491 | 0 | 0 | 100 | 100 | 100 |
| 1078delT | 500 | 1 | 1 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| G330X | 500 | 1 | 1 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| R334W | 500 | 6 | 1 | 0 | 493 | 0 | 0 | 100 | 100 | 100 |
| otal call | Positiv | calls (Variants | Negative calls (Wild Type) | Miscal | ମାସେ ଚନ୍ଦ୍ର ମଧ୍ୟ # | Positive | Negative | Overal | ||
| Mutation ommon Nam | per nutation | ລາວປັນຈ ເຖິງແມງດ | Cell LineSample: | SyntheticSamples | greement | Agreemen196 | Agreemer1961 | |||
| 1336K | 500 | T | 0 | ਪਰੇਰੇ | 0 | 0 | 100 | 100 | 100 | |
| 154insT | 500 | 0 | I | 0 | ਪਰੇਰੇ | 0 | 0 | 100 | 100 | 100 |
| R347H | ર૦૮ | 9 | โ | t | 492 | 0 | 0 | 100 | 100 | 100 |
| R347F | 500 | t | ਟ | 0 | 495 | 0 | 0 | IDT | 100 | 100 |
| R352C | 500 | S | 0 | 0 | 495ﺴﻨ | 0 | 0 | 100 | 100 | 100 |
| A455E | 500 | ਸ | て | 0 | ਪਰੇਵ | 0 | 0 | 100 | 100 | 100 |
| 466X (C->G | 500 | โ | 0 | T | 498r | 0 | i T T0 | 100 | 100 | 100 |
| 548del | ર૦૮ | โ | 0 | T | 498 | 0 | 0 | 100 | 100 | 00T |
| Q493) | ર૦૮ | t | ਟ | 0 | 494 | 0 | 100 | 100 | 100 | |
| 507de | ક્વવ | 7 | ਟ | 0 | 4 ਰੋਪ | 0 : 0 | 0 | 100 | 100 | 100 |
| 508de | 500 | 78 | රිට | 0 | 387 | 0 | 0 | 100 | 100 | 100 |
| 677delT | 500 | L | 0 | 0 | ਧਰੇਰੇ | 0 | 0 | 100 | 100 | 100 |
| V520F | 500 | て | 0 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| 717-1G> | 500 | b | I | 0 | 495 | 0 | 0 | 100 | 100 | 100 |
| G542) | 200 | ਟ ਦ | € | 0 | 485 | 0 | 0 | 100 | 100 | 100 |
| 549N | 500 | ਟ | ਟ | T | ਕਰੇ ਟ | 0 | 0 | 100 | 100 | 100 |
| S549R 1647T>G | 500 | ಕ | T | 0 | ૫૦૦ | 0 | 0 | 100 | 100 | 100 |
| GSS10 | నంద | 8 | દ | 0 | 489 | 0 | 0 | 100 | 100 | 100 |
| R553X | ડવ્યુ | 8 | ਟ | 0 | 490 | 0 | 0 | 100 | 100 | 100 |
| 4559. | 500 | 7 | 0 | T | વવાયુ | 0 | 0 | 100 | 100 | 100 |
| R5601 | 500 | 9 | T | 0 | ਖਰੇਤ | 0 | 0 | 100 | 100 | 100 |
| 812-1 G-> | ર૦ત | 0 | ਟ | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| Mutation(Common Name) | Total callspermutation | Positive calls (Variants) | SyntheticSamples | Negative calls(Wild Type) | # Miscalls | # NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | |
| ClinicalSamples | Cell LineSamples | |||||||||
| 1898+1G>A | 500 | 2 | 1 | 0 | 497 | 0 | 0 | 100 | 100 | 100 |
| 2143delT | 500 | 2 | 1 | 0 | 497 | 0 | 0 | 100 | 100 | 100 |
| 2183AA>G | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| 2184insA | 500 | 3 | 0 | 1 | 496 | 0 | 0 | 100 | 100 | 100 |
| 2184delA | 500 | 1 | 1 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| R709X | 500 | 1 | 0 | 2 | 497 | 0 | 0 | 100 | 100 | 100 |
| K710X | 500 | 3 | 0 | 0 | 497 | 0 | 0 | 100 | 100 | 100 |
| 2307insA | 500 | 3 | 0 | 2 | 495 | 0 | 0 | 100 | 100 | 100 |
| R764X | 500 | 1 | 0 | 2 | 497 | 0 | 0 | 100 | 100 | 100 |
| W846X | 500 | 0 | 1 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| 2789+5G>A | 500 | 9 | 1 | 0 | 490 | 0 | 0 | 100 | 100 | 100 |
| Q890X | 500 | 1 | 0 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| 3120G>A | 500 | 1 | 0 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| 3120+1G>A | 500 | 7 | 1 | 0 | 492 | 0 | 0 | 100 | 100 | 100 |
| 3272-26A>G | 500 | 0 | 1 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| R1066C | 500 | 6 | 0 | 0 | 494 | 0 | 0 | 100 | 100 | 100 |
| R1066H | 500 | 1 | 0 | 1 | 498 | 0 | 0 | 100 | 100 | 100 |
| W1089X | 500 | 4 | 0 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| Y1092X (C>A) | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| M1101K | 500 | 2 | 2 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| R1158X | 500 | 7 | 1 | 0 | 492 | 0 | 0 | 100 | 100 | 100 |
| R1162X | 500 | 5 | 1 | 0 | 494 | 0 | 0 | 100 | 100 | 100 |
| 3659delC | 500 | 4 | 1 | 0 | 495 | 0 | 0 | 100 | 100 | 100 |
| Mutation(Common Name) | Total callspermutation | Positive calls (Variants) | Negative calls(Wild Type) | # Miscalls | # NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | ||
| ClinicalSamples | Cell LineSamples | SyntheticSamples | ||||||||
| S1196X | 500 | 1 | 0 | 0 | 499 | 0 | 0 | 100 | 100 | 100 |
| 3791delC | 500 | 2 | 0 | 0 | 498 | 0 | 0 | 100 | 100 | 100 |
| 3849+10kbC>T | 500 | 11 | 2 | 0 | 487 | 0 | 0 | 100 | 100 | 100 |
| 3876delA | 500 | 6 | 1 | 0 | 493 | 0 | 0 | 100 | 100 | 100 |
| S1251N | 500 | 1 | 0 | 1 | 498 | 0 | 0 | 100 | 100 | 100 |
| 3905insT | 500 | 3 | 1 | 0 | 496 | 0 | 0 | 100 | 100 | 100 |
| W1282X | 500 | 9 | 1 | 0 | 490 | 0 | 0 | 100 | 100 | 100 |
| N1303K | 500 | 9 | 1 | 0 | 490 | 0 | 0 | 100 | 100 | 100 |
| CFTRdele22,23 | 500 | 1 | 0 | 1 | 498 | 1" | 0 | 100 | 99.8 | 99.8 |
| M1V | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q39X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 405+1 G->A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| E92K | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q98X | 500 | 0 | 0 | 2 | 498 | 0 | 0 | 100 | 100 | 100 |
| 457TAT->G | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 574delA | 500 | 0 | 0 | 2 | 498 | 0 | 0 | 100 | 100 | 100 |
| 711+3A>G | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 711+5 G->A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 712-1 G->T | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| H199Y | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q220X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 852del22 | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Mutation(Common Name) | Total callspermutation | ClinicalSamples | Positive calls (Variants)Cell LineSamples | SyntheticSamples | Negative calls(Wild Type) | # Miscalls | # NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) |
| T3381 | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| S341P | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1213delT | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1248+1G>A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1259insA | 500 | 0 | 0 | 2 | 498 | 0 | 0 | 100 | 100 | 100 |
| W401X(c.1202G>A) | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| W401X(c.1203G>A) | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1341+1G->A | 500 | 0 | 0 | 2 | 498 | 0 | 0 | 100 | 100 | 100 |
| 1461ins4 | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1525-1G->A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| S466X (C->A) | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| L467P | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| S489X | 500 | 0 | 0 | 2 | 498 | 0 | 0 | 100 | 100 | 100 |
| S492F | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q525X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1717-8G->A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| S549R(c.1645A>C) | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q552X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| R560K | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 1811+1.6kb A->G | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| E585X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
27
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iSeqDx Cystic Fibrosis System
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MiSeqDx Cystic Fibrosis System
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MiSeqDx Cystic Fibrosis System
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MiSeqDx Cystic Fibrosis System
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MiSeqDx Cystic Fibrosis System
| Overal | greemenಳಿಕ | 100 | 100 | 100 | 100 | 00T | 001 00T | 100 | 00T | 100 | 00T | 100 | 100 | 100 | 00T | 00T | 100 | 100 | 100 | 100 | 100 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Negative | Agreemer(%) | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | 100 | от | 100 | 100 | 00T | 100 | 100 | 100 | 100 | 100 | 100 |
| Positive | greemer(%) | 100 | 100 | 100 | 100 | 100 | 00 T | 100 | 100 | 100 | 100 | 100 | 100 | 00T | 100 | 00T | 00 I | 100 | 100 | 100 | 100 | 100 |
| siles ON # | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| Miscal | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0. ﻟ . | VO | 0 | 0 | 0 | 0 | 0 | |
| Negative calls (Wild Type) | ਥ ਰੇਰੇ | 498 | 498 | 498 | 198 | 498 | 499 | ਰੇਰੇ | ਕਰੇਰੇ | ਖਰੇਰੇ | ਰ ਰੇਰੇ | aa | 499 | ਥ ਰੇਰੇ | 499 | 499 | ਖਰੇਰੇ | 499 | ਧ ਰੇਖੇ | ਧਰੇਤੇ | 499 | |
| SyntheticSamples | โ | ਟ | ਟ | ट | ट | で | સ્ન | T | T | पु | :T | I | I | โ | I | T | I | T | I | T | · , i 'T | |
| calls (Variants) | Cell Linemples୧୮ | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0) 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| Positive | SamplesClinical | 0 | 0 | 0 | 0 | 0 | 0 | (0) | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | |
| Total calls | per mutatio | 500 | 500 | ടറവ | 200 | റ്ററ | నంది. సంర | ક્તભ | నం | నంది. వైద | ર૦૮ | રૂભવ | నంర | నం | રેજિ | 500 | 200 | ર૦૮ | ર૦૮ | 500 | ર૦૮ | SOC |
| Mutation | ommon Nam | 898+3A> | L732) | 347delG | 2585del | E822) | 622+1G> | E831X | R851) | 2711del | L927F | 59451 | 007delG | G9708 | 121-1G-> | 1065b | L1077P | 1092X (C>G | E1104) | היסטורים אימריקע א לפני נייני נייני מיני מ לפני נייני נייני ניי אמטרוני אינטער א אמטע מעניינע א |
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| 2 |
|---|
| g |
| -S1 |
| - |
| œm |
| n |
| Chamble Comment |
| ages of the program and contrôleant |
| . .I |
iSeqDx Cystic Fibrosis Syste
| Mutation(Common Name) | Total callspermutation | Positive calls (Variants) | Negative calls(Wild Type) | # Miscalls | # NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | ||
|---|---|---|---|---|---|---|---|---|---|---|
| ClinicalSamples | Cell LineSamples | SyntheticSamples | ||||||||
| 4005+1G->A | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 4016insT | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| Q1313X | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 4209TGTT>AA | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| 4382delA | 500 | 0 | 0 | 1 | 499 | 0 | 0 | 100 | 100 | 100 |
| PolyTG/PolyT | 19 | 17 | 2 | 0 | 0 | 0 | 0 | 100 | N/A | 100 |
| I506V* | 1 | 0 | 0 | 0 | 1 | 0 | 0 | N/A | 100 | 100 |
| I507V* | 1 | 0 | 0 | 0 | 1 | 0 | 0 | N/A | 100 | 100 |
| F508C* | 1 | 0 | 0 | 0 | 1 | 0 | 0 | N/A | 100 | 100 |
| Total | 67522 | 557 | 66965 | 1 | 0 | 100.00 | >99.99 | >99.99 | ||
| * The Sanger report listed the P205S variant as heterozygous for the clinical sample. A review of the Sanger trace data however indicated that the variant was in fact homozygous and incorrectly |
The Sanger report listed le P2055 var
as heterozygous for the
sam
reported. MiSeqDx reported the variant as homozygous
Jaus Jouel contamination
he original syntheic hetercozygous specient was determined when it was subsequently tested after it was re prepared, using the same plasmid, it would be detece A youtheric sample heterozygous for exon 8 was reported on the variant CFTR dele2, 23. Further investigation revealed that this result was likely from lov level contanimal
When R117H is positive, the PolyTG/PolyT variant is additionally reported
In the case of one homozygous F508del variant, three additional wild type bases (1506V, I507V, F508C) were additionally reported
able 2: Accuracy of the MiSeqDx Cystic Fibrosis 139-Variant Assay for 1506V, 1507V and F5086
| Variant(CommonName) | TotalCalls perVariant | Positive Calls (Variants) | NegativeCalls (WildType) | =Miscalls | = NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | |
|---|---|---|---|---|---|---|---|---|---|
| 1506V | 500 | ClinicalSamples7 | Cell LineSyntheticSamples0 | 493 | 0 | 0 | 100 | 100 | 100 |
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MiSeqDx Cystic Fibrosis System
| Variant(CommonName) | TotalCalls perVariant | Positive Calls (Variants) | NegativeCalls (WildType) | Miscalls= NoCalls | PositiveAgreement(%) | NegativeAgreement(%) | OverallAgreement(%) | ||
|---|---|---|---|---|---|---|---|---|---|
| 1507V | 500 | ClinicalSamples0 | Cell LineSamples1 | SyntheticSamples0 | 499 | 0 | 100 | 100 | 100 |
| F508C | 500 | ClinicalSamples1 | Cell LineSamples1 | SyntheticSamples0 | 498 | 0 | 100 | 100 | 100 |
Table 3. Accuracy of the MiSeqDx Cystic Fibrosis 139-Variant Assay for PolyTG/PolyT Variants
| Genotype | ClinicalSamples | CellLineSamples | SyntheticSamples | Miscalls | NoCalls | Accuracy |
|---|---|---|---|---|---|---|
| (TG)9(T)7/(TG)11(T)7 | 2 | 0 | 0 | 0 | 1 | 50.0 |
| (TG)9(T)9/(TG)10(T)7 | 1 | 0 | 0 | 0 | 0 | 100 |
| (TG)9(T)9/(TG)11(T)7 | 5 | 1 | 0 | 0 | 0 | 100 |
| (TG)9(T)9/(TG)11(T)9 | 1 | 0 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)10(T)7 | 25 | 8 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)10(T)9 | 39 | 16 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)11(T)5 | 2 | 0 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)11(T)7 | 72 | 11 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)12(T)5 | 1 | 0 | 0 | 0 | 0 | 100 |
| (TG)10(T)7/(TG)12(T)7 | 10 | 1 | 0 | 0 | 1 | 90.9 |
34
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iSeqDx Cystic Fibrosis System
Premarket Notificati
| 100 | 100 | 00 I | 100 | 100 | 92.3 | 83.3 | 00 I | 0.0 | 100 | 100 | 100 | 100 | 98.44 | a suala firm the complex |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 0 | 0 | 0 | 0 | 0 | լ. | 0 | በ | 0 | 0 | 0 | 0 | 0 | ప్రాంత | |
| 0 | 0 | 0 | 0 | 0 | 0 | L | 0 | C | 0 | 0 | 0 | 0 | ోగా | 0 |
| 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | -r'l-. | |
| 9 | 0 | 07 | 0 | ਨ। | ﮨﮯ | 0 | 8 | I | 0 | € | 0 | で | ||
| 1 | ഗ്ര | 9 ﻳ | ಕ್ಕೆ | tt, | g I | 9 | ਟ ਉ | Z | ਟ | 1 g | € | ಗಳ | 148 | |
| (TC)10(T)9/(TC)10(T)9 | G)10(T)9/(TG)11(T)5 | TG)10(T)9/(TG)11(T)7 | TG)10(T)9/(TG)11(T)9 | (G)10(T)9/(TG)12(T) | TG)10(T)9/(TG)12(T)7 | G)11(T)5/(TG)11(T)7 | (TG)11(T)7/(TG)11(T)7 | TG)11(T)7/(TG)11(T)9 | TG)11(T)7/(TG)12(T)5 | G)11(T)7/(TC)12(T)7 | TG)11(T)9/(TG)12(T) | (TC)12(T)7/(TC)12( | Total** | ( |
One of the discordant results was from the repoducibility study. The PolyTC/PolyT result for the sample was concertains
across all 18 replicates, but discordant with Sanger b
Samples were not retested
" The bala ample count for the Roy Crise est a synhuic sample (n=2) were built by bending linerized plannis with cell him
smules which were also part of the spresented from
{14}------------------------------------------------
Image /page/14/Picture/0 description: The image shows the word "illumina" in a simple, sans-serif font. The letters are black against a white background, creating a clear contrast. A small superscript symbol appears to the right of the final "a", adding a subtle detail to the logo.
Reproducibility - 139-Variant Assay
The reproducibility of the MiSeqDx Cystic Fibrosis System was determined through a blinded study using 3 trial sites and 2 operators at each site. Two well characterized panels of 46 samples each were tested by each of the operators at each site for a total of 810 calls per site. The panels contained a mix of genomic DNA from lymphoblastoid cell lines with known mutations in the CFTR gene as well as some leukocyte-depleted blood spiked with lymphoblastoid cell lines with known mutations in the CFTR gene. The blood samples were provided to allow incorporation of the extraction steps used to prepare gDNA that serves as the primary input for the assay workflow.
The sample pass rate, defined as the number of samples passing QC metrics on the first attempt, was 99.9%.
The genotype-level Positive Agreement for all variants was 99.77%. The Negative Agreement for all WT positions was 99.88% and the Overall Agreement for all reported positions was 99.88%. All test results are based on initial testing. No repeat testing was done for the reproducibility study.
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Table 4: Reproducibility Panel Variants
.
| Panel | Sample # | Sample Genotype | Variants | Total calls per site | Positive Agreeing calls (Variants) | Negative Agreeing calls (Wild type) | # Miscalls | # No Calls | Positive Agreement (%) | Negative Agreement (%) | Overall Agreement (%) | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Site 1 | Site 2 | Site 3 | Site 1 | Site 2 | Site 3 | |||||||||||
| A | 1 | S549N (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 2 | 1812-1 G>A (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 3 | Q493X/F50 8del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 4* | F508del/21 84delA (HET) | 810 | 12 | 12 | 12 | 797 | 798 | 798 | 0 | 1* | 100 | 100 | 100 | ||
| A | 5^ | Y122X/R115 8X (HET) | 810 | 12 | 10 | 12 | 798 | 665 | 798 | 0 | 135 | 94.44 | 94.44 | 94.44 | ||
| A | 6 | F508del/21 83AA>G (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 7 | R75X (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 8 | 1507del/F50 8del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 9** | F508del/W1 282X (HET) | 810 | 12 | 11 | 12 | 798 | 797 | 798 | 2* | 0 | 97.22 | 99.96 | 99.92 | ||
| A | 10* | F508del/32 72-26A>G (HET) | 810 | 12 | 11 | 12 | 798 | 797 | 798 | 2* | 0 | 97.22 | 99.96 | 99.92 | ||
| A | 11 | F508del/38 49+10kbC>T (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 12 | 621+1G>T/3 120+1G>A (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 13 | E60X/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 14 | M1101K (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 15 | M1101K (HOM) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 16 | F508del(HOM) | I506V,I507V,F508Cnotpresent | 828 | 6 | 6 | 6 | 822 | 822 | 822 | 0 | 0 | 100 | 100 | 100 | |
| A | 17 | F508del/3659delC(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 18 | R117H/F508del (HET) | (TG)10(T)9/(TG)12(T)5 | 816 | 18 | 18 | 18 | 798 | 798 | 798 | 0 | 0 | 100 | 100.00 | 100 | |
| A | 19 | 621+1G>T/711+1G>T(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 20 | G85E/621+1G>T (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 21 | A455E/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 22 | F508del/R560T (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 23 | F508del/Y1092X (C>A)(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 24 | N1303K(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 25 | G542X(HOM) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 26 | G542X(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 27 | G551D/R553X (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 28 | 3849+10kbC>T (HOM) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 29 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| A | 30 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 31 | 1717-1G>A(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 32 | R1162X(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 33 | R347P/G551D (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 34 | R334W(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 35 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| A | 36 | G85E (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 37 | I336K (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 39 | F508del/3849+10kbC>T(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 40 | 621+1G>T/3120+1G>A(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 41 | F508del/3659delC(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 42 | R117H/F508del (HET) | (TG)10(T)9/(TG)12(T)5 | 816 | 18 | 18 | 18 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | |
| A | 43 | G85E/621+1G>T (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 44 | A455E/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| A | 45 | N1303K(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| A | 46 | G551D/R553X (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 47 | 2789+5G>A(HOM) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 48 | CFTR dele2,3/F508del(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 49 | F508del/1898+1G>A(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 50 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 51 | F508del/2143delT(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 52 | 3876delA(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 53 | 3905insT(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 54 | 394delTT(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 55 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 56 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 57 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 58 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 59 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 60 | L206W(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 61 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 62 | G330X(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 63 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 64 | R347H(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 65 | 1078delT(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 66 | G178R/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 67 | S549R(c.1647T>G)(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 68 | S549N (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 69 | W846X(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 70 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 71 | E92X/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 72" | 621+1G>T/1154insTC(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 797 | 0 | 1" | 100 | 99.96 | 99.96 | ||
| B | 73 | G542X(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 74 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 75^ | F508del(HET) | 810 | 6 | 5 | 6 | 804 | 670 | 804 | 0 | 135 | 94.44 | 94.44 | 94.44 | ||
| B | 76 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 77 | 621+1G>T/A455E (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 78 | 1812-1 G->A (HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 79 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 | ||
| B | 80 | F508del/R553X (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 81 | F508del/G551D (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 82 | R347P/F508del (HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | ||
| B | 83 | R117H/F508del (HET) | (TG)10(T)9/(TG)12(T)S | 816 | 18 | 18 | 18 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 | |
| B | 84 | I507del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 | ||
| B | 85 | 2789+5G>A(HOM) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 |
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| B | 86" | CFTR dele2,3/F508del(HET) | 810 | 12 | 12 | 12 | 798 | 797 | 798 | 0 | 1" | 100 | 99.96 | 99.96 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| B | 87 | F508del/1898+1G>A(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 |
| B | 88 | WT | 810 | 0 | 0 | 0 | 810 | 810 | 810 | 0 | 0 | N/A | 100 | 100 |
| B | 89 | F508del/2143delT(HET) | 810 | 12 | 12 | 12 | 798 | 798 | 798 | 0 | 0 | 100 | 100 | 100 |
| B | 90 | 3905insT(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 |
| B | 91 | 394delTT(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 |
| B | 92 | F508del(HET) | 810 | 6 | 6 | 6 | 804 | 804 | 804 | 0 | 0 | 100 | 100 | 100 |
| Total | 74556 | 2209 | 221182 | 4 | 273 | 99.77 | 99.88 | 99.88 |
- The wild type location corresponding to the N1303K variant for one replicate resulted in a No Call due to insufficient coverage.
^ One replicate of samples 5 and 75 had a 0% call rate. Further investigation indicates that samples may not have been added to the sample plate prior to library preparation.
-
- Evidence indicates that samples 9 and 10 were likely switched by the operator prior to library preparation.
The wild type location corresponding to the M1V variant for one replicate of each of two samples resulted in a No Call due to insufficient coverage.
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| Variation (CommonName) | Variant Type | CFTR GeneRegion |
|---|---|---|
| polyTG/PolyT | Compound DIV* | Intron 9 |
| 2183AA>G | Compound DIV* | Exon 14 |
| CFTR dele2, 3 | DEL | Intron1-Intron3 |
| 1154insTC | DIV* | Exon 8 |
| 1507del | DIV* | Exon 11 |
| F508del | DIV* | Exon 11 |
| 2143delT | DIV* | Exon 14 |
| 3659delC | DIV* | Exon 22 |
| 3876delA | DIV* | Exon 23 |
| 394delTT | DIV in homopolymericregion* | Exon 3 |
| 1078delT | DIV in homopolymericregion* | Exon 8 |
| 2184delA | DIV in homopolymericregion* | . Exon 14 |
| 3905insT | DIV in homopolymericregion* | Exon 23 |
| E60X | SNV | Exon 3 |
| R75X | SNV | Exon 3 |
| G85E | SNV | Exon 3 |
| E92X | SNV | Exon 4 |
| R117H | SNV | Exon 4 |
| Y122X | SNV | Exon 4 |
| Table 4: Reproducibility Panel Variants | ||
|---|---|---|
| Variation (CommonName) | Variant Type | CFTR GeneRegion |
| 621+1G>T | SNV | Intron 4 |
| G178R | SNV | Exon 5 |
| 711+1G>T | SNV | Intron 5 |
| L206W | SNV | Exon 6 |
| G330X | SNV | Exon 8 |
| R334W | SNV | Exon 8 |
| I336K | SNV | Exon 8 |
| R347P | SNV | Exon 8 |
| R347H | SNV | Exon 8 |
| A455E | SNV | Exon 10 |
| Q493X | SNV | Exon 11 |
| 1717-1G>A | SNV | Intron 11 |
| G542X | SNV | Exon 12 |
| S549N | SNV | Exon 12 |
| S549R (c.1647T>G) | SNV | Exon 12 |
| G551D | SNV | Exon 12 |
| R553X | SNV | Exon 12 |
| R560T | SNV | Exon 12 |
| 1812-1 G->A | SNV | Intron 12 |
| 1898+1G>A | SNV | Intron 13 |
| W846X | SNV | Exon 15 |
| 2789+5G>A | SNV | Intron 16 |
| Variation (CommonName) | Variant Type | CFTR GeneRegion |
| 3120+1G>A | SNV | Intron 18 |
| 3272-26A>G | SNV | Intron 19 |
| Y1092X (C>A) | SNV | Exon 20 |
| M1101K | SNV | Exon 20 |
| R1158X | SNV | Exon 22 |
| R1162X | SNV | Exon 22 |
| 3849+10kbC>T | SNV | Intron 22 |
| W1282X | SNV | Exon 23 |
| N1303K | SNV | Exon 24 |
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DNA Extraction
Three commonly used, commercially available extraction methods representing magnetic bead extraction, alcohol precipitation and silica filter column isolation methods, were evaluated using K2EDTA anti-coagulated whole blood. A total of 14 unique blood samples were used in the study representing wild type and three mutant genotypes (3 samples with F508del, 1 sample with I506V, and 1 sample with D110H). The three DNA extraction methods were tested independently by 2 different operators who each performed 3 runs per extraction method. Each extraction was performed by each operator on different days. The DNA concentration and A260/A280 ratio of the extracted gDNA samples was determined using spectrophotometry. The total sample size for each extraction method in this study was 168 (14 samples x 2 operators/extraction method x 3 runs/operator x 2 replicates/extracted gDNA sample).
| Extraction Method | Number of samplestested | CallRate | Accuracy | Sample First PassRate* |
|---|---|---|---|---|
| Alcohol Precipitation | 168 | 100% | 100% | 100% |
| Silica Filter ColumnIsolation | 168 | 100% | 100% | 100% |
| Magnetic Bead Extraction | 168 | 100% | 100% | 100% |
- Percent of samples having call rate of >99% in first run
DNA input
The DNA input range of the Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay was evaluated by performing a serial dilution study using 14 representative DNA samples containing 16 unique CF variants. Each sample was tested in duplicate at 9 DNA input levels ranging from 1250 ng to 1 ng (1250 ng, 500 ng, 250 ng, 100 ng, 50 ng, 25 ng, 10 ng, 5 ng, and 1 ng). For determination of accuracy, sample genotypes were compared to bidirectional Sanger sequencing data and the deletions were compared to PCR assay. 1250 ng and 25 ng were identified as the upper and lower bound for DNA input respectively as they had >95% sample first pass rate with no incorrect calls (100% accuracy and call rate).
DNA inputs of 1250 ng, 250 ng, and 100 ng were further tested with 4 representative DNA samples and 20 replicates per DNA input level for each sample (n=420=80 samples), while the lower bound of 25 ng was tested with 14 samples, 20 replicates for each sample (n=1420=280 samples). The accuracy and sample first pass rate was 100% at all DNA input levels.
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Interfering Substances
To assess the impact of interfering substances on the MiSeqDx Cystic Fibrosis 139-Variant Assay, the performance of the assay was evaluated in the presence and absence of potential interferents. Eight whole blood specimens were tested in the study including 3 CF positive samples with unique genotypes. Four endogenous interfering substances (bilirubin, cholesterol, hemoglobin, and triglycerides) were tested by spiking them into blood specimens prior to DNA extraction. The concentration limits for each substance is shown in the following table. Additionally, to assess interference resulting from blood collection (short draw), EDTA was spiked into blood samples, and to assess interference resulting from sample preparation, the final wash buffer from a silica filter column isolation method was added to purified genomic DNA.
The MiSeqDx Cystic Fibrosis 139-Variant Assay achieved 100% call rate for all samples tested, and 100% reproducibility in genotype calls between samples in the presence and absence of interfering substances.
To access the impact of multiplexing index primer interference, a cross-contamination study using two samples, each with unique homozygous genotypes at 4 different genomic positions, and two respective index primers was performed. No change in variant calling was observed with contamination levels <40%. The sample genotype became heterozygous when contamination levels were ≥ 40%.
| TestSubstance | Total Number ofReplicates | Concentration Testedin Blood(Upper Limit) | Concentration Testedin Blood(Lower Limit) | CallRate |
|---|---|---|---|---|
| Bilirubin | 16 | 684 µmol/L | 137 µmol/L | 100% |
| Cholesterol | 16 | 13 mmol/L | 2.6 mmol/L | 100% |
| Hemoglobin | 16 | 2 g/L | 0.4 g/L | 100% |
| Triglyceride | 16 | 37 mmol/L | 7.4 mmol/L | 100% |
| EDTA | 16 | 7.0 mg/mL | 2.8 mg/mL | 100% |
No interference was observed from any of the endogenous or exogenous interferents.
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Sample Indexing
Sample index primers are used in the assay to assign a unique barcode to each sample DNA, allowing the ability to pool multiple samples together into a single sequencing run. A total of 96 samples indexes were tested using 8 unique DNA samples to verify the ability of the assay to consistently make a genotyping call for a given sample across different indexing primer combinations. Each sample was tested with 12 different indexing primer combinations. Sample results were compared against bidirectional Sanger sequencing data for all positions/variants except the 2 large deletions, which were confirmed using a duplex PCR assay. Reproducibility and accuracy were 100% for all sample/index primer combinations.
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Image /page/26/Picture/0 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo is a circular seal with the department's name around the perimeter. Inside the circle is an abstract symbol that resembles an eagle or bird in flight. The symbol is composed of three curved lines that suggest wings and a tail.
DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
November 19, 2013
ILLUMINA, INC. LEANNE M. KIVIHARJU SENIOR DIRECTOR, REGULATORY AFFAIRS 5200 ILLUMINA WAY SAN DIEGO CA 92122
Re: K124006
Trade/Device Name: Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay Regulation Number: 21 CFR 866.5900 Regulation Name: Cystic fibrosis transmembrance regulator (CFTR) gene mutation detection system Regulatory Class: II Product Code: PFR Dated: November 18, 2013
Received: November 19. 2013
Dear Ms. Kiviharju:
· 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. Ilisting of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class 11 (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA 's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act s requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of
{27}------------------------------------------------
Page 2-Ms. Kiviharju
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.
If you desire specific advice for your device on our labeling regulations (2) CFR Parts 801 and 809), please contact 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/MedicalDevices/ResourcesforYou/Industry/default.hym. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 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/deliault.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/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours.
Reena Philip -S
for Maria M. Chan, Ph.D.
Director
Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health
Center for Devices and Radiological Health
Enclosure
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DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration
Indications for Use
Form Approved: OMB No. 0910-0120 Expiration Date: December 31, 2013 See PRA Statement on last page.
510(k) Number (if known) K124006
Device Name
Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay
Indications for Use (Describe)
The Illumina MiSeqDx Cystic Fibrosis 139-Variant Assay is a qualitative in vitro diagnostic system used to simultaneously detect 139 clinically relevant cystic fibrosis disease-causing mutations and variants of the cystic fibrosis transmembrance regulator (CFTR) gene in genomic DNA isolated from human peripheral whole blood specimens. The variants include those recommended in 2004 by the American College of Medical Genetics (ACMG)) and in 2011 by the American College of Obstetricians and Gynecologists (ACOG)2. The test is intended for carrier screening in adults of reproductive age, in confirmatory diagnostic testing of newborns and children, and as an initial test to aid in the diagnosis of individuals with suspected cystic fibrosis. The results of this test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available laboratory and clinical information.
This test is not indicated for use for newborn screening, fetal diagnostic testing, pre-implantation testing, or for stand-alone diagnostic purposes.
The test is intended to be used on the Illumina MiSeqDx instrument.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY Concurrence of Center for Devices and Radiological Health (CDRH) (Signature) K124006 November 19, 2013
Donna M. Roscoe -S
§ 866.5900 Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation detection system.
(a)
Identification. The CFTR gene mutation detection system is a device used to simultaneously detect and identify a panel of mutations and variants in the CFTR gene. It is intended as an aid in confirmatory diagnostic testing of individuals with suspected cystic fibrosis (CF), carrier identification, and newborn screening. This device is not intended for stand-alone diagnostic purposes, prenatal diagnostic, pre-implantation, or population screening.(b)
Classification. Class II (special controls). The special control is FDA's guidance document entitled “Class II Special Controls Guidance Document: CFTR Gene Mutation Detection System.” See § 866.1(e) for the availability of this guidance document.