ILLUMINA MISEQDX CYSTIC FIBROSIS CLINICAL SEQUENCING ASSAY
Device Facts
| Record ID | K132750 |
|---|---|
| Device Name | ILLUMINA MISEQDX CYSTIC FIBROSIS CLINICAL SEQUENCING ASSAY |
| Applicant | Illumina, Inc. |
| Product Code | PFS · Immunology |
| Decision Date | Nov 19, 2013 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.5900 |
| Device Class | Class 2 |
Intended Use
The Illumina MiSeqDx™ Cystic Fibrosis Clinical Sequencing Assay is a targeted sequencing in vitro diagnostic system that re-sequences the protein coding regions and intron/exon boundaries of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene in genomic DNA isolated from human peripheral whole blood specimens collected in K₂EDTA. The test detects single nucleotide variants, and small InDels within the region sequenced, and additionally reports on two deep intronic mutations and two large deletions. The test is intended to be used on the Illumina MiSeqDx Instrument. The test is intended to be used as an aid in the diagnosis of individuals with suspected cystic fibrosis (CF). The test is most appropriate when the patient has an atypical or non-classic presentation of CF or when other mutation panels have failed to identify both causative mutations. The results of the test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available information including clinical symptoms, other diagnostic tests, and family history. This test is not indicated for use for stand-alone diagnostic purposes, fetal diagnostic testing, for pre-implantation testing, carrier screening, newborn screening, or population screening.
Device Story
The Illumina MiSeqDx Cystic Fibrosis Clinical Sequencing Assay is a targeted NGS-based IVD system. Input: genomic DNA from K2EDTA peripheral whole blood. Process: library preparation (amplification and indexing) followed by Sequencing By Synthesis (SBS) on the MiSeqDx instrument. Output: detection of SNVs, small InDels, two deep intronic mutations, and two large deletions in the CFTR gene. Used in clinical molecular genetics laboratories; results interpreted by board-certified clinical molecular geneticists. The device aids in diagnosing suspected CF cases, especially atypical presentations or cases where other panels failed. Benefits include comprehensive re-sequencing of CFTR protein-coding regions and boundaries to identify causative mutations, supporting clinical decision-making alongside patient history and symptoms.
Clinical Evidence
Bench testing only. Accuracy assessed using 500 samples (366 clinical, 68 cell line, 14 extraction study, 52 synthetic). Genotype-level positive agreement (PA) was 99.66% (100% excluding PolyTG/PolyT). Negative agreement (NA) >99.99%. Overall agreement (OA) >99.99%. Reproducibility study across 3 sites/2 operators showed 99.7% sample first-pass rate. Analytical studies confirmed performance across DNA inputs (25-1250 ng) and in the presence of interfering substances (bilirubin, cholesterol, hemoglobin, triglycerides, EDTA).
Technological Characteristics
Targeted NGS system using PCR-based library preparation and Sequencing By Synthesis (SBS). Analyzes CFTR gene protein-coding regions and intron/exon boundaries. Connectivity: MiSeqDx instrument. Software: Data analysis software for variant calling. Sterilization: Not applicable (reagents).
Indications for Use
Indicated for individuals with suspected cystic fibrosis (CF), particularly those with atypical/non-classic presentation or where prior mutation panels failed to identify causative mutations. Not for fetal, pre-implantation, carrier, newborn, or population screening.
Regulatory Classification
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.
Special Controls
*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.
Predicate Devices
- x-TAG Cystic Fibrosis 60 Kit v2 (K083845)
Related Devices
- K124006 — ILLUMINA MISEQDX CYSTIC FIBROSIS 139-VARIANT ASSAY · Illumina, Inc. · Nov 19, 2013
- K083294 — VERIGENE CFTR NUCLEIC ACID TEST AND VERIGENE CFTR POLYT NUCLEIC ACID TEST · Nanosphere, Inc. · Jul 24, 2009
Submission Summary (Full Text)
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K132750
#### 510(k) Summary 1.3
The following 510(k) summary was prepared in accordance with 21 CFR 807.92.
NOV 1 9 2013
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#### 510(k) Summary
#### GENERAL INFORMATION
Illumina Inc. Submitted by: 5200 Illumina Way San Diego, CA 92122 858-202-4500 (phone) 858-202-4600 (fax)
Company Contact: Bryan Schneider Associate Director, Regulatory Affairs 858-255-5228 (phone) bschneider@illumina.com . :
:
: ,
Date Prepared: November 18, 2013
#### DEVICE IDENTIFICATION
Assay: ..... ... ... ..
#### Trade or Proprietary Name:
| | Illumina MiSeqDx™ Cystic Fibrosis Clinical<br>Sequencing Assay |
|----------------------|-------------------------------------------------------------------------------------------------------------------------------|
| Common Name: | Sequencing by synthesis cystic fibrosis test |
| Classification Name: | CFTR (cystic fibrosis transmembrane conductance<br>regulatory) gene mutation detection (21 CFR 866.5900,<br>Product Code PFS) |
| Predicate Device: | x-TAG Cystic Fibrosis 60 Kit v2 (k083845) |
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#### DEVICE DESCRIPTION
The Illumina MiSeqDx Cystic Fibrosis Clinical Sequencing 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 Clinical Sequencing Assay
The Illumina MiSeqDx(TM) Cystic Fibrosis Clinical Sequencing Assay is a targeted sequencing in vitro diagnostic system that re-sequences the protein coding regions and intron/exon boundaries of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene in genomic DNA isolated from human peripheral whole blood specimens collected in K2EDTA. The test detects single nucleotide variants, and small InDels within the region sequenced, and additionally reports on two deep intronic mutations and two large deletions. The test is intended to be used on the Illumina MiSeqDx Instrument.
The test is intended to be used as an aid in the diagnosis of individuals with suspected cystic fibrosis (CF). The test is most appropriate when the patient has an atypical or non-classic presentation of CF or when other mutation panels have failed to identify both causative mutations. The results of the test are intended to be interpreted by a board-certified clinical molecular geneticist or equivalent and should be used in conjunction with other available information including clinical symptoms, other diagnostic tests, and family history. This test is not indicated for use for stand-alone diagnostic purposes, fetal diagnostic testing, for preimplantation testing, carrier screening, newborn screening, or population screening. i : . . . . . . . . .
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#### SUBSTANTIAL EQUIVALENCE
・・・・
| Characteristic | Illumina | Luminex (K083845) |
|------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Assay Name | Illumina MiSeqDx Cystic Fibrosis<br>Clinical Sequencing Assay | Luminex xTAG®Cystic<br>Fibrosis 60 Kit v2 |
| Intended<br>Use | The Illumina MiSeqDx Cystic<br>Fibrosis Clinical Sequencing<br>Assay is a targeted sequencing in<br>vitro diagnostic system that re-<br>sequences the protein coding<br>regions and intron/exon<br>boundaries of the Cystic Fibrosis<br>Transmembrane Conductance<br>Regulator (CFTR) gene in<br>genomic DNA isolated from<br>human peripheral whole blood<br>specimens collected in K2EDTA.<br>The test detects single nucleotide<br>variants, and small indels within<br>the region sequenced, and<br>additionally reports on two deep<br>intronic mutations and two large<br>deletions. The test is intended to<br>be used on the Illumina MiSeqDx<br>Instrument.<br><br>The test is intended to be used as<br>an aid in the diagnosis of<br>individuals with suspected cystic<br>fibrosis (CF).This assay is most<br>appropriate when the patient has | The xTAG® Cystic Fibrosis 60<br>kit v2 is a device used to<br>simultaneously detect and<br>identify a panel of mutations<br>and variants in the cystic<br>fibrosis transmembrane<br>conductance regulator<br>(CFTR) gene in human blood<br>specimens. The panel<br>includes mutations and<br>variants currently<br>recommended by the<br>American College of Medical<br>Genetics and American<br>College of Obstetricians and<br>Gynecologists<br>(ACMG/ACOG) plus some<br>of the world's most common<br>and North American<br>prevalent mutations. The<br>xTAG Cystic Fibrosis 60 kit<br>v2 is a qualitative<br>genotyping test which<br>provides information<br>intended to be used for<br>carrier testing in adults of |
| Confidential 4 | | |
| Characteristic | Illumina | Luminex (K083845) |
| | an atypical or non-classic<br>presentation of CF or when other<br>mutation panels have failed to<br>identify both causative mutations.<br>The results of the test are<br>intended to be interpreted by a<br>board-certified clinical molecular<br>geneticist or equivalent and<br>should be used in conjunction<br>with other available information<br>including clinical symptoms,<br>other diagnostic tests, and family<br>history. This test is not indicated<br>for use for fetal diagnostic testing,<br>for pre-implantation testing,<br>carrier screening, newborn<br>screening, or population<br>screening.<br>The test is intended to be used on<br>the Illumina MiSeqDx™<br>Instrument. | reproductive age, as an aid<br>in newborn screening, and in<br>confirmatory diagnostic<br>testing in newborns and<br>children.<br>The kit is not indicated for<br>use in fetal diagnostic or pre-<br>implantation testing. The kit<br>is also not indicated for<br>stand-alone diagnostic<br>purposes. |
| Assay type | Sequencing by synthesis test | Qualitative nucleic acid<br>multiplex test |
| Variants<br>Detected | Mutations and variants from the<br>protein coding regions and<br>intron/exon boundaries of the<br>CFTR gene, including two deep<br>intronic mutations and two large<br>deletions. | 60 CFTR mutations and 4<br>variants (benign<br>polymorphisms) |
| Characteristic | Illumina | Luminex (K083845) |
| Technology | PCR-based amplification of<br>regions of interest that are then<br>hybridized to a flow cell to allow<br>sequencing by synthesis | Multiplex PCR followed by<br>multiplex allele specific<br>primer extension for<br>genotyping, hybridized to<br>multiplex fluorescent<br>microparticles, detected by<br>flow cytometry. |
| Sample Type | Nucleic acid from K2EDTA<br>anticoagulated blood | Nucleic acid from whole<br>blood anticoagulated with<br>either EDTA or citrate. |
| Sample<br>Preparation | DNA extraction using validated<br>laboratory method | Same |
| Contra-<br>indications | This test is not indicated for use<br>for fetal diagnostic testing, for<br>pre-implantation testing, carrier<br>screening, newborn screening, or<br>population screening. | Not indicated for fetal<br>diagnostic testing, for pre-<br>implantation testing, or for<br>stand-alone diagnostic<br>purposes. |
| Assay<br>Controls | Positive and negative controls<br>required, not supplied | Negative controls required,<br>not supplied. Positive<br>controls recommended, not<br>supplied. |
| Instrument<br>System | MiSeqDx | Luminex 100 or 200 IS |
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#### MiSeqDx Cystic Fibrosis Clinical Sequencing Assay
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#### PERFORMANCE.CHARACTERISTICS
#### Accuracy
Accuracy of the Illumina MiSeqDx Cystic Fibrosis Clinical Sequencing 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
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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 were designed to include the genomic context of rare variants, and contained anywhere from 1 to 10 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.
For the MiSeqDx Cystic Fibrosis Clinical Sequencing Assay, a total of 5,206 positions were compared to the reference methods of Sanger bi-directional sequencing and PCR testing. The genotyping results for SNV and small InDel sites, including the PolyTG/PolyT region, were compared to Sanger bi-directional sequence analysis.
Two validated PCR based assays were 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 PCR 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 samples.
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
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MiSeqDx Cystic Fibrosis Clinical Sequencing Assay
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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 MiSeqDx Cystic Fibrosis Clinical Sequencing Assay had a genotype-level PA of 99.66% including the PolyTG/PolyT variants and 100%, excluding PolyTG/PolyT variants (Table 1). The NA for all wild types was >99.99% and the OA for all WT and variants was >99.99%. The PolyTG/PolyT variant PA was 98.44%. All results are based on initial testing. No repeat testing was done for this study.
Accuracy of the PolyTG/PolyT variants is demonstrated in Table 2.
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| Genotype<br>(Common<br>Name) | cDNA name<br>coordinate | cDNA name | Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Positive calls (Variants) | | | No<br>Calls | Miscalls | Positive<br>Agreement |
|---------------------------------------------------------|--------------------------------|-----------------|----------------------------------|----------------------------------|---------------------------|-------------------------|----------------------|-------------|----------|-----------------------|
| | | | | | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | | | |
| 117120141 | N/A | SNV | Exon1 | 25 | 3 | 0 | 0 | 0 | 100 | |
| 117120145 | N/A | SNV | Exon1 | 3 | 2 | 0 | 0 | 0 | 100 | |
| M1V | c.1A>G | SNV | Exon1 | 0 | 0 | 1 | 0 | 0 | 100 | |
| CFTR dele2, 3 | c.54-<br>5940_273+10250del21kb | Del | Intron1 | 4 | 1 | 0 | 0 | 0 | 100 | |
| R31C | c.91C>T | SNV | Exon2 | 3 | 1 | 0 | 0 | 0 | 100 | |
| Q39X | c.115C>T | SNV | Exon2 | 0 | 0 | 1 | 0 | 0 | 100 | |
| E60X | c.178G>T | SNV | Exon3 | 6 | 1 | 0 | 0 | 0 | 100 | |
| P67L | c.200C>T | SNV | Exon3 | 1 | 0 | 1 | 0 | 0 | 100 | |
| R74W | c.220C>T | SNV | Exon3 | 0 | 2 | 0 | 0 | 0 | 100 | |
| R74Q | c.221G>A | SNV | Exon3 | 2 | 0 | 0 | 0 | 0 | 100 | |
| R75X | c.223C>T | SNV | Exon3 | 3 | 1 | 0 | 0 | 0 | 100 | |
| R75Q | c.224G>A | SNV | Exon3 | 20 | 1 | 0 | 0 | 0 | 100 | |
| G85E | c.254G>A | SNV | Exon3 | 6 | 2 | 0 | 0 | 0 | 100 | |
| Genotype<br>(Common<br>Name<br>cDNA name<br>coordinate) | cDNA name | Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Positive calls (Variants) | | | | | | Positive<br>Agreement |
| | | | | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | No<br>Calls | Miscalls | | |
| 394delTT | c.262_263delTT | DIV | Exon3 | 3 | 1 | 0 | 0 | 0 | 100 | |
| 405+1G>A | c.273+1G>A | SNV | Intron3 | 0 | 0 | 1 | 0 | 0 | 100 | |
| 406-1G>A | c.274-1G>A | SNV | Exon4 | 4 | 0 | 0 | 0 | 0 | 100 | |
| E92K | c.274G>A | SNV | Exon4 | 0 | 0 | 1 | 0 | 0 | 100 | |
| E92X | c.274G>T | SNV | Exon4 | 0 | 1 | 1 | 0 | 0 | 100 | |
| Q98X | c.292C>T | SNV | Exon4 | 0 | 0 | 2 | 0 | 0 | 100 | |
| 444delA | c.312delA | DIV | Exon4 | 0 | 2 | 0 | 0 | 0 | 100 | |
| 457TAT>G | c.325_327delTATinsG | DIV | Exon4 | 0 | 0 | 1 | 0 | 0 | 100 | |
| D110H | c.328G>C | SNV | Exon4 | 1 | 0 | 1 | 0 | 0 | 100 | |
| R117C | c.349C>T | SNV | Exon4 | 4 | 0 | 0 | 0 | 0 | 100 | |
| R117H | c.350G>A | SNV | Exon4 | 17 | 2 | 0 | 0 | 0 | 100 | |
| Y122X | c.366T>A | SNV | Exon4 | 0 | 1 | 0 | 0 | 0 | 100 | |
| F143LfsX10 | c.425delT | DIV | Exon4 | 0 | 1 | 0 | 0 | 0 | 100 | |
| 574delA | c.442delA | DIV | Exon4 | 0 | 0 | 2 | 0 | 0 | 100 | |
| Q151K | c.451C>A | SNV | Exon4 | 1 | 0 | 0 | 0 | 0 | 100 | |
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| Genotype<br>(Common<br>Name<br>cDNA name<br>coordinate) | cDNA name | Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Positive calls (Variants) | | | No<br>Calls | Miscalls | Positive<br>Agreement | | |
|----------------------------------------------------------|---------------------------------------|-----------------|----------------------------------|----------------------------------|---------------------------|-------------------------|-------------------------|----------------------|-----------------------|-----------------------|-----------------------|
| 621+1G>T | c.489+1G>T | SNV | Intron4 | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | 0 | 0 | 100 | | |
| 621+3A>G | c.489+3A>G | SNV | Intron4 | 1 | 0 | 0 | 0 | 0 | 100 | | |
| 663delT | c.531delT | DIV | Exon5 | 1 | 0 | 1 | 0 | 0 | 100 | | |
| G178R | c.532G>A | SNV | Exon5 | 1 | 1 | 0 | 0 | 0 | 100 | | |
| 711+1G>T | c.579+1G>T | SNV | Intron5 | 3 | 1 | 0 | 0 | 0 | 100 | | |
| 711+3A>G | c.579+3A>G | SNV | Intron5 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 711+5 G->A | c.579+5G>A | SNV | Intron5 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 712-1 G->T | c.580-1G>T | SNV | Exon6 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| H199Y | c.595C>T | SNV | Exon6 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| P205S | c.613C>T | SNV | Exon6 | 1 | 0 | 1 | 0 | 0 | 100 | | |
| L206W | c.617T>G | SNV | Exon6 | 8 | 1 | 0 | 0 | 0 | 100 | | |
| A209S | c.625G>T | SNV | Exon6 | 0 | 1 | 0 | 0 | 0 | 100 | | |
| Q220X | c.658C>T | SNV | Exon6 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| L227R | c.680T>G | SNV | Exon6 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 852del22 | c.720_741delAGGGAGAA<br>TCATCATCACTAC | DIV | Exon6 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| Genotype<br>(Common<br>Name<br>cDNA name,<br>coordinate) | cDNA name | Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Positive calls (Variants) | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | No<br>Calls" | Miscalls | Positive<br>Agreement | |
| E279D | c.837A>T | SNV | Exon7 | 1 | 0 | 0 | 0 | 0 | 100 | | |
| R297Q | c.890G>A | SNV | Exon8 | 2 | 0 | 0 | 0 | 0 | 100 | | |
| 1078delT | c.948delT | DIV | Exon8 | 1 | 1 | 0 | 0 | 0 | 100 | | |
| L320V | c.958T>G | SNV | Exon8 | 1 | 0 | 0 | 0 | 0 | 100 | | |
| G330X | c.988G>T | SNV | Exon8 | 1 | 1 | 0 | 0 | 0 | 100 | | |
| R334W | c.1000C>T | SNV | Exon8 | 6 | 1 | 0 | 0 | 0 | 100 | | |
| I336K | c.1007T>A | SNV | Exon8 | 0 | 1 | 0 | 0 | 0 | 100 | | |
| T338I | c.1013C>T | SNV | Exon8 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 1154insTC | c.1022_1023insTC | DIV | Exon8 | 0 | 1 | 0 | 0 | 0 | 100 | | |
| S341P | c.1021T>C | SNV | Exon8 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| R347H | c.1040G>A | SNV | Exon8 | 6 | 1 | 1 | 0 | 0 | 100 | | |
| R347P | c.1040G>C | SNV | Exon8 | 3 | 2 | 0 | 0 | 0 | 100 | | |
| R352Q | c.1055G>A | SNV | Exon8 | 5 | 0 | 0 | 0 | 0 | 100 | | |
| Q359K/T360K | c.[1075C>A;1079C>A] | SNV | Exon8 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 1213delT | c.1081delT | DIV | Exon8 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| Genotype<br>(Common<br>Name | cDNA name<br>coordinate) | cDNA name | Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Positive calls (Variants) | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | No<br>Calls | Miscalls | Positive<br>Agreement |
| 1248+1G>A | c.1116+1G>A | SNV | Intron8 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 1259insA | c.1127_1128insA | DIV | Exon9 | 0 | 0 | 2 | 0 | 0 | 100 | | |
| W401X<br>(c.1202G>A) | c.1202G>A | SNV | Exon9 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| W401X<br>(c.1203G>A) | c.1203G>A | SNV | Exon9 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| 1341+1G->A | c.1209+1G>A | SNV | Intron9 | 0 | 0 | 2 | 0 | 0 | 100 | | |
| PolyTGPolyT | N/A | PolyTGPolyT | Intron9 | 369 | 79 | 52 | 3 | 4 | 98.60 | | |
| 1461ins4 | c.1329_1330insAGAT | DIV | Exon10 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| A455E | c.1364C>A | SNV | Exon10 | 4 | 2 | 0 | 0 | 0 | 100 | | |
| 1525-1G->A | c.1393-1G>A | SNV | Exon11 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| S466X (C->A) | c.1397C>A | SNV | Exon11 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| S466X (C->G) | c.1397C>G | SNV | Exon11 | 1 | 0 | 1 | 0 | 0 | 100 | | |
| L467P | c.1400T>C | SNV | Exon11 | 0 | 0 | 1 | 0 | 0 | 100 | | |
| V470M | c.1408G>A | SNV | Exon11 | 311 | 71 | 0 | 0 | 0 | 100 | | |
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| Variant<br>Type | CFTR<br>gene<br>region<br>(hg19) | Clinical<br>Samples | Cell<br>Line<br>Samples | Synthetic<br>Samples | No<br>Calls | Miscalls | Positive<br>Agreement |
|-----------------|----------------------------------|---------------------|-------------------------|----------------------|-------------|----------|-----------------------|
| DIV | Exon11 | 1 | 0 | 1 | 0 | 0 | 100 |
| SNV | Exon11 | 0 | 1 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 1 | 0 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 0 | 0 | 2 | 0 | 0 | 100 |
| SNV | Exon11 | 0 | 0 | 1 | 0 | 0 | 100 |
| SNV | Exon11 | 4 | 2 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 7 | 0 | 0 | 0 | 0 | 100 |
| DIV | Exon11 | 4 | 2 | 0 | 0 | 0 | 100 |
| DIV | Exon11 | 84 | 29 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 0 | 1 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 1 | 1 | 0 | 0 | 0 | 100 |
| DIV | Exon11 | 1 | 0 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 2 | 0 | 0 | 0 | 0 | 100 |
| SNV | Exon11 | 0 | 0 | 1 | 0 | 0 | 100 |
| SNV | Exon11 | 3 | 2 | 0 | 0 | 0 | 100 |
'r
{14}------------------------------------------------
| | greemer<br>Positive | 100 | 100 | 00 I | 00 T | 00 T | 100 | 00 I | 100 | 100 | 100 | 100 | 100 | 100 | 100 |
|---------------------------------------------------------|-----------------------------------------------------------------------------------|-----------------|----------------------------------|---------------------------|-------------------------|----------------------|-------------|-----------------|-----------------------|---------|----------|-----------|-----------|----------|---------|
| | Aiscall | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | ાદ) તાલુકાઓ પૈકીના એક એવા ગુજરાત રાજ્યના | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 |
| | yntheti< Samples | 0 | L | 0 | 0 | I | L | 0 | 0 | L | 0 | 0 | I | I | I |
| ositive calls (Variants | ាជាតិ (ទី) | ਟ | 0 | I | દ | 0 | ਨ | I | € | 0 | ਟ | 0 | 0 | 0 | 0 |
| | amples<br>Clinica | 9 | 0 | P | ਹ…
