Capstone Diagnostics8601 Dunwoody Place,Building 400, Suite 444

Atlanta, GA 30350www.capstonediagnostics.com

844-497-8851Laboratory Director: John Hanson, PhD

CLIA ID Number: 11D2073885

TEST PERFORMED

C-55 (Comprehensive Panel)Targeted next-generation sequencing was performed on this specimen. See Methods and Limitations section formore information.

VARIANT SUMMARYVariants Detected Genotype Phenotype Assessment

c.798_799delTT

BRCA1

p.S267fs*19

HeterozygousSusceptibility To FamilialBreast-ovarian Cancer

Type 1Pathogenic

VARIANT INTERPRETATION

p.S267fs*19

c.798_799delTT

Pathogenic

BRCA1 Interpretation: BRCA1 is a tumor suppressor responsible for repair of DNA double strandbreaks and maintenance of genomic stability [3]. Deletions, loss of heterozygosity, and loss-of-function mutations cause BRCA1 inactivation [7, 6].

RESULT

Pathogenic Variant Detected

PATIENT INFORMATION PROVIDER INFORMATION SPECIMEN

Patient Name: Sarah Doe

Date of Birth: Apr 10, 1985

Age: 33

Sex: Female

Ethnicity: Caucasian

Physician: Dr Jospeh Smith

Client: St Michael Hospital

Client ID: 123654

Accession ID: 10000520

Specimen Type: Buccal Swab

Collection Date: Aug 21, 2018

Date Accessioned: Aug 22, 2018

Date Reported: Sep 7, 2018

Page 1 of 3Capstone Diagnostics | 8601 Dunwood Place | Building 400 | Suite 444 | Atlanta, GA | 30350

An abnormality is detected that has been reported to be associated with an increased risk for hereditary cancer. This finding may help you and your doctor make more informed choices about your health care and develop a strategy for cancer prevention and monitoring.

C-55 (Comprehensive Panel)

Accession ID: 10000520Laboratory Director: John Hanson, PhD

CLIA ID Number: 11D2073885

GENES TESTEDAPC, ATM, AXIN1, BAP1, BARD1, BMPR1A, BRCA1, BRCA2, BRIP1, CDC73, CDH1, CDKN1C, CDKN2A, CHEK2, DICER1,DIS3L2, EPCAM, FH, FLCN, GPC3, GREM1, MEN1, MET, MLH1, MSH2, MSH6, MUTYH, NBN, NF1, PAKAR1A, PALB2,PMS2, POLD1, PTCH1, PTEN, RAD50, RAD51C, RAD51D, RB1, REQL4, RET, SDHA, SDHB, SDHC, SDHD, SMAD4,SMARC4A, STK11, SUFU, TP53, TSC1, TSC2, VHL, WT1

METHODS AND LIMITATIONSDNA extracted from patient’s saliva sample is quantified using Qubit 2.0 Fluorimeter (Life Technologies Inc.). Germlinemutations of APC, ATM, AXIN1, BAP1, BARD1, BMPR1A, BRCA1, BRCA2, BRIP1, CDC73, CDH1, CDKN1C, CDKN2A,CHEK2, DICER1, DIS3L2, EPCAM, FH, FLCN, GPC3, GREM1, MEN1, MET, MLH1, MSH2, MSH6, MUTYH, NBN, NF1,PAKAR1A, PALB2, PMS2, POLD1, PTCH1, PTEN, RAD50, RAD51C, RAD51D, RB1, REQL4, RET, SDHA, SDHB, SDHC,SDHD, SMAD4, SMARC4A, STK11, SUFU, TP53, TSC1, TSC2, VHL, WT1 genes are tested by sequencing the entirecoding region for each gene with 5 bp of padding at both ends. The panel consists of amplicons that cover 98% of allcoding regions of the genes listed above. Each sample template is tagged with a unique barcode for sample identificationbefore processing. Sequencing is performed on the Ion Torrent S5 (Life Technologies Inc.) using AmpliSeq and Ion Cheftechnologies, and Hi-Q View chemistry. Data obtained are analyzed using the Applied Biosystems Ion Torrent variantanalyzing software, which includes signal processing, base calling,quality score assignment, adapter trimming, alignment tohuman genome 19 reference (hg19), coverage analysis, and variant calling. Variant annotation is performed on the QIAGENClinical Insight software. Sanger sequencing confirmation is performed on all pathogenic, likely pathogenic, and variantof unknown significance samples through Eurofins Clinical Molecular Testing Services, LLC.

Based on validation studies, our method has a >95% sensitivity and specificity for detecting single nucleotide variants,insertions/deletions, and other frame shift mutations. However, some rare and novel genetic variations might not be detectedby this method. Genes other then those listed above that are associated with certain types of cancer are beyond the scope ofthis test.

QIAGEN Clinical Insight (QCITM) is a variant analysis, interpretation and decision support tool for research and clinical labsanalyzing human genetics data and is not intended to be used for diagnostic purposes. QCI Interpret software includes thefollowing underlying databases, data reference sets and tools; QIAGEN Clinical Insight-Interpret (5.3.20180727), IngenuityKnowledge Base (Rohan 180831.001), CADD (v1.3), Allele Frequency Community (2018-05-25), EVS (ESP6500SI-V2),JASPAR (2013-11), Ingenuity Knowledge Base Snapshot Timestamp (2018-08-31 11:07:19.0), Vista Enhancer hg18 (2012-07), Vista Enhancer hg19 (2012-07), Clinical Trials (Rohan 180831.001), PolyPhen-2 (v2.2.2), 1000 Genome Frequency(phase3v5b), ExAC (0.3.1), iva (Jun 26 11:55 iva-1.0.476233.jar), PhyloP hg18 (2009-11), PhyloP hg19 (2009-11), DbSNP(151), TargetScan (6.2), CentoMD (4.2), OMIM (May 26, 2017), gnomAD (2.0.1), BSIFT (2016-02-23), TCGA (2013-09-05),Clinvar (2018-04-06), DGV (2016-05-15), COSMIC (v84), HGMD (2018.1), SIFT4G (2016-02-23)

DISCLAIMERFalse positives and false negatives are possible. Such errors can be due to contamination, technical errors, andinterference from rare genetic variants. It is recommended to consider alternative methods before taking any clinicalaction. Consulting your results with a genetic counselor is highly recommended. This test was developed, and itsperformance characteristics determined by Capstone Diagnostics. The laboratory is regulated and accredited by CLIAand the Joint Commission (JCHO) as qualified to perform high-complexity testing. This test is used for clinicalpurposes. It should not be regarded as investigational or for research.

Page 2 of 3Capstone Diagnostics | 8601 Dunwood Place | Building 400 | Suite 444 | Atlanta, GA | 30350

C-55 (Comprehensive Panel)

Accession ID: 10000520Laboratory Director: John Hanson, PhD

CLIA ID Number: 11D2073885

SELECTED CITATIONS

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Crawford B, Adams SB, Sittler T, van den Akker J, Chan S, Leitner O, Ryan L, Gil E, van 't Veer L (2017) Multi-gene paneltesting for hereditary cancer predisposition in unsolved high-risk breast and ovarian cancer patients. Breast Cancer ResTreat. 2017 Jun;163(2):383-390. Epub 2017 Mar 9 (PMID: 28281021)

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Farmer H, McCabe N, Lord CJ, Tutt AN, Johnson DA, Richardson TB, Santarosa M, Dillon KJ, Hickson I, Knights C, MartinNM, Jackson SP, Smith GC, Ashworth A (2005) Targeting the DNA repair defect in BRCA mutant cells as a therapeuticstrategy. Nature. 2005 Apr 14;434(7035):917-21 (PMID: 15829967)

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Hwang SM, Lee KC, Lee MS, Park KU (2017) Comparison of Ion Personal Genome Machine Platforms for the Detection ofVariants in BRCA1 and BRCA2. Cancer Res Treat. 2018 Jan;50(1):255-264. Epub 2017 Apr 7 (PMID: 28392550)

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Schenk D, Song G, Ke Y, Wang Z (2017) Amplification of overlapping DNA amplicons in a single-tube multiplex PCR fortargeted next-generation sequencing of BRCA1 and BRCA2. PLoS One. 2017;12(7):e0181062. Epub 2017 Jul 12 (PMID:28704513)

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Walsh T, Casadei S, Coats KH, Swisher E, Stray SM, Higgins J, Roach KC, Mandell J, Lee MK, Ciernikova S, Foretova L,Soucek P, King MC (2006) Spectrum of mutations in BRCA1, BRCA2, CHEK2, and TP53 in families at high risk of breastcancer. JAMA. 2006 Mar 22;295(12):1379-88 (PMID: 16551709)

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Wooster R, Bignell G, Lancaster J, Swift S, Seal S, Mangion J, Collins N, Gregory S, Gumbs C, Micklem G (1995)Identification of the breast cancer susceptibility gene BRCA2. Nature. 1995 Dec 21-28;378(6559):789-92 (PMID: 8524414)

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