Multiplex ddPCR assay for screening copy number variations in BRCA1 gene

被引：44

作者：

Oscorbin, Igor ^{[1
,2
]}

Kechin, Andrey ^{[1
,2
]}

Boyarskikh, Uljana ^{[1
]}

Filipenko, Maxim ^{[1
,2
]}

机构：

[1] RAS, SB, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia

[2] Novosibirsk State Univ, Novosibirsk 630090, Russia

来源：

BREAST CANCER RESEARCH AND TREATMENT | 2019年 / 178卷 / 03期

关键词：

BRCA1; Digital PCR; Gene rearrangements; CNV; Multiplex amplification; DROPLET DIGITAL PCR; 22Q11.2 DELETION SYNDROME; SOMATIC MUTATIONS; OVARIAN-CANCER; DNA; SYSTEM; BREAST; CELL; QUANTIFICATION; REARRANGEMENTS;

D O I：

10.1007/s10549-019-05425-3

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Purpose Germinal and somatic rearrangements in BRCA1 gene play a significant role in carcinogenesis of breast and ovarian cancer. The present study is dedicated to the development of multiplex droplet digital PCR (ddPCR) assay for detecting large deletions and duplications in the BRCA1 gene. Methods In-house tetraplex ddPCR assay for BRCA1 gene analysis was used for testing of DNA samples with BRCA1 status. Results DNA specimens were purified from 24 individuals. The presence of BRCA1 rearrangements in samples was confirmed by a commercial MLPA-based kit. An amplitude-based multiplex ddPCR assay was developed: 8 multiplexes, each containing primers and probes to amplify 3 BRCA1 exons and 1 reference gene (ALB or RPP30). A novel assay demonstrated 100% concordance with the commercial MLPA-based kit, identifying 9 specimens with different deletions in BRCA1, 1 with duplication, and 14 with the wild-type BRCA1. Conclusions We have designed a simple, precise, and cost-effective assay for BRCA1 rearrangement testing, based on ddPCR. The developed assay is the first multiplex ddPCR-based test that provides results in accordance with MLPA and can be used for routine clinical screening.

引用

页码：545 / 555

页数：11

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