Analysis of BRCA2 loss of heterozygosity in tumor tissue using droplet digital polymerase chain reaction

作者：Cochran Rory L; Cravero Karen; Chu David; Erlanger Bracha; Toro Patricia Valda; Beaver Julia A; Zabransky Daniel J; Wong Hong Yuen; Cidado Justin; Croessmann Sarah; Parsons Heather A; Kim Minsoo; Wheelan Sarah J; Argani Pedram; Park Ben Ho^*

来源：Human Pathology, 2014, 45(7): 1546-1550.

DOI：10.1016/j.humpath.2014.03.013

• Droplet digital PCR
• LOH
• Familial breast cancer
• BRCA2
• VUS

Loss-of-heterozygosity (LOH) analysis of archival tumor tissue can aid in determining the clinical significance of BRCA variants. Here we describe an approach for assessing LOH in formalin-fixed, paraffin-embedded (FFPE) tissues using variant-specific probes and droplet digital polymerase chain reaction (ddPCR). We evaluated LOH in 2 related breast cancer patients harboring a rare missense BRCA2 variant of unknown clinical significance (c.6966G>T; M2322I). Conventional PCR followed by Sanger sequencing suggested a change in allelic abundance in the FFPE specimens. However, we found no evidence of LOH as determined by the allelic ratio (wild type variant) for BRCA2 in both patients' archival tumor specimens and adjacent normal control tissues using ddPCR. In summary, these experiments demonstrate the utility of ddPCR to quickly and accurately assess LOH in archival FFPE tumor tissue.

• 出版日期2014-7