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

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

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1546-1550
Number of pages5
JournalHuman Pathology
Volume45
Issue number7
DOIs
StatePublished - 2014

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Loss of Heterozygosity
Polymerase Chain Reaction
Paraffin
Formaldehyde
Neoplasms
Breast Neoplasms

Keywords

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

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Analysis of BRCA2 loss of heterozygosity in tumor tissue using droplet digital polymerase chain reaction. / Cochran, Rory L.; Cravero, Karen; Chu, David; Erlanger, Bracha; Toro, Patricia Valda; Beaver, Julia; Zabransky, Daniel J.; Wong, Hong Yuen; Cidado, Justin; Croessmann, Sarah; Parsons, Heather A.; Kim, Minsoo; Wheelan, Sarah; Argani, Pedram; Park, Ben Ho.

In: Human Pathology, Vol. 45, No. 7, 2014, p. 1546-1550.

Research output: Contribution to journalArticle

Cochran, RL, Cravero, K, Chu, D, Erlanger, B, Toro, PV, Beaver, J, Zabransky, DJ, Wong, HY, Cidado, J, Croessmann, S, Parsons, HA, Kim, M, Wheelan, S, Argani, P & Park, BH 2014, 'Analysis of BRCA2 loss of heterozygosity in tumor tissue using droplet digital polymerase chain reaction', Human Pathology, vol. 45, no. 7, pp. 1546-1550. https://doi.org/10.1016/j.humpath.2014.03.013
Cochran, Rory L. ; Cravero, Karen ; Chu, David ; Erlanger, Bracha ; Toro, Patricia Valda ; Beaver, Julia ; Zabransky, Daniel J. ; Wong, Hong Yuen ; Cidado, Justin ; Croessmann, Sarah ; Parsons, Heather A. ; Kim, Minsoo ; Wheelan, Sarah ; Argani, Pedram ; Park, Ben Ho. / Analysis of BRCA2 loss of heterozygosity in tumor tissue using droplet digital polymerase chain reaction. In: Human Pathology. 2014 ; Vol. 45, No. 7. pp. 1546-1550.
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