Droplet digital PCR for large genomic rearrangements detection: A promising strategy in tissue BRCA1 testing

Elisa De Paolis, Maria De Bonis, Paola Concolino, Angelo Piermattei, Anna Fagotti, Andrea Urbani, Giovanni Scambia, Angelo Minucci, Ettore Capoluongo, Ettore Domenico Capoluongo

Research output: Contribution to journalArticle

1 Citation (Scopus)


Background and aims: With the introduction of Olaparib as target therapy for High Grade Serous Ovarian Cancer (HGSOC) patients with germline and somatic BRCA1/2 mutations, the genetic test performed on tumor tissue has become important like the germline test. In somatic testing the evaluation of Large Genomic Rearrangements (LGRs) represents the main challenge. We describe a droplet digital PCR (ddPCR) assay for the evaluation of target BRCA1 LGRs on blood and formalin-fixed paraffin-embedded (FFPE)/Fresh Frozen Tissue (FFT) samples. Materials and methods: We analyzed blood, FFPE and FFT samples in a validation setting of n = 78 HGSOC patients. We applied the ddPCR to BRCA1 exons 2, 20 and 21 as some of the most common BRCA1 exons involved in LGRs in our cohort of patients. Results: The ddPCR custom assays allowed the identification of LGRs in all sample types, including FFPE specimens. Moreover, we were able to clearly detect LGRs accounted as somatic event. Conclusion: The introduction of ddPCR in a comprehensive workflow, encompassing both germline and somatic tests, represents an improvement in BRCA1/2 testing. ddPCR can overcome challenges related to BRCA testing, especially on FFPE analysis. Finally, ddPCR represents a promising alternative strategy to the established standard methods currently used in clinical setting.
Original languageEnglish
Pages (from-to)17-24
Number of pages8
JournalClinica Chimica Acta
Publication statusPublished - 2021


  • BRCA
  • Droplet digital PCR
  • High grade serous ovarian cancer
  • Large genomic rearrangement


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