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Multiplex detection of rare mutations by picoliter droplet based digital PCR

Aug 29, 2016

In cancer research, the accuracy of the technology used for biomarkers detection is remarkably
important. In this context, digital PCR represents a highly sensitive and reproducible method that could serve as an appropriate tool for tumor mutational status analysis. In particular, droplet-based digital PCR approaches have been developed for detection of tumorspecific mutated alleles within plasmatic circulating DNA. Such an approach calls for the development and validation of a very significant quantity of assays, which can be extremely costly and time consuming. Herein, we evaluated assays for the detection and quantification of various mutations occurring in three genes often misregulated in cancers: the epidermal growth factor receptor (EGFR), the v-Ki-ras2 Kirsten rat sarcoma viral oncogene
homolog (KRAS) and the Tumoral protein p53 (TP53) genes. In particular, commercial competitive
allele-specific TaqMan1PCR (castPCR™) technology, as well as TaqMan1and ZEN™assays, have been evaluated for EGFR p.L858R, p.T790M, p.L861Q point mutations and in-frame deletions Del19. Specificity and sensitivity have been determined on cell lines DNA, plasmatic circulating DNA of lung cancer patients or Horizon diagnostics reference standards. To show the multiplexing capabilities of this technology, several multiplex panels for EGFR (several three- and four-plexes) have been developed, offering new"ready-to-use" tests for lung cancer patients.

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RainDance Technologies

RainDance company’s ultra-sensitive genomic tools enable research of novel non-invasive liquid biopsy applications that should result in more accurate, reliable, cost-effective, and early detection of cancer and other inherited and infectious diseases.

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Capable of generating more than a billion reactions in a single day, the RainDrop system surpasses all existing technologies and establishes a new performance standard in sensitivity, quantitation and multiplexing.

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