Evaluation of droplet digital PCR and next generation sequencing for characterizing DNA reference material for KRAS mutation detection.

ABSTRACT: KRAS gene mutations are predictive markers of non-response to anti-epidermal growth factor receptor. An increasing number of techniques are being developed to detect KRAS mutations. To obtain consistent and comparable results, a traceable reference material (RM) is necessary for validation the routinely used method. However, a lack of reference methods is a main impediment for deriving traceability and measurement comparability. In this study, droplet digital PCR (ddPCR) and next generation sequencing (NGS) were evaluated. No cross- reactivity was detected with any of the probe by ddPCR. The measured fraction of KRAS mutant allele by ddPCR and NGS agreed with the prepared value by gravimetrical dilution (concordance (k) >0.95 and >0.93 for ddPCR and NGS, respectively). The reliable limit o

SUBMITTER: Dong L 

PROVIDER: S-EPMC6269532 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature