Improved detection of BRAF V600E using allele-specific PCR coupled with external and internal controllers.

ABSTRACT: Although traditional allele-specific PCR (tAS-PCR) is a common screening method for BRAF V600E mutations, its lower amplification specificity and mutation selectivity have limited its clinical applications. We hypothesize that these limitations are associated with the weaker specificities of allele-specific primers and the thermodynamic driving forces of DNA polymerase. We used three strategies to circumvent these limitations, namely, modifying allele-specific primers, introducing a competitive external allele-specific controller (i.e., cAS-PCR), and introducing a referenced internal positive controller in the cAS-PCR (i.e., rcAS-PCR). The amplification sensitivities and specificities were influenced by the position of the artificially introduced mismatched nucleotide in the allele-specifi

SUBMITTER: Yang Z 

PROVIDER: S-EPMC5653796 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature