A multiple detection method for distinguishing gene mutations based on melting curves of extended quenching probes.

ABSTRACT: Conventional PCR methods can detect only a few targets simultaneously and do not fulfill most clinical requirements, especially those for detecting plasma circulating DNA. By designing characteristic universal fluorescent probes, combining multiplex PCR with the invasive reaction, and analyzing the resulting differences in the melting curves formed by extension with double-stranded probes, we developed a new method to distinguish between three mutations in the same fluorescent channel and nine mutations in three fluorescent channels in a single tube. After optimization, this method was used to distinguish between 27 mutations using only three reactions, and mutations representing as low as 0.2%-0.5% of DNA could be detected, even when up to nine mutations were present at the same time. Tes

SUBMITTER: Jianping W 

PROVIDER: S-EPMC9712562 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature