The invention discloses primers, a
reagent kit and a method for detecting IDH1 (
isocitrate dehydrogenase 1) R132H
gene variation by the aid of ddPCR (droplet
digital polymerase chain reaction) technologies. The method includes extracting cfDNA (
cell-free deoxyribonucleic acid) in
peripheral blood; designing two probes and a pair of primers for IDH1R132H
mutant types and wild types; carrying out ddPCR amplification by the aid of the cfDNA used as a template; carrying out
data analysis on obtained amplification products to obtain absolute quantities and proportions of IDH1R132H
mutant genes. Thereagent kit for detecting the IDH1R132H
gene variation is based on the method. The primers, the
reagent kit and the method have the advantages that provided primer amplification fragments 83bp are particularly applicable to amplifying small-fragment
DNA samples such as
plasma free DNA and can be combined with the detection probes, accordingly, the IDH1R132H
gene variation with extremely low abundance can be detected from the cfDNA, the primers, the
reagent kit and the method are good in specificity and high in sensitivity, human IDH1R132H gene variation can be absolutely quantitatively detected by the aid of the primers, the reagent kit and the method, and important effects can be realized by the primers, the reagent kit and the method for guiding
clinical treatment and improving patientprognosis; the IDH1R132H gene variation can be detected only by the aid of the
peripheral blood without
tissue specimen collection, and accordingly the method is particularly suitable for patients intolerant to
tissue sampling.