Detection of isocitrate dehydrogenase 1 mutation with MGB dual fluorescent probe
Han Song
Lu Yu
Dong Tao
Yu Chunyong
Li Xiaoming
Cao Peng
Pan Dongsheng
Feng Sizhe
Abstract：Objective To establish a rapid minor groove binder (MGB) dual fluorescent probe method for the detection of the single nucleotide polymorphism (SNP) of isocitrate dehydrogenase 1 (IDH1) gene, and determine the sensitivity of the method and the consistency with the direct sequencing method. Methods The wild-type (R132) and mutant (H132) plasmids were constructed and used to optimize the MGB dual fluorescent probe method. The detection sensitivity of the IDH1 gene mutation was determined using the known proportion of wild-type and mutant plasmids as templates. The genotype DNA of 56 patients with glioma was analyzed by direct sequencing and MGB dual fluorescent probe respectively. The IDH1 gene mutation type was identified and compared. Results IDH1 mutation can be detected by MGB dual fluorescent probe method quickly and accurately. The positivity rate of mutation was 26.79％in direct sequencing and 30.36％ in MGB dual fluorescent probe method. Kappa test showed that the results of the two methods were consistent (P<0.001, K =0.956). Conclusions MGB dual fluorescent probe detection can detect the IDH1 gene mutation of glioma genomic DNA sensitively, reliably and rapidly, thus being suitable for clinical rapid molecular diagnostic analysis.
Keywords：gliomaisocitrate dehydrogenase 1MGB dual fluorescent probegene mutation
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 369-372 )