Rapid detection of GSTM1 polymorphisms using SYBR green I and melting curve analysis / 临床检验杂志

ABSTRACT

Objective To establish a method to detect the glutathione stransferase(GST) M1 gene polymorphisms rapidly and to apply preliminary for detection.Methods GSTM1 gene and internal control gene CYP1A1 were amplified by multi-PCR with appropriate fluorescent dye SYBR green I in the reaction system,The changes of fluorescence values were recorded from 65 ℃ to 95 ℃ by a rate of(0.1 ℃/s) when PCR was just completed.Results There were two peaks in the melting curve of GSTM1~+ genotype,but only a single peak occurred for the GSTM1-genotype.The temperatures of two peaks corresponded to the expected Tm.agarose gel electrophoresis analysis demonstrated that these peaks correspond to the bands of the predicted molecular size.The detection can be completed within an hour after DNA was extracted.Conclusion Melting curve analysis combined with SYBR green I is a easy,rapid,accurate methodfor detection of GSTM1 gene polymorphisms.