Establishment and evaluation of GSTM1 genotyping based upon high resolution melting curve
OBJECTIVE To detect the polymorphism of GSTM1 by high resolution melting(HRM)method and evaluate its accuracy to provide references for individualized dosing of antineoplastic and anti-tuberculosis drugs.METHODS Peripheral venous blood samples were collected from 94 hospitalized patients at three medical institutions from March 2019 to October 2019,including Sichuan Provincial People's Hospital.The inclusion criteria included an age range of(16-88)years,signing informed consent and receiving physical examination with retained blood samples.Genomic DNA was extracted from whole blood by cen-trifugal column method and the concentration and purity of extracted DNA were determined.HRM analysis was performed.Addi-tionally,33 samples were randomly selected from both GSTM1 null and GSTM1 non-null genotypes for verification through pyro-sequencing by Stata17 software.RESULTS DNA concentration in all 94 samples ranged 10-60 ng·μL-1 with a purity ratio of OD260/280 ratio(1.6-2.0).All samples underwent normal amplification reactions.Among them,GSTM1 non-null genotype was detected in 37 cases(39.36％)while GSTM1 null genotype in 57 cases(60.64％).And 33 randomly selected samples were verified by pyrophosphate sequencing.There were 17 cases(51.52％)of GSTM1 non-null genotype and 16 cases(48.48％)of GSTM1 null genotype.As compared with pyrophosphate sequencing,the results of HRM method were completely consistent.CONCLUSION This study has successfully established an experimental system and method for detecting GSTM1 by HRM method,which was used to detect two different genotypes of GSTM1 gene locus with high accuracy.It provided a rapid and accu-rate drug gene detection method for the optimization of drug regimen for patients with clinical tumors and tuberculosis.Moreover,it holded significant guiding implications for the clinical application of anti-tumor and anti-tuberculosis medications.
high resolution melting curveGSTM1drug genegenotyping
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