等位基因特异性PCR结合金磁微粒免疫层析检测膀胱癌患者尿液中TERT突变

Detection of TERT mutation in urine of patients with bladder cancer by allele specific PCR combined with gold magnetic particle immunochromatography

目的建立可检测人尿液DNA中端粒酶逆转录酶(TERT)启动子区域突变的方法。方法设计等位基因特异性PCR引物,利用温度梯度PCR确定等位基因特异性PCR程序,利用金磁微粒检测板进行扩增产物检测。收集34例膀胱癌患者的尿液,提取DNA,利用该文建立的等位基因特异性PCR结合金磁微粒免疫层析方法检测TERT启动子区域突变。结果建立了结合等位基因特异性PCR和金磁微粒免疫层析的TERT启动子区域C228T和C250T突变检测方法。34例膀胱癌患者尿液中8例患者存在C228T突变,1例患者存在C250T突变。结论该研究建立了TERT启动子区域C228T和C250T突变检测方法,可应用于膀胱癌患者尿液DNA突变检测。

Objective To establish a method for detection of telomerase reverse transcriptase(TERT)promoter region mutation in human urine DNA.Methods Allele specific PCR primers were designed,and the allele specific PCR program was determined by temperature gradient PCR.The amplified products were detected by gold magnetic particle detection plate.The urine samples of 34 patients with bladder cancer were collected and DNA was extracted.The mutation of TERT promoter region was detected by allele specific PCR combined with gold magnetic particle immunochromatography.Results A method for the detection of C228T and C250T mutations in TERT promoter region was established by combining allele specific PCR and gold magnetic particle immunochromatography.The results showed that 8 patients had C228T mutation and 1 patient had C250T mutation.Conclusion In this study,we established a method for detecting C228T and C250T mutations in TERT promoter region,which can be used to detect DNA mutations in urine of patients with bladder cancer.