焦磷酸测序检测分析BRAF基因突变方法的建立

Pyro-sequencing analysis for the detection of BRAF gene mutation

目的建立鼠类肉瘤病毒癌基因同源物B1(BRAF)基因V600E(T>A)突变位点的焦磷酸测序(PYRO)方法。方法提取100例结直肠癌患者的gDNA,采用Qigene pyro24测序仪进行BRAF基因的PYRO,并通过重复性试验和Sanger测序法验证方法的正确性。结果建立了BRAF基因V600E突变位点的PYRO方法,经重复性试验和Sanger测序验证,结果准确可靠,100例结直肠癌患者的标本中共检测出6例有基因突变。结论建立的BRAF基因突变的PYRO方法具有快速、简便、灵敏度和准确性高的优点,适用于科研和临床批量检测。

Objective To establish a pyro-sequencing analysis(PYRO)for the detection of V-raf murine sarcoma viral oncogene homolog B1(BRAF)gene mutation.Methods The gDNA was extracred from100colorectal cancer patients'tissue samples.PYRO BRAF for gene was performed through Qigene pyro24.The accuracy was validated by repeat test and Sanger sequencing.Results The PYRO for the detection of BRAF gene mutation was established.Repeat test and Sanger sequencing showed that the accuracy and reliability were good.The detected gene mutation in100colorectal cancer patients'tissue samples accounted for6%.Conclusions The established PYRO for the detection of BRAF gene mutation is rapid,simple,sensitive and accurate.