连接酶链反应结合荧光PCR法检测UGT1A1*28基因多态性

目的: 建立一种连接酶链反应结合荧光PCR的方法，检测结肠癌患者外周血UGT1A1*28基因多态性，并探讨其临床应用价值。方法: 设计连接酶荧光PCR引物和探针，首先通过连接酶反应，然后结合荧光PCR技术检测UGT1A1*28基因多态性，构建标准品评判该体系的特异性和敏感性，并检测50例结肠癌患者外周血标本，与DNA测序进行平行比较。结果： 此方法能准确地区分UGT1A1*28基因TA6/6、TA7/7及TA6/7基因型，且最低可检测5 ng人基因组DNA， 50例结肠癌患者外周血标本的检测结果与测序结果一致。 结论： 连接酶链反应结合荧光PCR可准确检测出UGT1A1*28基因多态性。

Detection of UGT1A1*28 gene polymorphisms by ligase chain reaction-PCR

Objective: To establish a detection method of UGT1A1*28 gene polymorphisms based on ligase chain reaction and fluorescent PCR(LCR PCR) and to evaluate efficacy of its clinical application. Methods: LCR PCR primers and probes were designed to detect the UGT1A1*28 gene polymorphisms, constructed standard products to evaluate sensitivity and specificity of this method. Peripheral blood samples of 50 colorectal cancer patients were detected and compared with data obtained by DNA sequencing.Results: The LCR PCR technique could be used to detect the TA6/6,TA7/7 and TA6/7 of UGT1A1*28 gene, the detection limit is 5 ng genome DNA, and the results of 50 clinical samples detected by this method were in full agreement with those of the DNA sequencing. Conclusion: LCR PCR method can accurately detect UGT1A1*28 gene polymorphisms.