变性高效液相色谱法筛选胶质瘤易感基因ERCC2的单核苷酸多态性

目的筛选和鉴定脑胶质瘤易感基因ERCC2——纠正鼠突变细胞切除修复缺陷基因的单核苷酸多态性（SNPs）。方法应用参考文献所报道的引物序列，PCR扩增179例胶质瘤患者肿瘤及血液标本和44例正常对照组血液标本ERCC2基因第22外显子及其邻近的部分内含子序列，采用DHPLC技术对扩增片段进行基因变异检测，将不同类型的PCR片段进行全序列测序并与参考序列对比分析。结果在此片段中验证了一个已知的SNP位点，初步鉴定了1个新的SNP位点，排除了1个已知在自种人存在的SNP位点和基因型。结论SNPs位点存在人种间差异；DHPLC预测结合全序列测序是一种高效、经济、简便、可靠的SNP筛选方法。

Screening single nucleotide polymorphisms in the predisposing gene-ERCC2 of gliomas by denaturing high-performance liquid chromatography

Objective To screen single nucleotide polymorphisims(SNPs)of ERCC2 gene at 19q13.3 by way of denaturing high-performance liquid chromatography(DHPLC).Methods Twenty-second exon and the adjacent part of the intron in the ERCC2 gene at 19q13.3 were isolated from the glioma and the blood samples of 179 patients with gliomas and 44 healthy control groups were amplified by PCR,and the products were analyzed by D1HPLC according to the primer sequence of the literature report.Some meaningful fragments were sequenced and compared with the sequences available from National Center for Biotechnology Information(NCBI) database.Results In this study,1 new SNP loci was preliminary identified and 1 known SNP loci and genotype was confirmed,moreover,1 known SNP loci and genotype,which occurred in white tumor patients,was eliminated.Conclusion SNPs loci has the race discrepancy.DHPLC is an effective,economical,and simple method with reliability for SNPs screening.