单链构象多态性分析和变性高效液相色谱分析技术检测hMLH1和hMSH2基因突变的比较  

Comparison of the sensibility and specificity between single-stranded conformation polymorphism and denaturing high-performance liquid chromatography in screening hMSH2 and hMLH1 gene mutations in hereditary non-polyposis colorectal cancer

作　　者：魏广辉[1] 赵博[1] 王振军[1] 

机构地区：[1]首都医科大学附属北京朝阳医院外科,100020

出　　处：《中华胃肠外科杂志》2008年第5期462-464,共3页Chinese Journal of Gastrointestinal Surgery

基　　金：国家自然科学基金（39970817）

摘　　要：目的比较单链构象多态性分析（SSCP）和变性高效液相色谱分析技术（DHPLC）应用于遗传性非息肉病性结直肠癌（HNPCC）分子遗传学诊断的灵敏性和特异性。方法应用PCR-SSCP和DHPLC方法分析经测序验证有序列变异的7个家系的所有成员的相应PCR扩增产物，分别计算SSCP和DHPLC的灵敏度和特异度。结果应用SSCP对hMLH1和hMSH2进行基因突变筛查的敏感性和特异性分别为51．6％和66．6％；而应用DHPLC进行基因突变筛查的敏感性和特异性则分别为100％和93．3％；两种方法进行基因突变筛查的结果差异有统计学意义（P〈0．05）。结论DHPLC进行基因突变筛查的敏感性和特异性均高于SSCP，是理想的分子遗传学检测方法。Objective To compare the sensibility and specificity between single-stranded conformation polymorphism （SSCP） and denaturing high-performance liquid chromatography （DHPLC） in screening hMSH2 and hMLH1 gene mutations for the diagnosis of hereditary non-polyposis coloreetal cancer （HNPCC）. Methods Seven Chinese HNPCC kindreds were collected. PCR-SSCP and DHPLC were used to screen the coding regions of hMSH2 and hMLH1 genes and the abnormal profiles were sequenced by a 377 DNA sequencer. Results Seven gene sequence variations of hMSH2 or hMLH1 were found. Among them, 4 variations were not found by SSCP, but by DHPLC. The sensibility of SSCP and DHPLC were 51.6% and 100% respectively, and the specificity were 66.6% and 93.3% respectively. Conclusion DHPLC has better sensibility and specificity in screening hMSH2 and hMLH1 gene mutation as compared to SSCP. DHPLC is an ideal method in the diagnosis of HNPCC.

关 键 词：变性高效液相色谱分析 多态现象 单链构象 MLH1 MSH2 突变分析 结直肠肿瘤 遗传性非息肉性 

分 类 号：R735.3[医药卫生—肿瘤]