变性高效液相色谱法筛查间隙连接蛋白2耳聋基因突变

Screening for GJB2 deafness gene mutations by denaturing high performance liquid chromatograph

目的 探讨变性高效液相色谱法 (DHPLC)在耳聋基因筛查中的应用价值。方法 应用DHPLC检测 6个家系的 4 8名个体 ,然后进行直接测序。结果 6个家系中A ,B ,C ,E ,F家系有间隙连接蛋白 2 (GJB2 )耳聋基因的变化 ,F家系表现为 2 35delC突变 ,B家系表现为 2 35delC和 2 32G→A杂合性突变。 79G→A(V2 7I)、341A→G (E1 1 4G)杂合性改变分别为 1 9例和 1 6例 ;纯合性改变分别为 1 0例和 6例。 2种方法的检测结果均为阳性者 30例 ,均为阴性者 1 0例 ;DHPLC阳性 ,测序阴性 1例 ;DHPLC阴性 ,测序阳性 2例。DHPLC方法的敏感性为 95 % ,特异性为 91 %。结论 DHPLC是一种高效、经济、简便。

Objective To investigate the mutations of GJB2 genes by DHPLC Methods Forty eight samples in six pedigrees were detected by both DHPLC and direct sequencing Results There were A,B,C,E,F pedigrees with the nucleotide changes in 6 pedigrees The 235delC was found in pedigree F, heterozygous mutations with 235delC and 232G→A in pedigree B The heterozygous mutations with 79G→A(V27I), 341A→G(E114G) were found in 19 cases and 16 cases, respectively The homozygous mutations were found in 10 cases and 6 cases, respectively The positive results detected by the both methods were 30 and the negative results were 10 The results, which were positive by DHPLC, but negative by sequencing, was found only one The results of negative by DHPLC , but positive by sequencing were found two These results showed the sensitivity was 95% and specificity was 91% for DHPLC detection of GJB2 mutations Conclusion DHPLC was a high sensitive and specific method for screening sequence variation in GJB2 gene, which was time and labor efficient