摘要目的 对一个中国汉族残毁性掌跖角化病家系进行GJB2基因突变检测,以明确其致病基因.方法 收集该家系5例患者、4例正常人和100例非该家系成员外周血和家系患者临床资料.提取基因组DNA,PCR扩增包括GJB2基因整个编码序列在内的1015 bp,扩增产物纯化后应用ABI PRISM3730XL自动测序仪直接双向测序,Sequencher 4.10.1Demo软件与基因组序列进行比对分析,查找有无突变基因.结果 该家系所有患者GJB2基因均存在一个杂合错义突变196G→C,导致第一细胞外区域(El)第66位天冬氨酸被组氨酸替代(即D66H),而家系中4例正常人和100例非家系成员正常人对照的DNA测序结果均未发现此突变.结论 GJB2基因中D66H错义突变是中国汉族人群中残毁性掌跖角化伴耳聋的致病原因之一.

abstractsObjective To detect the mutation of GJB2 gene in a Chinese family with Vohwinkel syndrome.Methods Clinical data were collected from 5 patients with Vohwinkel syndrome in a family,and blood samples were obtained from the 5 patients and 4 unaffected individuals in the family as well as from 100 normal human controls.Genomic DNA was extracted and subjected to PCR for the amplification of the entire encoding and flanking sequences of GJB2 gene (1015 bp) followed by bidirectional sequencing with the ABI PRISM 3730 automatic DNA sequencer.Finally,sequence alignment was carried out by using the software Sequencher 4.10.1 Demo.Results A heterozygous missense mutation 196G→C in the GJB2 gene,which resulted in the substitution of aspartic acid by histidine at codon 66 (D66H) in the first extracellular domain of the protein,was observed in all the patients of this family,but in none of the 4 unaffected individuals in this family or the 100 normal human controls.Conclusion The D66H missense mutation in the GJB2 gene may contribute to the occurrence of Vohwinkel syndrome in Chinese Han population.