利用PCR-SSCP技术检测中国汉族人PKD2基因的突变

Detection of polycystic kidney disease gene 2 mutations in the Hans by PCR-SSCP

目的：检测中国汉族人Ⅱ型多囊肾病基因PKD2的突变。方法：筛选临床确诊的26个中国汉族家系中31例常染色体显性遗传性多囊肾病（ADPKD）患者，提取外周血白细胞DNA，应用聚合酶链反应-单链构象多态性分析（PCR－SSCP），取异常条带标本进行核苷酸序列测定，判别PKD2外显子突变位置及类型。结果：以20例健康志愿者为对照，从31例患者中成功检测出2种突变。1种为无义突变，系PKD2外显子13的第2407位碱基由胞嚼陡置换为胸腺嘧啶，形成1个终止密码子；另1种为错义突变，系PKD2外显子4的第964位碱基由胞嘧啶置换为胸腺嘧啶，使编码氨基酸由精氨酸变为色氨酸。结论：本研究建立了PCR－SSCP直接检测我国汉族人PKD2突变方法，检测出2种基因突变，为今后开展ADPKD患者囊肿前诊断提供了实验基础。

Objective: To investigate the mutations in polycystic kidney disease gene 2 (PKD2) in the Hans. Methods:The genomic DNA of the white blood cells in patients with autosomal dominant polycystic kidney disease(ADPKD) was iso-lated, amplified by PCR, and analyzed by SSCP, DNA Samples from abnormal bands were Sequenced. Results: Comparedwith 20 healthy individuals,two kinds of mutations were identified from 31 ADPKD patients. One nonsense mutation wasfound in PKD2 exon 13 (2407C→T) where a stop codon was formed. The other was a missense mutation in PKD2 exon 4(964C→T),which led to its abnormal translation(322Arg→Trp). Conclusion: The methods of direct detection of the muta-tions in the Hans PKD2 are developed and used successfully in identifying 2 kinds of mutations in the Hans. The methods usedin the study will be he1pful to diagnose the ADPKD patients in advance of cyst formation.