两个中国常染色体显性遗传性多囊肾病家系的表型与基因型分析

An analysis for the phenotype and genotype of autosomal dominant polycystic kidney disease from two Chinese families

目的研究中国人多囊肾病基因1(polycystic kidney disease1gene,PKD1)突变的特点,检测基因突变位点。方法25例多囊肾患者,正常对照16名,扩增PKD1基因的第44、45外显子的基因片段,变性梯度凝胶电泳突变检测系统进行初筛,然后测序。结果发现1个移码突变(12431delCT)、1个无义突变(C12217T)、1个多态性(A50747C),突变检测率为8%(2/25)。结论检测到2个新的可能的致病突变1个移码突变(12431delCT)、1个无义突变(C12217T)。

Objective To scan for mutations of polycystic kidney disease 1 gene （PKD1） in Chinese population in order to find some features about Chinese patients and a better approach to detect mutations. Methods Twenty-five PKD-affected individuals from twenty-one unrelated genealogies and sixteen controls participated in the study, Thirty-five blood samples and six tissues were obtained after receiving informed consent and were in accordance with institutional ethical guidelines. Genomic DNA was isolated from ripheral blood using standard procedures. PCR amplification of genomic DNA was performed to generate the aimed fragments. Amplified fragments were analyzed by denaturing gradient gel electrophoresis （DGGE）. A GC clamp was attached to the 5＇ primer. After that, the abnormal fragments were sequenced on freshly amplified specific PCR products with the dideoxynucleotide chain temtination method. Sequencing was performed for all samples to evaluate DGGE. Results Aimed fragments of exons 44 and 45 were amplified. DGGE detected eleven abnormal PCR fragments. Two novel mutations were identified by sequencing, included one nonsense mutation （C12217T） and one fxameshift （12431delCT）. In addition, one polymorphism （A50747C） was identified. The mutation detection rate is 8% in our study. Conclusion Two novel pathogenic mutations were detected, including one nonsense mutation （C12217T） and one frameshift （ 12431delCF）.