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目的:PKD2基因在人群中以多种等位基因的形式存在和遗传,其中某些变异型等位基因所编码的多囊蛋白2功能发生改变或缺失,从而引起常染色体显性遗传多囊肾病(ADPKD)的发生。检测PKD2基因的变异情况具有重要的临床诊断意义。方法:针对文献报道的中国人群中检测发现的PKD2变异位点,设计检测探针及引物,并制备基因芯片,构建了基因片段的质粒作为标准模板。结果:通过实验优化,建立了样品荧光标记、基因芯片杂交与检测的条件及分型标准。通过对标准质粒模板和人外周血提取的基因组DNA模板的检测,验证了基因芯片检测结果的重复性和准确性。结论:本实验制备的基因芯片可以准确区分PKD2基因中的9个突变位点的变异情况。
PURPOSE: PKD2 gene exists and inherits in the form of multiple alleles in human population. The function of polycystin 2 encoded by some variant alleles is changed or absent, resulting in autosomal dominant polycystic kidney disease (ADPKD) occurred. Detection of PKD2 gene mutation has important clinical significance. Methods: According to the reported detection of PKD2 locus in Chinese population, the detection probes and primers were designed and the gene chip was constructed. The plasmid of gene fragment was constructed as a standard template. Results: Through the experiment optimization, the conditions of fluorescence labeling and gene chip hybridization and detection and the typing criteria were established. Through the detection of the standard plasmid template and the genomic DNA template extracted from human peripheral blood, the repeatability and accuracy of the gene chip test results were verified. Conclusion: The gene chip prepared in this experiment can accurately distinguish the variation of 9 mutations in PKD2 gene.