Stekrova Jitka, Reiterova Jana, Svobodova Stanislava, Kebrdlova Vera, Lnenicka Petr, Merta Miroslav, Viklicky Ondrej, Kohoutova Milada
Institute of Biology and Medical Genetics of the 1st Faculty of Medicine and General Teaching Hospital, Charles University, Prague, Czech Republic.
BMC Med Genet. 2009 Aug 17;10:78. doi: 10.1186/1471-2350-10-78.
Autosomal dominant polycystic kidney disease (ADPKD) is the most common hereditary renal disease. The disease is caused by mutations of the PKD1 (affecting roughly 85% of ADPKD patients) and PKD2 (affecting roughly 14% of ADPKD patients) genes, although in several ADPKD families, the PKD1 and/or PKD2 linkage was not found. Mutation analysis of the PKD1 gene is complicated by the presence of highly homologous genomic duplications of the first two thirds of the gene.
The direct detection of mutations in the non-duplicated region of the PKD1 gene was performed in 90 unrelated individuals, consisting of 58 patients with end-stage renal failure (manifesting before their 50th year of life) and 32 individuals from families where the disease was clearly linked to the PKD1 gene. Mutation screening was performed using denaturing gradient gel electrophoresis (DGGE). DNA fragments showing an aberrant electrophoretic banding pattern were sequenced.
In the non-duplicated region of the PKD1 gene, 19 different likely pathogenic germline sequence changes were identified in 19 unrelated families/individuals. Fifteen likely pathogenic sequence changes are unique for the Czech population. The following probable mutations were identified: 9 nonsense mutations, 6 likely pathogenic missense mutations, 2 frameshifting mutations, one in-frame deletion and probable splice site mutation. In the non-duplicated region of the PKD1 gene, 16 different polymorphisms or unclassified variants were detected.
Twenty probable mutations of the PKD1 gene in 90 Czech individuals (fifteen new probable mutations) were detected. The establishment of localization and the type of causal mutations and their genotype phenotype correlation in ADPKD families will improve DNA diagnosis and could help in the assessment of the clinical prognosis of ADPKD patients.
常染色体显性遗传性多囊肾病（ADPKD）是最常见的遗传性肾病。该疾病由PKD1基因（约85%的ADPKD患者受其影响）和PKD2基因（约14%的ADPKD患者受其影响）突变引起，不过在一些ADPKD家族中，未发现PKD1和/或PKD2基因的连锁关系。PKD1基因的突变分析因该基因前三分之二区域存在高度同源的基因组重复而变得复杂。
对90名无亲缘关系的个体进行了PKD1基因非重复区域突变的直接检测，其中包括58名终末期肾衰竭患者（发病年龄在50岁之前）和32名来自疾病与PKD1基因明确连锁家族的个体。采用变性梯度凝胶电泳（DGGE）进行突变筛查。对显示异常电泳条带模式的DNA片段进行测序。
在PKD1基因的非重复区域，在19个无亲缘关系的家族/个体中鉴定出19种不同的可能致病的种系序列变化。15种可能致病的序列变化在捷克人群中是独特的。鉴定出以下可能的突变：9个无义突变、6个可能致病的错义突变、2个移码突变、1个框内缺失和可能的剪接位点突变。在PKD1基因的非重复区域，检测到16种不同的多态性或未分类变异。
在90名捷克个体中检测到20种PKD1基因可能的突变（15种新的可能突变）。确定ADPKD家族中致病突变的定位、类型及其基因型与表型的相关性，将改善DNA诊断，并有助于评估ADPKD患者的临床预后。
