Abstract：
OBJECTIVE: Locus and allellic heterogeneity in polycystic kidney disease (PKD) is a great challenge in precision diagnosis. We aim to establish comprehensive methods to distinguish the pathogenic mutations from the variations in PKD1, PKD2 and PKHD1 genes in a limited time and lay the foundation for precisely prenatal diagnosis, preimplantation genetic diagnosis and presymptom diagnosis of PKD.
METHODS: Nested PCR combined with direct DNA sequencing were used to screen variations in PKD1, PKD2 and PKHD1 genes. The pathogenicity of de novel variations was assessed by the comprehensive methods including clinic data and literature review, databases query, analysis of co-segregation of the variants with the disease, variant frequency screening in the population, evolution conservation comparison, protein structure analysis and splice sites predictions.
RESULTS: 17 novel mutations from 15 Chinese Han families were clarified including 10 mutations in PKD1 gene and 7 mutations in PKHD1 gene. The novel mutations were classified as 4 definite pathogenic, 2 highly likely pathogenic, 4 likely pathogenic, 7 indeterminate by the comprehensive analysis. The results were verified the truth by the follow-up visits.
CONCLUSIONS: The comprehensive methods may be useful in distinguishing the pathogenic mutations from the variations in PKD1, PKD2 and PKHD1 genes for prenatal diagnosis and presymptom diagnosis of PKD. Our results also enriched PKD genes mutation spectrum and evolved possible genotype-phenotype correlations of Chinese Han population.
METHODS: Nested PCR combined with direct DNA sequencing were used to screen variations in PKD1, PKD2 and PKHD1 genes. The pathogenicity of de novel variations was assessed by the comprehensive methods including clinic data and literature review, databases query, analysis of co-segregation of the variants with the disease, variant frequency screening in the population, evolution conservation comparison, protein structure analysis and splice sites predictions.
RESULTS: 17 novel mutations from 15 Chinese Han families were clarified including 10 mutations in PKD1 gene and 7 mutations in PKHD1 gene. The novel mutations were classified as 4 definite pathogenic, 2 highly likely pathogenic, 4 likely pathogenic, 7 indeterminate by the comprehensive analysis. The results were verified the truth by the follow-up visits.
CONCLUSIONS: The comprehensive methods may be useful in distinguishing the pathogenic mutations from the variations in PKD1, PKD2 and PKHD1 genes for prenatal diagnosis and presymptom diagnosis of PKD. Our results also enriched PKD genes mutation spectrum and evolved possible genotype-phenotype correlations of Chinese Han population.
摘要：
目的：多囊肾病（PKD）中的基因座和等位基因异质性是精确诊断的巨大挑战。我们的目标是在有限的时间内建立全面的方法来区分PKD1，PKD2和PKHD1基因的致病突变，并为精确的产前诊断奠定基础。PKD的胚胎植入前遗传学诊断和畸形前诊断。
方法：套式PCR结合直接DNA测序筛选PKD1、PKD2和PKHD1基因变异。通过包括临床数据和文献综述在内的综合方法评估了新变异的致病性。数据库查询,变异与疾病的共分离分析，在人群中进行变异频率筛查，进化守恒比较，蛋白质结构分析和剪接位点预测。
结果：阐明了来自15个中国汉族家庭的17个新突变，其中PKD1基因突变10个，PKHD1基因突变7个。新的突变被分类为4个明确的致病性，2极可能致病，4可能致病，7不确定由综合分析。通过后续访问验证了结果的真实性。
结论：综合方法可能有助于将致病性突变与PKD1，PKD2和PKHD1基因的变异区分开来，用于PKD的产前诊断和预诊断。我们的结果还丰富了PKD基因的突变谱，并进化了中国汉族人群可能的基因型-表型相关性。
方法：套式PCR结合直接DNA测序筛选PKD1、PKD2和PKHD1基因变异。通过包括临床数据和文献综述在内的综合方法评估了新变异的致病性。数据库查询,变异与疾病的共分离分析，在人群中进行变异频率筛查，进化守恒比较，蛋白质结构分析和剪接位点预测。
结果：阐明了来自15个中国汉族家庭的17个新突变，其中PKD1基因突变10个，PKHD1基因突变7个。新的突变被分类为4个明确的致病性，2极可能致病，4可能致病，7不确定由综合分析。通过后续访问验证了结果的真实性。
结论：综合方法可能有助于将致病性突变与PKD1，PKD2和PKHD1基因的变异区分开来，用于PKD的产前诊断和预诊断。我们的结果还丰富了PKD基因的突变谱，并进化了中国汉族人群可能的基因型-表型相关性。
