Abstract:

Ｏｂｊｅｃｔｉｖｅ：Autosomal dominant polycystic kidney disease (ADPKD) is a common hereditary kidney disease,ADPKD is thought to be caused by mutations in the PKD1 and PKD2 genes,PKD1 is comprises 46 exons and coding region is about 13 kb,PKD2 has 15 exons and a coding region of about 3 kb. Traditional analysis methods are tedious and waste time. The purpose of the study was to establish a convenient and rapid detection method to detect the ADPKD mutation position and the type of mutation.
Ｍｅｔｈｏｄｏｌｏｇｙ：According to the clinical and laboratory examination were selected for the three unrelated Chinese Han ADPKD families. The probands were detected that using the next generation sequencing technology(NGS) for gene mutation analysis. The exons,splicing sites and 10 bp flanking intron sequences of PKD1 and PKD2 were captured by a gene chip. Variants(single nucleotide variants and small insertion) were identified using the GATK Genotyper.
Ｒｅｓｕｌｔｓ：The PKD sequence variants were checked by published in the Human Gene Mutation Database. NGS sequencing showed two novel frameshift mutations （c. 7318delG，p. Asp2440Thrfs*180）and nonsense mutations（c. 5884C>T，p. Gln1962*）in PKD1mutations,one nonsense mutations（c. 916C>T，p. Arg306*）has been reported in PKD2 gene mutation. PCR combined with Sanger sequence analysis confirmed the existence of the heterozygous mutations in the ADPKD families. The genotype and phenotype were cosegregation in the pedigree. Lead to PKD1 encoded amino acid shift code(p. Asp2440Thrfs*180) and nonsense mutations triggered PKD1(p. Gln1962*) and PKD2(p. Arg306*) protein in the early termination,respectively.
Ｃｏｎｃｌｕｓｉｏｎ：Using NGS technology,we established a rapid analysis of ADPKD genetic variation method,successfully applied to three families,and identified two novel mutations. NGS technology can be applied to the routine screening of suspected ADPKD and family members.