Mutational Analysis Of Gene In Fujian Province With Familial Hematuria

Part one: Mutational Analysis of COL4A5 in familial X-linked Alport syndrome in Fujian provinceObjective Alport syndrome(AS)is a hereditary nephritis characterized by hematuria,progressive renal failure,sensorineural deafness and ocular lesions.About 80% of patients with AS have the X-linked form of the disease.X-linked AS(XLAS)is caused by mutations in COL4A5 gene.The COL4A5 gene is large and has no hot spots mutation.It needs the high technical conditions of mutational analysis,so there is no report about mutational analysis of the COL4A5 gene in familial XLAS in Fujian province.This study aimed to examine mutations in the COL4A5 gene in familial XLAS in Fujian province.Methods Peripheral blood samples were collected for genetic analysis from the probands and parents of four families with XLAS in Fujian province.We studied as controls 63(26 males and 37 females)unrelated adult volunteers whose urinalyses were normal.The mutational analysis of COL4A5 was performed by polymerase chain reaction and direct DNA sequencing.Results Four COL4A5 variants,2768-11A>G,3925-1G>C,4688G>A and 5032T>C,were identified in the 3 prohands from 4 families with XLAS in Fujian province.One variant,3925-1G>C,was absent in 63 normal Chinese controls,wich is proved a COL4A5 mutation.It was a novel mutation in COL4A5.Three variants,2768-11A>G,4688G>A and 5032T>C,were found in NCBI db SNP;they are benign,pathogenic and pathogenic mutations,respectively.The rate of detecting mutations in COL4A5 is 75%(3/4)in familial XLAS in Fujian province.Conclusions The results demonstrate that COL4A5 mutations are a cause of familial XLAS in Fujian province.Our investigation supports the necessity of searching for mutations in COL4A5 in familial XLAS in Fujian province.Part two: Mutational analysis of causitive genes in familial hematuria in Fujian province using Haloplex target enrichment system kits for Hi Seq 2500 sequencingObjective Familial hematuria is caused by mutations in a group of genes that are COL4A1,COL4A3,COL4A4,COL4A5,COL4A6,MYH9 and CFHR5.Hematuria usually develops in early childhood and continues until late in life and until death,characterized by an unpredictable risk of progression to chronic kidney disease(CKD).There is no special treatment of patients with hematuria.The objective of this study was to design a next-generation sequencing(NGS)protocol enabling simultaneous detection of all possible variants in the seven genes.Methods Peripheral blood samples were collected for genetic analysis from 17 probands(7 males and 10 females)from families with hematuria in Fujian province.Age of onset is 1-18 years(average,4.35±4.14 years).We studied as controls 63(26 males and 37 females)unrelated adult volunteers whose urinalyses were normal.We designed haloplex target enrichment system kits for Hi Seq 2500 sequencing.Significant variants detected by NGS were confirmed by conventional Sanger sequencing.Results The seven genes,COL4A1,COL4A3,COL4A4,COL4A5,COL4A6,MYH9 and CFHR5,of 17 children with familial hematuria are sequencing using NGS.