中国南方汉族散发性激素耐药型肾病综合征儿童NPHS1基因突变分析

Mutational analysis of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome in Southern Chinese Han ethnic group

目的 研究我国南方汉族散发性激素耐药型肾病综合征（SRNS）儿童NPHS1基 因突变情况.方法 对象为40例南方汉族散发性SRNS儿童,50例尿检正常的汉族健康成年人作为对照.取受检者外周静脉血3ml,10％枸橼酸钠抗凝,提取基因组DNA.应用PCR方法扩增NPHS1基因全部29个外显子及其周围的部分内含子,对PCR产物直接进行DNA序列测定及突变分析.结果 6例存在NPHS1基因突变-928G ＞A （D310N）、2677A ＞G（ T893A）、2869G＞C（ V957L）、IVS8＋30C＞T、IVS21＋ 14G＞A、IVS25-23C ＞T和＊142T＞C,突变检出率为15％.在50例健康对照人群中未检测出这7个变异,其中2677A ＞G、IVS8 ＋30C ＞T、IVS21＋ 14G＞A、IVS25-23C＞T和＊142T＞C为新发现NPHS1基因突变.此外,还确定了13个已报道的NPHS1基因多态性-294C ＞T、349G ＞A、IVS3＋ 15C ＞T、IVS3 ＋61A ＞G、803G ＞A、IVS8＋68A ＞G、1339G ＞A、1802G ＞C、2223C ＞T、2289C ＞T、IVS24 ＋36C ＞T、3315G＞A和IVS27 ＋45C ＞T.结论 在40例南方汉族散发性SRNS患儿中,NPHS1基因突变检出率为15％,提示南方汉族散发性SRNS患儿存在NPHS1基因突变,对这类患儿需进行NPHS1基因突变分析.

Objective To elucidate the mutations of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome （SRNS） in Southern Chinese Han ethnic group.Methods Peripheral blood samples were collected for genetic analysis from 40 patients with sporadic SRNS and 50 healthy volunteers as control.Genomic DNA was isolated from peripheral blood leucocytes.Twenty-nine exons and exon-intron boundaries of the NPHS1 gene were amplified by polymerase chain reaction.Mutational analysis was performed by DNA sequencing directly.Results Seven variants,928G〉A（D310N）,2677A〉G （T893A）,2869G〉C （V957L）,IVS8＋30C〉T,IVS21＋14G〉A,IVS25-23C〉T and ＊142T〉C,of NPHS1 gene were found in 6 of 40 children with sporadic SRNS,whereas they were not found in 50 healthy controls.2677A 〉G,IVS8 ＋30C 〉T,IVS21 ＋14G〉A,IVS25-23C 〉T and ＊142T〉C were novel.Moreover,thirteen already reported NPHS1 polymorphisms,294C〉T,349G〉A,IVS3＋15C〉T,IVS3＋61A〉G,803G〉A,IVS8＋68A〉G,1339G 〉A,1802G 〉C,2223C 〉T,2289C 〉T,IVS24 ＋36C 〉T,3315G〉A and IVS27 ＋45C 〉T,were detected in some patients and controls. Conclusions NPHS1 mutations in 6 of 40 children with sporadic SRNS in Southern Chinese Han ethnic group （15%） are detected.NPHS1 mutations are existed in Southern Chinese children,so it is necessary to perform the mutation analysis of NPHS1 gene in those children patients.