先天性软骨发育不全家系成纤维细胞生长因子受体3基因突变检测与分析  

Detecting and Analyzing on Mutation of Fibroblost Growth Factor Recepter 3 Gene in A Congenital Achondroplasia Family

作　　者：黄艳梅[1] 郭利伟[1] 李端[2] 祁英杰[1] 杨保胜[3] 

机构地区：[1]新乡医学院基础医学院法医物证学教研室,河南新乡453003 [2]新乡医学院生命科学技术系发酵工程教研室,河南新乡453003 [3]新乡医学院基础医学院,河南新乡453003

出　　处：《实用儿科临床杂志》2009年第20期1571-1573,共3页Journal of Applied Clinical Pediatrics

基　　金：河南省青年骨干教师项目资助(2007);新乡医学院博士启动基金项目资助(2005)

摘　　要：目的检测和分析一先天性软骨发育不全(ACH)家系6人成纤维细胞生长因子受体3(FGFR3)基因突变。方法根据知情同意原则采集该家系成员6人血样标本,并以2例健康人作对照。应用单链构象多态性-聚合酶链反应(PCR-SSCP)和限制性扩增片段长度多态性(PCR-RFLP)方法,检测该家系各成员FGFR3基因3跨膜区1138位核苷酸突变情况。结果FGFR3基因1138位核苷酸位点扩增产物SSCP方法显示,ACH家系中先证者及其父亲检测到单链泳动变位,且异常带型一致,家系中其他成员及健康人无单链泳动变位,进一步采用SfeⅠ酶切后,先证者及其父亲检测到164bp、109bp和55bp3条片段,均存在FGFR3基因1138位核苷酸G→A杂合性突变,而家系其他成员及2例健康人PCR产物酶解后只出现164bp片段。该位点又采用MspⅠ酶切,家系中所有成员及健康无关个体仅检测到一条带,大小为164bp,即未发现G→C颠换突变。结论FGFR3跨膜区1138位核苷酸G→A转换突变可能为该家系ACH的主要发病原因。在该家系中,先证者父亲表现为新发突变,然后又遗传给其子导致ACH。Objective To detect and analyze the mutation of fibroblast growth factor receptor 3 （ FGFR3 ） gene among a family with congenital aehondroplasia（ACH）. Methods Six blood samples of family member in this pedigree were cellected according to the informed consent process for genetic research,and 2 unralted healthy human blood sample were taken as controls. The mutation at nucleotide position 1 138 on FGFR3 gene was detected by using Polymerase chain reaction and single -strand conformation polymorphism（ PCR -SSCP）and polymerase chain reaction and restriction endonuelease technology （PCR - RFLP） methods. Results Using PCR - SSCP method firstly, only the proband with ACH and his father in this family had the same abnormal band. The amplified products including 1 138 loci on FGFR3 gene further was analyzed by Sfe I digestion, 3 fragments including 164 bp,t09 bp and 55 bp were detected in the proband and his father again,and the other members in the family and 2 controls just showed 164 bp hand. It indicated that just ：2 patients （ proband and his father） showed heterozygous G→A transition mutation at nucleotide position 1 138 on the FGFR3 gene. The amplified products at 1 138 loci was also detected by Msp I digestion, just 1 band was observed in all members in this family and 2 controls. It showed that there was no G→C substitution at nueleotide position 1 138. Conclusions The G→A transition mutation at nueleotide position 1 138 in transmembrane domain of FGFR3 gene may be the main cause of achondroplasia in this family. In this pedigree, the proband showed＇s father a de novo mutation which was transferred to his child again.

关 键 词：软骨发育不全 成纤维细胞生长因子受体3 基因突变 家系 

分 类 号：R726.8[医药卫生—儿科]