CAJ | 학술논문

目的对我国诺里病一家系进行致病基因突变的鉴定,确定其家系中是否存在NDP基因突变.方法基因家系研究.发现一个3代均患有诺里病的家系,共13人,患者3人,现存患者仅先证者1人,为20月龄男孩.根据诺里病家族史、临床体征及眼科B超检查对患者进行临床诊断；采集该家系成员外周血标本,常规提取基因组DNA;设计并合成3对特异性引物,通过PCR扩增NDP基因的外显子及外显子与内含子交界区,PCR产物直接测序；通过PCR产物限制性内切酶分析和基因型分析对家系所有13名成员进行突变鉴定.结果基因测序结果显示先证者及其母亲均具有NDP基因突变c.220C＞ T(p.Arg74Cys),且先证者为该突变的半合子,其母为该突变的携带者；NDP基因突变鉴定表明家系中另外4个表型正常的个体(Ⅲ3、Ⅳ4、Ⅲ5和Ⅱ2)均为该突变的携带者.结论我国诺里病一家系中发现存在NDP错义突变c.220C＞T.
목적 대아국낙리병일가계진행치병기인돌변적감정,학정기가계중시부존재NDP기인돌변.방법 기인가계연구.발현일개3대균환유낙리병적가계,공13인,환자3인,현존환자부선증자1인,위20월령남해.근거낙리병가족사、림상체정급안과B초검사대환자진행림상진단；채집해가계성원외주혈표본,상규제취기인조DNA;설계병합성3대특이성인물,통과PCR확증NDP기인적외현자급외현자여내함자교계구,PCR산물직접측서；통과PCR산물한제성내절매분석화기인형분석대가계소유13명성원진행돌변감정.결과 기인측서결과현시선증자급기모친균구유NDP기인돌변c.220C＞ T(p.Arg74Cys),차선증자위해돌변적반합자,기모위해돌변적휴대자；NDP기인돌변감정표명가계중령외4개표형정상적개체(Ⅲ3、Ⅳ4、Ⅲ5화Ⅱ2)균위해돌변적휴대자.결론 아국낙리병일가계중발현존재NDP착의돌변c.220C＞T.
Objective To detect the pathogenic mutation in a Chinese family with Norrie disease.Methods Clinical diagnosis was based on familial history,clinical sign and B ultrasonic examination.Peripheral blood samples were obtained from all available members in a Chinese family with Norrie disease.Genomic DNA was extracted from lymphocytes by the standard SDS-proteinase K-phenol/chloroform method.Two coding exons and all intron-exon boundaries of the NDP gene were PCR amplified using three pairs of primers and subjected to automatic DNA sequence.The causative mutation was confirmed by restriction enzyme analysis and genotyping analysis in all members.Results Sequence analysis of NDP gene revealed a missense mutation c.220C ＞ T (p.Arg74Cys ) in the proband and his mother. Further mutation identification by restriction enzyme analysis and genotyping analysis showed that the proband was homozygote of this mutation.His mother and other four unaffected members ( Ⅲ3,Ⅳ4,Ⅲ5 and Ⅱ2) were carriers of this mutation.The mutant amino acid located in the C-terminal cystine knot-like domain,which was critical motif for the structure and function of NDP. Conclusion A NDP missense mutation was identified in a Chinese family with Norrie disease.