CAJ | 학술논문

目的探讨眼脑肾综合征一家系的OCRL基因致病突变筛查与检测。方法采集家系成员外周血及胎儿羊水并提取基因组DNA。在OCRL基因两侧选取微卫星标记进行连锁分析；对患者OCRL基因全部外显子进行PCR扩增和测序；对可能致病突变采用限制酶分析验证。利用连锁分析、直接测序及酶切分析，对胎儿进行产前基因检测。结果先证者及3名肯定携带者OCRL基因存在无义突变c.2032C--→T(p.R678X)；其他7名家系成员及胎儿均不携带此突变。结论 OCRL基因c.2032C→T突变是该家系出现眼脑肾综合征的原因；家系中胎儿不携带此致病突变。
목적 탐토안뇌신종합정일가계적OCRL기인치병돌변사사여검측。방법 채집가계성원외주혈급태인양수병제취기인조DNA。재OCRL기인량측선취미위성표기진행련쇄분석；대환자OCRL기인전부외현자진행PCR확증화측서；대가능치병돌변채용한제매분석험증。이용련쇄분석、직접측서급매절분석，대태인진행산전기인검측。결과 선증자급3명긍정휴대자OCRL기인존재무의돌변c.2032C--→T(p.R678X)；기타7명가계성원급태인균불휴대차돌변。결론 OCRL기인c.2032C→T돌변시해가계출현안뇌신종합정적원인；가계중태인불휴대차치병돌변。
Objective To identify the pathogenic mutation underlying Lowe syndrome in a Chinese family. Methods After obtaining written informed consent of all participating individuals, peripheral blood samples were collected from 11 family members, including one affected male and three obligate female carriers, and an amniotic fluid sample was obtained from a pregnant carrier female in the family. Genomic DNA was extracted using the standard SDS-proteinase K-phenoL/chloroform method. Linkage analysis was carried out using microsatellite markers flanking the OCRL gene. All OCRL exons and their flanking intronic sequences were PCR-amplified and sequenced for the proband. Restriction analysis was performed to confirm the pathogenic mutation. Prenatal genetic testing was carried out by combining haplotype analysis, DNA sequencing and restriction analysis. ResultsLinkage analysis showed that the affected male and three obligate carrier females shared a same haplotype. Sequence analysis in the proband revealed a nonsence mutation c. 2032C→T (p. R678X) in exon 18 of the OCRL gene. Restriction analysis showed that the mutation was present in the proband and three obligate carriers, but not in the other 7 available family members. This nonsense mutation was not found in the amniotic fluid sample. Conclusion A recurrent OCRL nonsence mutation was found to be the pathogenic mutation in the Chinese family and the fetus didn't carry the mutation.