CAJ | 학술논문

目的：探讨连接片段在缺失型Duchenne型肌营养不良症（Duchenne muscular dystrophy, DMD）携带者检测中的应用价值。方法对1个DMD家系和1例DMD散发病例进行研究。DMD家系中以确诊的患者Ⅲ2和待查携带者Ⅱ3为研究对象；散发病例以患者Ⅱ1及其母亲Ⅰ2为研究对象。通过外显子检测确定家系中患者Ⅲ2第31~43外显子缺失，散发病例患者Ⅱ1第45~54外显子缺失。然后以PCR步移法在相应内含子设计引物定位断裂点的位点，最后在尽量靠近断裂连接点处设计1对引物直接对患者及其待查女性亲属进行连接片段的PCR扩增。结果对上述DMD家系中待查女性Ⅱ3的检测结果为PCR扩增出与患者Ⅲ2一致的阳性片段，诊断其为DMD携带者。对散发病例患者母亲Ⅰ2的检测结果为PCR反应阴性，而对照的患者Ⅱ1扩增出阳性结果，排除其母亲为DMD携带者。结论利用常规PCR技术直接检测缺失型DMD患者的女性亲属是否带有连接片段，可以同样达到进行携带者检测的目的。该方法有别于目前DMD携带者检测中所使用的各种定量分析方法。
목적：탐토련접편단재결실형Duchenne형기영양불량증（Duchenne muscular dystrophy, DMD）휴대자검측중적응용개치。방법대1개DMD가계화1례DMD산발병례진행연구。DMD가계중이학진적환자Ⅲ2화대사휴대자Ⅱ3위연구대상；산발병례이환자Ⅱ1급기모친Ⅰ2위연구대상。통과외현자검측학정가계중환자Ⅲ2제31~43외현자결실，산발병례환자Ⅱ1제45~54외현자결실。연후이PCR보이법재상응내함자설계인물정위단렬점적위점，최후재진량고근단렬련접점처설계1대인물직접대환자급기대사녀성친속진행련접편단적PCR확증。결과대상술DMD가계중대사녀성Ⅱ3적검측결과위PCR확증출여환자Ⅲ2일치적양성편단，진단기위DMD휴대자。대산발병례환자모친Ⅰ2적검측결과위PCR반응음성，이대조적환자Ⅱ1확증출양성결과，배제기모친위DMD휴대자。결론이용상규PCR기술직접검측결실형DMD환자적녀성친속시부대유련접편단，가이동양체도진행휴대자검측적목적。해방법유별우목전DMD휴대자검측중소사용적각충정량분석방법。
Objective To investigate the value of the junction fragments between the breakpoints of introns in identifying deletional Duchenne muscular dystrophy (DMD) carriers. Methods A DMD family (including the index patient III2 and the suspected carrier II3) and a sporadic DMD case (including the patient II1 and his mother I2) were studied. The patient III2 of the DMD family was identified as having exons 31-43 deletion of the DMD gene, and the sporadic patient II1 had exons 45-54 deletion. A PCR-based genome-walking method was used to locate the breakpoints in the corresponding introns. The junction fragments of the patients and their female relatives waiting for a diagnosis were amplified by PCR with primers adjacent to the deletion junctions. Results PCR amplification yielded identical positive results for the female suspected carrier II3 of and the index patient of the DMD family, and the former was thus diagnosed as a carrier of DMD. PCR amplification of the sporadic patient’s mother I2 showed a negative result, but the patient II1 had a positive result, so that the patient's mother was excluded as being a carrier of DMD. Conclusion Routine PCR technique for detecting the junction fragments allows identification of carriers among female relatives of patients with deletional DMD.