多重PCR技术及定量PCR技术在缺失型Duchenne型肌营养不良患儿及携带者诊断中的应用研究  

Application of PCR technique for the diagnosis in DMD patients and carriers with the deletion of dystrophin gene.

作　　者：牛国辉[1] 麻宏伟[1] 姜俊[1] 吕峻峰[1] 卢瑶 王岳平[3] 

机构地区：[1]中国医科大学附属第二医院发育儿科,遗传研究室,沈阳110004 [2]中国医科大学实验设备处实验技术中心三部,110001 [3]中国医科大学附属第二医院中心实验室,110004

出　　处：《中国实用儿科杂志》2005年第4期224-226,共3页Chinese Journal of Practical Pediatrics

摘　　要：目的　建立一种简便、可靠的缺失型Duchenne型肌营养不良(DMD)患儿及携带者的基因检测方法,为DMD产前诊断的开展奠定基础。方法　首先采用1 8对引物多重PCR反应体系,确定2 0 0 1年3月至2 0 0 4年8月就诊于中国医科大学附属第二医院遗传门诊的48例DMD患儿dystrophin基因缺失情况,然后应用定量PCR方法对1 5例缺失型患儿母亲进行携带者检测。结果　48例DMD患儿中2 5例存在基因缺失,缺失率52 % ( 2 5/ 4 8) ,共涉及1 2个外显子,主要集中在44～52外显子,其中外显子49和50缺失频率最高;1 5例缺失型患儿母亲中检出8例携带者,其中肯定携带者检出率1 0 0 % ( 4 / 4 ) ,可疑携带者检出率3 6% ( 4 / 1 1 )。结论　多重PCR结合定量PCR方法能有效地检出缺失型DMD患儿及携带者,可为产前诊断提供十分重要的信息。Objective To establish a simple and reliable method to examine the detection of dystrophin gene in patients and carriers with Duchenne muscular dystrophy (DMD) to lay a foundation for prenatal diagnosis. Methods Multiplex polymerase chain reaction (mPCR) was used to detect the deletion of dystrophin gene in DMD patients; quantititative polymerase chain reaction (Q-PCR) was used to identify the carriers with the deletion of dystrophin gene. Results Different fragments of deletions were identified in 25 patients (52%), covering 12 of 79 exons of dystrophin gene and mainly distributed in exons 44～52, with most frequency in exon 49 and 50. 8 carriers were identified in 15 mothers whose sons had been confirmed the deletion of dystrophin gene. Positive rate was 100%(4/4) in obligate carriers and 36%(4/11) in possible carriers. Conclusion The combination of mPCR and Q-PCR can diagnose patients and carriers in DMD families with the deletion of dystrophin gene effectively, which may provide significent information for prenatal diagnosis.

关 键 词：肌营养不良 基因缺失 携带者 聚合酶链反应 

分 类 号：R72[医药卫生—儿科]