运用多重连接探针检测DMD  

Detection of DMD by Multiplex Ligation-depenclent Probe

作　　者：王小竹[1] 姚凤霞[1,2] 卢天兰[1,3] Nanbert Zhong 

机构地区：[1]北京大学医学遗传系,100083 [2]中国医学科学院基础医学研究所,中国协和医科大学医学遗传学系 [3]北京大学第六医院生化实验室 [4]北京大学医学遗传学系 [5]Department of Human Genetics,New York State Institute for Basic Research in Development Disabilities

出　　处：《医学研究杂志》2006年第8期98-100,共3页Journal of Medical Research

摘　　要：目的杜氏肌营养不良是是由于抗肌萎缩蛋白的缺失、重复及点突变所致的一种X-连锁隐性遗传性神经肌肉病。目前运用多重PCR检测此基因热点区可以检测大部分病人的缺失突变,然而多重PCR不能检测非热点区及重复突变,且不能定量分析拷贝数。运用多重连接探针扩增(multiplexligation-dependentprobeamplification,MLPA)检测DMD可为临床诊断及产前诊断提供依据。方法本实验采用的多重连接探针扩增(MLPA)能快速、准确、半定量地分析患者及携带者缺失与重复突变的拷贝数,且检测范围涉及整个基因。结果15例DMD患者9例是由于缺失突变所致,6例未检测到缺失突变及重复突变。其中7例缺失突变病人经“一步到位法”多重PCR验证,2例缺失突变病人经多重PCR未检测到的缺失突变。结论MLPA是一种快速、准确、简便检测缺失及重复突变的方法。利用MLPA能检测所有DMD患者及携带者的缺失和重复突变。Objective Duchenne muscular dystrophy （DMD） is one of the most common X - linked recessive neuromuscular degeneration diseases. It is caused by genetic defects of dystropin gene with deletion, duplication, or point mutation that results in clinical muscle fatigue and dystrophy. Usually, gene deletion of one or a few exons of dystrophin accounts for about 55% - 65% patients, duplication for about 5% - 10% patients and point mutation for 25%. Most of hot - spot deletion mutation of DMD can be detected by multiplex PCR and the point mutation can be detected by PCR/sequencing analysis, however, it remains a challenge to detect duplication. The recently developed MLPA （multiplex ligation -dependent probe amplification） is an efficient procedure that can accurately analyze the copy number and deletion mutation of whole dystropin gene. Methods A validation for simultaneous detection of entire dystropin gene was performed with two reactions. Both of which detect 39 and half cxons of dystrophin gene. Results Nine out of 15 patients with DMD were found to have deletion mutation in different exons of dystrophin gene. Among these 9 patients, 7 were found having deletion previously with multiplex PCR for mutation of hot - spot by Peking Union Medical University. Two patients who had not been found deletion by multiplex PCR were shown to have rare deletion at exon 18 or 43 in this study. Conclusions MLPA provides a simple, rapid and accurate method of simultaneously detecting homozygous, heterozygous deletions and duplication mutation in two single reactions for all exons of dystrophin gene, which may be applied into clinical molecular analysis for DMD.

关 键 词：DMD 基因缺失 多重连接探针扩增 

分 类 号：R516.8[医药卫生—内科学] R472[医药卫生—临床医学]