应用多重PCR-变性高效液相色谱技术检测大片段重复或缺失突变  

Denaturing high-performance liquid chromatography coupled with multiplex PCR for rapid detection of large duplications or deletions in patients with Duchenne muscular dystrophy and spinal muscular atrophy

作　　者：邹海强[1] 赵保健[3] 闫瑾[3] 韩炜[2] 熊美华[2] 彭凯润[1] 

机构地区：[1]广州军区广州总医院神经内科,510010 [2]广州军区广州总医院预防保健科,510010 [3]中国人民解放军空军航空医学研究所附属医院分子病理中心

出　　处：《中华医学遗传学杂志》2012年第6期686-689,共4页Chinese Journal of Medical Genetics

摘　　要：目的探讨多重PCR-变性高效液相色谱（PCR-denature high performance liquid chromatography，PCR—DHPLC）技术在假肥大型肌营养不良症和脊肌萎缩症外显子拷贝数异常检测中的应用价值。方法应用多重PCR-DHPLC方法筛查35例假肥大型肌营养不良症患者和6例脊肌萎缩症患者。选择阳性对照和阴性对照进行方法学可靠性检验。结果多重PCR-DHPLC可完全检测出全部阳性对照中重复或缺失片段。35例假肥大型肌营养不良症样本中检测出5例大片段重复，20例大片段缺失，突变检出率为71．4％。6例脊肌萎缩症患者均检测到第7外显子缺失。结论多重PCR-DHPLC分析系统可作为有效筛查大片段重复或缺失突变的检测方法。Objective To assess the value of multiplex PCR-denaturing high-performance liquid chromatography （PCR-DHPLC） method for screening large duplications or deletions in patients with Duchenne muscular dystrophy （DMD） and spinal muscular atrophy （SMA）. Methods DNA was extracted from peripheral venous blood samples from 35 DMD and 6 SMA patients. Large duplications or deletions were screened with multiplex PCR coupled with DHPLC method. The results were validated with testing of positive and negative controls. Results Known duplications or deletions in all controls were reliably detected with multiple PCR coupled with DHPLC. Large duplications or deletions were found in 71.4% of 35 DMD patients, which included 5 large duplications and 20 large deletions. For SMA patients, deletions of SMN1 exon 7 were detected in 16 samples. Conclusion Multiplex PCR coupled with DHPLC method is an effective and reliable method for detecting large genomic duplications or：deletions in patients with DMD or SMA.

关 键 词：变性高效液相色谱 多重聚合酶链反应 大片段重复或缺失突变 

分 类 号：R440[医药卫生—诊断学]