CAJ | 학술논문

目的评价基因组测序技术在诊断缺失型脊肌萎缩症(spinal muscular atrophy,SMA)中的应用.方法应用聚合酶链反应扩增100例SMA临床确诊患儿和110名对照样本的运动神经元存活基因(spinal muscular atrophy,SMN),基因组测序技术通过识别扩增片段中SMN1和SMN2的4个差异位点用来诊断SMN1的纯合缺失,并以多重连接探针扩增进行验证.结果 100例SMA样本中基因组测序显示差异位点仅有SMN2基因特有碱基峰,多重连接探针扩增检测示SMN1与SMN2拷贝数为0∶2或0∶3,表明SMN1基因纯合缺失.对照组中5例样本的差异位点仅为SMN1基因特有碱基峰,SMN1与SMN2拷贝数为2∶0,为SMN2纯合缺失.105例样本的差异位点均为杂合峰,SMN1与SMN2拷贝数为2∶2,未显示SMN1和SMN2的缺失.所有测序结果与MLPA检测结果一致.结论基因组测序技术在进行缺失型脊肌萎缩症的分子诊断中具有特异、准确、实用的特点.
목적 평개기인조측서기술재진단결실형척기위축증(spinal muscular atrophy,SMA)중적응용.방법 응용취합매련반응확증100례SMA림상학진환인화110명대조양본적운동신경원존활기인(spinal muscular atrophy,SMN),기인조측서기술통과식별확증편단중SMN1화SMN2적4개차이위점용래진단SMN1적순합결실,병이다중련접탐침확증진행험증.결과 100례SMA양본중기인조측서현시차이위점부유SMN2기인특유감기봉,다중련접탐침확증검측시SMN1여SMN2고패수위0∶2혹0∶3,표명SMN1기인순합결실.대조조중5례양본적차이위점부위SMN1기인특유감기봉,SMN1여SMN2고패수위2∶0,위SMN2순합결실.105례양본적차이위점균위잡합봉,SMN1여SMN2고패수위2∶2,미현시SMN1화SMN2적결실.소유측서결과여MLPA검측결과일치.결론 기인조측서기술재진행결실형척기위축증적분자진단중구유특이、준학、실용적특점.
Objective To detect homozygous deletions of survival motor neuron (SMN) gene with genomic DNA sequencing,and to assess the value of genetic testing for the diagnosis of spinal muscular atrophy (SMA).Methods Polymerase chain reaction (PCR) was used for amplifying SMN gene in 100 SMA patients and 110 controls.Four different bases (g.31957,g.32006,g.32154 and g.32269) between SMN1 and SMN2 within the amplified segments were identified with genomic DNA sequencing.Homozygous deletion of SMN1 or SMN2 was determined by the presence or absence of base peaks at such four sites.Multiplex ligation-dependent probe amplification (MLPA) was carried out to confirm the results of genomic DNA sequencing.Results In the 100 SMA samples,only SMN2-specific base peaks were detected at the four sites,for which the copy numbers of SMN1 and SMN2 was 0 ∶ 2 or 0 ∶ 3,suggesting homozygous deletion of SMN1 gene.By contrast,only SMN1-specific base peaks were detected in 5 samples,for which the ratio of SMN1 ∶ SMN2 was 2 ∶ 0,indicating homozygous deletion of SMN2.At four different sites,SMN1 /SMN2 heterozygous peaks were detected in the remaining 105 samples,for which SMN1 ∶ SMN2 was 2 ∶ 2,suggesting non-deletion of SMN1 or SMN2.The results of sequencing were consistent with those of MLPA.Conclusion Genomic DNA sequencing is a rapid,accurate and economic method for the diagnosis of homozygous deletion of SMA.