荧光定量PCR法应用于脊髓性肌萎缩症携带者筛查的研究

Study on the application of fluorescence quantitative PCR in screening carriers of spinal muscular atrophy

目的 通过对比TaqMan探针荧光定量PCR法与临床广泛应用的MLPA法对脊髓性肌萎缩症的检测效果,探索荧光定量PCR法用于人群致病基因携带者筛查的可行性。方法 收集2018年10月—2021年5月就诊于深圳市龙岗区妇幼保健院和广东省生殖医院的脊髓性肌萎缩症临床疑似病例、携带者及健康者的外周血标本共34例,应用TaqMan探针荧光定量PCR法进行检测,采用SMN1基因与参比序列相对定量确定SMN1基因拷贝数进行结果判读,以MLPA法作为本方法的参照体系进行双盲对照分析。结果 荧光定量PCR法可明确区分SMN1基因纯合缺失的患者、单拷贝数携带者及多拷贝数健康者的标本,与MLPA法检测结果全部相符。结论 荧光定量PCR法可对SMN1基因拷贝数进行相对定量检测,该方法可用于脊髓性肌萎缩症的人群携带者筛查。

Objective To compare the detection effect of TaqMan probe fluorescent quantitative PCR method and MLPA method on spinal muscular atrophy, and explore the feasibility of fluorescent quantitative PCR method for screening disease-causing gene carriers in the population. Methods Collected 34 cases of peripheral blood samples form clinical suspected cases of spinal muscular atrophy, carriers, and normal people from maternal and child health hospital of Shenzhen Longgang district and Guangdong provincial reproductive hospital from October 2018 to May 2021. The TaqMan probe fluorescence quantitative PCR method was used to detect the samples. The SMN1 gene and the reference sequence were used to determine the copy number of SMN1 gene relative and quantitatively for result interpretation, and the MLPA method was used as the reference system of this method for double-blind control analysis.Results Fluorescence quantitative PCR method could clearly distinguish specimens from patients with homozygous deletion of SMN1 gene, single copy number carriers, and multiple copy number normal people, the results of fluorescence quantitative PCR and MLPA method were all consistent. Conclusions The fluorescent quantitative PCR method could be used for the relative quantitative detection of SMN1 gene copy number. This method could be used for screening of SMA carriers in the population.