qPCR法检测脊髓性肌萎缩症基因的质控点初探

Topics of quality control of q PCR diagnostic reagent for gene detection of spinal muscular atrophy

脊髓性肌萎缩症(spinal muscular atrophy,SMA)是一种常染色体隐性遗传病,95%的SMA患者为运动神经元存活基因1(survival motor neuron 1,SMN1)外显子7和/或外显子8缺失。SMA基因检测有利于开展携带者筛查以判断受孕风险。荧光定量PCR法具有设备普及率高、速度快、通量大以及配套设施成熟等优势,但需对PCR扩增体系的各要素进行合理的质量控制。本文结合体外诊断试剂注册管理相关法规规章、技术审评要求,对用于SMA检测的荧光定量PCR产品的样本采集、核酸提取、引物/探针设计、扩增体系、适用机型等质量控制点进行讨论,以指导相关生产企业的研发和注册人员进行合理的产品设计开发及注册申报。

Spinal muscular atrophy(SMA)is an autosomal recessive hereditary disease,with which 95%of patients show exon 7 and/or exon 8 loss of SMN1 gene.The detection of SMA gene is beneficial to screening carriers in terms of pregnancy risk assessment.Fluorescence quantitative PCR(qPCR)method has the advantages of easy device access,fast speed,high-flux and mature facilities,but it needs a reasonable quality control on each element of PCR amplification system.In accordance with IVD registrations and technical review requirements,the topics of quality control of the qPCR diagnostic reagent for detection of SMA gene,including sample collection,nucleic acid extraction,primer/probe design,amplification system,applicable model,are discussed in this paper so as to enlighten R&D in relevant enterprises,optimize product design and guide RA to prepare product registration dossier.