荧光定量PCR筛查脊髓性肌萎缩症基因携带孕妇的效果观察

Effect of quantitative fluorescent PCR on screening pregnant women carrying spinal muscular atrophy gene

摘要

目的 观察荧光定量聚合酶链式反应(PCR)筛查脊髓性肌萎缩症(SMA)基因携带孕妇的效果。方法 回顾性分析2022年10月至2023年04月来我院自行做SMA筛查的600例孕妇的外周血标本,采用多重连接探针扩增技术(MLPA)和荧光定量PCR检测孕妇SMA基因携带情况,以MLPA技术检测结果为金标准,分析荧光定量PCR检测准确性。结果 MLPA分析检测显示:600例孕妇中,携带者共60例,其有50例运动神经元存活基因1(SMN1)第7号外显子和第8号外显子基因拷贝数均为1,9例仅SMN1第8号外显子基因拷贝数为1,1例仅SMN1第7号外显子基因拷贝数为1,540例SMN1第7号外显子和第8号外显子基因拷贝数均为2。荧光定量PCR检测结果显示:携带者共60例,其中SMN1第7、8号外显子均杂合缺失共50例,仅第七号外显子缺失1例,仅第8号外显子杂合缺失9例,第7、8号外显子均未缺失共540例。荧光定量PCR检测的灵敏度为100.00%,特异度为100.00%,准确率为100.00%,Kappa值为1。结论 实时荧光定量PCR与MLPA技术筛查SMA基因携带孕妇筛查效果一致且实时荧光定量PCR具有便捷和高效等优势,值得临床推广应用。

Abstract

Objective To evaluate the effectiveness of quantitative fluorescent polymerase chain reaction (PCR) in screening pregnant women for spinal muscular atrophy (SMA) gene carrier status. Methods A retrospective analysis was conducted on peripheral blood samples from 600 pregnant women who voluntarily underwent SMA screening at our hospital between October 2022 and April 2023. Multiplex ligation-dependent probe amplification (MLPA) and quantitative fluorescent PCR were used to detect SMA gene carrier status. MLPA served as the gold standard to assess the accuracy of quantitative fluorescent PCR. Results MLPA analysis identified 60 carriers among the 600 pregnant women, comprising 50 cases with 1 gene copy of both SMN1 exon 7 and exon 8, 9 cases with only SMN1 exon 8 heterozygous deletion, and 1 case with only SMN1 exon 7 heterozygous deletion, while 540 cases showed 2 gene copies of both exon 7 and exon 8 of SMN1. Quantitative fluorescent PCR indicated that there were 60 carriers, of which 50 cases were missing heterozygosity in exon 7 and exon 8 of SMN1, 1 case was missing heterozygosity in exon 8, 9 cases were missing heterozygosity in exon 8, and 540 cases were not missing exon 7 and exon 8. The sensitivity, specificity, accuracy rate and Kappa value of quantitative fluorescent PCR were 100.00%, 100.00%, 100.00% and 1 respectively. Conclusion The study confirmed that quantitative fluorescent PCR provides screening performance identical to MLPA for SMA carrier detection in pregnant women. With its additional advantages of operational efficiency and clinical convenience, quantitative fluorescent PCR emerges as a highly suitable alternative for routine screening applications in prenatal care settings.

关键词

孕妇 / 脊髓性肌萎缩症 / 荧光定量聚合酶链式反应 / 基因

Key words

pregnant women / spinal muscular atrophy / quantitative fluorescent polymerase chain reaction / gene

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基金资助

东莞市社会科技发展项目(20231800904412)