Abstract

Spinal muscular atrophy (SMA) is a common autosomal recessive disorder which has been considered as the second common cause of infant death, with an estimated prevalence of 1 in 10,000 live births. The disorder is caused by survival motor neuron 1 gene (SMN1) deficiency leading to limb weakness, difficult swallowing and abnormal breathing. Here, a fast and accurate method for SMA detection has been developed. Genomic DNA sample collected from whole blood, amniotic fluid, or dried blood spots can be analysed by using the Clarity™ Digital PCR (dPCR) System for determining the copy numbers of SMN1 and SMN2 genes. Two hundred and fourteen clinical samples determined by qPCR-based method were enrolled and used to establish the cut-off ranges for unaffected individual, SMA carrier and SMA patient categories. After setting the cut-off range for each group, 12 samples were analyzed by both dPCR-based method and MLPA (multiplex ligation-dependent probe amplification), the current testing golden standard for SMA, and 100% concordant results between the two testing methods were performed. CSB SMA Detection Kit combined with dPCR platform provides a robust and precise approach to distinguish unaffected individuals, SMA carrier and SMA patients. This rapid molecular diagnostic method can be adapted to pre-pregnancy eugenics inspection, prenatal testing as well as newborns screening and help physicians or genetic counselors to improve population SMA incidence.

摘要

脊髓性肌萎缩症（SMA）是一种常见的常染色体隐性遗传疾病，已被认为是婴儿死亡的第二大常见原因，估计每10,000名活产婴儿中有1名患病。该疾病是由存活运动神经元1基因（SMN1）缺乏引起的，导致肢体无力，吞咽困难和呼吸异常。在此，已开发出一种快速，准确的SMA检测方法。可以使用Clarity™数字PCR（dPCR）系统分析从全血，羊水或干血斑收集的基因组DNA样品，以确定SMN1和SMN2的拷贝数基因。纳入了基于qPCR的方法确定的214个临床样品，并用于确定未受影响的个体，SMA携带者和SMA患者类别的截断范围。设定每组的截止范围后，通过基于dPCR的方法和MLPA（多重连接依赖性探针扩增），SMA的当前检测黄金标准以及两种检测方法之间100％一致的结果进行分析，对12个样品进行了分析。被执行。CSB SMA检测套件与dPCR平台相结合，提供了一种强大而精确的方法来区分未受影响的个体，SMA携带者和SMA患者。这种快速的分子诊断方法可适用于孕前优生学检查，产前检查以及新生儿筛查，并帮助医生或遗传咨询师改善人群SMA的发生率。