Evaluation and comparison of three assays for molecular detection of spinal muscular atrophy

Abstract

AbstractBackground:Spinal muscular atrophy (SMA) is mainly caused by deletions in SMA-related genes. The objective of this study was to develop gene-dosage assays for diagnosing SMA.Methods:A multiplex, quantitative PCR assay and a CNVplex assay were developed for determining the copy number ofResults:The multiplex quantitative PCR (qPCR) assay had higher reproducibility. Intra-assay CVs were 3.01%–8.52% and inter-assay CVs were 4.12%–6.24%. The CNVplex assay had ratios that were closer to expected (0.49–0.5 for one copy, 1.03–1.0 for two copies, and 1.50–1.50 for three copies). Diagnostic accuracy rates for the two assays were 100%.Conclusions:The multiplex qPCR assay was a simple, rapid, and cost-effective method for routine SMA diagnosis and carrier screening. The CNVplex assay could be used to detect SMAs with complicated gene structures. The assays were reliable and could be used as alternative methods for clinical diagnosis of SMA.