A multiplex allele specific PCR capillary electrophoresis (mASPCR-CE) assay for simultaneously analysis of SMN1/SMN2/NAIP copy number and SMN1 loss-of-function variantsYunli Laion August 18, 2025 at 10:00 am

Clin Chem Lab Med. 2025 Aug 19. doi: 10.1515/cclm-2025-0175. Online ahead of print.

ABSTRACT

OBJECTIVES: Spinal muscular atrophy (SMA) is a severe inherited neuromuscular disorder with a high carrier frequency and incidence rate. An accurate molecular method for SMA genes is crucial in carrier screening, clinical diagnosis, outcome assessment and precision therapies.

METHODS: Comprehensively using the multiplex allele specific PCR (mASPCR) and capillary electrophoresis (CE), a novel single tube assay was developed to simultaneously determine the copy number of SMN1/SMN2/NAIP genes and five common loss-of-function variants in SMN1. A total of 383 genotype known subjects were detected to evaluate the accuracy, while 564 clinical random samples were double-blind detected with this assay and MLPA to assess the specificity and sensitivity.

RESULTS: This assay had high accuracy of 100 % consistency with the predetermined values in 383 genotype known subjects. Among 564 clinical random samples, the correlation between this assay and comparative method was 100 %, which showing high specificity and sensitivity.

CONCLUSIONS: This mASPCR-CE assay is easy to use and cost-effective, making it suitable for routine use in molecular screening and clinical diagnosis of SMA.

PMID:40824197 | DOI:10.1515/cclm-2025-0175

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