Establishment of a molecular diagnostic system for spinal muscular atrophy experience from a clinical laboratory in...

Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration of the anterior horn of the spinal cord. The disease gene survival motor neuron 1 (SMN1) is homozygously absent in approximately 95% of patients, and approximately 5% of patients are believed to have subtle mutations. Although methods for molecular diagnosis of SMA have been reported singly, no diagnostic methodological system to tackle different SMA cases has been reported. Thirty-two families affected by SMA enrolled into this study. Our system comprised PCR-restriction fragment length polymorphism and allele-specific PCR for homozygous deletion analysis of SMN1, multiplex ligation-dependent probe amplification analysis for the determination of the copy number of SMN1, and SMN1 subtle mutation analysis at both the transcript and genomic levels. In 23 families, 21 patients had a homozygous deletion of SMN1. The remaining two patients without a deletion had a single SMN1 copy containing the subtle mutations S230L and L228X, respectively. In nine families in whom samples from the index patients were unavailable, parents from eight families showed one SMN1 copy, and one parent in the remaining family showed two SMN1 copies, one being normal and the other carrying the subtle mutation 22_23insA. To our knowledge, our methodological system for the molecular diagnosis of SMA offers the most complete evaluation of family members affected by SMA at this time.