Rapid diagnosis of spinal muscular atrophy using High-Resolution Melting Analysis

Abstract

Background: Spinal muscular atrophy (SMA) is an autosomal recessive hereditary disorder caused by mutations of the survival motor neuron 1 (SMN1) gene. Recently, high-resolution DNA melting analysis (HRMA) with saturation LC Green dyes has become a powerful post-PCR technique for genotyping or mutation scanning. So far, no studies have applied HRMA to the molecular analysis of SMA.

Diagnosis of Spinal Muscular Atrophy Via High-Resolution Melting Analysis Symmetric Polymerase Chain Reaction Without Probe: A Screening Evaluation forSMN1Deletions and Intragenic Mutations

“…The difficulty in distinguishing patients lacking SMN1, from normal controls lacking SMN2, was one of the major factors attributing to the authors' final assessment contending the apparent inferiority of symmetric PCR-HRMA (Chen et al, 2009). In contrast, in the present study, we demonstrate that symmetric PCR-HRMA-with a simple change of primer sets-is an able and dexterous method which allows for distinguishing patients lacking SMN1 from normal controls, via discernible differences in melting curve patterns.…”

“…Recently, Chen et al (2009) evaluated the application of HRMA to screenings for deletion of SMN1 exon 7, and based on their results, the authors conclude that asymmetric PCR-HRMA is superior to symmetric PCR-HRMA. The difficulty in distinguishing patients lacking SMN1, from normal controls lacking SMN2, was one of the major factors attributing to the authors' final assessment contending the apparent inferiority of symmetric PCR-HRMA (Chen et al, 2009).…”

Newborn screening for spinal muscular atrophy: Anticipating an imminent need

“…4 High-resolution DNA melting analysis (HRMA) HRMA has been used to evaluate individuals for SMA in screening and diagnostic applications. 3,119,120 The difference in SMN1 and SMN2 copy number present in the specimens will create various unique HRMA profiles, distinguishing a heterozygous PCR product from a homozygous product. Therefore, the SMN1 homozygous deletions can be readily identified since these samples will generate the same profile independent of SMN2 copy number.…”

Copy Number Assessment by Competitive PCR with Limiting Deoxynucleotide Triphosphates and High-Resolution Melting

“…However, a single copy at each locus (carrier) cannot be distinguished from 2 copies at each locus (normal). The same concern occurs in the other melting method that uses an unlabeled probe (35 ). We used allele-specific duplex PCR with limiting dNTPs to independently assess the copy number of each gene.…”