A duplex allele-specific amplification PCR to detect SMN1 deletion.
Spinal muscular atrophy (SMA), the leading genetic cause of death in childhood, is an autosomal recessive neuromuscular disorder characterized by progressive muscle weakness, associated with deletions of the survival motor neuron (SMN) gene identified and mapped to chromosome 5q13. SMN is present in two highly homologous copies (SMN1 and SMN2). In the general population, normal individuals (noncarriers) have at least one telomeric (SMN1) copy, and 5% of them have no copies of SMN2. Approximately 95% of SMA patients carry homologous deletions of SMN1 exon(s) 7 (and 8). SMN1 and SMN2 exons 7 and 8 differ only by 1 bp each, and SMA diagnosis might be performed by single-strand conformational polymorphism, PCR amplification followed by restriction fragment length polymorphism (RFLP), multiple ligation-dependent probe amplification, or realtime PCR of SMNs exons 7 and 8. We developed a simpler and cost-effective method to detect SMN1 exon 7 deletion based on allele-specific amplification PCR.
Laboratório de Pediatria Clínica-Lim 36, Instituto da Criança-HCFMUSP, Rua Dr. Eneas de Carvalho Aguiar, São Paulo 05403-900, Brazil. patricia.pieri@icr.usp.br
Patrícia de Campos
Pde C
Pieri
Jeronimo de Alencar
Jde A
Nogueira
Maria Joaquina
MJ
Marques-Dias
Bernadete
B
Resende
Chong Ae
CA
Kim
Umbertina Conti
UC
Reed
Thelma Suely
TS
Okay
D000483:Alleles; D016022:Case-Control Studies; D017931:DNA Primers / Q000737:chemistry; D005091:Exons; D017353:Gene Deletion; D006801:Humans; D009134:Muscular Atrophy, Spinal / Q000175:diagnosis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016622:Silver Staining; D055533:Survival of Motor Neuron 1 Protein / Q000235:genetics*
D017931:DNA Primers; D055533:Survival of Motor Neuron 1 Protein