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Title: Mutation analysis in spinal muscular atrophy using allele-specific polymerase chain reaction
Authors: Kesari, Akanchha
Mukherjee, Monisha
Mittal, Balraj
Keywords: spinal muscular atrophy;allele-specific amplification;DNA diagnosis;prenatal diagnosis;SMN gene;mutation analysis;polymerase chain reaction
Issue Date: Dec-2003
Publisher: CSIR
Abstract: Polymerase chain reaction (PCR), followed by restriction digestion is universally used for molecular diagnosis of spinal muscular atrophy (SMA). In the present study, we have used a modified strategy based on amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) to develop a rapid and reliable method for mutation detection and prenatal diagnosis in SMA patients. The telomeric (SMN1) and centromeric (SMN2) copies of exon 7 of the survival motor neuron (SMN) gene were amplified by ARMS-PCR, using primers specific to SMN1 and SMN2 nucleotide sequence with the exonic mismatch G (for SMN1) and A (for SMN2) at the 3’ end. The PCR products were analyzed on agarose gels. All the patients who had homozygous deletion of exon 7 of SMN1 gene by conventional PCR-restriction fragment length polymorphism (PCR-RFLP) method showed the same deletion status by ARMS-PCR. This procedure showed a 100% concordance between PCR-RFLP and ARMS-PCR methods for the detection of SMN1/SMN2 status in patients with SMA. An artifact due to incomplete digestion is not a problem while using ARMS-PCR. The modified protocol is specific, rapid and highly reliable for use in prenatal diagnosis as well.
Page(s): 439-441
ISSN: 0301-1208
Appears in Collections:IJBB Vol.40(6) [December 2003]

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