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Mutation analysis in spinal muscular atrophy using allele-specific polymerase chain reaction

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Title Mutation analysis in spinal muscular atrophy using allele-specific polymerase chain reaction
 
Creator Kesari, Akanchha
Mukherjee, Monisha
Mittal, Balraj
 
Subject spinal muscular atrophy
allele-specific amplification
DNA diagnosis
prenatal diagnosis
SMN gene
mutation analysis
polymerase chain reaction
 
Description 439-441
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.
 
Date 2009-04-13T11:47:00Z
2009-04-13T11:47:00Z
2003-12
 
Type Article
 
Identifier 0301-1208
http://hdl.handle.net/123456789/3822
 
Language en_US
 
Publisher CSIR
 
Source IJBB Vol.40(6) [December 2003]