Evaluation of multiplex ligation dependent probe amplification as a tool for diagnosis and carrier detection in families with a dystrophinopathy

S. Salian, S. A. Vahab, H. Shah, A. Shukla, B. Ramamurthy, R. Shenoy, N. Kamath, J. Shenoy, K. Satyamoorthy, K. M. Girisha

Research output: Contribution to journalArticle

Abstract

Evaluation of multiplex ligation dependent probe amplification as a tool for diagnosis and carrier detection in families with a dystrophinopathy: We set out to evaluate multiplex ligation dependent probe amplification (MLPA) as a tool for diagnosis and carrier detection in families with a dystrophinopathy. Fifty three Indian families with provisional diagnosis of Duchene muscular dystrophy or Becker muscular dystrophy were evaluated by MLPA and multiplex polymerase chain reaction (PCR). Sanger sequencing was used to analyze the entire gene in one patient. Mothers were tested for carrier status whenever possible. Molecular analysis of DMD gene by combining MLPA and multiplex PCR yielded a mutation detection rate of 62% (33/53). Deletions were detected in 27/53 (51%) cases, duplications in 5/53 (9%) cases, a small deletion one case and Sanger sequencing detected a nonsense mutation in one case. Mutation was not detected in 36% (19/53) cases. Fifty six percent of mothers (9/16) were found to be carriers. MLPA helped io refine the results of multiplex PCR testing in 22 patients (5 duplications, 16 deletions and one small deletion). We also describe a situation where a deletion of single exon on MLPA (but not detected by multiplex PCR) was actually due to a deletion of two nucleotides in the probe ligation site. MLPA appears to score over multiplex PCR in diagnosis and carrier detection, specifically by detecting deletions and duplications that are not detected by traditional mu.

Original languageEnglish
Pages (from-to)449-460
Number of pages12
JournalGenetic Counseling
Volume27
Issue number4
Publication statusPublished - 2016

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Multiplex Polymerase Chain Reaction
Mothers
Duchenne Muscular Dystrophy
Muscular Dystrophies
Nonsense Codon
Mutation Rate
Genes
Ligation
Exons
Nucleotides

All Science Journal Classification (ASJC) codes

  • Genetics(clinical)

Cite this

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abstract = "Evaluation of multiplex ligation dependent probe amplification as a tool for diagnosis and carrier detection in families with a dystrophinopathy: We set out to evaluate multiplex ligation dependent probe amplification (MLPA) as a tool for diagnosis and carrier detection in families with a dystrophinopathy. Fifty three Indian families with provisional diagnosis of Duchene muscular dystrophy or Becker muscular dystrophy were evaluated by MLPA and multiplex polymerase chain reaction (PCR). Sanger sequencing was used to analyze the entire gene in one patient. Mothers were tested for carrier status whenever possible. Molecular analysis of DMD gene by combining MLPA and multiplex PCR yielded a mutation detection rate of 62{\%} (33/53). Deletions were detected in 27/53 (51{\%}) cases, duplications in 5/53 (9{\%}) cases, a small deletion one case and Sanger sequencing detected a nonsense mutation in one case. Mutation was not detected in 36{\%} (19/53) cases. Fifty six percent of mothers (9/16) were found to be carriers. MLPA helped io refine the results of multiplex PCR testing in 22 patients (5 duplications, 16 deletions and one small deletion). We also describe a situation where a deletion of single exon on MLPA (but not detected by multiplex PCR) was actually due to a deletion of two nucleotides in the probe ligation site. MLPA appears to score over multiplex PCR in diagnosis and carrier detection, specifically by detecting deletions and duplications that are not detected by traditional mu.",
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AU - Salian, S.

AU - Vahab, S. A.

AU - Shah, H.

AU - Shukla, A.

AU - Ramamurthy, B.

AU - Shenoy, R.

AU - Kamath, N.

AU - Shenoy, J.

AU - Satyamoorthy, K.

AU - Girisha, K. M.

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