Determination of Duchenne muscular dystrophy carrier status by single strand conformation polymorphism analysis of deleted regions of the dystrophin locus.

Richards, Robert I.; Friend, K

doi:10.1136/jmg.28.12.856

Cited by 6 publications

(2 citation statements)

References 14 publications

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“…Second, the standardization of the electrophoretic conditions of migration of the SSCP gels may have prevented the detection of base substitution in one of the 20 amplified fragments. Nevertheless, the conditions selected here were nearly optimal as we have been able to identify new polymorphisms in exon 3, 13, and 49 and to detect all the polymorphisms previously described either by Kiliman et al (1992) using the chemical cleavage method or by Richards et al (1991) and Zietkiewicz et al (1992), using the SSCP technique with amplification and electrophoretic conditions different from ours.…”

Section: Discussionmentioning

confidence: 74%

Base substitutions in the human dystrophin gene: Detection by using the single-strand conformation polymorphism (SSCP) technique

Tuffery¹,

Moine²,

Demaille³

et al. 1993

Hum. Mutat.

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We have established the experimental conditions to screen twenty regions of the dystrophin gene using the method of single-strand conformational polymorphism (SSCP) analysis. The aim of this study was to identify point mutations in patients with Duchenne or Becker muscular dystrophy (DMD or BMD) who have no gross DNA rearrangements detectable by Southern blot analysis or multiplex exon amplification. The investigation of thirteen patients using this procedure resulted in the detection of seven sequence polymorphisms (four identified in this study) that will be useful allelic markers in familial DNA analysis. Three rare sequence variants could be found (two of them being novel variants) but we were unable to demonstrate mutations that could be clearly sufficient to be responsible for the phenotype. This analysis confirmed the efficiency of the SSCP technique for the detection of nucleotide substitutions. Application of this approach to mutation or polymorphism detection to other exons of the gene will improve carrier and prenatal diagnosis.

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Section: Discussionmentioning

confidence: 74%

Base substitutions in the human dystrophin gene: Detection by using the single-strand conformation polymorphism (SSCP) technique

Tuffery¹,

Moine²,

Demaille³

et al. 1993

Hum. Mutat.

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“…In contrast to the mild allelic Becker muscular dystrophy (BMD), the Duchenne type is progressive and usually results in death during the second decade of life. The cDNA of the gene responsible for DMD/BMD has been cloned2 and sequenced.3 The gene product has been named 'dystrophin' and is encoded by an mRNA of 14 kb derived from at least 74 exons spread over 2300 kb of the X chromosome. 4 The extraordinary size of the gene causes extreme difficulties in molecular analysis.…”

mentioning

confidence: 99%

Identification of a new DMD gene deletion by ectopic transcript analysis.

Rininsland¹,

Hahn²,

Niemann-Seyde³

et al. 1992

Journal of Medical Genetics

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The detailed genetic analysis of the Duchenne/Becker muscular dystrophy gene is hindered by the large number of exons involved and their separation by huge introns. These problems can be overcome by the analysis of mRNA rather than genomic DNA and ectopic transcripts derived from peripheral blood lymphocytes provide a convenient source of material. Using reverse transcription and nested PCR, we show here a comprehensive strategy for the rapid and complete analysis of the coding sequences from complex genes and illustrate its potential by the identification of a hitherto undescribed single exon deletion.

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Determination of human alcohol dehydrogenase and acetaldehyde dehydrogenase genotypes by single strand conformation polymorphism in discontinuous buffer electrophoresis

et al. 1993

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Under appropriate conditions single strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products allows the detection of single base mutations in a given DNA fragment. We adapted this method for the routine determination of allele variants of human alcohol and acetaldehyde dehydrogenase without radioisotopic labeling. After PCR amplification of the selected exon, the DNA fragments were heat-denatured and loaded on a polyacrylamide gel containing glycerol. For electrophoresis a discontinuous buffer system was used with sulfate as leading ion and borate as trailing ion. The DNA bands were revealed by silver staining. Acrylamide concentrations, ionic strength and electrophoresis temperature were systematically investigated for each DNA fragment. The polymorphisms detected by SSCP were identical to those found by hybridization with 32P-labeled allele-specific oligonucleotides. This method avoids the use of radioactivity, is less expensive and simpler than the allele-specific oligonucleotide (ASO) methodology and thus particularly suited for routine analysis.

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Determination of Duchenne muscular dystrophy carrier status by single strand conformation polymorphism analysis of deleted regions of the dystrophin locus.

Abstract: The molecular characterisation of the dystrophin gene, mutations in which are responsible for X linked Duchenne

Cited by 6 publications

References 14 publications

Base substitutions in the human dystrophin gene: Detection by using the single-strand conformation polymorphism (SSCP) technique

Base substitutions in the human dystrophin gene: Detection by using the single-strand conformation polymorphism (SSCP) technique

Identification of a new DMD gene deletion by ectopic transcript analysis.

Determination of human alcohol dehydrogenase and acetaldehyde dehydrogenase genotypes by single strand conformation polymorphism in discontinuous buffer electrophoresis

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