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A Quantitative Polymerase Chain Reaction (PCR) Assay Completely Discriminates between Duchenne and Becker Muscular Dystrophy Deletion Carriers and Normal Females
- Publication Year :
- 1996
- Publisher :
- Elsevier Science Limited:Oxford Fulfillment Center, PO Box 800, Kidlington Oxford OX5 1DX United Kingdom:011 44 1865 843000, 011 44 1865 843699, EMAIL: asianfo@elsevier.com, tcb@elsevier.co.UK, INTERNET: http://www.elsevier.com, http://www.elsevier.com/locate/shpsa/, Fax: 011 44 1865 843010, 1996.
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Abstract
- Duchenne/Becker muscular dystrophy (DMD/BMD) is a severe X-linked myopathy. In 65% of the patients, the mutations responsible for the disease are macrodeletions in the dystrophin-encoding gene that can be identified with multiplex polymerase chain reaction (PCR) technology. We developed a method for quantitative PCR analysis of deletion carriers involving the use of phosphorimager-based scanning of radioactive-labelled PCR products. We calculated the ratios between the areas of two peaks, one corresponding to the deleted segments to be analysed and the other taken as a reference. In carriers, these ratios (R value) were always about half those obtained in normal females. The final diagnostic result, the diagnostic index (DI), is the ratio of the R values between the propositus and a normal subject. We also assessed the variability of each step of the procedure and the overall variability of the DI value, thus obtaining cut-off values that completely discriminated BMD/DMD deletion carriers from normal females. We were also able to classify, as either «carrier» or «normal», several females whose status was not identified with linkage analysis.
- Subjects :
- musculoskeletal diseases
X Chromosome
Duchenne muscular dystrophy
Pcr assay
Biology
Polymerase Chain Reaction
Muscular Dystrophies
Dystrophin
Genetic linkage
Reference Values
Multiplex polymerase chain reaction
medicine
Humans
Muscular dystrophy
Myopathy
Molecular Biology
Gene
DNA Primers
Sequence Deletion
Genetic Carrier Screening
Reproducibility of Results
Cell Biology
medicine.disease
Molecular biology
Real-time polymerase chain reaction
Regression Analysis
Electrophoresis, Polyacrylamide Gel
Female
medicine.symptom
Subjects
Details
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....6c48c7f2c42e8e98f1bb42639cbfdd3d