Document Detail


Single cell multiplex PCR amplification of five dystrophin gene exons combined with gender determination.
MedLine Citation:
PMID:  11331673     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Large deletions in the dystrophin gene account for > 60% of mutations responsible for Duchenne muscular dystrophy (DMD). We have developed a genetic test that can be used directly for the preimplantation genetic diagnosis (PGD) of a majority of couples at risk of transmitting DMD. The test, a double nested multiplex polymerase chain reaction assay for the amplification of exons 8, 19, 45, 47 and 51 allows the detection of over 70% of all DMD deletions. Amelogenin sequences on the X and the Y chromosomes were also co-amplified to provide a correlation between embryo gender and deletion status. The setting up of reliable single cell assays for preimplantation genetic diagnosis is delicate and time consuming. Assays have to be validated on a large number of single cells for each specific mutation to assess efficiency and accuracy before being applied clinically. The multiplex procedure permitted the validation of all tested loci in the same series of isolated lymphocytes rather than in separate series for each exon. One hundred single lymphocytes, 50 female and 50 male cells, were analysed with an overall amplification rate of 98% and an amplification failure of 2% per exon. We suggest that this test is reliable, easy to set up and much preferable to a mere sex determination with the selective transfer of female embryos.
Authors:
P F Ray; M Vekemans; A Munnich
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular human reproduction     Volume:  7     ISSN:  1360-9947     ISO Abbreviation:  Mol. Hum. Reprod.     Publication Date:  2001 May 
Date Detail:
Created Date:  2001-05-02     Completed Date:  2001-08-30     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9513710     Medline TA:  Mol Hum Reprod     Country:  England    
Other Details:
Languages:  eng     Pagination:  489-94     Citation Subset:  IM    
Affiliation:
Département de génétique and INSERM U393, IRNEM, Hopital Necker Enfants Malades, 75743 Paris Cedex 15, France. ray@necker.fr
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MeSH Terms
Descriptor/Qualifier:
Dystrophin / genetics*
Exons*
Humans
Polymerase Chain Reaction / methods
Sex Determination (Genetics)*
Chemical
Reg. No./Substance:
0/Dystrophin

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