Detection of @entity5345 and XXXX mutations in @entity0 families by a comprehensive two-stage screening procedure.

@entity1 @entity53007 @entity5344 @entity0 @entity5345 @entity72747 @entity2674 @entity377
We have developed a 2-stage protocol for @entity5345 and @entity2674 mutation screening from blood spot paper. Stage 1 screening was aimed to analyze @entity1 at highest risk for the most common disease-associated sequence variants listed in the BIC database. Accordingly, stage1 testing implied detection of 18 disease- associated @entity5345 and 9 @entity2674 mutations by adapting the 5' nuclease assay to heterozygote screening. For stage 2 screening, we applied the conformation sensitive gel electrophoresis (CSGE) method by adapting this technique to automated heteroduplex analysis of @entity5345 and @entity2674 using fragment scanning on an ABI 377 sequencing device. Of the 120 @entity1 with a family history of @entity5344 who took part in this study so far, 45 entered stage 1 testing. Disease-associated mutations were detected in 6 @entity1 by stage 1 testing (13%). For these @entity1 , the final result was available within 10 days. Mutation @entity53007 was found in 2 @entity1 . One @entity1 with mutation @entity72747 in @entity2674 reported only 1 sister with a multifocal bilateral @entity0 . New disease-associated mutations were detected in 2 of the 114 @entity1 who entered the stage 2 test (1.7%). Of particular interest was 1 @entity1 who was diagnosed with a @entity0 at age 39 and who had no family history of @entity0 . We conclude that pre-screening by 5' nuclease assay for the mutations most frequently seen in a given population represents a relatively effective first line of analysis. Subsequent detailed analysis by fluorescence conformation sensitive gel electrophoresis (F-CSGE) and fragment sequencing is a sensitive alternative to full @entity377 sequencing.
