Scanning by DOVAM-S detects all unique sequence changes in blinded analyses: evidence that the scanning conditions are generic.

摘要: The [detection of virtually all muta tions]-SSCP (DOVAM-S) is a highly sens i tive variant single strand conformation polymorphism (SSCP). Mutations in the fa c tor IX gene were used to find set fiv e SSCP conditions that detects al l mutations. A blinded analysis facto r patients with hemophilia B d tected 82 unique Since th method was developed and tested on gene, it possible condition s selected work more efficiently than other genes. To test gen eral applicability unde which DOVAM-S detected mutations this analyses performed human factor VIII ataxia-telang iectasia (ATM) Segments ampl fied individually, combined into groups 16 18 amplified segments electropho resed five different nondenaturing cond tions varying matrices, buffers, tempera tures additives. Blinded wer 92 samples from (factor gene) 19 sam ples A-T (ATM gene). Com bined an earlier 250 polymorphisms (180 are ) both analyses. For two , three four joint conditions, averag detection frequency ranged 77% – 97% 91% 100% 95% 100%, respectively . each genes, one mutation ma y have been missed if only used. With DOVAM-S, approximatel 500 kb autosomal sequence can b scanned gels tection within gion. 180 out uniqu changes implies at least 96.5% ( P=0.03) tions. detect 400 required termine scanning t 98.5%