SIMULTANEOUS ANALYSIS OF VARIOUS MUTATIONS ON THE 21-HYDROXYLASE GENEBY MULTI-ALLELE SPECIFIC AMPLIFICATION AND CAPILLARY GEL-ELECTROPHORESIS

A detailed study is presented on the detection of various known point mutations using polymerase chain reaction (PCR) based multi-allele spe cific amplification (MASA) in conjunction with capillary gel electroph oresis (CGE) separation. The resulting PCR products, corresponding to the individual mutations, are labeled with ethidium bromide during CGE separation, and detected by laser-induced fluorescence. MASA proved t o be a novel, fast and cost-effective method for simultaneous analysis of multiple known mutation sites, employing more than one allele spec ific primers in a single PCR reaction. It results in coexisting amplif ication of numerous DNA fragments differing in size, which are subsequ ently separated by CGE. In the present study, several point mutations were analyzed simultaneously by MASA-CGE on the 21-hydroxylase gene of a patient with congenital adrenal hyperplasia. (C) 1998 Published by Elsevier Science B.V.