Molecular analysis of the N-acetyltransferase 1 gene (NAT1*) using polymerase chain reaction-restriction fragment-single strand conformation polymorphism assay

One major interest to analyse the extent of N-acetyltransferase 1 (NAT1*) a llelic variation in the human population stems to a great extent from the p ossible association of interindividual differences in the metabolism of aro matic amines with certain chemically induced diseases, including cancer. Co nsidering the increasing number of mutations in the NAT1 gene that are dete cted, NAT1* genotyping using conventional polymerase chain reaction (PCR) r estriction fragment length polymorphism (RFLP) or allele-specific amplifica tion assays has become complicated. We developed a rapid and powerful strat egy allowing the full characterization of NAT1* alleles. This method, based on single-strand conformation polymorphism analysis of a unique PCR produc t encompassing the entire intronless NAT1* coding region along with additio nal flanking segments in the 5' and 3' untranslated regions, was then appli ed to DNA samples from 270 individuals. Nine NAT1* allelic variants, includ ing two novel (NAT1*28 and NAT1*29), and 15 different genotypes were identi fied, This approach could be advantageously used in epidemiological studies to provide more definite data on suspected associations between NAT1* geno type and certain pathological processes. Pharmacogenetics 10:293-300 (C) 20 00 Lippincott Williams & Wilkins.