Detection of known and new mutations in the thiopurine S-methyltransferase gene by single-strand conformation polymorphism analysis.

摘要: To detect mutations in the thiopurine S-methyltransferase gene (TPMT), we have developed a strategy based on single-strand conformation polymorphism (SSCP) analysis of amplified by polymerase chain reaction (PCR). The sensitivity method was first evaluated analyzing DNA samples from five individuals, including two high methylators (HMs), intermediate (IMs), and one deficient methylator (DM). TPMT alleles each these individuals had previously been characterized conventional PCR-based assays direct sequencing analysis. All were associated with particular shifts electrophoretic mobility fragments, allowing their identification. We further tested efficiency to new mutations. For this purpose, additional DNAs 15 IMs HMs submitted PCR-SSCP A total 7 characterized, alleles. one, termed TPMT*1A, harbors single mutation C-->T at nucleotide -178 exon 1 detected HM subject. second TPMT*7, T-->G transversion 681 10. This allele should be nonfunctional since it observed combination wild-type an methylator. conclude that could advantageously used fully characterize extent allelic variation locus populations thus improve our understanding genetic activity, which has considerable consequences for toxicity efficacy therapeutically important widely drugs.