ABSTRACT
OBJECTIVE: To establish a method to determine the genotypes of the three SNPs(-31T/C, -133C/T and -140T/C) of uridinediphosphateglucuronosyltransferase 1A3(UGT1A3)and to preliminarily analyze the genetic frequencies of different genotypes among healthy Chinese population. METHODS: The genotypes of the three SNPs(-31T/C, -133C/T and -140T/C) of 123 healthy Chinese subjects(89 males, 34 females) were screened by two-step allele specific amplification(two-step ASA) method. Firstly, a pair of primers of the first exon for the amplification of the genomic DNA were designed. And then two parallel reaction tubes were set up for each site which included a forward primer whose last base of 3'end pair with wild type or mutant type and an identical reverse prime for the amplification of the target gene. The amplification Results were examined to differentiate the genotypes. The validity was verified by direct sequencing. RESULTS: Among the 123 subjects, the frequencies of -31T and -31C were 0.776 and 0.224, of -133C and -133T were 0.935 and 0.065, of -140T and -140C were 0.915 and 0.085, respectively. Based on the genotyping Results, there were 59 cases of UGT1A3*1/*1, 1 case of UGT1A3*2/*2, 17 cases of UGT1A3*1/*2, 12 cases of UGT1A3*1/*4, and 1 case of UGT1A3*4/*4. CONCLUSION: There are genetic polymorphisms of UGT1A3 in healthy Chinese population and the mutation frequency is remarkable. The gene detection method established in this study is verified to be sensitive and specific.