A NOVEL GENOTYPING SYSTEM FOR ANALYZING THE NAT2 GENE USING WHOLE BLOOD AS STARTING MATERIAL DIRECTLY

摘要

The arylamines N-Acetyltransferase 2 (NAT2) has a role in the metabolism of many clinically important substances. The NAT2 polymorphisms have been reported to be associated with a number of cancers including childhood acute lymphoblastic leukemia (ALL). Several methods, such as PCR-RFLP and LightCycler - NAT2 Mutation Detection Kit have been established to genotype NAT2 alleles. However, these methods are time-consuming and expensive. Here we established a new genotyping procedure based on adapter-ligation mediated allele-specific amplification (ALM-ASA) to analyze four SNPs in the NAT2 gene from whole blood directly. The association of NAT2 genotypes with adult ALL was investigated by conducting a case-control study on 88 ALL patients and 136 health controls of Chinese origin. The SNPs 191G＞A, 481C＞T, 590G＞A and 857G＞A were successfully detected by ALM-ASA assay and the results were identical to that obtained by both Sanger's sequence and PCR-RFLP analysis. A genotype could be completed within 4 hours using whole blood sample as starting material directly. Genotypic frequencies obtained were consistent with previously published results. Compared to the control group, the frequencies of NAT2 genotypes and variants in ALL group showed no statistically significant difference. These results suggest that genetic polymorphisms in the NAT2 gene do not influence susceptibility to adult ALL. The ALM-ASA assay is rapid, inexpensive, easy-to-perform and amenable to automation and could facilitate the largescale genotypic analysis of NAT2.