A reverse dot blot method for genotyping of NAT2 in Chinese subjects / 中国药理学通报

ABSTRACT

Aim To establish a simple reverse dot blot m ethod (RDB) for detecting the genotype of NAT2 in Chinese people. Methods PCR was performed to obtain a biotin labeled DNA fragment. Allele specific oligonucleotide probes were spotted onto a nylon membrane. The DNA fragment hybridized with the membrane under stringent conditions. Finally, a nonradioactive colorimetric reaction was used to detect five mutants of NAT2. NAT2 genotypes of 48 patients with pulmonary tuberculosis were detected with RDB.Results The results obtained from RDB were in consistent with those from allele specific amplification. The NAT2 allele frequencies of *5, *6, *7 were 1.04%, 22.9% and 15.6%, respectively.Homozygous wildtype,heterozygous mutant and homozygous mutant subjects were 33.3%, 54.2% and 12.5%, respectively.Conclusion RDB method was proved to be accurate and convenient, it can be u sed in rational drug therapy.