Objective To explore and analyze the distribution of NAT2 gene in Han population in Shenzhen and provide the basis for treatment of NAT2-related metabolic diseases and cancer research. Method The study diluted the DNA sample(d0 ng/μl) as follows: 1 × 100, 1 × 101, 1 × 102, 1 ×103 , 1 × 104, to verify the sensitivity of detection of NAT2 gene polymorphism by real-time PCR. Mutation locus of 282,341,481, 590 and 857 of NAT2 in 554 normal samples were detected and genotyped by realtime PCR with Taqman probes. Forty-seven cases among 554 healthy controls were detected by DNA sequencing to verify the sensitivity, specificity and accuracy of this assay. Results The lowest detection limit was 10-4 ng/μl. The frequencies of each allele of NAT2 were: *4 allele 64. 6% (358/554), *5 allele 6. 3% (35/554), *6 allele 25. 3% ( 140/554), *7 allele 30. 0% ( 166/554), * 11 allele 0. 6% (3/554) The frequencies of main genotypes of NAT2 were as the follows : NAT2 *4/* 6 12.3% ( 68/554 ), *6/* 78. 3% (46/554), * 13/* 13 or * 12/* 12 or *4/* 4 7.6% (42/554), *4/* 7 8. 1% (45/554), * 7/* 1 312. 3% (68/554), * 12 7.2% (40/554), *6/* 13( * 12)5.6% (31/554). The samples with 7 genotypes account for 89. 6% (496/554) of the total cases. Rapid acetylation and Intermediate type accounting for 40. 5% (224/554) and 46. 7% (259/554) respectively were the main phenotypes of NAT2 in Han people in Shenzhen. In addition, the accuracy, specificity and sensitivity of the PCR were compared to direct DNA sequencing. The accuracy of 282, 341,481, 590 and 857 were 88.2% (30/34), 87.5% (7/8), 80. 0%(4/5), 100. 0% ( 22/22 ) and 93.8% ( 15/16 ) respectively. The specificities were 100. 0% ( 13/13 ),94. 9% ( 37/39), 100. 0% ( 42/42 ) , 96.0% ( 24/25 ) and 96. 8% ( 30/31 ) respectively. The sensitivities were 91.5% ( 43/47 ), 93.6% ( 44/47 ), 97.6% ( 46/47 ) 97.9% ( 46/47 ) and 95.7% ( 45/47 ),respectively. Conclusions The sensitivity, specificity and accuracy of real-time PCR established in this study is good. It can be widely used for clinical and scientific research. The major alleles of NAT2 of Han population in Shenzhen are * 4, *6, * 7. The most common slow acetylation genotype is *6/* 7. This study can provide a basis for NAT2-related metabolic diseases and cancer research. Key words: Arylamine N-acetyhransferase; Polymorphism,single nucleotide; Polymerase chain reaction