Objective In order to improve clinical treatment and STD control，we profiled the infection characteristics of herpes simplex virus （HSV） in sexually transmitted disease （STD） clinics in Guangzhou. Methods In 488 outpatients at Guangdong Provence Dermatology Hospital′s STD department，polymerase chain reaction（PCR）was utilized to identify HSV⁃1/2 DNA，with 286 cases also tested for HSV IgM/IgG antibodies using an enzyme ⁃linked immunosorbent assay （ELISA）. The characteristics of HSV infection were analyzed. Results Among the nucleic acid test results of 488 patients aged from 17 to 86，136 were positive for HSV⁃2，6 were positive for HSV ⁃1，and 1 was double positive for HSV ⁃1 and HSV ⁃2 among them，with a positive rate of 28.89%（141/488）. In the positive detection，the HSV⁃2 to HSV⁃1 ratio was 22.67∶1（136/6）. HSV⁃2 DNA was found in 85 of 351 male patients（24.22 %）and of 137 female patients（40.88%），which was significantly higher than men（P < 0.001）. There was no significant difference in the positive detection rate among age groups：< 30， 30 ~ 40，41 ~ 50，51 ~ 60 and > 60（P > 0.05）. In the male positive samples，the positive rates of samples taken from the glans prepuce，penis，scrotum，mound，urethra swab，anus，buttock were 70.59%（60/85），12.94%（11/85）， 11.76%（10/85），1.18%（1/85），1.18%（1/85）and 2.35%（2/85），respectively. In the female positive samples， the positive rate of samples from vulva，cervix，vagina，anus，buttock were 73.21%（41/56），7.14（4/56），16.07% （9/56）and 3.57%（2/56），respectively. The results of the serum antibodies tests were carried out at the same time in 286 of the 488 patients. Among the 68 HSV⁃2 DNA positive patients，the positive rates of HSV⁃2 IgM and HSV⁃2 IgG were 5.88%（4/68）and 66.18%（45/68），respectively. Conclusion HSV⁃2 is the most common cause of HSV infection，but HSV⁃1 is not uncommon. Suspicious patients should be tested for HSV⁃1/2 DNA. Male prepuce glans and female vulva are predilection sites for genital herpes. The combined detection of HSV⁃2 DNA and IgG antibody is helpful for the diagnosis of initial genital herpes and recurrent genital herpes.