Abstract: Objective To develop a multiplex fluorescent real-time PCR method and typing technique of herpes simplex virus for providing a rapid tool with a high sensitivity and specificity for the diagnosis of genital herpes and HSV typing. Methods By using the viral culture as a standard control, the Smart Cycler^® System as a laboratory platform and the fluorescence-labeled probe technique was used to develop a multiplex fluorescent real-time PCR for the detection of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) DNA in clinical specimens of patients with genital herpes. Results Both sensitivity and specificity of the multiplex fluorescent real-time PCR in detecting HSV-1 and HSV-2 DNA were 100%, whereas the sensitivity and specificity of viral culture were 70.1% and 100%, respectively. It took about 30 minutes to complete the whole process with the multiplex real-time PCR, and the detectable limit was as low as 1-5 copies of HSV DNA. Conclusions The multiplex fluorescent real-time PCR based on Smart Cycler^® System is an easy-to-operate and closed amplification system with high amplification efficacy. A single PCR test can provide a rapid, sensitive and accurate detection and typing of HSV in clinical practice.

Key words: Herpes genitalis, Herpesvirus 1, human, Herpesvirus 2, human, Polymerase chain reaction