Abstract： Objective To establish a multiple PCR method that can be used to spontaneously detect five kinds ofpathogens such as NG,MH,MG,CT and UU.Method With the fluorescence-quantitative PCR technique in conjunction with another detection technique as the golden standard,evaluation was conducted on the sensitivity,specificity,accuracy and repeatability on the detection of 5 kinds of STD pathogens using single-tube multiplex PCR.Result The sensitivity,specificity and match rate of the method ale 10-9fg/μl,100%,97.8%respectively,and the repeatability of 5 continuous days of 20 clinical specimens is good.Conclusion Single-tube multiplex PCR technique provides a new method to detect 5 kinds of STD pathogens.