Performance evaluation and application of a multiplex PCR capillary electrophoresis method for detecting nucleic acids of seven sexually transmitted pathogens.

Objective: To establish a rapid multiplex PCR capillary electrophoresis (MPCE) method for the simultaneous detection of seven sexually transmitted infection pathogens, and to evaluate its clinical utility. Methods: We designed specific primers and optimized multiplex PCR conditions. Capillary electrophoresis enabled fragment analysis. We assessed diagnostic performance, including sensitivity, specificity, anti-interference capacity, and repeatability. Using commercially available reagents as references, we evaluated the agreement of MPCE with 205 clinical samples. Kappa consistency test and McNemar test were used to evaluate the consistency and difference between this method and the reference methods. Results: The detection system was successfully constructed, with a detection time of three hours. It enabled simultaneous detection of seven common genital tract pathogens, including Conclusion: This study successfully established a MPCE-based assay for the simultaneous detection of seven common genital tract pathogens (CT, UU, MG, NG, MH, HSV-2, and UP) in a single reaction. The method demonstrated high sensitivity, excellent repeatability and strong agreement with commercial reference assays. With its high throughput and rapid turnaround time, this method is well-suited for clinical screening and large-scale epidemiological surveillance of genital tract infections.