Front Cell Infect Microbiol. 2026 May 18;16:1816857. doi: 10.3389/fcimb.2026.1816857. eCollection 2026.
ABSTRACT
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 Chlamydia trachomatis (CT), Ureaplasma urealyticum (UU), Mycoplasma genitalium (MG), Neisseria gonorrhoeae (NG), Mycoplasma hominis (MH), Herpes simplex virus type 2 (HSV-2), and Ureaplasma parvum (UP). The method demonstrated good specificity and anti-interference capacity, with a limit of detection (LOD) ranging from 325 to 900 copies/mL across all targets. Repeatability results showed minimal fragment length fluctuations (0.13-0.20 nt), and the coefficient of variation (CV) for log peak area ranged from 0.78% to 5.56%. Agreement between the developed method and commercial reference assays was good for all seven pathogens: CT, 94.93% (Kappa = 0.84); MG, 95.12% (Kappa = 0.88); NG, 96.59% (Kappa = 0.91); UP, 95.61% (Kappa = 0.88); MH, 97.07% (Kappa = 0.82); HSV-2, 100% (Kappa = 1.00); and UU, 91.71% (Kappa = 0.69). The positivity rates of MG and NG detected by this method were higher than those by the reference methods (P = 0.008 and P = 0.011, respectively), and there was no statistical difference for the other pathogens (P>0.05).
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.
PMID:42233157 | PMC:PMC13223153 | DOI:10.3389/fcimb.2026.1816857
