XÂY DỰNG QUY TRÌNH REAL-TIME PCR ĐA MỒI XÁC ĐỊNH 3 CHỦNG VI KHUẨN LÂY TRUYỀN QUA ĐƯỜNG TÌNH DỤC THƯỜNG GẶP: CHLAMYDIA TRACHOMATIS, NEISSERIA GONORRHOEAE, TREPONEMA PALLIDU

Abstract

Objective: To establish a multiplex real-time PCR assay for the simultaneous detection of three common sexually transmitted pathogens: Chlamydia trachomatis, Neisseria gonorrhoeae, and Treponema pallidum. Subjects and Methods: Three reference strains of C. trachomatis, N. gonorrhoeae, and T. pallidum. 39 clinical specimens of cervical swabs from women with vaginitis who visited Hanoi Obstetrics and Gynecology Hospital from August to December 2023. Results: The multiplex real-time PCR assay performed well within the annealing temperature range of 56°C to 62°C and annealing time of 20 to 60 seconds. The limit of detection (LOD) of the assay for all three pathogens was 10² copies/reaction with a 100% detection rate. At a concentration of 1 copy/reaction, the detection rate of N. gonorrhoeae was 95% and C. trachomatis was 100%. Comparative analysis showed complete concordance between the two methods in all 39 patients: 11 samples positive for N. gonorrhoeae, 18 samples positive for C. trachomatis, 10 samples negative for all three pathogens. Conclusion: The successfully developed multiplex real-time PCR assay is applicable for the simultaneous detection of C. trachomatis, N. gonorrhoeae, and T. pallidum with high efficiency and accuracy.