Abstract: To rapidly identify and diagnose mixed infections of common pathogens in the urogenital tract,including Mycoplasma hominis,Ureaplasma urealyticum,Mycoplasma genitalium,and Chlamydia trachomatis,a quadruple fluorescence quantitative PCR technique was employed.Specific primers and fluorescence probes were designed based on the conserved gene regions of the four pathogens,and the reaction system was systematically optimized (primers concentration,probes concentration,and annealing temperature).Performance verification was also conducted.The results showed that the quadruple fluorescence quantitative PCR detection technique exhibited excellent specificity and high repeatability,with a detection sensitivity of up to 1000 copies/mL.Compared with conventional PCR technology,the four-step fluorescence quantitative PCR technique significantly improved the diagnostic efficiency and accuracy of mixed infections,and was suitable for rapid differential diagnosis of clinical mixed infections.

Key words: Mycoplasma hominis, Ureaplasma urealyticum, Mycoplasma genitalium, Chlamydia trachomatis, quadruple fluorescence PCR detection technology