Detection of Treponema pallidum DNA polymerase I gene by real-time polymerase chain reaction

Treponema pallidum je gram-negativna bakterija spiralnog izgleda koja uzrokuje sustavnu bolest sifilis. Stopa sifilisa u svijetu je u porastu prvenstveno među muškarcima koji imaju spolne odnose s drugim muškarcima (MSM). Sifilis povećava opasnost od prijenosa HIV-a od dva do pet puta, a istovremena infekcija s oba uzročnika je česta. U Hrvatskoj je u 2020. g. zabilježen porast učestalosti sifilisa od 91,4% među MSM populacijom. Cilj ovog diplomskog rada bio je ispitati prisutnost gena za DNA polimerazu I (polA) bakterije T. pallidum u različitim uzorcima ispitanika s različitim stadijima sifilisa te ispitati pogodnost korištenja uzorka sline za molekularnu dijagnostiku T. pallidum. Istraživanje je provedeno na 48 uzoraka ispitanika populacije MSM od kojih je 28 imalo i HIV-infekciju. U 9/48 (18,8%) uzoraka detektiran je gen za DNA polimerazu I T.pallidum Najveći broj pozitivnih uzoraka su bili brisevi ždrijela (6/26, 23,1%) dok su ostala 3/15 (20%) pozitivna uzorka predstavljali uzorci sline. U 4/28 ispitanika s HIV-infekcijom detektirana je DNA T.pallidum. U ovom istraživanju se po prvi puta u Hrvatskoj koristila metoda molekularne detekcije koja se pokazala učinkovitom u dokazivanju prisutnosti uzročnika sifilisa.Treponema pallidum is a gram-negative spiral bacterium that causes systemic disease syphilis. The prevalence of syphilis worldwide is increasing primarily among men who have sex with other men (MSM). Syphilis increases the risk of HIV transmission from two to five times, while a coinfection with both of them at the same time is quite common. In 2020. a 91.4% increase in the frequency of syphilis was recorded in MSM population in Croatia. The object of this study was to examine the presence of the DNA polymerase I (polA) T. pallidum in different samples with different stages of syphilis and examine the suitability of saliva for molecular diagnosis of T. pallidum. The research was conducted on 48 samples of respondent MSM population, 28 of whom had a HIV infection. Treponema pallidum DNA polymerase I gene were detected in 9/48 (18,8%) samples. The largest number of positive samples were throat swabs (6/26, 23,1%), while the others 3/15 (20%) samples were samples of saliva. Treponema pallidum DNA was detected in 4/28 samples with HIV infection. In this research, conducted for the first time in Croatia, molecular detection method was used, which proved to be effective in proving the presence of the causative agent of syphilis