Comparison of PCR and Cultivation Methods to Determine the Incidence of Infections due to Mycoplasma Hominis and Mycoplasma Fermentans in Women Genitourinary Tract

Objective: In this study, in order to compare PCR and cultivation methods to determine the incidence of infections due to  M. hominis  and  M. fermentans  in women genitourinary tract  100 genital swabs and 100 urine samples obtained from women with genitourinary tract (GUT) infection were studied. Method:  Genital swab and urine samples were inoculated and transported with a selective mycoplasma  transport media. After incubation at 37°C for 18-24 hours 0.3 mL medium samples were transferred to the specific solid medium for mycoplasma.  The agar plates were incubated at the same atmosphere conditions (5% CO2 and 95% N2 at 37°) for 48-72 hours. Characteristic mycoplasma colonies were determined by staining with Dienneis stain and examined by x10 microscope objective. The genital swab and urine samples were also analyzed by a nested PCR protocol with genus specific MCGpF11, R23-TR, R16-2 and MCGR21 primers. Another PCR protocol was also performed in order to confirm the samples which have compatible target sequences for M. fermentans by using RW004-RW005 primers. On the other hand, all other mycoplasma positive amplicons were also digested with  VspI    in order to determine two DNA fragments (123bp and 113bp) which were compatible for M. hominis in tested samples. Results: Mycoplasma    strains were isolated from 26 (26%) genital swabs and 11 (11%) urine samples by using a selective mycoplasma isolation media. Totally 40 samples were found to be positive for mycoplasmas which consisted of target genomic sequences of  M. hominis  and  M. fermentans  in 37(37%) and 3(3%) samples respectively. Conclusions:  We  found that there could be an association with  M. hominis  (37%) and women with genital infection, also with  M. fermentans  (3%) and although the high specificity (100%) of cultivation, it has a low sensitivity (70.3%) and time consuming when compared with PCR . On the other hand, we concluded that, PCR is a sensitive and easily applicable protocol when genus specific primers are used for the diagnosis of mycoplasmas. Key words:  Mycoplasma hominis, mycoplasma fermentans, PCR, DNA, RFLP