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

Year 2001, Volume: 6 Issue: 2, 48 - 52, 25.02.2013

M.S. Serin C. Evruke F. Kibar F. Köksal

Abstract

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 Dienneís
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

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