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From the * Department of Microbiology and Research
Laboratory "Microorganismes et Pathologie Humaine," Habib
Bourguiba Hospital, Sfax, Tunisia; and the
Laboratory of Histology-Embryology and Biology of Reproduction, Medical
School, Sfax, Tunisia.
| Correspondence to: Dr R. Gdoura or Prof A. Hammami, Laboratory of Microbiology, Faculté de Médecine de Sfax, Avenue Magida Boulila, 3029, Sfax, Tunisia (e-mail: gdourar{at}yahoo.com) (Dr Gdoura) (e-mail: Adnene.hammami{at}rns.tn) (Prof Hammami). |
| Received for publication June 9, 2007; accepted for publication November 27, 2007. |
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Abstract |
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> 0.9) between the detection of Chlamydia
trachomatis, genital ureaplasmas, and Mycoplasma hominis in
semen and corresponding FVU specimens. Our findings also show a high
concordance (81.1%) and a good agreement ( = 0.79) between the
detection of Mycoplasma genitalium in both specimens. C
trachomatis, genital mycoplasmas, and genital ureaplasmas were found to
be widespread among infertile male patients in Tunisia, as shown by their
respective prevalences of 43.3%, 18.3%, and 14.4%. The mean values of seminal
volume, sperm concentration, sperm viability, sperm motility, sperm
morphology, and leukocyte count were not significantly related either to the
detection of C trachomatis DNA or to that of genital ureaplasma or
mycoplasma DNA in semen specimens. Using our in-house PCR, both semen and FVU
were found to be sensitive diagnostic specimens for the detection of C
trachomatis, ureaplasmas, and mycoplasmas. The FVU, a less invasive and
self-collected specimen, can serve as a marker for the presence of these
organisms in the genital tract and can be used as a reliable way of detecting
asymptomatic carriers of infection.
Key words: Male infertility, sperm quality
Various studies conducted in developed countries have demonstrated that C trachomatis, genital ureaplasma, and genital mycoplasma infections can lead to sterility (Upadhyaya et al, 1984; Paavonen and Wolner-Hassen, 1989; Vigil et al, 2002; Askienazy-Elbhar, 2005; Wang et al, 2006). In developing countries, however, infections caused by Chlamydia and Mycoplasma have not been well studied, and the prevalence of C trachomatis, genital ureaplasmas, and genital mycoplasmas among infertile couples has not been determined.
Asymptomatic infections of the male urogenital tract are difficult to detect. Screening with classical cell culture requires specimens from endourethral swabs, which are unacceptable to many asymptomatic men. For Chlamydia, the cell culture method cannot be used for semen and urine samples because of their cytotoxicity (Mardh et al, 1980), and is not sufficiently sensitive to rule out infections of accessory glands (Berger et al, 1978). Other routine tests for Chlamydia, such as enzyme immunoassays, may have a reduced sensitivity in asymptomatic individuals. Genital mycoplasma and ureaplasma colonizations are commonly diagnosed by culture. Cultures, however, are time-consuming as they require 2–5 days for Ureaplasma spp. and M hominis and up to 8 weeks for M genitalium, whereas nucleic acid amplification techniques can detect infectious agents in less than 8 hours.
A great deal of research has led to the development and evaluation of sensitive and specific diagnostic tests based on nucleic acid amplification tests for C trachomatis, M hominis, M genitalium, U urealyticum, and U parvum. The important aspect of these assays is their capacity to be used on less-invasive specimens, such as first void urine (FVU), which can be self-collected. Studies have shown that nucleic acid amplification tests performed on these less-invasive samples are able to detect as many or more infected patients than traditional swabs from the urethra or cervix and are more suitable methods for screening and diagnosing chlamydial and genital Mycoplasmataceae infections (Chernesky et al, 1994; Consentino et al, 2003; Gaydos et al, 2004; Jensen et al, 2004). However, one major problem with them is the presence of potential inhibitors in clinical specimens that can lead to false negative results.
The purposes of our study were: first, to compare semen specimens and FVU specimens from asymptomatic male members of infertile couples for the detection of C trachomatis, U urealyticum, U parvum, M hominis, and M genitalium infections using an in-house inhibitor-controlled polymerase chain reaction (PCR)–microtiter plate hybridization assay; and second, to determine the prevalence of those organisms in infertile men in Tunisia. We also investigated the relationship between those bacteria and semen quality.
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Patients and Methods |
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TopAbstractPatients and MethodsResultsDiscussionReferences |
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Specimens
Semen and corresponding FVU samples of the 104 patients were collected
after approval of our institutional review board and each patient's informed
consent. In the morning, when patients had not had oral intake for at least 8
hours, FVU was collected in a sterile plastic container and stored at
–20°C. After FVU had been collected, patients were instructed to
wash the penis, scrotum, and hands with soap and water, and then semen
specimens were obtained by masturbation and collected into standard containers
that had previously been shown not to have any cytotoxic effects on human
spermatozoa. All male partners had at least 3 days of sexual abstinence before
donation of semen.
Semen Analysis
Semen analysis was performed according to the methods outlined by the World
Health Organization (WHO,
1999) to determine the following variables: seminal volume, sperm
concentration, viability, total progressive motility (category [a + b]), rapid
progressive motility (category [a]), and normal morphology, as well as
leukocyte count. Azoospermia was defined by the absence of spermatozoa after
at least 2 consecutive spermiograms, oligospermia by a sperm concentration of
<20 x 106/mL, asthenospermia by sperm motility (category
[a + b]) <50%, necrospermia by sperm viability <50%, and teratospermia
by normal morphology <30%.
Peroxidase staining, being a practical and reliable method recommended by
the WHO for determining leukocytes in the semen, was employed to count and
differentiate leukocytes from immature germ cells (1999). Leukocytospermia was
defined by a concentration of leukocytes 
106/mL.
Detection of C trachomatis, Genital Mycoplasmas, and Genital Ureaplasmas in Semen Specimens by In-House PCR–Microtiter Plate Hybridization Assay
For each male patient, 200 µL of semen specimens and 1 mL of
corresponding FVU samples were used for DNA detection of C
trachomatis, genital mycoplasmas (M hominis and M
genitalium), and genital ureaplasmas (U urealyticum and U
parvum) by an amplification-based method according to the following
protocol.
Extraction of DNA by Cetyltrimethylammonium Bromide–Phenol-Chloroform/Isoamyl Alcohol Method— The precipitates from each 200 µL of semen specimens and 1 mL of corresponding FVU samples were harvested by centrifugation at 14 000 x g for 20 minutes. The precipitates were then treated with 5 µl of proteinase K (20 mg/mL) at 55°C for 2 hours in 600 µL of digestion buffer (30 µl of 10% sodium dodecyl sulfate and 570 µL of TE buffer [10 mM Tris-HC1 (pH 8), 1 mM EDTA]).
After homogenization, the samples were incubated in a solution of cetyltrimethylammonium bromide (CTAB)-NaCl (100 µL of 5 M NaCl and 80 µL of 10% CTAB) for 10 minutes at 65°C, and then mixed with 750 µL of chloroform–isoamyl alcohol (24:1 [vol/vol]) and centrifuged for 15 minutes at 14 000 x g in an Eppendorf centrifuge. The aqueous phase was separated, mixed with 750 µL of phenol-chloroform–isoamyl alcohol (25:24:1 [vol/vol/vol]), and centrifuged for 15 minutes at 14 000 x g in an Eppendorf centrifuge. The obtained aqueous phase was reseparated and mixed with an equal volume of isopropanol. The samples were left at –80°C for 1 hour and then centrifuged for 15 minutes at 14 000 x g. The DNA pellet was washed up once with 70% ethanol, air-dried, and dissolved in a final volume of 100 µLof TE buffer.
In-house PCR Assay— Initially, the extracted DNA was tested for human β-globin gene to check that there were no PCR inhibitors in the samples. Primers β-GPCO (5'-ACACAACTGTGTTCACTAGC-3') and β-GPCPO (5'-GAAACCCAAGAGTCTTCTCT-3') were used to amplify a 209-bp fragment of the human β-globin gene (Vogels et al, 1993). Samples found to be β-globin negative by PCR were retested after dilution 10-fold in distilled water. Samples shown to be β-globin–positive were then examined for the presence of C trachomatis, U urealyticum, U parvum, M hominis, and M genitalium.
A forward primer, T1 (5'-GGACAAATCGTATTTCGG-3'), and a reverse primer, T2 (5'-GAAACCAACTCTACGCTG-3'), were used to amplify an approximately 517-bp region of the cryptic plasmid of C trachomatis (Claas et al, 1990).
A forward primer, My-ins (5'-GTAATACATAGGTCGCAAGCGTTATC-3'), and 2 reverse primers, MGSO-2-Bi (5'-CACCATCTGTCACTCTGTTAACCTC-3') and UGSO-Bi (5'-CACCACCTGTCATATTGTTAACCTC-3'), were used to amplify an approximately 520-bp region of the 16S rRNA gene of mycoplasmas and ureaplasmas (Yoshida et al, 2003).
The PCR mixture, which was made up to 50 µL with sterile water, contained 1x PCR buffer (50 mM Tris-HCl [pH 8.3], 10 mM KCl, 5.0 mM (NH4)2SO4, and 2.0 mM MgCl2); 0.5 µM each My-ins and MGSO-2-Bi for detection of genital mycoplasmas, 0.5 µM each My-ins and UGSO-Bi for detection of genital ureaplasmas, or 0.5 µM each T1 and T2 for detection of C trachomatis; 0.2 mM each dATP, dCTP, and dGTP; 0.6 mM dUTP; 1.25 U of Go Taq DNA polymerase (Promega, Lyon, France); and 10 µL of prepared DNA solution. PCR was performed using the Gene-Amp PCR System 9700 (PerkinElmer, Waltham, Mass) under the following conditions: an initial cycle at 95°C for 5 minutes, followed by 40 cycles of denaturation at 94°C for 30 seconds, annealing at 50°C (T1/T2) or at 55°C (My-ins/MGSO-2-Bi or My-ins/UGSO-Bi) for 30 seconds, and elongation at 72°C for 45 seconds, with a final cycle at 72°C for 7 minutes. Each run PCR included a positive control (M hominis PG21, U urealyticum, or C trachomatis genotype E) and 2 negative controls (previously negatively tested samples and distilled water). The PCR products were then subjected to hybridization assays.
Twenty semen specimens from infertile men were used as controls and were blindly re-examined. Twelve semen specimens were used as positive controls; 5 of these were previously shown to be C trachomatis–positive by Cobas Amplicor CT/NG PCR (Gdoura et al, 2001b), and 4 and 3 were previously shown to be U urealyticum–positive and M hominis–positive, respectively, by culture (Mycoplasma IST; bioMérieux, Lyon, France). Eight semen specimens negative for these 3 bacteria were used as negative controls.
Hybridization— Amplified products of 517 bp of C trachomatis and of 520 bp of U urealyticum, U parvum, M hominis, and M genitalium were detected by using a molecular hybridization technique in a liquid phase assay (Argene, Varilhes, France) with species-specific internal biotinylated probes (Claas et al, 1990; Yoshida et al, 2003):
T3 (5'-biotin-CGCAGCGCTAGAGGCCGGTCTATTTATGAT-3') specific for C trachomatis, Uure-P4-Am (5'-biotin-GGCTCGAACGAGTCGGTGT-3') specific for U urealyticum, Upar-P6-Am (5'-biotin-GTCTGCCTGAATGGGTCGGT-3') specific for U parvum, Mhom-P10-Am (5'-biotin-GACACTAGCAAACTAGAGTTAG-3') specific for M hominis, Mgen-P3-Am (5'-biotin-TCGGAGCGATCCCTTCGGT-3') specific for M genitalium.
The amplified products were denatured in a denaturing solution for 10 minutes and then immobilized to a microtiter plate (DNA-BIND 1 x 8 strip well plates; Argene) as described in the manufacturer's instructions. Species-specific capture probe diluted in hybridization solution was added to the wells of each microtiter plate immobilized by the denatured amplicons, and hybridization was carried out at 37°C for 30 minutes. Following hybridization, the wells were washed 4 times (soaking for 30 seconds) with a washing solution. One hundred microliters of ready-to-use conjugate were added per well and incubated for 15 minutes at room temperature. Wells were washed as described above. One hundred microliters of substrate (tetramethylbenzidine) were added per well and incubated for 30 minutes at room temperature in the darkness. The reaction was stopped by adding 100 µL of stop solution to each well, and the optical density (OD) at 450 nm was measured.
Samples with an OD value of greater than the cutoff value (OD of negative control + 0.15) + 10% of cutoff value were considered positive, as suggested by the manufacturer.
Statistical Analysis
All data were collected using standardized forms and were analyzed using
SPSS 13.0 software (SPSS Inc, Chicago, Ill).
To assess the agreement between the detection of C trachomatis, U
urealyticum, U parvum, M hominis, and M genitalium in semen and
corresponding FVU specimens, we used (nominal scale variables) as
proposed by Landis and Koch
(1977). Guidelines for the
interpretation of were as follows: < 0.20, poor agreement;
= 0.21–0.40, fair agreement; = 0.41–0.60, moderate
agreement; = 0.61–0.80, good agreement; =
0.81–1.00, very good agreement.
Age, seminal volume, sperm concentration, sperm viability, sperm motility, sperm morphology, and leukocyte counts are presented as mean values with standard deviation (SD).
Semen characteristics were compared among C trachomatis semen-positive and semen-negative groups, ureaplasma semen-positive and semen-negative groups and mycoplasma semen-positive and semen-negative groups.
The statistical significance of the differences between groups regarding age, seminal volume, sperm concentration, percentage of viability, percentage of progressive motility, percentage of rapid progressive motility, percentage of normal morphology, and leukocyte counts was assessed using the Mann-Whitney test and the Kruskal-Wallis nonparametric analysis of variance test. Fisher's exact test was used to test the statistical significance of the differences between groups regarding proportions of oligospermia, necrospermia, asthenospermia, teratospermia, and leukocytospermia.
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Results |
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TopAbstractPatients and MethodsResultsDiscussionReferences |
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= 0.901).
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Detection of Genital Ureaplasma DNA in Semen and Corresponding FVU Specimens by PCR
Genital ureaplasma DNA was detected in 19 (18.3%) of the semen or FVU
specimens. Genital ureaplasma–positive men had a median age of 39.1
years; this was not statistically different from the median age of the
negative men (36.6 years; P = .1).
U urealyticum DNA and U parvum DNA were detected in 16
(15.4%) and in 3 (2.9%) of semen and corresponding FVU specimens respectively
(Table 1). Thus, the
concordance between the detection of genital ureaplasma DNA in semen specimens
and corresponding FVU specimens of infertile men was 100%. A very good
agreement between the 2 specimens was found ( = 1).
Detection of Genital Mycoplasma DNA in Semen and Corresponding FVU Specimens by PCR
A total of 15 (14.4%) of the 104 patients were positive for genital
mycoplasmas in semen or FVU specimens. There was no significant difference
between the median age of men with or without genital mycoplasma infections
(37.4 years vs 37 years; P = .8).
M hominis DNA was detected in 10 (9.6%) of semen specimens and in
11 (10.6%) of corresponding FVU samples
(Table 1). Ten patients were
positive in both semen and FVU specimens and 1 was positive in the FVU
specimen only (Table 1). Thus,
the concordance between the detection of C trachomatis DNA in semen
specimens and the corresponding FVU of infertile men was 99%. A very good
agreement between the 2 specimens was found ( = 0.947).
M genitalium DNA were detected in 5 (4.8%) of both semen and
corresponding FVU specimens (Table
1). Four patients were positive in both FVU and semen specimens,
whereas 1 was positive in the semen specimen only, and 1 was positive in the
FVU specimen only (Table 1).
The concordance between the detection of M genitalium DNA in semen
specimens and the corresponding FVU of infertile men was 81.1%. Therefore, a
good agreement between the 2 specimens was found ( = 0.790).
C trachomatis, Genital Ureaplasma, and Genital Mycoplasma Infections and Semen Quality
A standard semen analysis showed that 10.6% (11/104) of specimens were
azoospermic. The remaining were asthenospermic (96.8%), teratospermic (91.4%),
oligospermic (32.3%), necrospermic (20.4%), and leukocytospermic (17.2%). The
prevalence of M genitalium DNA in semen samples was significantly
higher in azoospermic patients than in nonazoospermic patients (27.3% vs 2.1%
respectively, P = .008; Table
2).
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A summary of abnormalities in seminal fluid of nonazoospermic patients with C trachomatis, genital ureaplasma, or genital mycoplasma DNA in semen samples is shown in Table 2. Oligospermia, necrospermia, asthenospermia, teratospermia, and leukocytospermia were not significantly related to the presence of C trachomatis, genital ureaplasma, or genital mycoplasma DNA in semen samples.
The mean values of seminal volume, sperm concentration, sperm viability, total progressive sperm motility, rapid progressive sperm motility, sperm morphology, and leukocyte counts were significantly related neither to the detection of C trachomatis DNA nor to that of genital ureaplasma or mycoplasma DNA in semen specimens (Table 3).
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Discussion |
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TopAbstractPatients and MethodsResultsDiscussionReferences |
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Because asymptomatically infected individuals may shed fewer organisms (Witkin, 2002), and in order to make tests easier and with a higher specificity, nucleic acid amplification tests such as PCR may be the techniques of choice for C trachomatis, genital ureaplasma, and genital mycoplasma assessment in asymptomatic male partners.
The main purpose of the current study was to evaluate the relative performance of FVU compared with semen specimens for the detection of C trachomatis, U urealyticum, U parvum, M hominis, and M genitalium in asymptomatic male members of infertile couples. Thus, we used in-house PCR–microtiter plate hybridization assays that can facilitate the detection of C trachomatis and the identification of U urealyticum, U parvum, M hominis, and M genitalium in semen and FVU specimens. Initially, we tested extracted DNA of each semen and FVU samples for human β-globin gene to check that there were no PCR inhibitors in the samples. In a routine diagnostic setting, the internal processing control is important for providing a high sensitivity and specificity (Mahony et al, 1992), which is particularly important when dealing with sexually transmitted infections. In our study, known positive and negative control specimens were also blindly re-examined to confirm the sensitivity and specificity of the assays.
In this study, we found a very high concordance (>95%) and a very good
agreement ( > 0.8) between the detection of C trachomatis
DNA, genital ureaplasma DNA, and M hominis DNA in semen and
corresponding FVU specimens. A high concordance (81.1%) and a good agreement
( = 0.79) were also found between the detection of M
genitalium DNA in both specimens. Using our in-house PCR and sample
preparation methods, FVU was found to be a sensitive diagnostic specimen for
these pathogens. Several studies have shown that nucleic acid amplification
tests performed on FVU samples are able to detect as many or more infected
patients than traditional swabs from the urethra or cervix or from semen
(Chernesky et al, 1994;
Pannekoek et al, 2003;
Gaydos et al, 2004;
Hamdad-Daoudi et al, 2004;
Jensen et al, 2004). In some
cases, we found discrepancies between the detection of C trachomatis
DNA and genital mycoplasma DNA in semen and corresponding FVU specimens. The
presence of C trachomatis DNA and genital mycoplasma DNA in FVU
samples and its absence in semen specimens may indicate an asymptomatic
urethral infection. The detection of C trachomatis in 4 patients and
M genitalium in 1 patient only in semen may indicate that these
organisms are harboured in the epididymis or seminal vesicles.
Various studies conducted in industrialized countries proved the high prevalence of C trachomatis, genital ureaplasmas, and genital mycoplasmas among male partners of infertile couples and its important role in some cases in infertility (Upadhyaya et al, 1984; Paavonen and Wolner-Hassen, 1989; Vigil et al, 2002; Askienazy-Elbhar, 2005; Wang et al, 2006). Yet, in developing countries, the situation is not always clear.
Our study demonstrated that C trachomatis seems to be widespread among male partners of infertile couples in Tunisia, as shown by its high prevalence (43.3%). Our findings confirm our previous reports (Gdoura et al, 2001a,b) and are generally similar to those reported in studies conducted in developed countries (Vigil et al, 2002; Askienazy-Elbhar, 2005). Our results revealed also that genital mycoplasmas and genital ureaplasmas seem to affect a large number of infertile male patients, as disclosed by their rates of 18.3% and 14.4% respectively. These data are comparable with those reported in previous studies (Tully et al, 1981; Andrade-Rocha, 2003; Knox et al, 2003). U urealyticum was the most prevalent species detected (15.4%) in the current study. The prevalence of U urealyticum in the semen samples of male infertile patients in the literature varies from 5% to 42% (De Jong et al, 1990; Andrade-Rocha, 2003; Knox et al, 2003; Rosemond et al, 2006; Wang et al, 2006). This wide range might be explained by the diversity of detection methods used for characterizing the studied populations. Most of the previous reported studies have discussed the role of ureaplasmas in male infertility without discriminating between U urealyticum and U parvum (Naessens et al, 1986; Bornman et al, 1990; De Jong et al, 1990). In our study, we used the PCR–microtiter plate hybridization assay for facilitating the identification of U urealyticum, U parvum, M hominis, and M genitalium in semen specimens. By this method, U parvum was detected in 2.9% of semen samples. The prevalence of this species in our study was lower than that reported by Knox et al (2003) (2.9% vs 19.2%).
In the literature, M hominis has been associated with bacterial vaginosis, pelvic inflammatory disease, postpartum fever, and postabortal fever, as well as a number of gynecologic infections (Soffer et al, 1990; Yoshida et al, 2003). However, its role in nongonoccocal urethritis and in infertility is rarely investigated (Pannekoek et al, 2000). The prevalence of M hominis in our study was comparable to that reported by Andrade-Rocha (2003) but higher than that found by Rosemond et al (2006).
M genitalium is gaining increasing interest as a pathogen in male urethritis. Several studies (Björnelius et al, 2000; Totten et al, 2001) have shown a significant association between the presence of M genitalium DNA and symptoms and/or signs of urethritis (recently reviewed by Jensen et al, 2004). Hitherto, M genitalium has seldom been investigated in semen of infertile men. In our study, the prevalence of M genitalium in male partners of infertile couples was higher than that reported by Kjaergaard et al (1997) (4.8% vs 0.9%). This difference might be explained by the use of different methods for the detection of this bacterium. We have used PCR, which is more sensitive than a culture and facilitates the detection of M genitalium in clinical samples (Yoshida et al, 2003).
In addition to the determination of the prevalence of C trachomatis, genital mycoplasmas, and genital ureaplasmas in the male partners of infertile couples, we tried to evaluate the relationship between the detection of these pathogens in semen specimens and sperm quality.
Several lines of evidence suggest a role of C trachomatis in the etiology of male infertility. However, no direct link has been demonstrated (Witkin et al, 1993; Dieterle et al, 1995; Bollmann et al, 1998). In the current study, no relationship has been found between the detection of C trachomatis DNA in semen specimens and sperm abnormalities (concentration, sperm motility, sperm viability, sperm morphology, and leukocyte count). This result was in accordance with several studies (Gregoriou et al, 1989; Soffer et al, 1990; Dieterle et al, 1995; Eggert-Kruse et al, 1996, 1997; Weidner et al, 1996; Habermann and Krause, 1999; Gdoura et al, 2001b). However, some other studies have shown that infection is associated with poorer semen quality (Custo et al, 1989; Wolff et al, 1991; Witkin et al, 1995; Cengiz et al, 1997).
Reports are controversial about the effects of genital mycoplasma and ureaplasma infections or colonizations on semen andrology variables (Potts et al, 2000; Knox et al, 2003; Sanocka-Maciejewska et al, 2005; Wang et al, 2006). Previous studies have established that the presence of mycoplasma and ureaplasma in sperm specimens has no real effect on the semen quality (Soffer et al, 1990; Kjaergaard et al, 1997; Andrade-Rocha, 2003; Knox et al, 2003). Conversely, a relationship between U urealyticum and semen characteristics has been documented in literature (Potts et al, 2000; Sanocka-Maciejewska et al, 2005; Wang et al, 2006). In the present study, we failed to demonstrate any correlation between the abnormalities in seminal fluid and the detection of genital ureaplasma DNA or the detection of genital mycoplasma DNA in semen specimens. We found only a significantly higher prevalence of M genitalium in azoospermic patients than in nonazoospermic patients. Recent investigations seem to show that the presence of mycoplasmas reflects a silent colonization rather than an infection in infertile patients (Pannekoek et al, 2000), although the attachment and invasiveness of mycoplasmas towards human sperm cells has been demonstrated in vitro (Diaz-Garcia et al, 2006; Baczynska et al, 2007). These findings suggest that M genitalium may have an effect on the genital tract, leading to an inflammatory obstructive process, rather than on functional characteristics of semen.
In conclusion, using our in-house PCR and sample preparation methods, both semen and FVU were found to be sensitive diagnostic specimens for the detection of C trachomatis, genital ureaplasma, and genital mycoplasma DNA. The FVU, a less invasive and self-collected specimen, can be used as a marker for the presence of these organisms in the genital tract and as a reliable way of detecting asymptomatic carriers of infection.
The results of our study indicated that our in-house PCR–microtiter plate hybridization method provides a rapid and effective measure to detect human C trachomatis, genital mycoplasmas, and genital ureaplasmas, which is useful for etiologic and epidemiologic studies of these pathogens. Our results also demonstrated that C trachomatis, genital mycoplasmas, and genital ureaplasmas seem to be widespread among male partners of infertile couples in Tunisia, but no evidence of the effect of these pathogens on semen parameters was found.
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Acknowledgments |
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