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The prevalence of urogenital mycoplasmas in men with NGU in Upper Silesia (Poland) was studied. Mycoplasmas were detected in 36.7% men (Ureaplasma parvum and Mycoplasma genitalium were found in 30% and 16.7% respectively). Urealyticum urealyticum was not detected. We suggest including M. genitalium in the diagnostic scheme for nongonococcal urethritis (NGU).

Background: Periodic etiological surveillance of sexually transmitted infection (STI) syndromes is required to validate treatment algorithms used to control STIs. However, such surveys have not been performed in Zimbabwe over the past decade., Methods: A cross-sectional study design was used to determine the prevalence of the key STI etiological agents causing male urethral discharge (MUD). Urethral swab specimens were collected for molecular analysis and Neisseria gonorrhoeae isolation from consenting men 18 years and older who presented with MUD to the 12 clinics in Harare, Zimbabwe, between November 2010 and May 2011. A validated in-house multiplex polymerase chain reaction assay was used to detect the presence of N. gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and Mycoplasma genitalium. Gonococci were cultured on selective media, and antimicrobial susceptibilities were determined locally for ciprofloxacin, kanamycin, ceftriaxone, and cefixime using Etest strips, and minimum inhibitory concentrations were reported using defined breakpoints., Results: Among 130 participants, N. gonorrhoeae was the most frequent pathogen detected (106; 82.8%), followed by C. trachomatis (15; 11.7%), M. genitalium (6; 4.7%), and T. vaginalis (2; 1.6%). Four (6.1%) of the 66 gonococci isolated were resistant to fluoroquinolones, whereas all viable isolates were susceptible to kanamycin, cefixime, and ceftriaxone., Conclusions: Gonorrhea is the most important cause of MUD in men in Harare, and our study highlights the emergence of fluoroquinolone-resistant N. gonorrhoeae. Further STI surveys are required in other regions of Zimbabwe to obtain a nationally representative picture of gonococcal burden and antimicrobial resistance among MUD patients.

Background: Previous studies have identified Mycoplasma genitalium as a cause of urethritis in men. As there is no New Zealand data, a case-control study was conducted to determine whether this organism is a significant cause of urethritis in men presenting to Auckland Sexual Health Service., Methods: Enrollment for the study commenced in March 2006 and finished in February 2008. Inclusion criteria for cases of non-gonococcal urethritis were onset of urethritis symptoms within one month confirmed by urethral Gram staining showing >or=10 polymorphonuclear leucocytes per high-powered field. Controls were men presenting during the same time period for asymptomatic sexual health screening. All participants were tested for Neisseria gonorrhoeae, M. genitalium, and Chlamydia trachomatis. Information regarding symptoms, sexual behaviour and treatment was collected using a standard questionnaire., Results: We recruited 209 cases and 199 controls with a participation rate of 96%. The prevalence of C. trachomatis and M. genitalium in non-gonococcal urethritis cases was 33.5% and 10% respectively. Co-infection with these organisms was uncommon (1.9%). C. trachomatis and M. genitalium were diagnosed in 4% and 2% of controls, respectively, and both infections were detected significantly less often than in the cases (P < 0.0001, P < 0.005). Cases were more likely to report inconsistent condom use, multiple sexual contacts and not having sexual intercourse in the last week (P = 0.03, P = 0.02, P = 0.03). A past history of non-gonococcal urethritis was a significant predictor of current symptoms (P < 0.0001)., Conclusions: This is the first study to investigate M. genitalium infection in New Zealand men. Our results confirm that M. genitalium is a cause of non-gonococcal urethritis in men presenting to our service.

Mycoplasmas comprise a big group of organisms consisting of one hundred eighty species which are found in nature as parasites of humans, other mammals, reptiles, fishes and plants, or living as commensals. The group commonly referred to as genital mycoplasmas comprise species most often found in the genitourinary tract of sexually active adults as common commensal inhabitants, or pathogens which can cause many different infections. The species we are most interested in this work are called: Mycoplasma genitalium, Mycoplasma hominis and Ureaplasma urealyticum. There is no doubt that they can cause non-gonococcal urethritis (NGU) in both men and women and bacterial vaginosis (BV), cervicitis, endometritis. Infection can spread to the upper port of female genital tract and it can lead to pelvic inflammatory disease, or if it happens during pregnancy--to chorioamnionitis and further pregnancy complications. Even though mycoplasmas have been known and described since 1898, the problem of their morbidity and the possible influence they have on human fertility is still not clear. Similar to research from 30 years ago, connecting Chlamydia trachomatis with infertility, new scientific work as well as the dynamic development of diagnostics procedure, especially more common use of PCR method, may be helpful in discovering the potential role genital mycoplasmas play in infertility.

Detection and quantification of Mycoplasma genitalium were evaluated in 83 patients with urethritis (group 1), 60 patients with urethral symptoms but no urethritis (group 2), and 50 asymptomatic men (group 3). Quantification of M. genitalium was carried out using real-time polymerase chain reaction (PCR) analysis of first-pass urine samples. The rate of detection of M. genitalium was significantly higher in group 1 than in groups 2 and 3 (P<.0001). The mean observed concentration of M. genitalium was 1.2x10(4) equivalent genomes/mL of urine (range, 50 to 8x10(4) equivalent genomes/mL). Analysis of M. genitalium load in serial urine samples collected before and after the administration of antibacterial treatment showed an association between clinical and microbiological responses, with a shift to negative PCR results in symptom-free patients. Our results illustrate the usefulness of monitoring the M. genitalium load in evaluating the susceptibility of M. genitalium to antibacterial treatment.

In order to determine the colonisation patterns of several Mycoplasma species in homosexual men, urethral, oral and rectal specimens from 10 homosexual men with acute non-gonococcal urethritis (NGU) and 18 without NGU were examined using sensitive methods. Mycoplasma hominis and Ureaplasma urealyticum existed in both groups, which is in keeping with previous studies of heterosexual men. Mycoplasma genitalium was detected in the rectum of both NGU-positive and NGU-negative men and in the urethra of one man with chlamydia-negative NGU, but not in those without urethritis. Mycoplasma fermentans was found in the throat and rectum only and Mycoplasma penetrans in all three anatomical sites. In contrast, Mycoplasma pirum was found in the rectum only, that is, in 5 of the 28 men studied. Infrequent examination of this site is a possible explanation for previous failures to detect Mycoplasma pirum at a mucosal surface.

Aims: To design and validate a polymerase chain reaction (PCR) assay targeting the 16S rRNA gene of Mycoplasma genitalium., Methods: Primers were designed that were complementary to the 16S rRNA gene sequence of M genitalium. After optimisation of the reaction conditions, the PCR was tested against nine M genitalium strains, a dilution series of M genitalium DNA, and a panel of common microorganisms. The PCR was also challenged in parallel with a published assay against 54 urine specimens from men with urethritis., Results: The expected 341 bp product was produced on amplification of material from all M genitalium strains and from none of the other microorganisms tested. The lower limit of detection was 50 genome copies. The new assay detected M genitalium DNA in nine of 54 men with urethritis, in comparison with eight positive specimens detected with the alternative PCR., Conclusions: This novel PCR targeting the M genitalium 16S rRNA gene has been optimised and now provides a sensitive and specific alternative or addition to the available MgPa gene targeting assays.

The aetiological importance of Chlamydia (C.) trachomatis in non-gonococcal urethritis (NGU) is undisputed. Mycoplasma (M.) genitalium has been shown to be strongly associated with NGU and with mucopurulent cervicitis and also with acute endometritis independent of C. trachomatis. In this prevalence study we examined 946 patients, 445 women and 501 men, attending the STD clinic, for M. genitalium as well as C. trachomatis and Neisseria gonorrhoeae. M. genitalium was detected in urethral samples from 17 (13.4 per cent) of 127 men with both symptoms and signs of urethritis and from 2 (1.3 per cent) of 160 men without (p < 0.001). Corresponding figures for M. genitalium in the women were 15 (11 per cent) of 136 women with symptoms and signs of urethritis or cervicitis compared to 3 (2.2 per cent) of 139 women without (p = 0.005). Examinations of partners of female and male index patients indicated that M. genitalium is sexually transmitted. Some M. genitalium infected patients had a history of irregular vaginal bleeding, lower genital tract pain, epididymitis and arthritis. Investigation of the aetiological role of M. genitalium in salpingitis, epididymitis and sexually acquired arthritis is urgently needed.

We report the results of a study of Mycoplasma genitalium (detected with a modified polymerase chain reaction [PCR] assay) in men with urethritis and in asymptomatic control subjects at a sexually transmitted diseases clinic in New Orleans. Data for 97 men with urethritis and 184 asymptomatic men were available for analysis. M. genitalium infection rates in symptomatic and asymptomatic men who were negative for Chlamydia trachomatis and Neisseria gonorrhoeae were 25% and 7%, respectively (P=.006). M. genitalium coinfection rates among men with chlamydial and gonococcal urethritis were 35% and 14%, respectively. Men with M. genitalium urethritis resembled those with C. trachomatis in that both groups were younger and more likely to experience milder urethral symptoms. Among men with urethritis, the sensitivities of PCR of urine and swab specimens for the detection of M. genitalium were 87% and 91%, respectively. M. genitalium is associated with nongonococcal urethritis in this population.

By using a TaqMan assay we monitored longitudinal changes in Mycoplasma genitalium loads in five men with recurrent M. genitalium-positive nongonococcal urethritis. We observed regrowth of M. genitalium persisting in hosts after treatment and a possible association of the increase in the M. genitalium load with emergence of symptoms and signs of nongonococcal urethritis in four of these patients.

The occurrence of Neisseria gonorrhoeae, Chlamydia trachomatis and Mycoplasma genitalium was determined by molecular techniques in urine specimens from 182 black South African men who had symptoms and/or overt signs of urethritis. Eighty-six (47.3%) of these men were infected with N. gonorrhoeae. There were 185 men without overt evidence of urethritis, 16 (8.6%) of whom were also infected with N. gonorrhoeae. Of the 96 men who had non-gonococcal urethritis, 14 (14.6%) were infected with C. trachomatis, 16 (16.7%) with M. genitalium and only one with both microorganisms. In comparison, 15 (8.9%) of 169 men without overt urethritis and without N. gonorrhoeae were infected with C. trachomatis and 15 (8.9%) with M. genitalium, proportions that were about half the size of those in the group with overt urethritis.

We developed a TaqMan-based real-time PCR assay for quantifying Mycoplasma genitalium. This assay is able to specifically quantify concentrations of the M. genitalium 16S rRNA gene ranging from 10(7) to 10 copies/reaction. Using the TaqMan assay, we quantified the M. genitalium 16S rRNA gene in first-pass urine of men with urethritis and asymptomatic men who were positive for M. genitalium by PCR- and phylogeny-based assay. Of 130 men with gonococcal urethritis (GU), five were positive for M. genitalium. The mycoplasma load for each specimen was <5 x 10 copies/ml. Of 84 men with chlamydial non-GU (CNGU), seven were positive for M. genitalium. One man had an M. genitalium load of <5 x 10 copies/ml, and six men had loads ranging from 1.1 x 10(7) to 2.7 x 10(2) copies/ml. Of 86 men with nonchlamydial NGU (NCNGU), 17 were positive for M. genitalium. The mycoplasma loads for these men ranged from 3.3 x 10(6) to 2.3 x 10(2) copies/ml. Of 76 asymptomatic men, only two were positive for M. genitalium. For these men, the loads were 2 x 10(2) and <5 x 10 copies/ml. The patients with NGU had significantly higher concentrations of M. genitalium in their first-pass urine than did men with GU (P < 0.01) or asymptomatic men (P < 0.05). In addition, M. genitalium loads were significantly higher in men with NCNGU than those in asymptomatic men (P < 0.05). The quantitative assessment of M. genitalium loads by the TaqMan assay will provide useful information for understanding the pathogenicity of this mycoplasma in the urogenital tract.

Purpose: We reviewed findings on the pathogenic role of Mycoplasma genitalium in nongonococcal urethritis and the treatment of men with M. genitalium positive nongonococcal urethritis., Materials and Methods: We reviewed literature selected from peer reviewed journals listed in MEDLINE and from resources cited in those articles from 1967 to January 2001., Results: M. genitalium was first isolated from 2 men with nongonococcal urethritis and thereafter it was shown to cause urethritis in subhuman primates inoculated intraurethrally. This mycoplasma has been detected significantly more often in patients with acute nongonococcal urethritis, particularly in those with nonchlamydial nongonococcal urethritis, than in those without urethritis. The prevalence of M. genitalium positive nonchlamydial nongonococcal urethritis is 18.4% to 45.5% of all nonchlamydial nongonococcal urethritis cases. In addition, the persistence of M. genitalium in the urethra after antimicrobial chemotherapy is associated with persistent or recurrent nongonococcal urethritis. M. genitalium is highly susceptible to tetracycline, macrolide and some new fluoroquinolones. The regimen of 100 mg. doxycycline orally twice daily for 7 days, which is recommended for chlamydial nongonococcal urethritis, seems to be effective for M. genitalium positive nongonococcal urethritis, although clinical data to substantiate this regimen are limited., Conclusions: The various results reported to date tend to support the proposition that M. genitalium is a pathogen of nongonococcal urethritis. However, currently diagnostic methods for this important mycoplasma are not available in clinical practice. Because of the possible association of the posttreatment presence of M. genitalium in the urethra with persistent or recurrent nongonococcal urethritis, eradication of this mycoplasma from the urethra is essential for managing M. genitalium positive disease. However, clinical data on treating M. genitalium positive nongonococcal urethritis are extremely limited. Thus, further studies are required to develop new diagnostic methods that would be available in clinical settings and establish a new treatment algorithm for nongonococcal urethritis, including M. genitalium positive disease.

Mycoplasmas are the smallest free-living organisms, widespread in nature. Several mycoplasma species have been isolated from humans. For 6 of them: Mycoplasma hominis, Ureaplasma urealyticum, M. primatum, M. genitalium, M. spermatophilum and M. penetrans, the genital tract is the main site of colonization. This review is concentrated on the role of mycoplasmas as sexually transmitted agents, with the emphasis to M. genitalium infections. M. hominis and U. urealyticum are isolated from the genital tract of healthy men and women with considerable frequency. The biological features (attachment properties, possible intracellular location) and experimental inoculation studies of M. genitalium indicate that this mycoplasma has pathogenic potential. Data from case-control studies, looking at men with non-gonococcal urethritis and women with cervicitis, have revealed that M. genitalium behave similarly to Chlamydia trachomatis and have revealed that carriage of M. genitalium and C. trachomatis is usually independent of one another. M. genitalium could be considered as a potential cause of sexually transmitted urethritis in men, including men with persistent or recurrent urethritis. More studies are expected to ascertain the role of M. genitalium in the female genital tract. Evidence-based data are needed to decide whether current non-gonococcal infection treatment principles are applicable or not for M. genitalium infections.

Background: Tip-mediated cytadherence in Mycoplasma genitalium requires the structural and functional stability of the P140 adhesin, its operon-related protein P110, and the high molecular weight protein MG218 (190-kDa). Disruption mutants of mg218 unable to express MG218 exhibit both a non-cytadhering phenotype and P140/P110 instability, while disruption mutants that synthesize a truncated MG218 (160 kDa) retain the stability of P140/P110 and are >95% cytadhering. However, the origin of the MG218 truncated protein in these mutants is unclear. Therefore, we attempted to identify the origin of the truncated MG218 protein and to evaluate whether this truncated protein possessed the C-terminal part of MG218. In addition, we used spontaneous mutants lacking P140 to assess the role of MG218 in the stability of P140/P110., Methods: RNA from M. genitalium mutant producing truncated MG218 was subjected to primer extension analysis to identify the origin of expression of truncated MG218. Extracts of this mutant were examined for the presence of the C-terminal region of MG218 by immunoblot. In addition, pulse-chase analysis was performed to assess the role of MG218 in the stability of P140/P110 in spontaneous p140 mutants., Results: Primer extension analysis identified a transcriptional start point adjacent to the gentamycin-resistance gene in disrupted mg218 mutants. Antibodies directed against the C-terminal region (amino acids 1651-1666) of MG218 bound to truncated MG218 protein from mutants. Spontaneous p140 mutants subjected to pulse chase analysis indicated that solely class I mutants exhibited instability of P140/P110 in the presence of intact MG218., Conclusions: Expression of truncated MG218 in M. genitalium mg218 mutants appears to be due to the presence of a putative promoter upstream to the point of mg218 disruption; this truncated protein possesses the C-terminal region of MG218. However, pulse chase results from spontaneously arising, non-cytadhering P140-deficient M. genitalium mutants suggest that the stability of P140 and P110 requires not only MG218 but also additional factors.

Some strains of mycoplasmas and ureaplasmas (family Mycoplasmataceae) are associated with nongonococcal urethritis (NGU) or other genitourinary infections. We have developed a rapid and reliable method of identifying the presence and prevalence of mycoplasmas and ureaplasmas in men with NGU. This method is based on the amplification of a part of the 16S rRNA gene by PCR and phylogenetic analysis. A portion of the 16S rRNA gene from 15 prototype strains was amplified with a set of common primers, and their nucleotides were sequenced. The nucleotide sequence of the V4 and V5 regions was analyzed by the neighbor-joining method. The 15 prototype strains were grouped into three distinct clusters, allowing us to clearly segregate the strains into distinct lineages. To determine the prevalence of these pathogens among patients with NGU, this protocol was tested with 148 urine samples. Amplifications were observed for 42 samples, and their nucleotide sequences were analyzed along with those of the 15 prototype strains. The phylogenetic tree thus constructed indicated that 15 of the 42 formed a cluster with Mycoplasma genitalium. Among the remaining specimens, 2 formed a cluster with Mycoplasma hominis, 19 with Ureaplasma urealyticum, and 5 with Ureaplasma parvum; the remaining sample contained both M. genitalium and U. urealyticum. This phylogeny-based identification of mycoplasmas and ureaplasmas provides not only a powerful tool for rapid diagnosis but also the basis for etiological studies of these pathogens.

Background: Most patients with recurrent symptomatic nongonococcal urethritis receive negative test results for Chlamydia trachomatis and Ureaplasma urealyticum, and the cause of such recurrence usually is unknown., Goal: To assess the association of Mycoplasma genitalium with recurrent nongonococcal urethritis., Study Design: In this study, 72 men with nongonococcal urethritis were treated with levofloxacin. Before and after treatment, symptoms and signs were assessed and first-pass urine was examined for C trachomatis, M genitalium, U urealyticum, and Mycoplasma hominis by polymerase chain reaction-based assays., Results: In 6 of 45 men who had no symptoms and no evidence of inflammation after treatment, nongonococcal urethritis recurred. Of these 6 men, 5 had positive test results for M genitalium before levofloxacin treatment, which remained positive afterward. After the second treatment for recurrent nongonococcal urethritis, one man was still had a positive test result for the mycoplasma and experienced a subsequent recurrence., Conclusions: This study suggests that the persistence of M genitalium in the urethra may be associated with recurrence of nongonococcal urethritis.

Chlamydia trachomatis and Neisseria gonorrhoeae are universally acknowledged as urethral pathogens, yet the etiology in the majority of cases of urethritis is unclear. Our case-control study assessed the association of Mycoplasma genitalium, Ureaplasma urealyticum, and other potential pathogens with acute nongonococcal urethritis (NGU) in heterosexual men presenting to an urban sexually transmitted diseases clinic. M. genitalium was detected in 27 (22%) of 121 NGU case patients and in 5 (4%) of 117 control subjects (P<.01). Although C. trachomatis was detected in 36 (30%) of 121 NGU case patients and in 4 (3%) of 117 control subjects (P<.01), only 3 men with NGU were infected with both C. trachomatis and M. genitalium. U. urealyticum was not associated with NGU. By multivariate analyses, controlling for age, race, history of prior urethritis, and chlamydial infection, M. genitalium was associated with a 6.5-fold increased risk of urethritis (95% confidence interval, 2.1-19.5), which supports a role of this organism in the etiology of NGU.

Objective: To determine the etiological role of pathogens other than Neisseria gonorrhoeae and Chlamydia trachomatis in urethral discharge in West African men., Methods: Urethral swabs were obtained from 659 male patients presenting with urethral discharge in 72 primary health care facilities in seven West African countries, and in 339 controls presenting for complaints unrelated to the genitourinary tract. Polymerase chain reaction analysis was used to detect the presence of N. gonorrhoeae, C. trachomatis, Trichomonas vaginalis, Mycoplasma genitalium, and Ureaplasma urealyticum., Findings: N. gonorrhoeae, T. vaginalis, C. trachomatis, and M. genitalium--but not U. urealyticum--were found more frequently in men with urethral discharge than in asymptomatic controls, being present in 61.9%, 13.8%, 13.4% and 10.0%, respectively, of cases of urethral discharge. Multiple infections were common. Among patients with gonococcal infection, T. vaginalis was as frequent a coinfection as C. trachomatis. M. genitalium, T. vaginalis, and C. trachomatis caused a similar clinical syndrome to that associated with gonococcal infection, but with a less severe urethral discharge., Conclusions: M. genitalium and T. vaginalis are important etiological agents of urethral discharge in West Africa. The frequent occurrence of multiple infections with any combination of four pathogens strongly supports the syndromic approach. The optimal use of metronidazole in flowcharts for the syndromic management of urethral discharge needs to be explored in therapeutic trials.

Two hundred and thirty-three men and 85 women visiting STD clinics in western Sweden between April 1997 and March 1998 were examined for Mycoplasma genitalium and Chlamydia trachomatis. The bacteria were identified by the polymerase chain reaction (PCR) technique. Three women (3.5%) and 18 men (7%) were positive for M. genitalium. Seventeen (14%) of the 115 men with urethritis were infected but only one of the men was without urethritis. After treatment with tetracyclines for 10 days, one woman and 8 of the 13 men still harboured M. genitalium. M. genitalium-infected men did not have more life-time partners than other men visiting STD clinics. More men positive for M. genitalium gave a history of previous urethritis but the difference was not significant.

Urethral swab specimens obtained from 101 men attending an STD clinic were examined for the presence of Mycoplasma genitalium by polymerase chain reaction (PCR) amplification. Fifty patients had non-gonococcal urethritis (NGU), and 51 patients were included as controls without urethritis. M. genitalium DNA was detected in 13 (26%) of the urethritis patients and in 5 (10%) of the control patients (P=0.06). No patient positive for M. genitalium had a simultaneous chlamydial infection. Thus, in the 36 patients with non-chlamydial NGU, the prevalence of M. genitalium infection was 36% (P=0.007 compared with controls). All patients with M. genitalium positive urethritis had a high grade urethritis defined as >10 polymorphonuclear cells per high power microscopical field. Compared with the control group, those with M. genitalium positive urethritis had more often had a history of urethritis than had those with chlamydial NGU or those with M. genitalium negative, non-chlamydial NGU.

Background: Mycoplasma genitalium is regarded as a potential pathogen of the human urogenital tract based on prevalence findings of several European studies., Goal: To determine the prevalence of M genitalium in urethral specimens of symptomatic patients with nongonococcal urethritis and from asymptomatic patients attending a sexually transmitted disease clinic in Milan, and to verify the clinical efficacy of M genitalium eradication by antibiotic treatment., Study Design: From May 1998 to late April 1999, a routine analysis for M genitalium by DNA amplification (polymerase chain reaction) was performed in patients attending the Institute of Dermatological Science in Milan. The authors examined urethral swabs from 178 symptomatic and 23 asymptomatic males. M genitalium-positive patients were clinically and microbiologically tested after treatment with either doxycycline or azithromycin., Results: Among males with nongonococcal urethritis, M genitalium was detected in 14.0% of patients as the only agent; in 15.1% of patients in association with Chlamydia trachomatis and/or Ureaplasma urealyticum; and in 1 asymptomatic patient. In all symptomatic M genitalium-positive patients, antibiotic treatment eradicated the infection and cured clinical symptoms., Conclusion: These data reveal the high prevalence of M genitalium in symptomatic patients, the rarity of asymptomatic carriers, the high susceptibility to antibiotic treatment, and the clinical efficacy of M genitalium eradication. Moreover, data confirm the etiologic role of M genitalium in inflammatory processes of the human urogenital tract in the Mediterranean area.

Purpose: We attempted to detect Mycoplasma genitalium in urethral swab specimens by a polymerase chain reaction based assay to determine the prevalence of M. genitalium in patients with urethritis., Materials and Methods: We examined a total of 171 Japanese men who presented to our hospital from February 1995 through January 1997. Of these men 150 had symptoms and signs compatible with acute urethritis and 21 had no symptoms or signs of urethritis. Urethral swab specimens were used to culture Neisseria gonorrhoeae, to detect Chlamydia trachomatis by an enzyme immunoassay and to detect M. genitalium by a polymerase chain reaction based assay., Results: Gonococcal urethritis was diagnosed in 74 symptomatic men, and nongonococcal urethritis was diagnosed in 76 symptomatic men. Of the 74 cases of gonococcal urethritis 3 (4.1%) were positive for M. genitalium, and 14 (18.9%) were positive for C. trachomatis. Of the 76 cases of nongonococcal urethritis 10 (13.2%) were positive for M. genitalium, and 42 (55.2%) were positive for C. trachomatis. While only 1 of the 42 cases with chlamydial nongonococcal urethritis (2.4%) was positive for M. genitalium, 9 of the 34 chlamydia negative nongonococcal urethritis cases (26.5%) were positive for the mycoplasma. In contrast, all 21 cases men were negative for N. gonorrhoeae, M. genitalium, and C. trachomatis., Conclusions: The prevalences of M. genitalium in patients with gonococcal urethritis and nongonococcal urethritis who attended our clinic were 4.1 and 13.2%, respectively. M. genitalium was detected significantly more often in men with nongonococcal urethritis than in asymptomatic men. In addition, its prevalence in men with chlamydia negative nongonococcal urethritis (26.5%) was significantly greater than in those with chlamydia positive nongonococcal urethritis (2.4%). These findings suggest that M. genitalium may be associated with the development of nongonococcal urethritis independent of C. trachomatis.

The use of PCR assays as a fast and reliable method is constantly improving and easing microbiological diagnosis. We used a polymerase chain reaction (PCR) assay designed to detect Mycoplasma genitalium and Chlamydia trachomatis in urethral swab samples of 56 males with urethritis and 44 asymptomatic patients as a control group. The PCR assay provides an amplification of target sequence within MgPa (M. genitalium protein attachment) gene. Results indicated that M. genitalium was present in 6 (10.7%) patients with urethritis and none in the control group. Eleven of 56 (17.8%) patients were positive for Chlamydia trachomatis when tested by an outer membrane protein primer-based PCR. The amplified DNA fragments were homogeneous as shown by restriction enzyme analysis and found to be consistent with the published sequences. The PCR assay employed was as reliable as the cultural method in detecting C. trachomatis in the urethral swabs of patients with urethritis (100% of sensitivity when compared with the cultural method) and it has been revealed as an essential method for detection of M. genitalium.

Urethral swab specimens collected from 108 male Japanese patients with acute nongonococcal urethritis (NGU) and from 50 Japanese men without NGU were examined for the presence of Mycoplasma fermentans, Mycoplasma penetrans, and Mycoplasma pirum by means of polymerase chain reaction-based assays. These mycoplasmas were not detected in any of the specimens, which suggests that they are unlikely to have a pathogenic role in acute NGU.

Mycoplasma genitalium is a human mycoplasma species which, on the basis of detection by PCR, has been incriminated as a cause of nongonococcal urethritis. Previously, only two strains from the urogenital tract and five strains from extragenital sites have been isolated. We have developed a method for the isolation of this fastidious microbe. M. genitalium from PCR-positive urethral specimens was initially propagated in Vero cell cultures grown in serum-free medium supplemented with Ultroser HY serum substitute. Growth was monitored by PCR. The M. genitalium strains grown in cell cultures could subsequently be subcultured in modified Friis's FF broth medium. Several passages in broth medium were required before growth on agar medium was attained. A total of 11 urethral specimens positive for M. genitalium by PCR from male patients with urethritis were investigated. Six strains were adapted to growth in broth medium, and four of these strains were cloned. Three specimens were overgrown by other mycoplasmas during propagation in the cell cultures. In only two PCR-positive specimens was propagation of M. genitalium unsuccessful. The use of cell culture combined with PCR monitoring of mycoplasmal growth may prove to be more widely applicable for the isolation of other fastidious mollicutes.

A polymerase chain reaction (PCR)-based assay was developed for the detection of Mycoplasma hominis. This assay generates a 152-bp PCR product which was part of an initial 471-bp M. hominis genomic DNA fragment. The 471-bp DNA fragment was shown by hybridization analysis to be unique for M. hominis. The PCR assay can amplify as few as 18 molecules of target DNA. This diagnostic assay offers potential for wide clinical application as it is rapid and can be successfully performed on crude sample preparations from a variety of media or biopsies. The use of this assay should aid in defining the aetiologic and pathologic roles played by M. hominis and thereby benefit patients.

Background: To compare and evaluate two commercial methods for Mycoplasma hominis and Ureaplasma urealyticum with the isolation in agar A7 medium., Method: One hundred and twenty three vaginal and cervical swabs in women prostitutes, and urethral swabs in men, were studied for Mycoplasma hominis and Ureaplasma urealyticum isolation in agar A7 medium (BioMérieux; France), Mycoplasma IST (BioMérieux, France) and MycoFast ALL-IN (Unipath, U.K.)., Results: Of all 123 samples studied, it was isolated 24 Mycoplasma hominis (19.5%) and 56 Ureaplasma urealyticum (45.5%) in agar A7. The sensitivity was 100% for Mycoplasma hominis and Ureaplasma urealyticum in Mycoplasma IST versus 29.2% and 50% in MycoFast ALL-IN, both respectively. Specificity was lower for Ureaplasma urealyticum (59.7%) in Mycoplasma IST than in MycoFast ALL-IN (100%). It was found a tetracycline resistance of 15% and 12.5% for Ureaplasma urealyticum and Mycoplasma hominis, both respectively., Conclusions: The use of commercial methods permits to count the genital mycoplasmas, being Mycoplasma IST better than MycoFast ALL-IN for Mycoplasma hominis isolation. Both methods have limitations for Ureaplasma urealyticum and they require the use of agar A7. Mycoplasma IST could be a good method to survey tetracycline resistance.

Mycoplasma genitalium causes urethritis in non-human primates, but studies on its pathogenicity in man have been hampered by the difficulty in isolating this oragnism in culture. We have used a specific polymerase chain reaction to examine the role of M. genitalium in non-gonococcal urethritis (NGU). Oligonucleotide primers were used to amplify a 281 bp of 140-KDa adhesin gene of M. genitalium. A characteristic PCR product was amplified, when M. genitalium DNA was template for the PCR. No amplified product was detected in Mycoplasma pneumoniae DNA, Mycoplasma hominis DNA or other bacterial DNAs. M. genitalium DNA was detected in urethral swabs from 17 (14.9%) of 114 men with NGU. Three (9.1%) of the 33 men with Chlamydia-positive NGU and 14 (17.3%) of the 81 with Chlamydia-negative NGU were positive for M. genitalium DNA, but 29 men without urethritis were negative. The prevalence of M. genitalium in NGU and in Chlamydia-negative NGU was significantly higher than that in the normal control. These findings suggest that M. genitalium would be a cause of NGU.

The original two strains of Mycoplasma genitalium were isolated from the human urogenital tract. No other strains have been isolated from this site since then. We have recently succeeded in propagating a third strain from a urogenital specimen from a patient with urethritis in Vero cell cultures. By electron microscopy mycoplasmas were demonstrated intracellularly in about 10% of the examined Vero cells. Various stages of penetration into the cells could be observed. The flask-shaped organisms seemed to penetrate into the cells by the tip-end which included a rodlike structure. The intracellular location of normal mycoplasmas were in membrane-bound vacuoles very close to the nucleus, occasionally together with a few disintegrated organisms. In a few cells additional material was entangling the mycoplasmas in the cytoplasmic vacuoles. The potential for intracellular survival of M. genitalium may help the organism to evade the defence mechanisms of the human body. This trait may be considered a pathogenic property which supports the presumption that M. genitalium has clinical importance.

Background: Male urethritis may be caused by mycoplasmas. Since Mycoplasma genitalium has previously been isolated from the urethra of two men with non-gonococcal urethritis (NGU), it was the aim of the study further to elucidate its role by measuring the prevalence of this organism in men with NGU., Material and Methods: The polymerase chain reaction was used. Two different sequences of the gene coding for the main adhesin MgPa were amplified. Urethral, rectal, and throat samples from 99 male sexually transmitted disease (STD) patients with and without urethritis were studied., Results: M genitalium DNA was demonstrated in 17/99 (17%) of the urethral swabs, but in none of the rectal and throat swabs. Significantly more patients with urethritis (13/52) were positive for M genitalium DNA than were patients without urethritis (4/47) (p < 0.03). In those with urethritis M genitalium DNA was found more often in Chlamydia trachomatis negative NGU (12/34) than in those with chlamydial NGU (1/14) (p = 0.05). Attempts to culture M genitalium from the PCR positive specimens were unsuccessful., Conclusion: M genitalium DNA was found significantly more often in male STD patients with non-chlamydial NGU than in men with chlamydial urethritis (p = 0.05) and in men without urethritis (p = 0.003), suggesting that M genitalium may be a cause of NGU. M genitalium DNA was not demonstrated in any of the throat or rectal swabsindicating that the urogenital tract is probably the primary site of infection or colonisation of this species.

The incidence of Chlamydia trachomatis, Ureaplasma urealyticum, Mycoplasma hominis, Neisseria gonorrhoeae and Trichomonas vaginalis was studied in men with urethritis. Out of 150 examined men 48.7% had the positive isolation of U. urealyticum, 26.0% C. trachomatis, 22.7% N. gonorrhoeae, 18.7% M. hominis and in one (0.7%) patient T. vaginalis was found. None of the above mentioned microorganisms was detected in 24.7% of examined men. In 43.3% cases only one agent was isolated. In 23.3% of the men the combination of two agents, in 8.0% the combination of three and in 0.7% even the combination of four studied microorganisms was observed. C. trachomatis was most frequently observed in combination with N. gonorrhoeae (15 cases) and U. urealyticum (14 cases). M. hominis and U. urealyticum occurred simultaneously in 22 examined men.N.gonorrhoeae was most frequently found together with U. urealyticum (16 cases). Concerning the occurrence of other bacteria and yeasts, no significant difference was found between the groups positive and negative for the above mentioned microorganisms.

The efficacy of ofloxacin, a new quinolone derivate, was tested against that of erythromycin in a prospective double-blind trial in patients with non-gonococcal urethritis (NGU) with special reference to the occurrence of Mycoplasma hominis and Ureaplasma urealyticum. 188 male NGU patients were randomized to treatment with either ofloxacin 200 mg b.i.d. or erythromycin 500 mg b.i.d. for seven days. Before treatment eight (4.3%) patients, five in the erythromycin group and three in the ofloxacin group, were M. hominis positive. At follow-up day 8 and 15 after start of treatment all five in the erythromycin group and two in the ofloxacin group were still positive. U. urealyticum was recovered in 16 patients (8.5%) before treatment. One patient was still positive in the erythromycin group when examined day 15, whereas all patients were negative in the ofloxacin group at both follow-up controls. Clinically, the efficacy of treatment day 15 was 77.4% in the erythromycin group and 84.3% in the ofloxacin group. The difference was not significant. Side-effects occurred in 38.5% in the erythromycin group and in 21.3% in the ofloxacin group. This difference is significant. Ofloxacin is effective in the treatment of NGU in males and is an alternative to conventional antibiotic treatment.

162 women were investigated. Group I consisted of 85 women, who were partners to men with non-gonococcal urethritis (NGU) or presented macroscopic signs of cervicitis; patients who had harbored Neisseria gonorrhoeae were excluded from the study. Group II was a control group of 77 women without any complaints from the urogenital tract and with normal findings at pelvic examination. All the women were tested for infection with Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, and Candida albicans. In group I, chlamydiae and mycoplasmas were recovered in 44% and 36%, respectively, the corresponding figures for the control group (group II) being 5% and 19%. The difference is highly significant. No such difference between the two groups was found for ureaplasmas. Sixteen percent of the patients in group I were positive for C. albicans; 12% were positive in group II. Fifty per cent of asymptomatic NGU-partners were chlamydia-positive, and about one-third of patients with either dysuria or vaginal discharge harbored the organism. No difference in the isolation frequency of mycoplasmas was observed between asymptomatic partners to male NGU carriers and women with increased vaginal discharge, whereas the organism was isolated more frequently from patients with dysuria. Fifty-nine per cent of patients with cervicitis were chlamydia-positive, compared with 30% of patients with normal cervical appearance and normal vaginal discharge. Samples obtained from the cervix were more often positive than samples from the urethra. In conclusion, if samples can be taken from only one of the two sites in patients with lower genital tract infection, the cervix is the optimal sampling site.

Today, 12 years after the identification of Eaton agent as a mycoplasma, much new information on this infection has accumulated. Better methods have become available, which will be helpful in determining whether other mycoplasmas might be human pathogens, e.g., some ureaplasmas in nongonococcal urethritis. Experimental mycoplasma infections in birds and animals have revealed common features with immune complex diseases in man. They may serve, therefore, as useful models for human mycoplasma disease. However, the most important finding in recent mycoplasma research has been the isolation of mycoplasma viruses. Viruses belonging to three different groups have been discovered. It will be interesting to study whether the virus determines the pathogenicity of mycoplasmas or not.

A new mycoplasma, serologically distinct from all other known mycoplasmas, was isolated from urethral specimens from two of thirteen men with non-gonococcal urethritis. Repeatable isolation and propagation was accomplished by use of a special culture medium. The organisms adhered to glass or plastic, erythrocytes, and monkey kidney cells. This property appears to be associated with surface material restricted to the area of a terminal structure of the flask-shaped mycoplasmas. Although the data are insufficient to implicate the new mycoplasmas in human disease, the fact that they are unique, extremely fastidious, and have adherence properties, has stimulated efforts to assess their pathogenicity and possible role in human urogenital disease.

Major advances have occurred the past few years in the cultivation of a number of new, fastidious mollicutes--events which can be traced directly to successful efforts to develop culture media for the expanding group of helical mollicutes (spiroplasmas) inhabiting plants and arthropods. A description of cultivation techniques successful in primary isolation of three unusual mollicutes, representing new mycoplasmas from man and animals and a new spiroplasma from ticks, emphasizes some important factors in recovery of wall-less prokaryotes with special cultural requirements. Vigorous efforts to understand the distribution of spiroplasmas in plant and insect hosts also led to the cultivation of new, non-helical mollicutes. Preliminary characterization of a number of these new agents offers strong evidence for a unique and distinct Acholeplasma and Mycoplasma flora of both plants and insects.

The criteria that need to be fulfilled before regarding a mycoplasma as a cause of non-gonococcal urethritis (NGU) are outlined. Of the seven mycoplasmas that have been isolated from the human genitourinary tract, most cannot be considered as contenders for causing NGU. Although there is no evidence to support an etiological role for Mycoplasma hominis, it may be unwise to ignore this mycoplasma in view of its known pathogenicity in other situations. The cumulative weight of evidence indicates that strains of Ureaplasma urealyticum (ureaplasmas) cause NGU in some patients. The reason for their occurrence in the urethra of some men without disease needs to be established. Ureaplasmas do not seem to cause post-gonococcal urethritis. The role in NGU of M. genitalium, newly discovered in the male urethra, is unknown, but its biological features, morphological appearance, and ability to cause genital disease in marmosets suggest that it may be pathogenic for man.

Urethral specimens from 726 patients with nongonococcal urethritis (NGU) were examined for Chlamydia trachomatis, Ureaplasma urealyticum, and Mycoplasma hominis. Chlamydiae were isolated from 35.9% of ureaplasma-positive patients and from 36.5% of ureaplasma-negative patients. Ureaplasmas were isolated from 52.5% of chlamydia-positive patients and from 53.1% of chlamydia-negative patients, an observation which contrasts with that of some workers who have suggested that ureaplasmas are significantly associated with chlamydia-negative NGU. Furthermore, the numbers of ureaplasmas isolated from patients who did or did not harbour chlamydiae were not significantly different nor was there a particular association of ureaplasmas with chlamydia-negative NGU in patients experiencing their first episode of disease. In addition, M. hominis was not isolated more frequently from those from whom chlamydiae were or were not isolated. The only significant associations were the isolation of M. hominis from patients who were ureaplasma-positive and of ureaplasmas from those who were M. hominis-positive. These findings do not necessarily mitigate against ureaplasmas being responsible for some cases of chlamydia-negative NGU.

The roles of Mycoplasma hominis and Ureaplasma urealyticum in the pathogenesis of nongonococcal urethritis (NGU) remain controversial. Isolation and serologic studies and results of antibiotic therapy do not provide good evidence that M. hominis causes NGU. The results of some studies of therapy and of some experiments in which humans were inoculated with U. urealyticum indicate that this organism may cause some cases of NGU. However, U. urealyticum is ubiquitous and apparently nonpathogenic at other genital sites; thus there are serious difficulties in deciding its role in NGU. Chronic prostatitis is a poorly defined clinical entity. At present there is no convincing evidence that either M. hominis or U. urealyticum is involved in its pathogenesis.

184 urethral swabs from patients with unspecific urethritis were taken. Ureaplasma urealyticum was isolated with 86 (46.74%), and mycoplasma hominis with 26 (14.13%) patients. In the control group of 156 examinees u. urealyticum was isolated in 43 (27.56%), and m. hominis in 13 (8.33%) cases. U. urealyticum with patients was isolated in a significantly larger number than with the control group (p less than 0.05) which shows possible part of this mycoplasma in the etiology of unspecific urethritis.