Association of human papillomavirus and Chlamydia trachomatis with intraepithelial alterations in cervix samples

The influence of different infectious agents and their association with human papillomavirus (HPV) in cervical carcinogenesis have not been completely elucidated. This study describes the association between cytological changes in cervical epithelium and the detection of the most relevant aetiological agents of sexually transmitted diseases. Samples collected from 169 patients were evaluated by conventional cytology followed by molecular analysis to detect HPV DNA, Chlamydia trachomatis, herpes simplex virus 1 and 2,Neisseria gonorrhoeae, Mycoplasma genitalium, Trichomonas vaginalis, andTreponema pallidum, besides genotyping for most common high-risk HPV. An association between cytological lesions and different behavioural habits such as smoking and sedentariness was observed. Intraepithelial lesions were also associated with HPV and C. trachomatis detection. An association was also found between both simple and multiple genotype infection and cytological changes. The investigation of HPV and C. trachomatisproved its importance and may be considered in the future for including in screening programs, since these factors are linked to the early diagnosis of patients with precursor lesions of cervical cancer.

Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

[Detection of Chlamydia trachomatis and Mycoplasma genitalium in semen samples of infertile men using multiplex PCR]

Chlamydia trachomatis is the most common bacterial sexually transmitted infection in the world, but the effect of this infection on male fertility is still controversial. Despite reports of interaction between Mycoplasma genitalium and sperm, this pathogen in semen samples of infertile men is less studied. We studied, the prevalence of Chlamydia trachomatis and Mycoplasma genitalium infection in infertile men. Among attending Avicenna Infertility Center, 120 men who had abnormal semen analysis tests were selected and the samples were taken. After detailed analysis of semen quality, DNA was extracted from each sample by chelex. Samples were evaluated for these two pathogens by multiplex PCR. Results were statistically analyzed. Chlamydia trachomatis and Mycoplasma genitalium was detected in 23/3% and 12/5% of the samples, respectively. Although, Mycoplasma genitalium infection rises by increasing [P=0.640] and decreasing in age of first sexually activity [P=0.203], and also positive cases of Chlamydia trachomatis infection showed increase regarding age increase [P=0.619] and age decrease in first sexually activity [P=0.511], but these differences were not statistically significant. All in all, regarding to the increased prevalence of Chlamydia trachomatis infection compared with the only similar study in Iran and high prevalence of Mycoplasma genitalium infection in infertile men, this assessment was done. A multiplex PCR protocol rapidly and simultaneously identify these organisms in comparison with uniplex from clinical samples. Based on our results screening for Chlamydia trachomatis and Mycoplasma genitalium infection among infertile men seems to be valuable

Mycoplasmateceae species are not found in fallopian tubes of women with tubo-peritoneal infertility

BACKGROUND: The role of mycoplasmas on the development and sequelae of pelvic inflammatory disease remains controversial. The objective of the present study is to correlate directly the presence of Mycoplasmateceae through polimerase chain reaction (PCR) determinations in cervix and Fallopian tubes of infertile patients with tubo-peritoneal factor diagnosed through laparoscopy. METHODS: Thirty patients with tubo-peritoneal infertility and 30 normal fertile patients were included in the study; cervical samples and tubal flushings were obtained during laparoscopy. PCR determinations for the detection of genetic material of Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealiticum, Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis in cervix and tubal flushings were performed. RESULTS: No Mycoplasmataceae species as "only" microorganisms were found in tubal flushings of tubo-peritoneal infertility patients, whereas three (10%) fertile patients with normal tubes were positive for mycoplasma presence. This difference was not significant (p = 0.237). Among the 30 patients suffering from tubal infertility diagnosed through laparoscopy, Mycoplasmatecae species were not detected in the Fallopian tubes by PCR determinations, while in normal tubes from fertile patients these and other microorganisms could be found without distorting tubal anatomy. CONCLUSION: Mycoplasmateceae species were not detected in Fallopian tubes of women with tubo-peritoneal infertility.

Simultaneous and rapid differential diagnosis of Mycoplasma genitalium and Ureaplasma urealyticum based on a polymerase chain reaction-restriction fragment length polymorphism.

Objectives: The aim of this investigation was to simultaneously detect and differentiate Mycoplasma genitalium and Ureaplasma urealyticum in female patients suffering from genital complications by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP). Materials and Methods : Genital swabs were taken from 210 patients. They were transported to the laboratory in phosphate-buffered saline. For PCR, samples were analysed with genus-specific MyUu-R and MyUu-F primers. This primer set, which was originally designed in our laboratory, amplified a 465 bp fragment (M. genitalium) and a 559 bp fragment (U. urealyticum). Samples containing a band of the expected sizes for the Mycoplasma strains were subjected to digestion with a restriction endonuclease enzyme of TaqI and Cac8I. Results: Of the 210 samples, a total of 100 (47.6%) samples were found to be positive for Mycoplasmas (seven M. genitalium isolates, 3.3%; and 89 U. urealyticum isolates, 42.4%), and coinfections with both species were detected in four samples (1.9%). The PCR-RFLP results showed that M. genitalium and U. urealyticum are different by enzyme patterns. Conclusion: PCR-RFLP offers a rapid and easily applicable protocol to simultaneous detection and differentiation of M. genitalium and U. urealyticum from clinical samples when specific primers and restriction enzymes are used.

Association of genital mycoplasmas including Mycoplasma genitalium in HIV infected men with nongonococcal urethritis attending STD & HIV clinics.

Background & objectives: Acute nongonococcal urethritis (NGU) is one of the commonest sexually transmitted infections affecting men. The role of genital mycoplasmas including Mycoplasma genitalium in HIV infected men with NGU is still not known. The aim of this study was to determine the isolation pattern/detection of genital mycoplasma including M. genitalium in HIV infected men with NGU and to compare it with non HIV infected individuals. Methods: One hundred male patients with NGU (70 HIV positive, 30 HIV negative) were included in the study. Urethral swabs and urine samples obtained from patients were subjected to semi-quantitative culture for Mycoplasma hominis and Ureaplasama urealyticum, whereas M. genitalium was detected by PCR from urine. The primers MgPa1 and MgPa3 were selected to identify 289 bp product specific for M. genitalium. Chalmydia trachomatis antigen detection was carried out by ELISA. Results: M. genitalium and M. hominis were detected/isolated in 6 per cent of the cases. M. genitalium was more common amongst HIV positive cases (7.1%) as compared to HIV negative cases (3.3%) but difference was not statistically significant. Co-infection of C. trachomatis and U. urealyticum was found in two HIV positive cases whereas, C. trachomatis and M. hominis were found to be coinfecting only one HIV positive individual. M. genitalium was found to be infecting the patients as the sole pathogen. Interpretation & conclusions: Patients with NGU had almost equal risk of being infected with M. genitalium, U. urealyticum or M. hominis irrespective of their HIV status. M.genitalium constitutes one of the important causes of NGU besides other genital mycoplasmas.

Diagnóstico de Mycoplasma genitalium por amplificación de los genes MgPa y ARN ribosomal 16S / Diagnosis of Mycoplasma genitalium by MgPa and rRNA 16S gene amplification

OBJETIVO: El microorganismo Mycoplasma genitalium se ha relacionado con la uretritis no gonocócica (UNG). La técnica de PCR se ha convertido en el principal método de detección de este patógeno. En consecuencia, debe aplicarse un método de diagnóstico mediante la amplificación de fragmentos de ADN por la técnica PCR. MATERIAL Y MÉTODOS: Se seleccionaron los cebadores MGF-MGR y MgPaF-MgPaR, complementarios de los genes de ARNr 16S y MgPa de M. genitalium, respectivamente. Se efectuaron ensayos de especificidad y sensibilidad y se estudiaron muestras clínicas. RESULTADOS: La PCR con cada grupo de cebadores utilizado fue específica sólo para M. genitalium y la sensibilidad fue mayor con el grupo de cebadores MGF-MGR. En el estudio de 34 muestras clínicas, 18.5 por ciento fue positivo a M. genitalium y se encontró un mayor número de muestras positivas al utilizar los cebadores MgPaF-MgPaR. CONCLUSIONES: Debe aplicarse en la práctica clínica el diagnóstico de M. genitalium mediante la amplificación del ADN por PCR en los pacientes con UNG.

OBJECTIVE: Mycoplasma genitalium has been associated with nongonococcal urethritis (NGU). Diagnosis by PCR has become the primary detection method for this organism. Thus, diagnosis by DNA amplification using the PCR technique should be utilized. MATERIAL AND METHODS: GMF/GMR and MgpF/MgpR primer pairs, complementary to the M. genitalium 16S rRNA and MgPa genes, respectively, were selected. Specificity and sensibility assays were conducted and clinical samples were studied. RESULTS: The PCR with each primer pair was specific only for M. genitalium, and the sensibility was higher with the GMF/GMR primers. In the study of 34 clinical samples, 18,5 percent were positive for M. genitalium, with more positive samples when the MgpF/MgpR primers were used. CONCLUSIONS: DNA amplification by PCR should be applied in clinical practice to the diagnosis of M. genitalium in patients with NGU should using.

Detección de Mycoplasma genitalium y correlación con manifestaciones clínicas en una población del estado Zulia, Venezuela / Mycoplasma genitalium detection and correlation with clinical manifestations in population of the Zulia State, Venezuela

Diverse studies demonstrate an association between Mycoplasma genitalium and urogenital pathologies. The aim of this study was to investigate the prevalence of M. genitalium in patients attending gynecological evaluation in private clinics (n = 172). DNA amplification assays of the genes 16S rRNA and MgPa were utilized. The prevalence of M. genitalium in the study population was 7.5 percent. M. genitalium was detected in 12.1 percent and 4.1 percent of the symptomatic and asymptomatic patients, respectively (p = 0.047). The infection was diagnosed in patients with cervicitis (17.2 percent) and mucopurulent secretion (16.6 percent) and the highest prevalence of infections was registered in the 31-40 years age group. No significant association between the presence of M.genitalium and individual clinical manifestations or the patients age was showed (p > 0.05). The high prevalence of M. genitalium infections, mostly in patients with clinical manifestations showed in this study, warrants the application of diagnostic strategies in the population to investigate the clinical meaning of these microorganisms and to reevaluate therapeutic schemes against non-gonococcal and non-chlamydial infections.

Diversos estudios demuestran una asociación entre Mycoplasma genitalium y patologías urogenitales. El objetivo de este trabajo fue investigar la prevalencia de infecciones por M. genitalium en pacientes atendidas en clínicas privadas (n = 172). Se utilizaron ensayos de amplificación de genes 16S rARN y MgPa. La prevalencia de M. genitalium en esta población fue 7,5 por ciento. Mycoplasma genitalium fue detectado en 12,1 y 4,1 por ciento) de las pacientes sintomáticas y asintomáticas, respectivamente (p = 0,047). La infección se diagnosticó en pacientes con cervicitis (17,2 por ciento) y con secreción mucopurulenta (16,6 por ciento) y la mayor prevalencia de infecciones se registró en el grupo etario de 31 a 40 años. No se encontró asociación significativa entre la presencia de M. genitalium y manifestaciones clínicas individuales o edad de las pacientes (p > 0,05). La alta prevalencia de infecciones por M. genitalium, principalmente en pacientes con manifestaciones clínicas demostrada en este estudio, demanda la aplicación de estrategias diagnósticas en la población para investigar el significado clínico de estos microorganismos y reevaluar esquemas terapéuticos contra infecciones no gonocóccicas y no clamidiales.

Avaliação da taxa de isolamento de micoplasmas do trato urogenital por um método comercial em comparação com meios de cultura produzidos no próprio laboratório / Assessing the rate of isolation of mycoplasma treatment of urogenital by a business method in compared with media culture produced in itself laboratory

Este trabalho procurou avaliar a taxa de isolamento de Mycoplasma hominis e Ureaplasma urealiticum/U. parvum emamostras de conteúdo vaginal de 84 mulheres atendidas em um laboratório de analises clínica de Blumenau – SC, por dois meios de cultura de procedência diferentes. Os resultados demonstraram 50,0 % de positividade para Ureaplasma urealiticum/U. parvum e 20,24% de positividade para Micoplasma hominis por cultura em meio feito em nosso laboratório (P< 0,0001) e 16,67% de positividade para U. urealiticum e 8,33% de positividade para M. hominis por cultura em meio comercial (P< 0,0001). As amostras que foram cultivadas por meio próprio e obtiveram resultado positivo foram também analisadas por diluições seriadas para identificar a quantidadedestes microrganismos expresso em C.C.U. (color changing units). Os resultados demonstraram que 52,94% destas amostras apresentaram C.C.U. maior que 104 para M. hominis e 42,86% apresentaram C.C.U. maior que 103 para U. urealyticum, sendo estesos níveis considerados clinicamente significativos para que a presença destes microrganismos seja implicada na etiologia dos sintomas. No presente trabalho discutiu-se as possíveis causas desta diferença de positividade entre os dois métodos utilizados.

Aplicación de método molecular en la detección de Mycoplasma genitalium en hombres y en mujeres embarazadas / Molecular detection of Mycoplasma genitalium in men and pregnant women

Mycoplasma genitalium es un patógeno oportunista del tracto genital. En el hombre es causa de uretritis, en tanto que en mujeres ha sido implicado en la etiología de cervicitis y de enfermedad inflamatoria pelviana (EIP). El objetivo de este estudio fue determinar la prevalencia de M. genitalium en pacientes masculinos con uretritis y en muestras vaginales de mujeres embarazadas. Se obtuvo muestras de secreción uretral en 37 pacientes con uretritis y de muestras vaginales de 50 consecutivas mujeres embarazadas, determinándose la presencia de M. genitalium mediante reacción de polimerasa en cadena (RPC). Las muestras de secreción uretral fueron también evaluadas en busca de Chlamydia trachomatis, Neisseria gonorrhoeae y Ureaplasma sp en tanto que en las de origen vaginal se investigó la microbiota y presencia de micoplasmas de tipo genital. Veintitrés casos fueron clasificados como uretritis no gonocóccica (UNG) y 14 como enfermedad gonocóccica. M. genitalium fue detectado en 3 de 23 (13,04 por ciento) varones con UNG; en dos casos asociado a Ureaplasma sp, y en un paciente como agente único. C. trachomatis fue detectado en 7 pacientes con UNG y en uno con gonorrea. Ureaplasma sp fue aislado en 13 (35,1 por ciento) pacientes, 8 casos de UNG y en 5 con gonorrea. El microorganismo fue detectado también en 6 (15 por ciento) de 40 mujeres; en 5 casos en presencia de microbiota normal (score de Nugent 0-3), y en un caso en presencia de vaginosis bacteriana. Ureaplasma spp fue aislado en las seis muestras positivas. En conclusión, este estudio demuestra que M. genitalium debe ser también considerado en la etiología de la UNG así como en el tracto genital inferior en la mujer embarazada, en presencia de una microbiota vaginal normal.