Prevalence of Sexually Transmitted Chlamydia trachomatis and Neisseria gonorrhoeae Infections in an Asymptomatic Population in Dakar, Senegal

Introduction: Sexually transmitted infections (STIs) represent a major public health problem. Chlamydia trachomatis and Neisseria gonorrhoeae infections are often asymptomatic, thus leading to a high risk of transmission in subjects with risky behaviors. The objective of this study was to determine the prevalence of these 2 pathogens in an asymptomatic population. Methodology: A retrospective, cross-sectional, descriptive study was conducted in the medical biology laboratory of the Pasteur Institute of Dakar over a period of 23 months in asymptomatic patients who were seen as part of a travel check-up. A first-draft urine sample was collected and tested for C. trachomatis and N. gonorrhoeae by molecular biology techniques. Data entry and statistical analysis were performed by Excel 2010 and SPSS 2.0 respectively. Results: A total of 5012 patients were included and the overall prevalence of STIs related to these 2 pathogens was 3.8% (194/5012). The prevalences of C. trachomatis and N. gonorrhoeae were 2.7% (137/5012) and 1.0% (55/5012), respectively. The age group most affected was [20-29 years] with 58.4% (80/137; p=0.0001) for C. trachomatis and 45.5% (25/55; p=0.471) for N. gonorrhoeae. Co-infection with these two germs was observed in 0.3% (18; p=0.001) of patients. Conclusion: STIs with C. trachomatis and/or N. gonorrhoeae can be asymptomatic and continue the chain of transmission. Thus, for a better prevention of STIs due to these pathogens, it is important to screen, educate and sensitize the populations considered at risk.

Perspectivas en investigación: células HEp-2 infectadas con EB's de Chlamydia trachomatis serovar 2 (VR-902B) / Chlamydia trachomatis: Research Perspectives. HEp-2 Cells Infected with EB's from Chlamydia trachomatis serovar 2 (VR-902B): Research Perspectives

Resumen Chlamydia trachomatis (C. trachomatis) es una bacteria Gram negativa inmóvil, caracterizada por ser un microorganismo intracelular obligado y por poseer un ciclo reproductivo en el que puede distinguirse una forma infecciosa extracelular metabólicamente inerte (cuerpo elemental - EB's), y una forma no infecciosa intracelular y activa (cuerpo reticulado - RB's). C trachomatis se caracteriza por causar infección en humanos, está relacionada con enfermedades de transmisión sexual e infecciones oculares; por lo que puede conllevar a secuelas de interés, si no se da un tratamiento oportuno. El objetivo de este estudio fue optimizar el modelo de infección de C. trachomatis en células HEp-2 con cuerpos elementales (EB's) de C. trachomatis serovar L2. Inicialmente, se establecieron las condiciones para el crecimiento adecuado de las células HEp-2 en tiempo y con una confluencia del 90%, para continuar con la optimización de un protocolo de infección. La infección fue confirmada a partir de la coloración con Giemsa permitiendo evaluar características morfológicas tanto de las células HEp-2 sin infectar e infectadas, y así mismo, de los cuerpos elementales de C. trachomatis. Finalmente, se corroboró la infección con la técnica de inmunofluorescencia directa que detecta la proteína de membrana MOMP de C. trachomatis. Tras los ensayos realizados se evidenció la presencia de cuerpos elementales próximos y dentro del citoplasma celular, así como células vacuoladas y daño celular causado por la infección.

Abstract Chlamydia trachomatis (C. Trachomatis) is a Gram negative unmoving bacterium, characterized by being an obligate intracellular microorganism and having a reproductive cycle in which a metabolically inactive extracellular infectious form (elementary body - EB's) can be distinguished from an intracellular active and non-infectious form (reticulated body - RB's). C trachomatis is characterized by causing infection in humans, is related to sexually transmitted diseases and eye infections, so it can lead to sequelae of interest if timely treatment is not given. The objective of this study was to optimize the infection model of C. trachomatis in HEp-2 cells with elementary bodies (EB's) of C. trachomatis serovar L2. Initially, the conditions for the adequate growth of HEp-2 cells were established in time and with a confluence of 90%, to continue with the optimization of an infection protocol. The infection was confirmed from the staining with Giemsa allowing to evaluate morphological characteristics of both uninfected and infected HEp-2 cells and also of the elementary bodies of C. trachomatis. Finally, the infection was corroborated with the direct immunofluorescence technique, that detects the C. trachomatis MOMP membrane protein. After the tests were performed, the presence of elementary bodies nearby and within the cellular cytoplasm was evidenced, as well as vacuolated cells and cellular damage caused by the infection.

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.

Urogenital Tract Infection with Chlamydia trachomatis among Women Attended at Different Units in a Referral Hospital in Spain.

Aims: The objective of this study was to estimate the prevalence and characteristics of Chlamydia trachomatis infections in a group of women visiting different Units in a referral Hospital from Spain Study Design: This was a hospital retrospective and descriptive study for the presence of C. trachomatis in endocervical, vaginal and urine swabs obtained from consecutive sexually active women attendees at different Units. Also their medical records were reviewed. Retrospective ethical approval was granted by the Ethical Committee of Clinical Investigation of Principality of Asturias. Place and Duration of Study: Units of Gynecology, Obstetrics and Infertility of Hospital Universitario Central de Asturias, between January 2007 to December 2011. Methodology: We included 1997 symptomatic and asymptomatic unselected women (mean age 29.1 range 18 to 45 years) who were evaluated for urogenital chlamydial infection. Results: The overall prevalence of C. trachomatis was 6.3%. The C. trachomatis infection had the highest prevalence among the age group below 25 years of age (n=30, 7.5%). Genotypes E, G and D constitute 89.4% of the genotyped strains. Infections with genotypes G and F were more often (n=31, 42%) associated with clinical manifestations that suggest cervical infection and genotype E was observed more frequently (n=17, 85%) in asymptomatic women. Conclusions: In our study, similar prevalence rates between both symptomatic and asymptomatic women, under 25 years, were found. Self-collected vaginal swabs are an appropriate alternative for routine diagnosis of C. trachomatis infection. The findings of this work highlighted the need for a possible Chlamydia screening program, offered especially in younger women. Delays in seeking a diagnosis and treatment among asymptomatic females can result in increased transmission of this bacterium and its serious consequences for women reproductive health.

Determinación de IgG contra Chlamidya trachomatis en mujeres con artritis de la Ciudad de Bogotá D.C. Un estudio piloto / Determination of IgG in women infected with Chlamydia trachomatis and reactive- arthritis from Bogota, Colombia: a pilot study

Objetivo. Evaluar la presencia de IgG contra C. trachomatis en mujeres entre 18 a 30 años con diagnóstico de artritis de la ciudad de Bogotá D.C. Métodos. Los grupos de estudio están compuestos por un grupo de casos AR (mujeres con diagnóstico de artritis reumatoide y reactiva) y un grupo control. Se obtuvieron muestras de suero y se realizó la determinación cuantitativa de IgG contra C. trachomatis, la determinación PCR (proteína C reactiva) y FR (factor reumatoide). Resultados. Las variables de talla y peso descritas en la población de estudio, muestran un comportamiento homogéneo, por lo cual no interfieren en los resultados obtenidos. Las medias se compararon por los test Mann Whitney y t test no pareado. El porcentaje de resultados positivos en la determinación cualitativa de PCR fueron de: 36,6% para el grupo de casos y 14,5% para el grupo control. En la determinación cualitativa de FR el porcentaje de resultados positivos fueron: 48,8% para el grupo de casos y 5,3% para el grupo control. Finalmente, el porcentaje de resultados positivos en la determinación de anticuerpos IgG específicos contra C trachomatis fue de 2,4% para el grupo de casos y 22,4% para el grupo de controles. Las variables de PCR y FR mostraron mayor frecuencia de resultados positivos en el grupo de casos, en comparación con el grupo control, lo cual se correlaciona con la existencia de procesos inflamatorios propios del desarrollo de artritis. Sin embargo el grupo control presentó mayor frecuencia de anticuerpos IgG específicos contra C. trachomatis en comparación con el grupo de casos. Estos resultados hacen necesario evaluar a futuro, el comportamiento de las pacientes del grupo control con resultados positivos de IgG contra C. trichomatis que permitirían observar una posible aparición de síntomas relacionados con artritis.

Objective. Evaluate the presence of IgG in women infected with C. trachomatis, and diagnosed with arthritis, aged 18-30 years, in Bogota, Colombia. Methods. This project is composed of two study groups: Firstly, female cases diagnosed with rheumatoid reactive-arthritis and secondly, a control group. Quantitative determination of IgG in serum samples from patients with C. trachomatis. Were determined also, PCR determination (C reactive protein) and RF (rheumatoid factor) were performed. The statistical analysis included the comparison of means, using Mann Whitney and unpaired t test. Results. Patient samples that were positive by PCR: 36.6% for the female case group, and 14.5% for the control group. In the qualitative determination of RF positives, results were: 48.8% for the case group and 5.3% for the control group. Finally, the percentages of positive results for IgG antibodies determination against C. trachomatis were: 2.4% for the case group and 22.4% for the control group. PCR and FR variables showed higher frequency of positive results in the female case group compared with the control group, which correlates with the presence of inflammatory processes, where it is typical in the development of arthritis. However, the control group had increased frequency of specific IgG antibodies against C. trachomatis, when it is compared with the case female group. These results need to be evaluated in the future, where, the control group that tested positive for IgG against C. Trachomatis, would allow to observe the possible apparition of symptoms related to arthritis.

Sero prevalence of C.trachomatis in patient of Bad Obstetric History (BOH).

Background : Bad obstetric history (BOH) implies previous unfavorable fetal outcome. Causes of BOH may be genetic, hormonal, abnormal maternal immune response and maternal infection. C.trachomatis is the most common bacterial sexually transmitted infection worldwide. Also it has been associated with adverse outcomes of pregnancy. The identification of C. trachomatis during pregnancy and its early treatment prior to delivery would therefore be an important strategy to prevent multiple complications in mother. Serology might act as a convenient tool for diagnosis. Methods:139 pregnant patients having history of BOH as study group and 30 normal pregnant as control group were tested by Indirect ELISA for detection of anti C.trachomatis IgM and IgG. Results: Anti C.trachomatis IgM was detected in 21 (15.11%) patients and anti C.trachomatis IgG was detected in 33 (23.74%) patients. Interpretation and Conclusion: Higher numbers of positive patients were found among younger age group. Statistical analysis shows that serological detection of anti C.trachomatis IgM and IgG antibodies for detection of current and past infection by C.trachomatis is significant in BOH patients.

Diagnosis and follow-up of genital chlamydial infection by direct methods and by detection of serum IgG, IgA and secretory IgA.

Purpose: To determine the prevalence of Chlamydia trachomatis infection in a high-risk population by direct and indirect methods and to evaluate the diagnosis of secretory immunoglobulin A (sIgA). Patients and Methods: Urethral or endocervical specimens from 78 patients (48 females and 30 males) were examined by cell culture, direct fluorescence assay, PCR Cobas Amplicor (Roche Molecular Diagnostics), and sIgA was detected by the recombinant lipopolysaccharide (LPS)-enzyme-linked immunoassay (rELISA). Serum from each patient was also obtained and analysed for the presence of IgG and IgA antibody by in-house microimmunofluorescence (MIF) and by the rELISA method (Medac, Hamburg, Germany). Results: The overall C. trachomatis prevalence determined by direct methods was 28%. The detection of sIgA antibodies was significantly higher in the group of patients with a positive direct detection (50%) than in the group of negative direct detection (10.7%). The Chlamydia-specific IgA antibodies were detected by the rELISA in 40.9 and 53.6% of group I (positive direct detection) and group II patients (negative direct detection), respectively. The species-specific IgA antibodies were detected by the MIF method in 18.2 and 16.1% of group I and II patients, respectively. Chlamydia genus-specific IgG antibodies were detected by the rELISA in 86.4 and 83.9% of group I and group II patients and, C. trachomatis specific IgG were present in 81.8 and 73.2% of group I and group II patients, respectively, as assessed by the MIF test. Conclusion: Combining the positive direct methods and/or positive sIgA antibody results from cervical or urethral specimens had an indication of current C. trachomatis infection.

Infecção clamidiana feminina e masculina / Male and female chlamydial infection

Nos últimos anos os conceitos em DST têm evoluído (doenças sexualmente transmissíveis). A ideia de que as DST acometiam apenas mulheres e homens de vida promíscua não se aplica mais. Os portadores de DST, hoje em dia, são pessoas de vida cotidiana normal e não têm obrigatoriamente múltiplos parceiros sexuais. Por outro lado, os quadros clínicos clássicos e sintomáticos das DST têm sido sobrepostos por infecções assintomáticas, como a infecção clamidiana. A atualização objetiva a revisão do tema e destacar importantes aspectos diagnósticos e terapêuticos da infecção clamidiana. Os autores enfatizam a importância do diagnóstico precoce e tratamento adequado, visando não apenas acabar com a infecção propriamente dita, mas também prevenir sequelas, evitar complicações e interromper a cadeia epidemiológica. Considerando a importância e os agravos da infecção clamidiana, o rastreio para C. trachomatis em populações de risco (gestantes, adolescentes, pessoas com outras DST) e precedendo cirurgias ginecológicas deveria ser implantado na rotina de todos os serviços preocupados em prevenir esta doença e, principalmente, evitar os seus agravos.

Frecuencia de anticuerpos IgA e IgM anti Chlamydia trachomatis: en mujeres embarazadas procedentes de una consulta prenatal Cumaná, Estado Sucre, Venezuela, marzo-junio de 2006 / Frequency of IgA and IgM anti Chlamydia trachomatis antibodies: in pregnant women coming from a prenatal consult in Cumana, State of Sucre, Venezuela, march-june of 2006

Con el propósito de establecer la frecuencia de anticuerpos IgA e IgM anti-C-trachomatis en mujeres embarazadas se realizó un estudio en 84 mujeres con esa condición, en edades comprendidas entre 14 y 43 años, que acudieron a la consulta prenatal, del Servicio Autónomo Hospital Universitario “Antonio Patricio de Alcalá”, en Cumaná, estado Sucre, Venezuela, durante el período marzo-junio de 2006. Para ello se obtuvieron 84 muestras de suero para la determinación de anticuerpos IgA e IgM anti C-trachomatis a través del método de inmunoabsorción ligado a enzimas ELISA (Diagnostic Automation INC). Del total de muestras analizadas 16 (19,05 por ciento) y 55 (65,48 por ciento) resultaron positivas para la determinación de anticuerpos IgA e IgM anti C-trachomatis respectivamente. No se encontró asociación entre la presencia de estos anticuerpos con la edad de las pacientes, aunque el mayor número de pacientes positivas se ubicó en el intervalo de edades comprendidas entre 14 a 23 años. Asimismo al asociarse las manifestaciones clínicas genitales con la presencia de anticuerpos IgA e IgM anti C- trachomatis no se encontraron valores estadísticamente significativos. Por lo anteriormente expuesto se concluye que la infección genital por Chlamydia trachomatis en mujeres embarazadas es extremadamente frecuente, de manera especial en las edades comprendidas entre 24 a 33 años, y ocurre habitualmente en forma asintomática con las graves repercusiones que esto acarrea a la paciente, al feto y a su pareja.

In order to establish the frequency of IgA and IgM anti-C. Trachomatis antibodies in expectant women, a study was made of 84 women between the ages of 14 and 43, who attended prenatal consults in the Autonomous Service at the University Hospital “Antonio Patricio of Alcalá,” Cumaná, State of Sucre, during the March-June period, 2006. 84 serum samples were obtained to determine IgA and IgM anti-C. trachomatis antibodies using the immunoabsorption method connected to ELISA enzymes (Diagnostic Automation INC). Of the total samples studied, 16 (19.05 percent) and 55 (65.48 percent) resulted positive for the IgA and IgM anti-C. trachomatis antibodies, respectively. No association was found between the presence of these antibodies and the age of the patients, although the greater number of positive patients was in the 14 to 23 year age interval. Likewise, no statistically significant values were found between the association of clinical genital manifestations and the presence of IgA and IgM anti C- trachomatis antibodies; therefore, it was shown that Chlamydia trachomatis is presented asymptomatically in most cases. Conclusions were that genital infection by Chlamydia trachomatis in pregnant women is extremely frequent, especially for ages between 24 and 33 years, and it occurs habitually in an asymptomatic form with the serious repercussions that this produces on the patient, the fetus and the partner.

Detección de Chlamydia trachomatis en orina por LCR: aplicación del método de mezcla de muestras biológicas / Ligase chain reaction testing of pooled urine specimens to diagnose Chlamydia trachomatis infection

Objective. The purpose of this study was to assess the utility and validity of pooling urine samples for molecular diagnosis of Chlamydia trachomatis infection. Material and methods. Of 1,220 urine samples collected from Mexican female and male adolescents, 305 pools were composed of fourth individual samples each, based on a calculation of optimal pool size. These were processed by ligase chain reaction (LCR) for the detection of C. trachomatis. Positive and gray-zone pools were reanalyzed individually. Cost savings were calculated comparing actual costs of testing to the cost that would have been incurred testing all 1,220 samples individually. Results.Pools results were: 56 positive, 19 gray-zones and 230 negative. Following individual retesting of positive and gray-zone pools, 59 cases of C. trachomatis infection were identified (4.8% prevalence). Thus, a total of 601 LCR tests were performed, for a 50.4% savings considering only the direct cost of the test. Conclusions.Our experience shows that sample pooling is both a reliable and convenient tool for CT surveillance in our setting. It should be considered in other similar settings where limited resources constraint surveillance of STIs.

Objetivo. Evaluar la validez y conveniencia de la estrategia de la mezcla de muestras de orinas para el diagnóstico molecular de Chlamydia trachomatis (CT). Material y métodos. A partir de 1,220 muestras de orina recolectadas de jóvenes de uno y otro sexos, se conformaron 305 mezclas con cuatro alícuotas de muestras individuales, previo cálculo del tamaño óptimo de la mezcla. A continuación se determinó la presencia de ácidos nucleicos de clamidia en esas mezclas, mediante el método de reacción en cadena de la ligasa. Las mezclas positivas o en zona gris fueron reanalizadas de manera individual (cuatro pruebas adicionales). El número final de pruebas realizadas se comparó con el total de pruebas que se habrían efectuado individualmente. Resultados. Del total de mezclas analizadas, 230 resultaron negativas, 56 fueron positivas y 19 más se ubicaron en zona gris. Una vez reanalizadas de manera individual las mezclas positivas y las de zona gris, se obtuvieron 59 muestras de orina positivas a clamidia (prevalencia de 4.81%). De esta manera, el número total de pruebas efectuadas fue de 605 en contraste con las 1,220 que tendrían que haberse hecho si se hubieran procesado las muestras individualmente, es decir, que se logró un ahorro de 50.5% del costo directo del reactivo de diagnóstico. Conclusiones. La metodología aplicada mostró ser tanto confiable como conveniente en el entorno mexicano para llevar a cabo vigilancia epidemiológica de la infección por CT. Dado lo anterior, esta metodología podría ser considerada en otros entornos en los que la falta de recursos limita la vigilancia de las infecciones de transmisión sexual.