Implementación del algoritmo reverso para el cribado de sífilis en un laboratorio clínico / Implementation of the reverse screening syphilis algorithm in a clinical laboratory

Resumen Introducción: La sífilis sigue siendo un problema de salud pública en todo el mundo; la precisión de las pruebas de diagnóstico es fundamental para el éxito de su control. Actualmente, hay dos enfoques para el diagnóstico serológico de la sífilis: el algoritmo tradicional y el algoritmo reverso. Objetivo: Analizar las ventajas y desventajas en la implementación del cribado para sífilis con el algoritmo reverso en un laboratorio clínico de pacientes ambulatorios. Materiales y Métodos: Se realizó un estudio de corte transversal analizando 246 sueros reactivos en el cribado sobre un total de 14.700 solicitudes de serología para sífilis. Se utilizaron los ensayos ARCHITECT SyphilisTP, V.D.R.L. y FTA-Abs. Resultados: De los 246 sueros reactivos por ARCHITECT Syphilis TP, 129 fueron reactivos y 117 no reactivos con V.D.R.L., éstos últimos resultaron 97 reactivos y 20 no reactivos por FTA-Abs, sugiriendo falsos positivos (0,13%). Se detectaron dos casos de infección primaria, no detectados con V.D.R.L y un caso de infección primaria en una gestante con un valor alto S/CO y V.D.R.L. de 1 dils. Conclusiones: Entre las ventajas de utilizar el algoritmo reverso se encontró mayor sensibilidad en la detección de sífilis primaria; automatización, trazabilidad, interpretación objetiva y resultados concluyentes.

Background: Syphilis remains a public health concern worldwide, the accuracy of diagnostic tests is critical for its successful control. Currently, there are two approaches to the diagnosis of syphilis using serological tests: the traditional algorithm and the reverse algorithm. Aim: The goal of this study was to analyse the advantages and disadvantages in the implementation of the syphilis reverse-screening algorithm in an outpatient clinical laboratory. Methods: An observational cross-sectional study was carried out analyzing 246 reactive sera from a total of 14700 requests for syphilis serology. Chemiluminescent assay ARCHITECT Syphilis TP, V.D.R.L. and FTA-Abs were performed. Results: Among 246 reactive sera by ARCHITECT Syphilis TP, 129 were reactive and 117 were non-reactive by V.D.R.L. the last mentioned resulted in 97 reactive and 20 non-reactive by FTA-Abs, suggesting false positives (0.13%). Two patients with primary infection were detected, that were not detected by V.D.R.L. and one pregnant woman with primary infection with a high value S/CO and V.D.R.L.:1 dils. Conclusions: Among the advantages of using a reverse algorithm were greater sensitivity in the detection of patients with primary syphilis; automation, complete traceability of the samples; objective interpretation and conclusive results.

Serorreacción y prevalencia de sífilis en donantes de un banco de sangre de Barranquilla, Colombia / Seroreactivity and prevalence of syphilis in donors at a blood bank in Barranquilla, Colombia

Resumen Introducción. La sífilis es una enfermedad de interés en salud pública por sus elevadas tasas de morbilidad y mortalidad. Objetivo. Determinar la serorreacción y la seroprevalencia de sífilis según las variables sociodemográficas de los donantes de un banco de sangre del distrito de Barranquilla, Colombia, durante 2015 y 2016. Materiales y métodos. Se hizo un estudio descriptivo de corte transversal basado en los resultados de las pruebas treponémicas y no treponémicas. Se analizaron las variables sociodemográficas de la población estudiada y se hizo un análisis univariado en el que se determinaron las frecuencias absoluta y relativa de cada una de las variables categóricas. Se determinó la serorreacción a Treponema pallidum y la prevalencia de la infección activa. Se utilizó la prueba de ji al cuadrado de Pearson para evaluar las diferencias entre las proporciones. Resultados. Se encontró una serorreacción de 1,86 % para la infección previa con T. pallidum y una prevalencia de 0,93 % para la infección activa, las cuales fueron más altas en hombres adultos y en adultos mayores, viudos, desempleados y personas residentes en otros municipios del departamento de Atlántico diferentes de Barranquilla y su área metropolitana. Se encontró una asociación significativa entre la sífilis y las variables de sexo y ocupación. Conclusión. Se registró una serorreacción elevada a T. pallidum en donantes de sangre, comparada con el promedio nacional. Se encontró asociación entre la sífilis, y las variables sociodemográficas de sexo y ocupación, principalmente.

Abstract Introduction: Syphilis is a public health concern given its high impact on morbidity and mortality. Objective: We aimed to determine the association of syphilis seroreactivity and seroprevalence with sociodemographic variables of donors at a blood bank in the district of Barranquilla, Colombia, during 2015 and 2016. Materials and methods: We conducted a descriptive cross-sectional study based on the results of the treponemal and nontreponemal tests and the sociodemographic variables of the study population. We performed a univariate analysis to determine the absolute and relative frequencies for each categorical variable. We determined the seroreactivity against Treponema pallidum and the prevalence of active syphilis infection, and we used Pearson's chi-square test to evaluate the differences between the proportions. Results: We found a seroreactivity of 1.86% in individuals with previous T. pallidum infection, and a prevalence of 0.93% in those with active T. pallidum infection. These values were higher in adult men and older adults, widowers, the unemployed, and people living in municipalities of the Department of Atlántico other than Barranquilla and its metropolitan area. The incidence of syphilis infection showed a significant association with sex and occupation. Conclusion: In comparison to the national average, syphilis seroreactivity was high among blood donors. There was an association between syphilis infection incidence and sociodemographic variables such as sex and occupation.

Cerebrospinal fluid examination may be useful in diagnosing neurosyphilis in asymptomatic HIV+ patients with syphilis / El examen del líquido céfalo raquídeo puede ser útil en el diagnóstico de neurosífilis en pacientes VIH + asintomáticos con sífilis

ABSTRACT Lumbar puncture in neurologically asymptomatic HIV+ patients is still under debate. There are different criteria for detecting neurosyphilis through cerebrospinal fluid (CSF), especially in cases that are negative through the Venereal Disease Research Laboratory (VDRL), regarding cellularity and protein content. However, a diagnosis of neurosyphilis can still exist despite negative VDRL. Treponema pallidum hemagglutination assay (TPHA) titers and application of the TPHA index in albumin and IgG improve the sensitivity, with a high degree of specificity. Thirty-two patients were selected for this study. VDRL was positive in five of them. The number of diagnoses reached 14 when the other techniques were added. It was not determined whether cellularity and increased protein levels were auxiliary tools in the diagnosis. According to our investigation, CSF analysis using the abovementioned techniques may be useful in diagnosing neurosyphilis in these patients.

RESUMO La punción lumbar (PL) en pacientes VIH+ neurológicamente asintomáticos es controversial. Existen diferentes criterios para detectar en el líquido cefalorraquídeo (LCR) neurosífilis (NS): el examen Venereal Disease Research Laboratory (VDRL) en primer lugar, en caso de negatividad: la celularidad y el tenor de proteinas. Sin embargo el diagnóstico de NS puede ser sostenido a pesar de la negatividad de las técnicas mencionadas. La titulación del Treponema pallidum hemagglutination assay (TPHA) y la aplicación del índice de TPHA en Albúmina e Ig G mejoran la sensibilidad asociando elevado grado de especificidad. 32 pacientes fueron seleccionados para este estudio, el VDRL fue positivo en 5. El diagnóstico se elevó a 14 cuando se sumaron el resto de las técnicas. No se evidenció que la celularidad y el aumento de proteínas fueran herramientas auxiliares para el diagnóstico. De acuerdo a nuestro trabajo el estudio del LCR con las técnicas señaladas puede ser de utilidad en el diagnóstico de NS en estos pacientes.

Soroprevalência e fatores associados ao Vírus da Imunodeficiência Humana (HIV) e sífilis em presidiários do Estado de Pernambuco, Brasil / Seroprevalence and factors associated with human immunodeficiency virus (HIV) and syphilis in inmates in the state of Pernambuco, Brazil

Os detentos apresentam maior vulnerabilidade ao HIV e outras Infecções Sexualmente Transmissíveis (IST) devido a fatores de risco como: compartilhamento de seringas e sexo desprotegido. O objetivo do presente trabalho foi determinar a soroprevalência e fatores de risco para o HIV-1/2 e sífilis entre presidiários do sexo masculino em Caruaru, Pernambuco, Brasil. Foi realizado um corte transversal no período de maio a julho de 2011, no qual 1097 detentos de uma penitenciária de Caruaru foram avaliados, por meio de entrevista e coleta de amostra de sangue para realização dos testes. A prevalência de infecção pelo HIV foi de 1,19% e de sífilis 3,92%. A associação com a infecção pelo HIV mostrou-se estatisticamente significante com uso de drogas injetáveis, homossexualismo e condição de transfusão (p < 0,05). Em relação à soropositividade para sífilis, fatores relacionados à vida sexual mostraram-se estatisticamente significante (p < 0,05). A população carcerária constitui um grupo de alto risco para as doenças pesquisadas. As taxas de prevalência aqui identificadas indicam a necessidade de implantação de programas de prevenção para que as mesmas não venham a se disseminar no âmbito desta população.

Prison inmates are more vulnerable to HIV and other Sexually Transmitted Infections (STIs) due to risk factors such as needle sharing and unprotected sex with homosexuals. The aim of this work was to determine the seroprevalence and risk factors associated with the human immunodeficiency virus (HIV-1/2) and syphilis among male inmates in Caruaru, State of Pernambuco, Brazil. A cross-sectional study was performed between May and July 2011, when 1,097 inmates at a prison in Caruaru were assessed by means of interviews and blood sample collection for performing the respective tests. The prevalence was 1.19% for HIV infection and 3.92% for syphilis. HIV infection showed a statistically significant association (p <0.05), with injected drug use, homosexuality and blood transfusions. With respect to HIV status and syphilis, factors related to sex life were statistically significant (p <0.05). The prison population is a high risk group for the diseases investigated. The prevalence rates identified indicate the need to implement prevention programs, helping to contain such diseases in this particular population group.

Sexually transmitted infections [syphilis] in long distance truck drivers

To find out the frequency of sexually transmitted infections [Syphilis] in long distance truck drivers [LDTDs]. This study was conducted in the department of Microbiology Shaikh Zayed Hospital Lahore on 199 long distance truck drivers. Presence of syphilis was detected by rapid plasma reagin and enzyme link immunosorbent assay for treponema pallidum syphilis. 10.5% long distance truck drivers showed syphilis positive by enzyme link immunosorbent assay and 20.1% by rapid plasma reagin. Number of cases missed by rapid plasma regain were 03 [1.5%]. Sensitivity, specificity, positive predictive value and negative predictive value of rapid plasma reagin compared with enzyme link immunosorbent assay were 85%, 87%, 42% and 98% respectively. Enzyme Link immunosorbent assay syphilis is more accurate for diagnosis of syphilis than rapid plasma reagin.

Molecular detection of Treponema pallidum sp. pallidum in blood samples of VDRL-seroreactive women with lethal pregnancy outcomes: a retrospective observational study in northern Brazil / Detecção molecular do Treponema pallidum sp. pallidum em amostras de sangue de mulheres sororeativas ao VDRL com resultado letal da gravidez: um estudo observacional retrospectivo no norte do Brasil

INTRODUCTION: Although control measures of maternal and congenital syphilis are available in Brazil, difficulties exist within the healthcare network in providing a laboratory diagnosis of the infection during the prenatal period. The objective of this study was to confirm the presence of Treponema pallidum by PCR in women with positive VDRL serology and lethal pregnancy outcomes, i.e., abortion, stillbirth and neonatal death. METHODS: A retrospective study was conducted on VDRLseroreactive women with lethal pregnancy outcomes admitted to the Fundação Santa Casa de Misericórdia do Pará (FSCM-PA) between January and July 2004. Serum samples and DNA from whole blood were obtained at the time of screening by the VDRL test. These samples were analyzed by IgG ELISA, IgM FTA-Abs and simple PCR (polA). RESULTS: During the study period, 0.7% (36/4,912) of women with lethal pregnancy outcomes presented a positive VDRL test. The polAgene was amplified in 72.7% (24/33) of these women, with 55.6% (20/36) and 94.4% (34/36) presenting IgM and IgG antibodies against T. pallidum, respectively. Comparison of these results showed a significant difference, with agreement between the PCR and IgM FTA-Abs results, suggesting that maternal syphilis was an active infection. No basic cause of death of the conceptus was reported in 97.2% (35/36) of cases. Among women who were submitted to the VDRL test during the prenatal period, only four of the nine seroreactive patients underwent treatment. CONCLUSIONS: The high frequency of syphilis in the group studied indicates the fragility of the service of infection diagnosis, treatment and monitoring, compromising epidemiological control.

INTRODUÇÃO: Apesar das medidas de controle da sífilis materna e congênita estarem disponíveis no Brasil, existem dificuldades da rede em prover o diagnóstico laboratorial da infecção durante o pré-natal. O objetivo deste estudo foi confirmar a presença do Treponema pallidum pela PCR em mulheres com sorologia positiva ao VDRL e com resultado letal da gravidez, isto é, aborto, natimorto e neomorto. MÉTODOS: Estudo retrospectivo realizado em mulheres VDRL-sororeativas com resultado negativo da gravidez, admitidas na Fundação Santa Casa de Misericórdia do Pará FSCM-PA entre janeiro e julho de 2004. As amostras de soro e DNA de sangue total foram obtidas no mesmo período da triagem pelo VDRL. Estas amostras foram analisadas pelo ELISA IgG, FTA-Abs IgM e PCR simples (polA). RESULTADOS: Durante o período de estudo, 0,7% (36/4.912) das mulheres com resultado letal da gravidez apresentaram VDRL positivo. O genepolA foi amplificado em 72,7% (24/33) destas mulheres,com 55,6% (20/36) e 94,4% (34/36) apresentando anticorpos tipo IgG e IgM contra o T. pallidum, respectivamente. A comparação destes resultados mostrou uma diferença estatística significativa, sendo que os resultados da PCR versus FTA-Abs Ig Mmostraram-se concordantes, sugerindo que a sífilis materna era uma infecção ativa. A causa básica de morte dos conceptos não foi relatada em 97,2% (35/36) dos casos. Entre as mulheres que foram submetidas ao VDRL no pré-natal, somente quatro das nove soropositivas receberam tratamento. CONCLUSÕES: A elevada frequência de sífilis no grupo de estudo indica a fragilidade do serviço no diagnóstico, tratamento e monitoramento da infecção, comprometendo o controle epidemiológico.

Profile of Anti-Tp47 antibodies in patients with positive serology for syphilis analized by western blot

In Brazil, syphilis is still a great problem of public health. Serological test is essential for syphilis diagnosis and the current trend is the use of recombinant antigen in the treponemal tests, due to its confirmed higher sensibility and specificity. The purpose of the present study was to analyze the profile of anti-Tp47 antibodies in patients with positive serology for syphilis. One hundred positive sera samples were analyzed by Western Blot (WB) technique, using the recombinant antigen (rTp47). Ten of them did not present antibodies against the fraction rTp47, the results were confirmed by WB using native T. pallidum antigen. All ten samples had antibodies against the fractions Tp17 and Tp15 and presented low reactivity in VDRL, negative results or title below than 1:4. Considering that VDRL is used for therapeutic monitoring due to seroreversion of nontreponemal antibodies in response to the treatment, and that some studies reported loss of treponemal antibodies after treatment, we could speculate if these ten samples are cases of serological memory from patients previously treated for syphilis. In addition, although several features state the Tp47 fraction as one of the major antigenic components, based on our results we point out to the importance of including other antigenic proteins such as Tp17 and Tp15 in addition to Tp47 in tests for serological screening of syphilis.

Analysis of Positive Results in Mediace Rapid Plasma Reagin and Treponema pallidum Latex Agglutination as the Automated Syphilis Test / 대한진단검사의학회지

BACKGROUND: We compared the results of automated and quantitative methods for the diagnosis of syphilis, Mediace Rapid Plasma Reagin (RPR) and Mediace Treponema pallidum Latex Agglutination (TPLA) (Sekisui Chemical Co., Ltd, Japan) with those of conventional methods. METHODS: Sera from 3,896 persons who had health checkups between December 2005 and November 2006 were included in the evaluation of positive rates and biological false positives (BFP) for Mediace RPR and TPLA. In addition, 134 patients' sera positive for automated Mediace RPR or TPLA were tested for VDRL and TPHA. Discrepancies between TPLA and TPHA results were confirmed by the RecomBlot Treponemal IgG/IgM (Mikrogen GmbH, Germany). Automated Mediace RPR and TPLA were performed using the Hitachi 7600 chemistry autoanalyzer (Hitachi, Japan). Samples with positive Mediace RPR and negative TPLA results were defined as BFP. RESULTS: Positive rate of automated Mediace RPR was 0.23% (9/3,896). BFP of the Mediace RPR was 0.18%. Positive rate of automated TPLA was 1.62% (37/2,284). Among the 134 patients' sera, 33 (24.6%) showed a discrepancy between conventional VDRL and automated Mediace RPR results: Among 31 Mediace RPR(+)/VDRL(-) sera, 13 were positive and 18 were negative for TPLA. The remaining 2 sera of discrepancy with Mediace RPR(-)/VDRL(+) were all positive for TPLA. There were seven sera that showed a discrepancy between automated TPLA and TPHA results: Two sera with Mediace RPR(+)/TPLA(-)/TPHA(+) showed negative recomBlot Treponemal IgG/IgM results, and among five sera with TPLA(+)/TPHA(-), three demonstrated IgG or IgM by recomBlot Treponemal IgG/IgM. CONCLUSIONS: The results of comparison data demonstrated that automated TPLA results had a high concordance with recomBlot Treponemal IgG/IgM results. Moreover, there are additional advantages of automated methods such as quantitative detection, low infection risk, and no influence by human handling.

Comparative evaluation of three different treponemal enzyme immunoassays for syphilis.

BACKGROUND: To reduce human errors and subjective interpretation, automation is currently a trend. However, replacing any tests with automation must first be validated. OBJECTIVE: Evaluate the EIA tests performance characteristics of three commercially available enzyme immunoassays; Enzygnost Syphilis (Dade Behring Ltd), Syphilis EIA 480 (Newmarket Laboratory Ltd) and ICE* Syphilis (Abbott Murex). MATERIAL AND METHOD: Three thousand and fifty-five serum samples were obtained from all workers who came for physical check ups before working abroad at the physical check-up unit of the out- patient department at Siriraj Hospital between February and August, 2001. Serum specimens known to be positive with VDRL and TPHA or FTA-ABS tests were included in the present study. RESULTS: Of all the samples, 2953 were from workers who came for physical check ups; 102 were selected from known specimens positive with the Venereal Disease Research Laboratory test (VDRL) and Treponema Pallidum Hemagglutination Assay (TPHA) or Fluorescent Treponemal Antibody ABSorption (FTA-ABS) test. A true positive result was determined when the sample was reactive either with two out of three enzyme immunoassays and TPHA or FTA-ABS, or both TPHA and FTA-ABS. A true negative result was determined when the aforementioned were absent. The sensitivity and specificity of Enzygnost Syphilis, Syphilis EIA 480 and ICE* Syphilis were 100% and 97.89%, 100% and 99.59%, and 99.1% and 99.76%, respectively. The results suggest that the specificity of Enzygnost Syphilis is the lowest among these three enzyme immunoassays; the price is also the cheapest. The decision to replace an existing test depends not only on the performance characteristics but also on other factors such as cost effectiveness, turnaround time, instrument maintenance, etc. The present study shows performance characteristics, whereas an economic evaluation is only briefly mentioned regarding a hospital's decision in making test selection. CONCLUSION: Among the three commercial kits, the specificity of Enzygnost Syphilis was the lowest. However, the replacement of any existing test depends greatly on the purpose of the individual laboratory whereas performance characteristics will provide us with an appropriate economic evaluation.

Sífilis materna y congénita en dos hospitales mexicanos: evaluación de una prueba diagnóstica rápida / Maternal and congenital syphilis in two Mexican hospitals: Evaluation of a rapid diagnostic test

Objective. To estimate the prevalence of maternal and neonatal syphilis, to assess the usefulness of a rapid treponemic diagnostic test, and to evaluate the frequency of screening for syphilis during prenatal care in Mexican women. Material and methods. This was a cross-sectional study that included 1,322 women interviewed in two hospitals (Hospital General in Cuernavaca, Morelos, and Hospital de la Mujer in Mexico City). Women answered a questionnaire on reproductive background, exposure to sexually transmitted infections and prenatal care. In order to diagnose syphilis, a rapid Determine TP test and a latex-VDRL test were used among all participating women. Positive cases were confirmed using FTA-ABS test. Results. Prevalence of serologically active syphilis was 0.3% (4/1322). Using as comparison standard a combination of VDRL and FTA-ABS tests, the Determine TP test had a sensitivity and specificity of 100%. Two newborns with positive FTA-ABS IgM, one negative with FTA-ABS IgM and one first-trimester abortion were found from seropositive women. Prenatal serum screening for syphilis was conducted in only 6.9% of women included in the study. Conclusions. Although the prevalence of maternal syphilis is relatively low, results show that if the situation found in the hospitals studied is shared by other hospitals in Mexico, a systematic screening for maternal syphilis would help to diagnose more cases of this infection than the number reported in official figures. Screening of syphilis in pregnant women using rapid tests may help in the prevention of congenital syphilis.

Objetivos. Estimar la seroprevalencia de sífilis materna y neonatal, analizar la utilidad de una prueba diagnóstica treponémica rápida y estudiar la frecuencia de escrutinio serológico de sífilis durante el embarazo. Material y métodos. Se realizó un estudio transversal que incluyó a 1,322 mujeres entrevistadas en dos hospitales (Hospital General de Cuernavaca y Hospital de la Mujer de la ciudad de México). Las mujeres contestaron un cuestionario sobre antecedentes ginecoobstétricos, de exposición a infecciones de transmisión sexual y de atención prenatal. Para el diagnóstico de sífilis se empleó en primera instancia la prueba rápida Determine TP a todas las participantes, posteriormente se aplicó de la misma manera la prueba de VDRL-látex. Los casos positivos se confirmaron con la prueba FTA-ABS. Resultados. La prevalencia de sífilis materna serológicamente activa fue de 0.3% (4/1322). Utilizando como pruebas de referencia la combinación de VDRL y FTA-ABS, Determine TP mostró una sensibilidad y una especificidad de 100%. De las mujeres seropositivas nacieron dos neonatos con FTA-ABS IgM positiva, en otra con FTA-ABS IgM negativa y la restante tuvo un aborto en el primer trimestre. En el transcurso del control prenatal la realización de la prueba de anticuerpos reagínicos por VDRL fue de 6.9% en la muestra de mujeres analizadas. Conclusiones. Si bien la frecuencia observada de sífilis materna es relativamente baja, los resultados encontrados en este estudio sugieren que la prevalencia de sífilis es mayor a la informada de manera global por la Dirección General de Epidemiología. El escrutinio sistemático de sífilis con pruebas rápidas en embarazadas, coadyuvaría a la prevención de la sífilis congénita.

Seroprevalencia de sífilis en mujeres embarazadas en San Luis Potosí / Seroprevalence of syphilis in pregnant women in San Luis Potosí

OBJETIVO: Determinar la seroprevalencia de sífilis en mujeres embarazadas. MATERIAL Y MÉTODOS: Encuesta seroepidemiológica en 1 857 mujeres que acudieron para la atención del parto a un hospital general de la ciudad de San Luis Potosí. RESULTADOS: Se diagnosticó sífilis en cinco (0.27 por ciento) mujeres al momento del parto. Los factores maternos asociados con una probabilidad superior de presentar sífilis incluyeron mayor edad materna, mayor número de embarazos previos y vivir en unión libre con su pareja. CONCLUSIONES: El número de recién nacidos expuestos a sífilis durante el embarazo, en la ciudad de San Luis Potosí, está subestimado. Los resultados de este estudio sustentan la necesidad de identificar, al momento del parto, a madres infectadas con sífilis.

The Effects of Treponema pallidum on Human Dendritic Cells

Cell mediated immune responses play a prominent role in syphilis, which is caused by Treponema pallidum. The role of dendritic cells (DC) in the syphilitic infection is not well understood in human. In the present study, we studied interaction of T. pallidum with DC, generated from human peripheral blood mononuclear cells with GM-CSF and IL-4. After adding T. pallidum for 16 hours to immature DC at culture day 7, the change of surface antigens on DC was monitored by flow cytometry, the amount of IL-12 in culture supernatant of DC was measured by ELISA and T cell stimulatory capacity of DC was checked in mixed lymphocyte reaction (MLR). We have observed an efficient phagocytosis of T. pallidum by electron microscopy as early as 2 hours after addition of T. pallidum to DC. Interaction of DC with T. pallidum resulted in increased surface expression of CD83 which was proportionally increased according to the number of T. pallidum. Expressions of CD80, CD86 and HLA-DR on DC were slightly increased. The amount of IL-12 in the culture supernatant of DC was increased (1, 099pg/ml) after the addition of T. pallidum. T. pallidum-infected DC also displayed enhanced T cell stimulatory capacity in MLR. As seen from the above, we observed phagocytosis of T. pallidum by DC as early as 2 hours after addition of T. pallidum to DC and found that T. pallidum can stimulate DC maturation which mean that DC modulate an protective immune response during T. pallidum infection.

Prevalencia y factores asociados con sífilis y herpes genital en dos grupos de población femenina / Prevalence and factors associated with syphilis and genital herpes in two female population groups

OBJETIVO: Evaluar la frecuencia de marcadores de infección por el VHS-2 y Treponema pallidum, y su relación con algunas características de las mujeres que acuden a consulta ginecológica. MATERIAL Y MÉTODOS: Durante 1994 y 1995 se estudiaron 388 y 448 mujeres en sendos hospitales, el primero en Cuernavaca, Morelos, México, y el segundo en la Ciudad de México. Las participantes proporcionaron una muestra de sangre para identificar, a través de la técnica de Western blot y las pruebas de VDRL y FTA-ABS, anticuerpos específicos contra los microrganismos mencionados; asimismo, contestaron un cuestionario sobre sus características sociodemográficas y de comportamiento sexual. Los datos se analizaron con los paquetes estadísticos SPSS y EGRET. RESULTADOS: Las frecuencias de anticuerpos contra el VHS-2 fueron 28.3 por ciento, para las mujeres del primer hospital, y 18.1 por ciento para las del segundo. En el caso de anticuerpos contra T pallidum las frecuencias fueron de 2.3 por ciento y 1.1 por ciento, respectivamente. La edad, el estado civil, la escolaridad y el número de compañeros sexuales de las mujeres estudiadas estuvieron asociados con los marcadores de infección por el VHS-2. CONCLUSIONES: Se encontraron frecuencias bajas de infección por los microrganismos estudiados en ambos grupos de mujeres. La infección por el VHS-2 estuvo asociada a periodos de exposición, comportamiento sexual y nivel socioeconómico de las mujeres.

Novo teste de immunoblot (IB-rTp) com antígenos recombinantes de Treponema pallidum para o diagnóstico da sífilis / Novel immunoblot treponemal assay using recombinat antigens for immunodiagnosis of syphilis

A sífilis ainda é um problema de saúde pública, apesar dos esforços empreendidos para a sua prevençäo e a existência de tratamento eficiente. Os testes sorológicos säo fundamentais para o diagnóstico da doença. O presente trabalho descreve um novo teste de immunoblot )IB-rTp), utilizando três antígenos recombinantes de Treponema pallidum (GST-rTp47, GST-rTp17 e GST-rTp15), além da glutathiona S-tranferase (GST) purificada. FOram testadas 114 amostras séricas (53 de pacientes com sífilis, 50 de indivíduos sadios e 11 de pacientes com outras doenças) verificou-se concordância de 100 por cento em o IB-rTp e os testes treponêmicos convencionais, que utilizam antígeno nativo. A reatividade, quando presente, foi especificada contra a fraçäo treponêmica de cada proteína de fusäo. O teste IB-rTp apresentou alta especificidade; é de fácil execuçäo, näo requerendo absorçäo prévia do soro, sendo de utilidade como teste confirmatório de sífilis

Syphilitic seroreactivity among the Thai population aged 50 years and above: value of mass screening.

The present study is concerned with the evaluation of routine serologic screening for syphilis among the general population aged 50 and above living in a suburban area of Bangkok, Thailand. Among 941 subjects aged 50 years and above recruited in a cohort study named the CERB study, blood of 798 subjects was obtained and their sera were tested for syphilis using the passive particle agglutination test for detection of antibodies to Treponema pallidum (TPPA) and the rapid plasma reagin (RPR). Of 798 subjects, 132 (16.5%) were reactive to serum TPPA. Their mean age (standard deviation) was 61.4 (8.4) years. Fifty-six subjects (42.4%) were male. Prevalence of syphilitic seroreactivity increased with age (p = 0.004). Prevalence of syphilitic seroreactivity among male and female subjects was 18.3% and 15.4% respectively (p = 0.001). Prevalence of syphilitic seroreactivity among subjects with formal education (14.4%) was significantly lower than that among subjects without formal education (26.5%) (p = 0.001). Of 132 subjects, 10 (7.6%) had and a rapid plasma reagin (RPR) titer of 1:16 or higher. None of them reported symptoms of primary or secondary syphilis during the initial survey. Of 125 subjects who were interviewed in depth during the revisit survey, 24 subjects had known about their syphilitic infection prior to this study. Among these 24 subjects, 4 had not received any treatment for syphilis and 1 used herbal medicine. Therefore, 106 subjects (84.4%) were untreated or inadequately treated. These findings suggest the value of mass screening for syphilis among the Thai population aged 50 years and above.

Virulent Treponema pallidum 47 kDa antigen regulates the expression of cell adhesion molecules and binding of T-lymphocytes to cultured human dermal microvascular endothelial cells

Perivasculitis and endothelial cell abnormalities are prominent histopathologic features of syphilis. Various cutaneous lesions are the main clinical features of syphilis. We examined whether Treponema pallidum 47 kDa antigen regulates the expression of cell adhesion molecules on human dermal microvascular endothelial cells (HDMEC) and the regulation of T-lymphocytes binding to HDMEC. Using immunofluorescence flow cytometry and enzyme-linked immunosorbent assay (ELISA), we demonstrated that T. pallidum upregulated the expression of adhesion molecules, including intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1) and E-selectin. The 47 kDa antigen of T. pallidum also activated endothelium as measured by the upregulation of the expression of adhesion molecules on HDMEC, and it also promoted an increased adherence of T-lymphocytes to HDMEC. The expressions of ICAM-1 and VCAM-1 on HDMEC and the adherence of T-lymphocytes to HDMEC were inhibited by treatment with anti-TNF-alpha antibody or anti-IL-1alpha antibody. These results show that T. pallidum or T. pallidum-specific 47 kDa antigen are capable of stimulating HDMEC to increase the expression of ICAM-1, VCAM-1 and E-selectin and thereby, promote the adherence of T-lymphocytes. The whole process may play an important role in the immunopathogenesis of syphilis and it is likely that TNF-alpha and IL-1alpha are involved.

Analysis of Treponema pallidum recombinant antigens for diagnosis of syphilis by Western blotting technique

Os antigenos recombinantes de Treponema pallidum GST-rTp47, GST-rTp17 e GST-rTp15, produzidos em fusao com glutationa S-transferase (GST) em E. coli, foram analisados quanto ao potencial diagnostico da sifilis pela tecnica de Western blotting. Foram testadas 53 amostras, sendo 25 pacientes em diferentes estagios clinicos da sifilis, com resultados positivos no teste treponemico classico; 25 amostras procedentes de doadores de banco de sangue, com sorologia negativa e 3 de pacientes com doenca sexualmente transmissivel nao relacionado a sifilis. Todas as amostras de pacientes com sifilis apresentaram alta reatividade com o antigeno GST-rTp17...