Detection of HPV DNA in vaginal samples self-collected by women living with HIV treated through the Brazilian public health system: Prevalence and analysis of risk factors

ABSTRACT Background: Women living with human immunodeficiency virus (HIV) (WLWH) are more likely to be infected with the oncogenic human papillomavirus (HPV). We assessed the prevalence of high-risk (HR) (16/18/31/33/35/39/45/51/52/56/58/59/68/73/82), probable high-risk (pHR) (26/53/66), and low-risk (LR) (6/11/40/42/43/44/54/61/70) HPV types and their associated risk factors. Methods: This cross-sectional study of WLWH aged 18-64 years included one laboratory and eight HIV-specialty healthcare facilities in the pilot network. Descriptive statistics were used to assess sociodemographic and behavioral characteristics. Adjusted analyses were conducted to evaluate risk factors associated with HR and/or pHR HPV infection in WLWH. Results: From May/2021 to May/2022, 1,914 (92.5%) WLWH participated in the pilot study and had valid HPV-DNA results of self-collected vaginal samples. The median age of the participants was 45 years, 60.1% had ≥ 9 years of schooling, 80.5% were ≤ 18 years at first sexual intercourse, and 51.7% had > 4 sexual partners throughout life. The prevalence of any HPV type, HR HPV, pHR HPV, and LR HPV was 65.8%, 49.6%, 16.7%, and 40.0%, respectively. Age was inversely associated with pHR and/or HR-HPV (p < 0.001), and education level was inversely associated with HR-HPV (p = 0.003) types. Any HR or pHR was associated with being single (p = 0.029) and exchanging sex for drugs (p = 0.037). Conclusions: The prevalence of HPV, especially HR HPV, among WLWH is high in Brazil, highlighting the need for HPV screening in this population. Self-collection of vaginal samples is an important strategy for increasing testing access.

Syphilis, history, science, and arts: syphilis history calendar / Sífilis, história, ciência e artes: calendário da história da sífilis / Sífilis, historia, ciencia y artes: calendario de la historia de la sífilis

Introduction: Syphilis is a very old disease involved by many mysteries. From its origins to the reason why even having simple and effective diagnosis and treatment for more than half a century, it remains, in 2021, as a public health problem practically all over the world. Be it in high-developing countries as well as in developing ones. Objective: To relate and describe, with dating, by day, month and year, in a calendar model, events and important characters that involve studies and quotes about syphilis. Methods: We used the prior personal knowledge of health professionals and researchers, who have worked in the field of sexually transmitted diseases for decades, combined with the search on the Internet, often on Wikipedia and Google, about events and characters involving subjects and publications about syphilis. Results: After several analyses and researches, we listed 68 facts and characters from around the world, including several Brazilians. Conclusion: At a time (2020 and 2021) where the subject, worldwide, on a daily basis, is SARS-CoV-2/COVID-19, this unprecedented publication on facts and characters involving syphilis can be a milestone so that, in addition to technical and scientific knowledge, the academic community realizes the anthropological, sociological, and cultural wealth that involves an entity that is much more than an infectious disease, such as syphilis.

Introdução: Sífilis é uma doença muito antiga envolvida por muitos mistérios. Desde a sua origem até o porquê de mesmo tendo diagnóstico e tratamento simples e eficazes há mais de meio século, se mantém, em 2021, como um problema de saúde pública em praticamente todo o mundo. Seja em países de alto desenvolvimento como naqueles em desenvolvimento. Objetivo: Relacionar e descrever, com datação, por dia, mês e ano, em um modelo de calendário, acontecimentos e personagens marcantes que envolvem estudos e citações sobre a sífilis. Métodos: Usamos o conhecimento prévio pessoal, de profissionais de saúde, de pesquisadores, com atuações na área das doenças sexualmente transmissíveis há décadas, aliado a busca na internet, muitas das vezes em Wikipédia e Google, sobre acontecimentos e personagens que envolvem assuntos e publicações sobre sífilis. Resultados: Depois de várias análises e pesquisas relacionamos 68 fatos e personagens de âmbito mundial, sendo vários brasileiros. Conclusão: Em uma época (2020 e 2021) onde o assunto, em todo o mundo, diuturnamente, é SARS-CoV-2/COVID-19 esta inédita publicação sobre fatos e personagens que envolvem sífilis pode ser um marco para que, além de conhecimentos técnicos e científicos, a comunidade acadêmica se dê conta da riqueza antropológica, sociológica, cultural que envolve uma entidade que é muito mais que uma doença infectocontagiosa, como a sífilis.

Introducción: La sífilis es una enfermedad muy antigua envuelta por muchos misterios. Desde sus orígenes hasta el motivo por el cual aun teniendo un diagnóstico y tratamiento sencillo y eficaz desde hace más de medio siglo, sigue siendo, en 2021, un problema de salud pública prácticamente en todo el mundo. Tanto en países de alto desarrollo como en países en desarrollo. Objetivo: Relacionar y describir, con datación, por día, mes y año, en un modelo de calendario, eventos y personajes importantes que involucran estudios y citas sobre la sífilis. Métodos: Utilizamos el conocimiento personal previo de profesionales e investigadores de la salud, que han trabajado en el campo de las enfermedades de transmisión sexual durante décadas, combinado con la búsqueda en Internet, a menudo en Wikipedia y Google, sobre eventos y personajes que involucran temas y publicaciones sobre sífilis. Resultados: Después de varios análisis e investigaciones, enumeramos 68 hechos y personajes de todo el mundo, incluidos varios brasileños. Conclusión: En un momento (2020 y 2021) donde el tema, a nivel mundial, a diario, es el SARS-CoV-2/COVID-19, esta publicación sin precedentes sobre hechos y personajes relacionados con la sífilis puede ser un hito para que, además Para el conocimiento técnico y científico, la comunidad académica se da cuenta de la riqueza antropológica, sociológica y cultural que involucra a una entidad que es mucho más que una enfermedad infecciosa, como la sífilis.

Brazilian protocol for sexually transmitted infections 2020: syphilis diagnostic tests

Abstract The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. This article presents direct tests to detect Treponema pallidum in lesions and algorithms that combine treponemal and non-treponemal antibody tests to assist in syphilis diagnosis, with the aim of contributing to the efforts of health service managers and health professionals in qualifying health care. The article also covers the use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring the treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The critical role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

An investigation about chronic prostatitis in ankylosing spondylitis

Abstract Background: Chronic prostatitis has been a common disease reported with high frequency in ankylosing spondylitis (AS) even from decades ago. Infectious (Chlamydia trachomatis) or non-infectious (uric acid) prostatitis can hypothetically trigger vertebral inflammation in AS. This study aimed to assess the features of chronic prostatitis in patients with AS compared to healthy controls. Methods: A cross-sectional study including male patients with AS and healthy controls who agreed to undergo a prostate examination was conducted. Structured clinical interviews, prostate physical examinations, and cytological, biochemical, and microbiological tests on urinary samples collected before and after standardized prostatic massage (pre- and post-massage test) were performed. Results: Ninety participants (45 AS patients, mean age: 52.5 ± 10.0 years, with longstanding disease, 12.4 ± 6.9years, and 45 controls, mean age: 52.8 ± 12.1 years) were included. National Institutes of Health - Chronic Prostatitis Symptom Index (NIH-CPSI) scores were similar in the AS and control groups (4.0 [1.0-12.0] vs. 5.0 [1.0—8.5], p = 0.994). The frequencies of symptoms of chronic prostatitis (NIH-CPSI Pain Domain ≥4) were also similar in both groups (23.3% vs. 22.7%, p = 0.953). Results of polymerase chain reaction tests for Chlamydia trachomatis were negative in all tested urinary samples, and uric acid concentrations and leukocyte counts were similar in all pre- and post-massage urinary samples. Conclusions: In this study, chronic prostatitis occurred in male patients with AS, but its frequency and characteristics did not differ from those found in the healthy male population of similar age.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis / Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: pruebas de diagnóstico de sífilis

As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: infecção pelo papilomavírus humano (HPV) / Brazilian Protocol for Sexually Transmitted Infections 2020: human papillomavirus (HPV) infection / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: infección por papilomavirus humano (PVH)

O artigo aborda a infecção pelo papilomavírus humano (human papillomavirus, HPV), tema constitutivo do Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis, publicado pelo Ministério da Saúde. Tal documento foi elaborado com base em evidências científicas e validado em discussões e consenso entre especialistas. São apresentados aspectos epidemiológicos e clínicos, bem como orientações para os gestores e profissionais de saúde no diagnóstico, tratamento e prevenção da infecção pelo HPV. Este tema representa importante problema de saúde pública, haja vista essa infecção sexualmente transmissível ser a mais prevalente no mundo, capaz de desencadear o processo oncogênico do câncer do colo uterino, além de possibilitar a ocorrência de verrugas anogenitais. Neste artigo, são apresentadas informações importantes para o conhecimento do HPV, estratégias de ação para a prevenção e controle da infecção, uma assistência de qualidade e tratamento efetivo da doença.

This article addresses human papillomavirus (HPV) infection, this being one of the topics covered by the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health. The Protocol and Guidelines have been developed based on scientific evidence and validated in discussions with specialists. This article presents epidemiological and clinical aspects, as well as guidelines for health service managers and health workers about diagnosing and treating people with papillomavirus infection. This theme is an important public health problem, since it is the most prevalent sexually transmitted infection in the world, capable of triggering the oncogenic process of cervical cancer, as well as the possibility anogenital warts occurring. Important information is presented for gaining knowledge about HPV, as well as action strategies for infection prevention and control, provision of quality care and effective treatment of the disease.

El tema del papilomavirus humano (PVH) es uno de los capítulos del Protocolo Clínico y Conductas Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual, publicado por el Ministerio de Salud de Brasil. Este documento fue desarrollado en base a evidencia científica y validado en discusiones con especialistas. Se presentan aspectos epidemiológicos y clínicos, así como guías para gestores y profesionales de la salud en el diagnóstico, tratamiento y prevención de la infección por papilomavirus. Este tema constituye uno de los principales problemas de salud pública, pues además de ser la infección transmitida sexualmente más prevalente en todo el mundo, puede tener implicaciones para el proceso oncogénico del cáncer de cuello uterino y la posibilidad de presentar verrugas anogenitales. Se presenta información sobre estrategias de acciones de prevención y control, las cuales son importantes para entender el problema, ofrecer asistencia de calidad y tratamiento efectivo.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: testes diagnósticos para sífilis / Brazilian Protocol for Sexually Transmitted Infections 2020: syphilis diagnostic tests / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: pruebas de diagnóstico de sífilis

Resumo As recomendações de testes diagnósticos para investigação da sífilis compõem o Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis e o Manual Técnico para Diagnóstico de Sífilis, publicados pelo Ministério da Saúde do Brasil. Tais recomendações foram elaboradas com base em evidências científicas e discussões com painel de especialistas. Visando contribuir com gestores e profissionais de saúde na qualificação da assistência, este artigo apresenta o uso dos exames diretos para detecção de Treponema pallidum em lesões, assim como algoritmos que combinam testes imunológicos treponêmicos e não treponêmicos para auxiliar no diagnóstico da sífilis. O artigo também apresenta o uso dos testes não treponêmicos para investigação de neurossífilis e orientações para interpretação do título dos anticorpos não treponêmicos no monitoramento do tratamento e diagnóstico de sífilis congênita, bem como as perspectivas futuras de inovações em diagnóstico. Ressalta-se, além disso, o importante papel dos testes rápidos imunocromatográficos treponêmicos para a saúde pública e o enfrentamento da sífilis.

Abstract The recommendations for diagnostic tests for investigating syphilis are part of the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections and the Technical Manual for Syphilis Diagnosis, published by the Brazilian Ministry of Health. These recommendations were developed based on scientific evidence and discussions with a panel of experts. Aiming to contribute to the efforts of health service managers and health professionals in qualifying health care, this article presents the use of direct tests to detect Treponema pallidum in lesions, as well as algorithms that combine treponemal and non-treponemal antibody tests to assist in the diagnosis of syphilis. The article also covers use of non-treponemal tests to investigate neurosyphilis and guidelines for interpreting non-treponemal antibody titers in monitoring treatment and diagnosis of congenital syphilis, as well as prospects for innovations in diagnosis. The important role of rapid immunochromatographic treponemal tests for public health and for addressing syphilis is also highlighted.

Resumen Las recomendaciones de las pruebas de diagnóstico de sífilis forman parte del Protocolo Clínico y Directrices Terapéuticas para la Atención Integral a las Personas con Infecciones de Transmisión Sexual y del Manual Técnico para el Diagnóstico de Sífilis, publicados por el Ministerio de Salud de Brasil. Estos documentos fueron preparados en base a la evidencia científica y discusiones con especialistas. Con el objetivo de contribuir con los administradores y profesionales de la salud pública en la calificación de la atención, este artículo presenta el uso de pruebas directas para la detección de Treponema pallidum en lesiones, además de algoritmos que combinan pruebas treponémicas y no treponémicas para ayudar al diagnóstico. También se presentan las pruebas no treponémicas para investigar la neurosífilis y orientaciones para la interpretación de títulos de anticuerpos no treponémicos para monitorear la respuesta al tratamiento y diagnosticar la sífilis congénita, así como las perspectivas futuras para las innovaciones diagnósticas. También se enfatiza el uso de pruebas inmunocromatográficas treponémicas rápidas como una herramienta importante para la salud pública y para el enfrentamiento de la sífilis.

Protocolo Brasileiro para Infecções Sexualmente Transmissíveis 2020: infecção pelo papilomavírus humano (HPV) / Brazilian Protocol for Sexually Transmitted Infections 2020: human papillomavirus (HPV) infection / Protocolo Brasileño para Infecciones de Transmisión Sexual 2020: infección por papilomavirus humano (PVH)

Resumo O artigo aborda a infecção pelo papilomavírus humano (human papillomavirus, HPV), tema constitutivo do Protocolo Clínico e Diretrizes Terapêuticas para Atenção Integral às Pessoas com Infecções Sexualmente Transmissíveis, publicado pelo Ministério da Saúde. Tal documento foi elaborado com base em evidências científicas e validado em discussões e consenso entre especialistas. São apresentados aspectos epidemiológicos e clínicos, bem como orientações para os gestores e profissionais de saúde no diagnóstico, tratamento e prevenção da infecção pelo HPV. Este tema representa importante problema de saúde pública, haja vista essa infecção sexualmente transmissível ser a mais prevalente no mundo, capaz de desencadear o processo oncogênico do câncer do colo uterino, além de possibilitar a ocorrência de verrugas anogenitais. Neste artigo, são apresentadas informações importantes para o conhecimento do HPV, estratégias de ação para a prevenção e controle da infecção, uma assistência de qualidade e tratamento efetivo da doença.

Abstract This article addresses human papillomavirus (HPV) infection, this being one of the topics covered by the Clinical Protocol and Therapeutic Guidelines for Comprehensive Care for People with Sexually Transmitted Infections, published by the Brazilian Ministry of Health. The Protocol and Guidelines have been developed based on scientific evidence and validated in discussions with specialists. This article presents epidemiological and clinical aspects, as well as guidelines for health service managers and health workers about diagnosing and treating people with papillomavirus infection. This theme is an important public health problem, since it is the most prevalent sexually transmitted infection in the world, capable of triggering the oncogenic process of cervical cancer, as well as the possibility anogenital warts occurring. Important information is presented for gaining knowledge about HPV, as well as action strategies for infection prevention and control, provision of quality care and effective treatment of the disease.

Resumen El tema del papilomavirus humano (PVH) es uno de los capítulos del Protocolo Clínico y Conductas Terapéuticas para la Atención Integral a Personas con Infecciones de Transmisión Sexual, publicado por el Ministerio de Salud de Brasil. Este documento fue desarrollado en base a evidencia científica y validado en discusiones con especialistas. Se presentan aspectos epidemiológicos y clínicos, así como guías para gestores y profesionales de la salud en el diagnóstico, tratamiento y prevención de la infección por papilomavirus. Este tema constituye uno de los principales problemas de salud pública, pues además de ser la infección transmitida sexualmente más prevalente en todo el mundo, puede tener implicaciones para el proceso oncogénico del cáncer de cuello uterino y la posibilidad de presentar verrugas anogenitales. Se presenta información sobre estrategias de acciones de prevención y control, las cuales son importantes para entender el problema, ofrecer asistencia de calidad y tratamiento efectivo.

Molecular profiling of drug resistant isolates ofMycobacterium tuberculosis in the state of Santa Catarina, southern Brazil

Drug resistance is a global threat and one of the main contributing factors to tuberculosis (TB) outbreaks. The goal of this study was to analyse the molecular profile of multidrug-resistant TB (MDR-TB) in the state of Santa Catarina in southern Brazil. Fifty-three MDR Mycobacterium tuberculosisclinical isolates were analysed by spoligotyping and a partial region of therpoB gene, which is associated with rifampicin resistance (RMP-R), was sequenced. Some isolates were also distinguished by their mycobacterial interspersed repetitive units (MIRU). S531L was the most prevalent mutation found within rpoBin RMP-R isolates (58.5%), followed by S531W (20.8%). Only two MDR isolates showed no mutations withinrpoB. Isolates of the Latin American Mediterranean (LAM) family were the most prevalent (45.3%) found by spoligotyping, followed by Haarlem (9.4%) and T (7.5%) families. SIT106 was found in 26.4% of isolates and all SIT106 isolates typed by MIRU-12 (5 out of 14) belong to MIT251. There was a high correlation between the S531W mutation and the LAM family mainly because all SIT2263 (LAM9) isolates carry this mutation. Among isolates with the S531W mutation in rpoB MIRU demonstrates a cluster formed by four isolates (SIT2263 and MIT163) and very similar profiles were observed between eight of the nine isolates. Better characterisation of TB isolates may lead to new ways in which to control and treat TB in this region of Brazil.

External quality assessment in the voluntary counseling and testing centers in the Brazilian Amazon using dried tube specimens: results of an effectiveness evaluation

INTRODUCTION : In 2011, the Brazilian Ministry of Health rolled out a program for the external quality assessment of rapid human immunodeficiency virus (HIV) tests using the dried tube specimen (DTS) method (EQA-RT/DTS-HIV). Our objective was to evaluate the implementation of this program at 71 voluntary counseling and testing centers (VCTCs) in the Brazilian Legal Amazonian area one year after its introduction. METHODS : Quantitative and qualitative study that analyzed secondary data and interviews with healthcare workers (HCWs) (n=39) and VCTC coordinators (n=32) were performed. The assessment used 18 key indicators to evaluate the three dimensions of the program's logical framework: structure, process, and result. Each indicator was scored from 1-4, and the aggregate results corresponding to the dimensions were expressed as proportions. The results were compared to the perceptions of the HCWs and coordinators regarding the EQA-RT/DTS-HIV program. RESULTS: The aggregate scores for the three dimensions of structure, process, and result were 91.7%, 78.6%, and 95%, respectively. The lowest score in each dimension corresponded to a different indicator: access to Quali-TR online system 39% (structure), registration in Quali-TR online system 38.7% (process), and VCTC completed the full process in the program's first round 63.4% (result). Approximately 36% of the HCWs and 52% of the coordinators reported enhanced trust in the program for its rapid HIV testing performance. CONCLUSIONS: All three program dimensions exhibited satisfactory results (>75%). Nevertheless, the study findings highlight the need to improve certain program components. Additionally, long-term follow-ups is needed to provide a more thorough picture of the process for external quality assessment. .

Identificação por PCR-multiplex de espécies de Mycobacterium / Identification of Mycobacterium species by multiplex PCR assay

Reação em Cadeia da (La reacción en cadena de la) Polimerase Multiplex (PCR- multiplex) é um (es un) método diagnostico potencialmente útil para identificação rápida e acurada (y exacta) de Mycobacterium tuberculosis (TB) e de micobactérias não-tuberculosas (MNT) clinicamente relevantes. Este sistema multi-iniciadores (multiprimers) é capaz de identificar o (el) antígeno alfa de 32-kDa presente na (en la) maioria das (de las) espécies do gênero Mycobacterium, a sequência de inserção IS6110 pertencente ao complexo (al complejo) TB e as sequências espécie-específicas do (del) gene mtp 40 de Mycobacterium tuberculosis. Cento e oito a mostras (Ciento ocho muestras) (escarro [esputo] e extra-pulmonares) oriundas do Setor de Tuberculose do Laboratório Central de Saúde Pública de Santa Catarina (LACEN/SC) foram processadas e analisadas no Laboratório de Biologia Molecular e Micobactérias (LBMM) da Universidade Federal de Santa Catarina, Florianópolis, Brasil, entre janeiro e junho de 2011, representando 30% dasa mostras (de las muestras) positivas/ano no (al año en el) LACEN/SC. Dos (Entre los) 108 isolados (aislamientos), 80% foram identificados como TB e 20% como MNT. Os resultados da PCR multiplex foram comparados com os resultados do PRA-hsp65 (PCR do gene hsp65 seguida por análise de restrição enzimática) e demonstraram 100% de concordancia (índice de concordância kappa 1). A PCR multiplex é uma ferramenta (es una herramienta)...

Mycobacteria mobility shift assay: a method for the rapid identification of Mycobacterium tuberculosis and nontuberculous mycobacteria

The identification of mycobacteria is essential because tuberculosis (TB) and mycobacteriosis are clinically indistinguishable and require different therapeutic regimens. The traditional phenotypic method is time consuming and may last up to 60 days. Indeed, rapid, affordable, specific and easy-to-perform identification methods are needed. We have previously described a polymerase chain reaction-based method called a mycobacteria mobility shift assay (MMSA) that was designed for Mycobacterium tuberculosis complex (MTC) and nontuberculous mycobacteria (NTM) species identification. The aim of this study was to assess the MMSA for the identification of MTC and NTM clinical isolates and to compare its performance with that of the PRA-hsp65 method. A total of 204 clinical isolates (102 NTM and 102 MTC) were identified by the MMSA and PRA-hsp65. For isolates for which these methods gave discordant results, definitive species identification was obtained by sequencing fragments of the 16S rRNA and hsp65 genes. Both methods correctly identified all MTC isolates. Among the NTM isolates, the MMSA alone assigned 94 (92.2%) to a complex or species, whereas the PRA-hsp65 method assigned 100% to a species. A 91.5% agreement was observed for the 94 NTM isolates identified by both methods. The MMSA provided correct identification for 96.8% of the NTM isolates compared with 94.7% for PRA-hsp65. The MMSA is a suitable auxiliary method for routine use for the rapid identification of mycobacteria.

Herbalists, traditional healers and pharmacists: a view of the tuberculosis in Ghana

This paper is the result of a visit by Brazilian researchers to Ghana, with the aim of improving understanding of the relationship between traditional healers and conventional health practices, specifically in relation to tuberculosis. Through this exploratory visit, this group of researchers promoted by the Edital Pro Africa (CNPq) had an opportunity to learn about, reflect on, and discuss the different social, economic and cultural realities and contexts that have led to the different health conditions and forms of healthcare in Ghana. Besides the direct relationship between the social and economic conditions of the country and the health of its population, it was also concluded that there is a clear distancing, in the Ghanaian reality, between the traditional healers and the conventional system, in terms of culture and modes of operation, each constituting isolated systems with little or no collaboration between them.The visit enabled us to see the difficulties involved in managing TB, including diagnosis, treatment, monitoring and co-infection with HIV. The majority of patients with TB only go to hospital after several attempts at self-medication, due to the non-specificity of the principal symptoms, and also to the trust in the traditional medicine. Initiatives to encourage research into medicinal plants in Ghana are seeking partnerships with developed countries, but not always with clear or secure national interests. For the traditional healers, there are high hopes that the information gathered by researchers from the local universities, on the plants and traditional methods they use, will result in affirmation and recognition of their practices, but they complain strongly that they receive no feedback on the research carried out.

Cervical cytology, HPV DNA and mRNA - a comparative study in 162 patients / Citologia cervicovaginal, DNA HPV e RNAm - estudo comparativo em 162 pacientes

In contrast to the general improvement of the socioeconomic status of the Brazilian population, pathologies that are characteristic of poor health assistance persist. Among those, cervical cancer (CC) is emblematic; it still presents a persistently high incidence. Objective: to compare the performance of cervical cytology to HPV DNA and mRNA detection methods in 162 patients undergoing routine gynecological clinical practice. Methods:a total of 162 patients attended during routine gynecological examination in a private clinic in Florianópolis, Santa Catarina, Brazil, had cervical samplescollected and processed for cytopathological and molecular tests, conventional PCR and NASBA. Positive samples positive for HPV DNA were submittedto Type-Specific PCR (TS-HPV PCR). Patients with altered smears were submitted to colposcopy and biopsy. Results: among the 162 samples, 19.8%(32/162) had altered smears, being 4/32 classified as ASC-H, 9/32 as ASC-US, 9/32 as LSIL and 10/32 as HSIL. Biopsies revealed nine cases of CIN I,nine CIN II and one CIN III, while seven were negative for cervical neoplasia. Overall, HPV DNA was detected in 38.3% (62/162) of the samples and HPV E6/E7 mRNA expression was found in 13.6% (22/162). Using TS-HPV PCR, HPV 16 was the most frequent type, found in 8% of the samples (5/62).Considering CIN2+ the gold-standard, cytology had 38.5% of specificity. Sensitivity and specificity of HPV-DNA PCR and NASBA were, respectively,100% and 60%; 18.7% and 68.7%. Conclusion: mRNA E6/E7 expression was not a highly specific or sensitive marker for prevalent cervical disease while HPV DNA may be used for cervical cancer screening only in conjunction to more specific adjuvant tests.

Em contraste com a melhora geral da situação socioeconômica da população brasileira, patologias que são características de uma deficiente assistência à saúde persistem. Entre elas, o câncer cervical (CC) é emblemático, ainda apresentando uma persistente alta incidência. Objetivo: avaliaro desempenho da citologia e de métodos de detecção de DNA e RNAm de HPV em 162 pacientes submetidas a prática clínica ginecológica de rotina.Métodos: cento e sessenta e duas pacientes atendidas em uma clínica particular de Florianópolis, Santa Catarina, Brasil, tiveram amostras cervicais coletadas e processadas para estudo citopatológico e molecular; PCR convencional e NASBA. Amostras positivas para o DNA do HPV foram submetidas àPCR tipo-específica (PCR HPV-TE). Resultados: entre as 162 amostras, 19,8% (32/162) apresentaram esfregaços alterados, sendo 4/32 classificadas comoASC-H, 9/32 como ASC-US, 9/32 como LSIL e 10/32 como HSIL. Biópsias revelaram nove casos de NIC I, nove casos de NIC II e um caso de NIC III. ODNA do HPV foi detectado em 38,3% (62/162) das amostras. Expressão de E6/E7 (RNAm) foi encontrada em 13,6% (22/162) das amostras. Utilizando a PCR tipo-específica (HPV-TE), o HPV 16 foi o tipo mais frequente, encontrado em 8% (5/62) das amostras HPV+. Considerando NIC 2+ o padrão-ouro,a especificidade da citologia foi de apenas 38,5%, enquanto a sensibilidade e a especificidade da PCR DNA e RNAm foram, respectivamente, 100% e 60%;18,7% e 68,7%. Conclusão: a expressão de E6/E7 RNAm não se mostrou um marcador altamente específico ou sensível para doença cervical prevalente,enquanto o DNA HPV pode ser utilizado para rastreamento apenas em conjunto com testes adjuvantes mais específicos.

Diagnosis of the pulmonary tuberculosis by polymerase chain reaction: a comparative study between HIV - positive and - negative individuals

This study was performed to assess the efficiency of polymerase chain reaction (PCR) directly from sputum for the diagnosis of pulmonary tuberculosis by comparison between HIV-positive and HIV-negative individuals. Sputum samples were collected from hospitalized patients admitted with a clinical diagnosis of pulmonary tuberculosis, and subjected to smear microscopy, culture on LJ medium and detection of M. tuberculosis by PCR. Sensitivity, specificity, and predictive values (positive and negative) were calculated using smear and/or culture at day 42 as the gold standard, by comparing the yield in HIV-positive and HIV-negative individuals. Regardless of serostatus, the technique's yield had 62% sensitivity, 70% specificity, 79% positive predictive value, 50% negative predictive value, and 65% accuracy. HIV-negative had 64% sensitivity, 74% specificity, 75% positive predictive value, 63% negative predictive value, and 68% accuracy. HIV-positive had 59% sensitivity, 33% specificity, 87% positive predictive value, 10% negative predictive value, and 56% accuracy. The PCR showed a higher yield in HIV-negative individuals compared to HIV-positive individuals.

Micobactérias: epidemiologia e diagnóstico / Mycobacteria: epidemiology and diagnosis

Estima-se que um terço da população mundial esteja infectada com Mycobacterium tuberculosis, oque resulta em 2 milhões de mortes anualmente. No mundo, são registrados mais de 8 milhões de novos casos de tuberculose (TB) por ano e o Brasil ocupa o décimo nono lugar dentre os 23 países detentores da maior carga de TB. Os fatores determinantes para o controle desta doença incluem a detecção rápida, a terapia adequada e os meios para que sejam evitadas futuras transmissões. O diagnósticoconvencional (baciloscopia e cultura do microrganismo) apresenta limitações quanto ao tempo de execução e à operacionalidade, visto que o resultado pode levar até 60 dias para ser liberado.Portanto, a importância da detecção precoce do bacilo se torna fundamental para o bloqueio da cadeia de transmissão da TB. Já as micobacterioses, doenças causadas pelas micobactérias não tuberculosas, também estão provocando um importante impacto em razão do aumento dos surtos de infecções cirúrgicas. A identificação rápida e específica destes microrganismos é importante para o diagnóstico e a consequente escolha do tipo de tratamento do paciente, que está diretamente relacionada com a espécie. Portanto, o conhecimento dos agentes etiológicos das doenças causadaspelas micobactérias e o diagnóstico sensível e específico permitem o tratamento adequado e,consequentemente, o bloqueio da cadeia de transmissão da TB e o controle dos surtos relacionados às micobactérias não tuberculosas.

It is estimated that one third of the world population is infected with Mycobacterium tuberculosis, resulting in 2 million deaths annually. More than 8 million new cases of tuberculosis (TB) are registered per year worldwide, and Brazil ranks 19th among the top 23 countries with the highest rates of TB. The determining factors for the control of this disease include early detection, appropriate therapy and measuresfor avoiding transmission. The conventional diagnoses (smear and microorganism culture) have time limitations for implementation and operation, since the result may take up to 60 days to be released. Therefore, early detection is critical for blockingthe chain of TB transmission. Mycobacterial diseases caused by nontuberculous mycobacteria are also having a major impact due to increased outbreaks of surgical infections. Thereby, the rapid and specific identification of microorganisms is important for the diagnosis, which will determine the type of treatment (treatment according to species). Knowledge of etiologic agents of mycobacterial diseases, as well as sensitive and specific diagnosis allows proper treatment by blocking the chain of TB transmission and controlling nontuberculous mycobacteria outbreaks.

Prevenção do câncer cervical: associação da citologia oncótica a novas técnicas de biologia molecular na detecção do papilomavírus humano (HPV) / Cervical cancer prevention: association of new techniques of molecular biology with the oncotic cytology in detection of human papillomavirus (HPV)

O câncer do colo uterino é conhecido como uma das causas mais frequentes de óbito na população feminina em todo o mundo. A infecção persistente por papilomavírus humano (HPV) é o principal fator de risco para o câncer cervical e suas lesões precursoras. Objetivo: avaliar os métodos diagnósticos do câncer cervical: citologia oncótica, DNA do HPV pela PCR, detecção do RNAm das proteínas E6 e E7 dos HPVs de alto risco, para acrescentar novos marcadores na detecção do HPV em amostras com citologia alterada. Métodos: foram avaliados os resultados da citologia oncótica de3.000 pacientes, atendidas no Laboratório Vital, Nonoai - RS, de março de 2009 a janeiro de 2010, classificando-as conforme a faixa etária e codificação de Bethesda, 2001. Foram selecionados dois grupos: o grupo A incluiu pacientes com citologia prévia alterada e o grupo B, pacientes com citologia sem anormalidades. Foram coletadas 47 amostras, 22 do grupo A e 25 do grupo B. Três amostras do grupo B apresentaram inibidores da PCR e foram excluídas.A análise dos dados considerou 22 amostras em cada grupo. A detecção do HPV foi feita pela reação em cadeia da polimerase (PCR) utilizando-se os primers consenso MY9/11 e GP5+/6+. O RNAm das proteínas E6 e E7 foi detectado pelo método Real-Time NASBA. Resultados: nas amostras do grupo A, houve detecção do DNA/HPV pela PCR em 14 (64%) amostras, e RNAm das proteínas E6 e E7 do HPV em seis (27%) amostras. Todas as 22 amostras do grupoB foram negativas nos métodos moleculares. Conclusão: neste estudo, a citologia prévia alterada associada a testes moleculares indica a necessidade de monitoramento ou intervenção terapêutica das pacientes. Assim, conclui-se que a citologia oncótica associada à detecção do DNA do HPV pela PCR e à expressão das oncoproteínas E6 e E7 é a alternativa para identificar precocemente pacientes com elevado risco de desenvolver o câncer cervical.

Worldwilde, the cervical cancer is known as one of the most frequent cause of death in female population. Persistent infection with oncogenic human papillomavirus (HPV) is the main risk factor for cervical cancer and its precursor lesions. Objective: to evaluate the diagnostic methods of cervical cancer: oncotic cytology, HPV DNA by PCR, detection of mRNA E6 / E7 proteins of high-risk HPV, to add new markers in the detection of HPV.Methods: were evaluated 3,000 results of oncotic cytology from patients referred to Vital Laboratory, Nonoai- RS, Brazil, from March 2009 to January2010. The results were grouped according age and Bethesda classification, 2001. Group A included patients with previous altered cytology and GroupB patients without abnormalities in cytology. To molecular tests were collected 47 samples, 22 from group A and 25 from group B. Three samples from group B presented PCR inhibitors and were excluded. The detection of HPV was made by polymerase chain reaction (PCR) using the consensus primers MY9/11 and GP5 +/6 +. The mRNA E6 / E7 proteins were detected by real-time NASBA. Results: in samples of group A, DNA/HPV was detected in 14(64%) samples, and mRNA E6 / E7 proteins of HPV was detected in 6 (27%) samples. All 22 samples of group B were negative to molecular methods.Conclusion: in this study, the previous altered cytology associated with molecular methods indicates the need for monitoring or therapeutic intervention of patients. The oncotic cytology associated with the detection of HPV DNA by PCR and the expression of oncoproteins E6 / E7 are alternatives to identify early lesions in patients with high risk of developing cervical cancer.

Intrahospital spread of carbapenem-resistant Pseudomonas aeruginosa in a University Hospital in Florianópolis, Santa Catarina, Brazil / Disseminação Intrahospitalar de Pseudomonas aeruginosa em Hospital Universitário de Florianópolis, Santa Catarina, Brasil

INTRODUCTION: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) has been isolated with increasing frequency in Brazilian hospitals. Since June 2003, its detection in a teaching hospital in the city of Florianópolis, Brazil, has increased. This study aimed to investigate the minimal inhibitory concentration (MIC), presence of Metallo-β-lactamase (MβL) and a possible clonal relationship among the isolates. METHODS: The study included 29 CRPA and seven isolates with reduced susceptibility. The MIC was determined by agar-dilution. Detection of MβL was performed by Double Disk Sinergism (DDS) and Combined Disk (CD). The MβL gene was verified by PCR and nucleotide sequence analysis. Epidemiological typing was performed by pulsed-field gel electrophoresis. RESULTS: Among the 29 carbapenem-resistant isolates, polymyxin B presented 100 percent susceptibility and piperacillin/tazobactam 96.7 percent. Seventeen (62 percent) strains were verified as clonal (A clone) and among these, six isolates indicated phenotypically positive tests for MβL and harbored the blaSPM-1 gene. The first CRPA isolates were unrelated to clone A, harbored blaIMP-16 and were phenotypically positive only by CD. CONCLUSIONS: The spread of a high-level of resistance clone suggests cross transmission as an important dissemination mechanism and has contributed to the increased rate of resistance to carbapenems. This study emphasizes the need for continuous surveillance and improved strategies.

INTRODUÇÃO: O isolamento de Pseudomonas aeruginosa resistente aos carbapenêmicos (PARC) tem sido cada vez mais frequente nos hospitais brasileiros. O presente estudo investigou a concentração inibitória mínina (CIM), a presença de metalo-β-lactamases (MβL), e uma possível relação clonal entre PARC isoladas entre junho de 2003 a junho de 2005, em um hospital escola na cidade de Florianópolis, Brasil. MÉTODOS: O estudo incluiu 29 PARC e sete isolados com suscetibilidade reduzida. A CIM foi determinada por diluição em ágar. A detecção de MβL foi realizada por sinergismo de duplo disco (SDD) e disco combinado (DC). Genes para MβL foram pesquisados por PCR e confirmados pela análise da sequência de nucleotídeos. A tipagem epidemiológica foi realizada por gel de eletroforese em campo pulsátil. RESULTADOS: Entre os 29 isolados resistentes aos carbapenêmicos, 100 por cento apresentaram suscetibilidade a polimixina B, e 96,7 por cento a piperacilina/tazobactam. Dezessete (62 por cento) destes isolados pertenciam a um mesmo clone (clone A); entre estes, seis isolados apresentaram testes fenotípicos positivos para MβL e carreavam o gene blaSPM-1. O primeiro isolado PARC não foi relacionado ao clone A, carreava o gene blaIMP-16 e foi fenotipicamente positivo somente por DC. CONCLUSÕES: A propagação de um clone com alto nível de resistência sugere a transmissão cruzada como um importante mecanismo de disseminação e tem contribuído para o aumento nos níveis de resistência aos carbapenêmicos. Este estudo enfatiza a necessidade de vigilância contínua e melhoramento nas estratégias de controle de infecção nesta instituição.

Quality of sputum in the performance of polymerase chain reaction for diagnosis of pulmonary tuberculosis

SETTING: faster alternative techniques are required to improve the diagnosis and control of pulmonary tuberculosis. OBJECTIVE: To evaluate the sample quality in the performance of PCR for diagnosis of pulmonary tuberculosis. METHOD: during one year, sputum samples were collected from 72 pulmonary tuberculosis patients and 12 non-tuberculosis controls, which were admitted to the Nereu Ramos hospital, Florianópolis city, Brazil. The samples were subjected to Ziehl-Neelsen-stained sputum smear microscopy and Lowestein-Jensen medium culture, which were defined as gold standard tests for mycobacteria, and polymerase chain reaction (PCR). Those samples that presented more than 40 percent of viable cells and less than 25 percent of epithelial cells were defined as high quality samples. RESULTS: PCR showed sensitivity of 55.6 percent, specificity of 41.7 percent, positive predictive value of 85.1 percent, negative predictive value of 13.5 percent, and accuracy of 53.6 percent. High quality samples showed sensitivity of 72.4 percent, specificity of 50 percent, positive predictive value of 91.3 percent, negative predictive value of 20 percent, and accuracy of 69.7 percent. Low quality samples showed sensitivity of 44.2 percent, specificity of 37.5 percent, positive predictive value of 79.2 percent, negative predictive value of 11.1 percent, and accuracy of 43.1 percent. CONCLUSION: use of high quality samples improved significantly the PCR performance, especially on their sensitivity and positive predictive values.