Diversidade genética dos vírus linfotrópicos de Células T do tipo 1 na Região Metropolitana de Belém

The human T-cell lymphotropic virus type 1 (HTLV-1) is a DELTARETROVIRUS which was first isolated from a blood sample of a patient with african american cutaneous T-cell lymphoma, in the mid 1970s; Hodgkin was later classified as leukemia / lymphoma adult T-cell (LLcTA) severe disease that affects T lymphocytes Subsequently, the virus was also associated with tropical spastic paraparesis / HTLV-associated myelopathy (HAM / TSP), character chronic and progressive causing damage mainly at the thoracic spinal cord. Their genetic diversity varies according to the region studied, and its mutation rate is very low (about 1% per thousand years) compared to other viruses. The objective of this study was to assess the genetic diversity of HTLV-1 in the metropolitan area of Belém, as well as the frequency of genotypes and the comparison of samples obtained with the sequences available in databases such as GenBank. We used blood samples collected from patients enrolled in the NMT / UFPA between the period January 2010 to December 2013. Then these samples were subjected to DNA extraction and amplification of HTLV PX region, using oligonucleotides inciciadores specific, from the PCR in two steps: internal and external (nested PCR). Positive samples then were subjected to enzymatic digestion of TaqI enzyme so that we could identify and differentiate the HTLV 1 and 2. Next, we performed amplification of the 5 'LTR region also by nested PCR positive samples, which, then were subjected to genetic sequencing. Using the method of maximum likelihood, we constructed a phylogenetic tree for the group display in clades of sequences in question. A Bayesian analysis (molecular clock) to visualize the evolutionary profile of the samples was also performed. The tests identified 78 samples positive for HTLV-1, of these, 44 were sequenced. The aa genotype was common in 100% of samples; the different subtypes had the following range rate and mutations per site per year: a- 2.10 -3; b- 2,69 . 10 -2; c- 6,23 . 10 -2; d- 3,08 . 10 -2; e- 6 . 10 -2; f- 1,78 . 10 -3; g- 2,2 . 10 -2 changes per site per year. It was then revealed a low genotypic diversity and a high genetic stability of the samples positive for HTLV-1 in the region.O Vírus Linfotrópico de células T Humano do tipo 1 (HTLV-1) é um Deltaretrovírus que foi isolado pela primeira vez a partir de uma amostra de sangue de um paciente afro-americano com linfoma cutâneo de células T, em meados de 1970; o linfoma foi posteriormente classificado como leucemia/linfoma de células T do adulto (LLcTA) doença severa que acomete os linfócitos T. Posteriormente, o vírus também foi associado à paraparesia espática tropical/mielopatia associada ao HTLV, (PET/MAH), de caráter crônico e progressivo que causa danos principalmente ao nível da medula espinhal torácica. Sua diversidade genética varia de acordo com a região estudada, e sua taxa de mutação é muito baixa (cerca de 1% a cada mil anos) se comparada a de outros vírus. O objetivo deste trabalho foi verificar a diversidade genética do HTLV-1 na região metropolitana de Belém, bem como a freqüência dos seus genótipos e a comparação das amostras obtidas com as sequências disponíveis em bancos de dados como o GenBank. Foram utilizadas amostras de sangue coletadas de pacientes atendidos pelo NMT/UFPA entre o período de janeiro de 2010 à dezembro de 2013. Em seguida, tais amostras foram submetidas à extração do DNA e à amplificação da região PX do HTLV, utilizando-se oligonucleotídeos inciciadores específicos, a partir da técnica de PCR em duas etapas: interna e externa (Nested-PCR). As amostras positivas, posteriormente, foram submetidas à digestão enzimática da enzima TaqI, para que pudéssemos identificar e diferenciar os HTLV 1 e 2. Em seguida, foi realizada a amplificação da região 5’LTR também por Nested-PCR das amostras positivas, que, posteriormente, foram submetidas ao seqüenciamento genético. Utilizando o método de máxima verossimilhança, foi construída uma árvore filogenética para a visualização do agrupamento em clados das sequências em questão. Uma análise bayesiana (relógio molecular) para a visualização do perfil evolutivo das amostras também foi realizada. Os testes identificaram 78 amostras positivas para HTLV-1, destas, 44 foram sequenciadas. O genótipo aA foi frequente em 100% das amostras; os diferentes subtipos obtiveram a seguinte taxa de diversidade e mutações por sítio por ano: a- 2.10 -3; b- 2,69 . 10 -2; c- 6,23 . 10 -2; d- 3,08 . 10 -2; e- 6 . 10 -2; f- 1,78 . 10 -3; g- 2,2 . 10 -2 mutações por sítio por ano. Foi revelada então uma baixa diversidade genotípica e uma alta estabilidade genética entre as amostras positivas para HTLV-1 na região

Primera detección de coronavirus humano asociado a infección respiratoria aguda en la Región Norte de Brasil

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Os coronavírus humanos (CoVh) são responsáveis por ocasionar doenças respiratórias e entéricas, sendo associados às infecções agudas e graves do trato respiratório. O objetivo deste trabalho foi identificar a ocorrência dos CoVh em pacientes com infecção respiratória aguda (IRA) na Cidade de Belém, Estado do Pará, Brasil, atendidas pelo programa de vigilância do vírus da influenza do Instituto Evandro Chagas. No período compreendido entre agosto de 2009 a março de 2011, amostras de aspirado da nasofaringe ou swab combinado (nasal e oral) obtidas de 308 pacientes com diagnóstico clínico de IRA, foram laboratorialmente investigadas na busca de evidências de infecção pelo CoVh. A metodologia adotada envolveu quatro etapas principais: a) extração do RNA viral (RNAv) do espécime clínico; b) execução da técnica de reação em cadeia mediada pela polimerase precedida de transcrição reversa (RT-PCR); c) eletroforese em gel de agarose dos produtos gerados; e d) sequenciamento genético de nucleotídeos. O processamento laboratorial de todos os espécimes revelou positividade para CoVh em uma amostra clínica dos pacientes investigados. A análise filogenética da amostra positiva permitiu classificá-la como pertencente ao subtipo OC43 de CoVh. O presente achado representa a primeira comprovação laboratorial da circulação deste agente viral na Região Amazônica. Estudos adicionais são necessários para um maior conhecimento da etiologia e epidemiologia deste agente viral em casos de IRA que ocorrem na Região Norte do Brasil.Human coronaviruses (HCoVs) are responsible for causing respiratory and enteric diseases associated with severe acute respiratory tract infections. The aim of this study was to identify the occurrence of HCoVs in patients with acute respiratory infection (ARI) in Belém, Pará State, Brazil, assisted by the surveillance program of influenza viruses of Instituto Evandro Chagas. From August 2009 to March 2011, samples of nasopharyngeal aspirate or nasal and oral swabs obtained from 308 patients with clinical diagnosis of ARI were investigated in the laboratory in order to identify the HCoVs infection. The methodology involved four main steps: a) viral RNA (RNAV) extracted from the clinical specimen; b) application of reverse transcription polymerase chain reaction (RT-PCR); agarose electrophoresis gel of the generated products; and d) DNA sequencing. The laboratory processing of all specimens were positive for HCoVs in a clinical sample of patients investigated. The positive sample was classified as belonging to the subtype OC43 of HCoVs by using the phylogenetic analysis. This finding represents the first laboratory confirmation of this viral agent circulation in the Amazon Region. Additional studies are needed for a better understanding of the etiology and epidemiology of this viral agent in cases of ARI occurring in Northern Brazil

Gastroenterite por rotavírus do grupo A em Leopardus tigrinus e Leopardus pardalis (Carnivora: Felidae) órfãos na Amazônia Oriental

Fundação para a Ciência e a Tecnologia (Portugal), through the research project GHTM– UID/Multi/04413/2013; and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Brazil), for the Ph.D. scholarship of Áurea Martins Gabriel.Universidade Nova de Lisboa. Instituto de Higiene e Medicina Tropical. Lisboa, Portugal / Universidade Federal do Pará. Instituto de Ciências Biológicas. Programa de Pós-Graduação em Biologia de Agentes Infecciosos e Parasitários. Belém, PA, BrasilUniversidade Federal Rural da Amazônia. Laboratório de Patologia Animal. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Centro Nacional de Primatas. Ananindeua, PA, Brasil / Fundação Oswaldo Cruz. Instituto de Ciência e Tecnologia em Biomodelos. Rio de Janeiro, RJ, Brasil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Laboratórios de Vírus Gastroentéricos. Ananindeua, PA, BrasilUniversidade Federal do Pará. Núcleo de Medicina Tropical. Laboratório de Biologia Molecular e Celular. Belém, PA, Brasil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Laboratórios de Vírus Gastroentéricos. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Ciência, Tecnologia, Inovação e Insumos Estratégicos. Instituto Evandro Chagas. Laboratórios de Vírus Gastroentéricos. Ananindeua, PA, Brasil.Rotaviruses are highly infectious and transmitted by the fecal-oral route, representing a leading cause of diarrheal deaths in children in developing countries. Additionally, it causes significant economic impacts as an enteropathogen among domestic animals. This report presents clinical and diagnostic findings in two cases of rotavirus (RV) infection involving orphaned Leopardus tigrinus (Schreber, 1775) and Leopardus pardalis (Linnaeus, 1758), representing the first registered group A RV (RVA) infection in these species. Both felids were rescued in Pará State, Amazon Region, by the Brazilian Institute of Environment and Renewable Natural Resources and treated in an intensive care ward in a public Environmental Park of Belém City. After adaptation up to the 40th day of the quarantine period, these animals had no typical symptoms of acute, fulminant gastroenteritis. The collected samples were examined at the Virology Laboratory of Instituto Evandro Chagas. RVA antigen was detected in the blood and fecal samples of L. tigrinus by enzyme-linked immunosorbent assay and immunochromatography. RVA-positive samples were subjected to polyacrylamide gel electrophoresis and reverse transcriptase polymerase chain reaction for VP4 and VP7 genes using conventional set primers but with negative results. Stools were examined for ova and parasites yielding negative results and for intestinal endoparasites, yielding negative results. The animals died within a few days following a clinical exacerbation, therefore unresponsive to treatment. Necropsies and histopathological analysis were performed at the Laboratory of Veterinary Pathology of Universidade Federal Rural da Amazônia. Despite the pathologic findings typical of RV infection hemorrhagic enteritis was unusual if presumed etiology is considered

High prevalence of sexual infection by human papillomavirus and Chlamydia trachomatis in sexually-active women from a large city in the Amazon region of Brazil.

BackgroundThe Human Papillomavirus (HPV) and Chlamydia trachomatis are the most prevalent Sexually Transmitted Infections (STIs) worldwide, and are associated cervical cancer and pelvic inflammatory disease, respectively. However, 80% of women testing positive are asymptomatic. In the Amazon region, young women, in particular, are widely exposed to the infections and their consequences.ObjectivesDetermine the prevalence of sexual infection by HPV and C. trachomatis in young, sexually-active women treated at a university health program in a large city of the Brazilian Amazon region.MethodsWe amplified the L1 gene of HPV. We amplified ompA gene of C. trachomatis by nested PCR, and the study participants filled in a questionnaire on their social, epidemiological, and reproductive health characteristics. The data were analyzed using the Odds Ratio, to evaluate the degree of association of these variables with the observed infections.ResultsThe prevalence of infection by HPV was 15.5% (47/303). This infection was recorded in 32.2% of the women of less than 25 years of age (OR:3.02 [CI95%] = 1.32-6.92; p = 0.014), 17.9% of the single women (OR: 2.41 [CI95%] = 1.22-4.75; p = 0.014), 23.8% of the women that reported having first sexual intercourse at less than 15 years of age (OR: 2.22 [CI95%] = 1.16-4.23; p = 0.021), 20% of those that reported having had more than one sexual partner during their lifetime (OR: 3.83 [CI95%] = 1.56-9.37; p = 0.003), and in 28.3% that use oral contraceptives (CI95% = 1.33-5.43; p = 0.008). The prevalence of sexual infection by C. trachomatis was 4.6% (14/303), and this bacterium was present in 16.1% of the young women of less than 25 years of age (OR: 2.86 [CI95%] = 1.33-5.43; p = 0.008).ConclusionsWe found a high prevalence of HPV in young, unmarried women who started their sex lives early, who had several sexual partners in their lives and who used oral contraceptives. The prevalence of C. trachomatis was high only in young women. Our data are in accordance with other studies in Brazil and in the world and may serve to base the formulation of diagnostic and screening measures for these infections in women in the Amazon

High prevalence of sexual infection by human papillomavirus and Chlamydia trachomatis in sexually-active women from a large city in the Amazon region of Brazil

Background The Human Papillomavirus (HPV) and Chlamydia trachomatis are the most prevalent Sexually Transmitted Infections (STIs) worldwide, and are associated cervical cancer and pelvic inflammatory disease, respectively. However, 80% of women testing positive are asymptomatic. In the Amazon region, young women, in particular, are widely exposed to the infections and their consequences. Objectives Determine the prevalence of sexual infection by HPV and C. trachomatis in young, sexually-active women treated at a university health program in a large city of the Brazilian Amazon region. Methods We amplified the L1 gene of HPV. We amplified ompA gene of C. trachomatis by nested PCR, and the study participants filled in a questionnaire on their social, epidemiological, and reproductive health characteristics. The data were analyzed using the Odds Ratio, to evaluate the degree of association of these variables with the observed infections. Results The prevalence of infection by HPV was 15.5% (47/303). This infection was recorded in 32.2% of the women of less than 25 years of age (OR:3.02 [CI95%] = 1.32–6.92; p = 0.014), 17.9% of the single women (OR: 2.41 [CI95%] = 1.22–4.75; p = 0.014), 23.8% of the women that reported having first sexual intercourse at less than 15 years of age (OR: 2.22 [CI95%] = 1.16–4.23; p = 0.021), 20% of those that reported having had more than one sexual partner during their lifetime (OR: 3.83 [CI95%] = 1.56–9.37; p = 0.003), and in 28.3% that use oral contraceptives (CI95% = 1.33–5.43; p = 0.008). The prevalence of sexual infection by C. trachomatis was 4.6% (14/303), and this bacterium was present in 16.1% of the young women of less than 25 years of age (OR: 2.86 [CI95%] = 1.33–5.43; p = 0.008). Conclusions We found a high prevalence of HPV in young, unmarried women who started their sex lives early, who had several sexual partners in their lives and who used oral contraceptives. The prevalence of C. trachomatis was high only in young women. Our data are in accordance with other studies in Brazil and in the world and may serve to base the formulation of diagnostic and screening measures for these infections in women in the Amazon

Moderate endemicity of the human T-lymphotropic virus infection in the metropolitan region of Belém, Pará, Brazil

The spread of the HTLV infection in families living in the metropolitan area of Belém, Pará, Brazil, and the lack of studies in the general population requires studies to better understand its prevalence in the region. Methods: An anti-HTLV-1/HTLV-2 antibodies test was carried out on random adults in public places in Belém between November 2014 and November 2015. A proviral DNA test detected if the person was infected, and then a clinical evaluation and an intrafamilial investigation were carried out. Results: Of the 1059 individuals being investigated, 21 (2.0%) had seroreagent samples, 15 (1.4%) had HTLV-1, 5 (0.5%) had HTLV-2, and proviral DNA was undetectable in one case. The mean age of the infected people (57.2) was higher than that of those that were uninfected (46.2) (p = 0.0010). The prevalence of infection increased with age, especially in individuals with a family income equal to or less than a minimum wage. Intrafamilial transmission seems to have occurred in all of the families being studied. Among the patients with HTLV-1, 30% (3/10) already had some symptom related to the infection. Discussion: The increase in prevalence rates according to age may be due to late seroconversion of a previously acquired infection, or the cumulative risk of new infections, especially in women. Conclusion: There was a moderate prevalence of the HTLV infection among adult individuals from the metropolitan area of Belém, with a predominance of HTLV-1. This infection was associated with low income and increasingly older women. It also presented intrafamily spread and negligence in the diagnosis of associated diseases.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoPROEX/UFPAA disseminação da infecção pelo vírus linfotrópico-T humano (HTLV) em famílias da área metropolitana de Belém, Pará, Brasil, e a ausência de estudos na população em geral requisitam investigações que esclareçam melhor a sua prevalência na região. Metodologia: Foi realizada pesquisa de anticorpos antiHTLV-1/HTLV-2 em indivíduos adultos transeuntes de logradouros públicos de Belém, entre novembro de 2014 e novembro de 2015. A infecção foi confirmada por pesquisa de DNA proviral e foi realizada avaliação clínica e investigação intrafamiliar dos infectados. Resultados: Dos 1.059 indivíduos investigados, 21 (2,0%) apresentaram amostras sororeagentes, 15 (1,4%) confirmados para HTLV-1, 5 (0,5%) para HTLV-2 e o DNA proviral foi indetectável em 1 caso. A média de idade dos infectados (57,2) foi maior que a dos não infectados (46,2) (p = 0,0010). A infecção aumentou com a idade e se destacou nos indivíduos com renda familiar menor ou igual a um salário mínimo. A transmissão intrafamiliar parece ter ocorrido em todas as famílias investigadas. Dentre os portadores de HTLV-1, 30% (3/10) já apresentavam algum sintoma relacionado à infecção. Discussão: O aumento da infecção de acordo com a idade pode ocorrer por soroconversão tardia de infecção pré-adquirida ou pelo risco cumulativo de novas infecções, sobretudo em mulheres. Conclusão: A infecção por HTLV demonstrou moderada prevalência na população estudada, com predomínio do HTLV-1. Essa mostrou-se associada à baixa renda e ao aumento da idade das mulheres. Também apresentou disseminação intrafamiliar e negligência no diagnóstico das doenças associadas.NOBRE, A. F. S.; PEREIRA, C. C. C. UFPA/NM

Moderada endemicidade da infecção pelo vírus linfotrópico-T humano na região metropolitana de Belém, Pará, Brasil

RESUMO: Introdução: A disseminação da infecção pelo vírus linfotrópico-T humano (HTLV) em famílias da área metropolitana de Belém, Pará, Brasil, e a ausência de estudos na população em geral requisitam investigações que esclareçam melhor a sua prevalência na região. Metodologia: Foi realizada pesquisa de anticorpos anti-HTLV-1/HTLV-2 em indivíduos adultos transeuntes de logradouros públicos de Belém, entre novembro de 2014 e novembro de 2015. A infecção foi confirmada por pesquisa de DNA proviral e foi realizada avaliação clínica e investigação intrafamiliar dos infectados. Resultados: Dos 1.059 indivíduos investigados, 21 (2,0%) apresentaram amostras sororeagentes, 15 (1,4%) confirmados para HTLV-1, 5 (0,5%) para HTLV-2 e o DNA proviral foi indetectável em 1 caso. A média de idade dos infectados (57,2) foi maior que a dos não infectados (46,2) (p = 0,0010). A infecção aumentou com a idade e se destacou nos indivíduos com renda familiar menor ou igual a um salário mínimo. A transmissão intrafamiliar parece ter ocorrido em todas as famílias investigadas. Dentre os portadores de HTLV-1, 30% (3/10) já apresentavam algum sintoma relacionado à infecção. Discussão: O aumento da infecção de acordo com a idade pode ocorrer por soroconversão tardia de infecção pré-adquirida ou pelo risco cumulativo de novas infecções, sobretudo em mulheres. Conclusão: A infecção por HTLV demonstrou moderada prevalência na população estudada, com predomínio do HTLV-1. Essa mostrou-se associada à baixa renda e ao aumento da idade das mulheres. Também apresentou disseminação intrafamiliar e negligência no diagnóstico das doenças associadas

Low genetic diversity of the Human T-cell Lymphotropic Virus (HTLV-1) in an endemic area of the Brazilian Amazon basin

<div><p>The Human T-cell Lymphotropic Virus (HTLV-1) is a <i>Deltaretrovírus</i> that was first isolated in the 1970s, and associated with Adult T-cell Leucemia-Lymphoma (ATLL), and subsequently to Tropical Spastic Paraparesis-Myelopathy (TSP/HAM). The genetic diversity of the virus varies among geographic regions, although its mutation rate is very low (approximately 1% per thousand years) in comparison with other viruses. The present study determined the genetic diversity of HTLV-1 in the metropolitan region of Belém, in northern Brazil. Blood samples were obtained from patients at the UFPA Tropical Medicine Nucleus between January 2010 and December 2013. The DNA was extracted and the PX region of the HTLV was amplified using nested PCR. The positive samples were then digested using the Taq1 enzyme for the identification and differentiation of the HTLV-1 and HTLV-2. The 5’LTR region of the positive HTLV-1 samples were amplified by nested PCR, and then sequenced genetically. The phylogenetic analysis of the samples was based on the maximum likelihood method and the evolutionary profile was analyzed by the Bayesian approach. Overall, 78 samples tested positive for HTLV-1, and 44 were analyzed here. The aA (cosmopolitan-transcontinental) subtype was recorded in all the samples. The following evolutionary rates were recorded for the different subtypes–a: 2.10−³, b: 2.69. 10−², c: 6.23. 10−², d: 3.08. 10−², e: 6. 10−², f: 1.78. 10−³, g: 2.2. 10−² mutations per site per year. The positive HTLV-1 samples tested in the present study were characterized by their low genetic diversity and high degree of stability.</p></div

Phylogenetic analysis of the 744 bps nucleotide sequence corresponding to the 5’LTR region of the HTLV-1, based on the maximum likelihood method with 10,000 bootstrap replicates.

<p>The samples analyzed in the present study are shown in red, and all other were obtained from GenBank (<a href="https://www.ncbi.nlm.nih.gov/genbank" target="_blank">https://www.ncbi.nlm.nih.gov/genbank</a>).</p

Additional file 1: of Stability of the HTLV-1 glycoprotein 46 (gp46) gene in an endemic region of the Brazilian Amazon and the presence of a significant mutation (N93D) in symptomatic patients

The research ethics committee. Source: Study protocol, 2015, Table S1. Description of values of AICM applied in the statistics (Bayesian inference) in the study data. Source: Study protocol, 2016. (PDF 1549 kb