Distribuição dos genótipos do vírus GB-C (HGV) em indivíduos da cidade de São Paulo, Brasil

Há na literatura vários estudos filogenéticos e de distribuição de genótipos do chamado "Vírus GB-C" ou da "Hepatite G", mais conhecido pela dupla sigla "GBV-C/HGV". Ocorre que, em sua grande maioria, estas pesquisas foram realizadas com amostras de grupos ligados epidemiologicamente e não com indivíduos representativos da população geral. O presente estudo é uma continuação do primeiro trabalho no Brasil feito com este tipo de amostragem. Trata-se de análise filogenética e distribuição genotípica do GBV-C/HGV a partir de amostras isoladas dentre mais de 1.000 indivíduos da cidade de São Paulo. Para tanto, um fragmento de 728 pares de base da região 5' não-codificadora (5´NCR) do genoma viral, de 24 amostras, foi sequenciado e submetido à analise filogenética. Foram identificados os genótipos 1, 2a e 2b nas respectivas freqüências: 8,3% (2/24), 50% (12/24) e 41,7% (10/24). Concluindo, São Paulo apresenta uma distribuição de genótipos semelhante à publicada para outros estados e regiões do Brasil, endossando a idéia de que os tipos 1 e 2 teriam vindo com os africanos e europeus, respectivamente, e portanto estariam na população do país desde a sua formação.There has been several studies worldwide on phylogenetics and genotype distribution of the GB-virus C / Hepatitis G virus (GBV-C/HGV). However, in their great majority, those investigations were based on some epidemiologically linked group, rather than on a representative sampling of the general population. The present is a continuation of the first study in Brazil with such a population; it addresses the GBV-C/HGV phylogenetics and genotype distribution based on samples identified among more than 1,000 individuals of the city of São Paulo. For this purpose, a 728 bp fragment of the 5´non-coding region (5´NCR) of the viral genome, from 24 isolates, was sequenced and subjected to phylogenetic analysis. Genotypes 1, 2a and 2b were found at 8.3% (2/24), 50% (12/24) and 41.7% (10/24), respectively. In conclusion São Paulo displays a genotype distribution similar to the published data for other States and Regions of Brazil, endorsing the notion that types 1 and 2 would have entered the country with African and European people, respectively, since its earliest formation

Polymerase chain reaction targeting 16S ribosomal RNA for the diagnosis of bacterial meningitis after neurosurgery

OBJECTIVES: Bacterial and aseptic meningitis after neurosurgery can present similar clinical signs and symptoms. The aims of this study were to develop and test a molecular method to diagnose bacterial meningitis (BM) after neurosurgery. METHODS: A 16S ribosomal RNA gene PCR-based strategy was developed using artificially inoculated cerebrospinal fluid (CSF) followed by sequencing. The method was tested using CSF samples from 43 patients who had undergone neurosurgery and were suspected to suffer from meningitis, and from 8 patients without neurosurgery or meningitis. Patients were classified into five groups, confirmed BM, probable BM, possible BM, unlikely BM, and no meningitis. RESULTS: Among the samples from the 51 patients, 21 samples (41%) were culture-negative and PCR-positive. Of these, 3 (14%) were probable BM, 4 (19%) were possible BM, 13 (62%) were unlikely BM, and 1 (5%) was meningitis negative. Enterobacterales, non-fermenters (Pseudomonas aeruginosa and Acinetobacter baumannii), Staphylococcus haemolyticus, Granulicatella, Variovorax, and Enterococcus cecorum could be identified. In the group of patients with meningitis, a good agreement (3 of 4) was observed with the results of cultures, including the identification of species. CONCLUSION: Molecular methods may complement the diagnosis, guide treatment, and identify non-cultivable microorganisms. We suggest the association of methods for suspected cases of BM after neurosurgery, especially for instances in which the culture is negative

HCV Genotypes, Characterization of Mutations Conferring Drug Resistance to Protease Inhibitors, and Risk Factors among Blood Donors in São Paulo, Brazil

Background: Hepatitis C virus (HCV) infection is a global health problem estimated to affect almost 200 million people worldwide. the aim of this study is to analyze the subtypes and existence of variants resistant to protease inhibitors and their association with potential HCV risk factors among blood donors in Brazil.Methods: Repeat anti-HCV reactive blood donors are systematically asked to return for retest, notification, and counseling in which they are interviewed for risk factors for transfusion-transmitted diseases. We analyzed 202 donors who returned for counseling from 2007 to 2010 and presented enzyme immunoassay-and immunoblot-reactive results. the HCV genotypes and resistance mutation analyses were determined by the direct sequencing of the NS5b and NS3 regions, respectively. the HCV viral load was determined using an in-house real-time PCR assay targeting the 5'-NCR.Results: HCV subtypes 1b, 1a, and 3a were found in 45.5%, 32.0%, and 18.0% of the donors, respectively. the mean viral load of genotype 1 was significantly higher than that of the genotype 3 isolates. Subtype 1a was more frequent among young donors and 3a was more frequent among older donors. Protease inhibitor-resistant variants were detected in 12.8% of the sequenced samples belonging to genotype 1, and a higher frequency was observed among subtype 1a (20%) in comparison to 1b (8%). There was no difference in the prevalence of HCV risk factors among the genotypes or drug-resistant variants.Conclusions: We found a predominance of subtype 1b, with an increase in the frequency of subtype 1a, in young subjects. Mutations conferring resistance to NS3 inhibitors were frequent in treatment-naive blood donors, particularly those infected with subtype 1a. These variants were detected in the major viral population of HCV quasispecies, have replicative capacities comparable to nonresistant strains, and could be important for predicting the response to antiviral triple therapy.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundacao Pro-Sangue/Hemocentro de São PauloFundacao Prosangue Hemoctr São Paulo, São Paulo, BrazilUniversidade Federal de São Paulo, Infect Dis Div DIPA, São Paulo, BrazilUniv São Paulo, Fac Med, Discipline Med Sci, São Paulo, BrazilHCFMUSP, Dept Pathol, LIM Lab Medice Lab 03, São Paulo, BrazilUniv Sao Joao del Rei, Divinopolis, MG, BrazilUniv São Paulo, Fac Med, Dept Infect Dis, São Paulo, BrazilUniversidade Federal de São Paulo, Infect Dis Div DIPA, São Paulo, BrazilWeb of Scienc

Nova estimativa da prevalência da infecção pelo vírus "TT" (TTV) em populações de baixo e alto risco de São Paulo, Brasil

A prevalência da infecção pelo vírus "TT" (TTV) foi investigada pela técnica da Reação da Polimerase em Cadeia (PCR) em grupos considerados de baixo risco (doadores de sangue e crianças/adolescentes saudáveis) e de alto risco de exposição parenteral (hemofílicos); todos provenientes da cidade de São Paulo. Oligonucleotídeos empregados como primers, homólogos à região não traduzível (UTR) do genoma viral, mostraram-se muito mais universais, revelando frequências muito mais altas em ambos os grupos ( >; ou = 81%) do que os primers anteriormente utilizados, baseados na região genômica traduzível "N22" (doadores de sangue, 5,5%, e hemofílicos, 42,3%). O "PCR-UTR" também revelou um perfil interessante em crianças/adolescentes saudáveis: alta prevalência nos primeiros anos de vida e queda significativa em meninos adolescentes. O "PCR-N22", por sua vez, apresentou alta frequência em hemofílicos que receberam derivados de sangue fresco (58%) relativa àqueles que foram tratados com fatores de coagulação submetidos à inativação viral (9,4%) e doadores de sangue (5,5%).The prevalence of TT virus (TTV) infection was investigated by Polymerase Chain Reaction (PCR) in low- (blood donors and healthy children/adolescents) and high-risk (hemophiliacs) groups from São Paulo, Brazil. Primers based on the untranslated region (UTR) of the viral genome proved to be much more ubiquitous, leading to much higher frequencies for both groups ( >; or = 81%) than the earlier N22-PCR directed to the open reading frame 1 (blood donors, 5.5%, and hemophiliacs, 42.3%). The UTR-PCR also revealed an interesting profile for healthy children/adolescents: very high prevalence at the early years and significant decrease in male teenagers. The N22-PCR, in turn, demonstrated higher frequency in hemophiliacs treated with fresh blood products (58%), than in those treated with virus-inactivated clotting factors (9.4%) and blood donors (5.5%)

Yellow fever vaccine viremia following ablative BM suppression in AML

Univ São Paulo, Sch Med, Dept Infect & Parasit Dis, São Paulo, BrazilHosp Sirio Libanes, São Paulo, BrazilUniv São Paulo, Sch Med, Div Clin Immunol & Allergy, São Paulo, BrazilFundacao Prosangue Hemoctr São Paulo, São Paulo, BrazilUniversidade Federal de São Paulo, Infect Dis Div DIPA, São Paulo, BrazilFundacao Oswaldo Cruz, Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Infect Dis Div DIPA, São Paulo, BrazilWeb of Scienc

Plasma Cytokine Profile in Tropical Endomyocardial Fibrosis: Predominance of TNF-a, IL-4 and IL-10

Background: the participation of immune/inflammatory mechanisms in the pathogenesis of tropical endomyocardial fibrosis (EMF) has been suggested by the finding of early blood and myocardial eosinophilia. However, the inflammatory activation status of late-stage EMF patients is still unknown.Methodology/Principal findings: We evaluated pro- and anti-inflammatory cytokine levels in plasma samples from late stage EMF patients. Cytokine levels of Tumor Necrosis Factor (TNF)-alpha, Interferon (IFN)-gamma, Interleukin (IL)-2, IL-4, IL-6, and IL-10 were assayed in plasma samples from 27 EMF patients and compared with those of healthy control subjects. All EMF patients displayed detectable plasma levels of at least one of the cytokines tested. We found that TNF-alpha, IL-6, IL-4, and IL-10 were each detected in at least 74% of tested sera, and plasma levels of IL-10, IL-4, and TNF-alpha were significantly higher than those of controls. Plasma levels of such cytokines positively correlated with each other.Conclusions/Significance: the mixed pro-and anti-inflammatory/Th2circulating cytokine profile in EMF is consistent with the presence of a persistent inflammatory stimulus. On the other hand, the detection of increased levels of TNF-alpha may be secondary to the cardiovascular involvement observed in these patients, whereas IL-4 and IL-10 may have been upregulated as a homeostatic mechanism to buffer both production and deleterious cardiovascular effects of pro-inflammatory cytokines. Further studies might establish whether these findings play a role in disease pathogenesis.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Univ São Paulo, Sch Med, Inst Heart InCor, Immunol Lab, São Paulo, BrazilUniv São Paulo, Sch Med, Div Clin Immunol & Allergy, São Paulo, BrazilUniv São Paulo, Sch Med, Inst Heart InCor, Cardiomyopathy Unit, São Paulo, BrazilProSangue Fdn, São Paulo, BrazilInst Investigat Immunol, INCT, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Div Immunol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Immunol & Parasitol, Div Immunol, São Paulo, BrazilWeb of Scienc

Frequency and genotypic distribution of GB virus C (GBV-C) among Colombian population with Hepatitis B (HBV) or Hepatitis C (HCV) infection

<p>Abstract</p> <p>Background</p> <p>GB virus C (GBV-C) is an enveloped positive-sense ssRNA virus belonging to the <it>Flaviviridae </it>family. Studies on the genetic variability of the GBV-C reveals the existence of six genotypes: genotype 1 predominates in West Africa, genotype 2 in Europe and America, genotype 3 in Asia, genotype 4 in Southwest Asia, genotype 5 in South Africa and genotype 6 in Indonesia. The aim of this study was to determine the frequency and genotypic distribution of GBV-C in the Colombian population.</p> <p>Methods</p> <p>Two groups were analyzed: i) 408 Colombian blood donors infected with HCV (n = 250) and HBV (n = 158) from Bogotá and ii) 99 indigenous people with HBV infection from Leticia, Amazonas. A fragment of 344 bp from the 5' untranslated region (5' UTR) was amplified by nested RT PCR. Viral sequences were genotyped by phylogenetic analysis using reference sequences from each genotype obtained from GenBank (n = 160). Bayesian phylogenetic analyses were conducted using Markov chain Monte Carlo (MCMC) approach to obtain the MCC tree using BEAST v.1.5.3.</p> <p>Results</p> <p>Among blood donors, from 158 HBsAg positive samples, eight 5.06% (n = 8) were positive for GBV-C and from 250 anti-HCV positive samples, 3.2%(n = 8) were positive for GBV-C. Also, 7.7% (n = 7) GBV-C positive samples were found among indigenous people from Leticia. A phylogenetic analysis revealed the presence of the following GBV-C genotypes among blood donors: 2a (41.6%), 1 (33.3%), 3 (16.6%) and 2b (8.3%). All genotype 1 sequences were found in co-infection with HBV and 4/5 sequences genotype 2a were found in co-infection with HCV. All sequences from indigenous people from Leticia were classified as genotype 3. The presence of GBV-C infection was not correlated with the sex (p = 0.43), age (p = 0.38) or origin (p = 0.17).</p> <p>Conclusions</p> <p>It was found a high frequency of GBV-C genotype 1 and 2 in blood donors. The presence of genotype 3 in indigenous population was previously reported from Santa Marta region in Colombia and in native people from Venezuela and Bolivia. This fact may be correlated to the ancient movements of Asian people to South America a long time ago.</p

Transfusion-Acquired HIV: History, Evolution of Screening Tests, and Current Challenges of Unreported Antiretroviral Drug Use in Brazil

Prevention of HIV acquisition by blood transfusion from its emergence to the present day is reviewed, and current challenges are delineated. The experience of Funda&ccedil;&atilde;o Pr&oacute;-Sangue/Hemocentro de S&atilde;o Paulo, Brazil, is highlighted in the quest for improvements in blood safety and the evolution of increasingly sensitive and specific screening tests. Concerns and establishing stringent criteria in the screening of potential blood donors are emphasized, and the current criteria for identifying and deferring candidates at high risk of acquiring sexually transmitted diseases are summarized. Future challenges relate to the identification of donors with unreported use of antiretroviral drugs for prophylaxis against possible HIV exposure or for treatment of an HIV infection whose viral expression is undetectable by current analyses. There is a need to better understand the motivation of HIV-exposed donors and to educate them about the risk of transfusion-mediated HIV transmission despite having low or undetectable viral loads. In situations in which traditional HIV RNA or antibody detection assays remain negative, more sensitive analyses are needed to identify potential donors at risk for HIV transmission

Evaluation of Primary Resistance to HIV Entry Inhibitors Among Brazilian Patients Failing Reverse Transcriptase/Protease Inhibitors Treatment Reveal High Prevalence of Maraviroc Resistance-Related Mutations

Entry inhibitor is a new class of drugs that target the viral envelope protein. This region is variable; hence resistance to these drugs may be present before treatment. The aim of this study was to analyze the frequency of patients failing treatment with transcriptase reverse and protease inhibitors that would respond to the entry inhibitors Enfuvirtide, Maraviroc, and BMS-806. The study included 100 HIV-1 positive patients from one outpatient clinic in the city of Sao Paulo, for whom a genotype test was requested due to treatment failure. Proviral DNA was amplified and sequenced for regions of gp120 and gp41. A total of 80 could be sequenced and from those, 73 (91.3%), 5 (6.3%) and 2 (2.5%) were classified as subtype B, F, and recombinants (B/F and B/C), respectively. CXCR4 co-receptor use was predicted in 30% of the strains. Primary resistance to Enfuvirtide was found in 1.3%, following the AIDS Society consensus list, and 10% would be considered resistant if a broader criterion was used. Resistance to BMS-806 was higher; 6 (7.5%), and was associated to non-B strains. Strikingly, 27.5% of samples harbored one or more mutation among A316T, I323V, and S405A, which have been related to decreased susceptibility of Maraviroc; 15% of them among viruses predictive to be R5. A more common mutation was A316T, which was associated to the Brazilian B strain harboring the GWGR motif at the tip of V3 loop and their derivative sequences. These results may be impact guidelines for genotype testing and treatment in Brazil.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[04/15856-9]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)[09/52381-2