Anti-A and anti-A,B monoclonal antisera with high titers favor the detection of A weak phenotypes.

OBJECTIVES: This study aimed to evaluate the reactivity and the titers of commercial anti-A and anti-A,B antisera in the detection of A weak antigen expression in human red blood cells. BACKGROUND: Commercial monoclonal antisera for ABO phenotyping are useful reagents allowing the identification of the four main ABO phenotypes (A, B, AB, and O). However, the reactivity of these commercial reagents can not be evident when the A or B antigens are weakly expressed, and these antisera have low titers. METHODS/MATERIALS: Six samples from blood donors and five samples from patients with ABO forward and reverse discrepant phenotyping were evaluated. The ABO phenotyping was carried out with different commercial monoclonal anti-A and anti-A,B antisera under different temperatures, using test tubes and gel column agglutination. RESULTS: Monoclonal anti-A antisera with titers less than 256 and anti-A,B with titers less than 128 failed to detect the weak expression of A antigen in 73% and 67% of the A weak phenotypes, respectively. Titres equal to or higher than 2048 (anti-A) and 1024 (anti-A,B) showed better reactivity, independent of the cell clone. CONCLUSION: Our data indicate that anti-A and anti-A,B antisera with high titers give better reactivity with red blood cells carrying A weak antigen expression.

Origin, tempo, and mode of the spread of DENV-4 Genotype IIB across the state of São Paulo, Brazil during the 2012-2013 outbreak.

BACKGROUND: Dengue virus type 4 (DENV-4) was first reported in Brazil in 1982 and since then no more cases were detected again in Brazil until 2010, when the virus was reintroduced. Over the following years, the virus spread to several Brazilian states and resulted in about 1,400,000 dengue cases, in 2013. The largest number of cases were documented in the Southeast macro-region. OBJECTIVES: To determine the phylogeography of DENV-4 Genotype IIB strains isolated during the epidemics in 2012-2013 in São Paulo, Brazil, we aimed to contextualise the contribution of viruses sampled in different localities across the overall movement of DENV-4 in Brazil. METHODS: Based on the envelope gene sequences retrieved from GenBank, we employed a Bayesian phylogeographic approach to assess the spatiotemporal dynamics of DENV-4 Genotype IIB in São Paulo, Brazil. FINDINGS: The dispersal dynamics of DENV-4 Genotype IIB in Brazil indicated Rio de Janeiro and Mato Grosso states as the most likely routes toward São Paulo before the 2012-2013 outbreak. Likewise, Guarujá and São José do Rio Preto facilitated viral spread and transmission to other localities in the South and Southeast macro-regions in Brazil. CONCLUSIONS: The spread pattern of DENV-4 Genotype IIB strains across the country supports two independent introductions of the virus in São Paulo in a short period of time. Furthermore, São Paulo appears to have played a pivotal role in the dissemination of DENV-4 to other locations in Brazil.

Viral Load and Cytokine Response Profile Does Not Support Antibody-Dependent Enhancement in Dengue-Primed Zika Virus-Infected Patients.

Background: The pathogenesis of severe dengue disease involves immune components as biomarkers. The mechanism by which some dengue virus (DENV)-infected individuals progress to severe disease is poorly understood. Most studies on the pathogenesis of severe dengue disease focus on the process of antibody-dependent enhancement (ADE) as a primary risk factor. With the circulation of Zika virus (ZIKV) in DENV-endemic areas, many people infected by ZIKV were likely exposed to DENV. The influence of such exposure on Zika disease outcomes remains unknown. Methods: We investigated whether patients previously exposed to DENV exhibited higher viremia when exposed to a subsequent, heterologous dengue or Zika infection than those patients not previously exposed to dengue. We measured viral loads and cytokine profile during patients' acute infections. Results: Neither dengue nor Zika viremia was higher in patients with prior DENV infection, although the power to detect such a difference was only adequate in the ZIKV analysis. Of the 10 cytokines measured, only 1 significant difference was detected: Levels of interleukin 1ß (IL-1ß) were lower in dengue-infected patients who had experienced a previous dengue infection than patients infected with dengue for the first time. However, power to detect differences between groups was low. In Zika-infected patients, levels of IL-1ß showed a significant, positive correlation with viral load. Conclusions: No signs of ADE were observed in vivo in patients with acute ZIKV infection who had prior exposure to DENV.

A real-time RT-PCR for rapid detection and quantification of mosquito-borne alphaviruses.

Mosquito-borne alphaviruses are widely distributed throughout the world, causing important human illnesses. Therefore, the development of methods to enable early diagnosis of infections by alphavirus is essential. We show here the development and evaluation of a quantitative real-time RT-PCR using genus-specific primers to the nsP1 viral gene of all mosquito-borne alphaviruses. The specificity and sensitivity of the assay were tested using seven alphaviruses and RNA transcribed from Venezuelan equine encephalitis virus. The detection limits of real-time RT-PCR ranged from 10 to 76 copies per ml. The melting temperature (TM) values for amplification of the alphavirus genomes were 83.05 °C and 85.28 °C. Interestingly, the assay suggested the possibility the arthritogenic alphaviruses with TM peaks of 84.83 to 85.28 °C and encephalitic alphaviruses of 83.34 °C to 84.68 °C could be discriminated both diseases. Real-time RT-PCR may prove very useful for the screening and preliminary diagnosis in outbreaks and surveillance studies as well as for measuring the viral load in pathogenesis studies.

A real-time reverse transcriptase polymerase chain reaction for detection and quantification of Vesiculovirus.

Vesiculoviruses (VSV) are zoonotic viruses that cause vesicular stomatitis disease in cattle, horses and pigs, as well as sporadic human cases of acute febrile illness. Therefore, diagnosis of VSV infections by reliable laboratory techniques is important to allow a proper case management and implementation of strategies for the containment of virus spread. We show here a sensitive and reproducible real-time reverse transcriptase polymerase chain reaction (RT-PCR) for detection and quantification of VSV. The assay was evaluated with arthropods and serum samples obtained from horses, cattle and patients with acute febrile disease. The real-time RT-PCR amplified the Piry, Carajas, Alagoas and Indiana Vesiculovirus at a melting temperature 81.02 ± 0.8ºC, and the sensitivity of assay was estimated in 10 RNA copies/mL to the Piry Vesiculovirus. The viral genome has been detected in samples of horses and cattle, but not detected in human sera or arthropods. Thus, this assay allows a preliminary differential diagnosis of VSV infections.

Assessing Oncologists' Attitudes Concerning Comprehensive Genomic Profiling in Stage IV Lung Adenocarcinoma in Brazil.

Introduction: Advances in comprehensive genomic profiling (CGP) of lung adenocarcinomas (LUADs) led to personalized treatment for patients. This study evaluated medical oncologists' attitudes toward CGP in a scenario where sponsored funding for CGP was available. Methods: We designed an online survey assessing CGP use and treating physicians' confidence, composed of three self-confidence domains, which are as follows: confidence in interpreting CGP results, confidence in treating oncogenic-driven LUAD, and confidence in managing tyrosine kinase inhibitor adverse events. The survey was distributed to medical oncologists who treat lung cancer in Brazil. Comparisons between groups were performed using the chi-square or Fisher's exact test. Univariable and multivariable (adjusted OR) analyses were performed. Results: Among 104 respondents who treat patients with lung cancer, 55% were from the Southeast region, 28% had high lung cancer clinical load, and 33% had in-house molecular testing. More than half (51%) of the participants request CGP systematically to stage IV LUAD. As for provider confidence, 67% stated being confident in all three domains: 76% confident in interpreting CGP, 84% confident in treating oncogenic-driven LUAD, and 81% in managing tyrosine kinase inhibitor adverse events. Providers' confidence was associated with systematically requesting CGP to stage IV LUAD (p = 0.013). After controlling for the variables of interest, systematic requesting CGP for stage IV LUAD revealed a significant association with the provider's confidence (adjusted OR = 0.35, p = 0.028, 95% CI: 0.14-0.84). The major challenge for properly requesting CGP was the long turnaround time and the fear of treatment delays. Conclusions: Even though CGP for stage IV LUAD in Brazil is fully sponsored, only half of the oncologists in our survey systematically request it.. Requesting CGP was associated with providers' confidence. Improving access and promoting providers' awareness of CGP utility is necessary to increase CGP use and better inform treatment decisions.

Performance of CDC Trioplex qPCR during a dengue outbreak in Brazil.

BACKGROUND: In recent years real­time reverse transcription polymerase chain reaction (real-time RT-PCR) has become a leading technique for nucleic acid detection and quantification of flaviviruses, including Dengue virus (DENV). Trioplex real-time RT-PCR has the advantages of providing the concurrent detection of Zika virus (ZIKV), DENV, and Chikungunya virus (CHIKV) RNA in human serum. OBJECTIVE: This study sought to compare the sensitivity and specificity of the Trioplex real-time RT-PCR assay to those provided by CDC DENV TaqMan® RT-qPCR assay and conventional PCR when used for DENV detection in the context of a dengue epidemic. STUDY DESIGN: We analyzed 1656 serum samples from symptomatic patients with acute febrile disease for 5 days less between December 2018 and May 2019. The samples were tested using the various PCR-based assays. RESULTS: Of the 1656 serum samples analyzed, 713 (43%) were laboratory-confirmed as arboviruses: 99.86% (712/713) were confirmed as DENV and 0.14% (1/713) were confirmed as ZIKV. Next, 590 samples were selected, and of these, 331 samples (56.1%) were determined to be positive (Ct < 38) and 259 samples (43.9%) were determined to be negative (Ct > 38) using the Trioplex real-time RT-PCR assay. The multiplex method found that the test exhibits 95% sensitivity and 100% specificity. CONCLUSION: This evaluation demonstrates the capacity of the Trioplex real-time RT-PCR assay to detect DENV at a high sensitivity and specificity in a geographic area with a current dengue outbreak and a lower co-circulation of other arboviruses - such as ZIKV and CHIKV, and the results prove it´s applicability as clinical screening test that can serve as a confirmatory test.

The role of lipids in the inception, maintenance and complications of dengue virus infection.

Dengue fever is a viral condition that has become a recurrent issue for public health in tropical countries, common endemic areas. Although viral structure and composition have been widely studied, the infection phenotype in terms of small molecules remains poorly established. This contribution providing a comprehensive overview of the metabolic implications of the virus-host interaction using a lipidomic-based approach through direct-infusion high-resolution mass spectrometry. Our results provide further evidence that lipids are part of both the immune response upon Dengue virus infection and viral infection maintenance mechanism in the organism. Furthermore, the species described herein provide evidence that such lipids may be part of the mechanism that leads to blood-related complications such as hemorrhagic fever, the severe form of the disease.

Zika detection: comparison of methodologies.

Many countries in the Americas have detected local transmission of multiple arboviruses that cause febrile illnesses. Therefore, laboratory testing has become an important tool for confirming the etiology of these diseases. The present study aimed to compare the sensitivity and specificity of three different Zika virus detection assays. One hundred serum samples from patients presenting with acute febrile symptoms were tested using a previously reported TaqMan® RT-qPCR assay. We used a SYBR® Green RT-qPCR and a conventional PCR methodologies to compare the results. Of the samples that were determined to be negative by the TaqMan® RT-qPCR assay, 100% (Kappa=0.670) were also found to be negative by SYBR® Green RT-qPCR based on Tm comparison; however, 14% (Kappa=0.035) were found to be positive by conventional PCR followed by agarose gel electrophoresis. The differences between the ZIKV strains circulating worldwide and the low viremia period can compromise diagnostic accuracy and thereby the accuracy of outbreak data. Therefore, improved assays are required to improve the diagnosis and surveillance of arbovirus.

Clinical, laboratory and virological data from suspected ZIKV patients in an endemic arbovirus area.

BACKGROUND: The emergence of Zika virus (ZIKV) presents new challenges to both clinicians and public health authorities. Overlapping clinical features between the diseases caused by ZIKV, dengue (DENV) and chikungunya (CHIKV) and the lack of validated serological assays for ZIKV make accurate diagnosis difficult. Brazilian authorities largely rely on clinical and epidemiological data for the epidemiological and clinical classifications of most ZIKV cases. OBJECTIVE: To report the laboratory and clinical profiles of patients diagnosed with Zika fever based only on clinical and epidemiological data. STUDY DESIGN: We analyzed 433 suspected cases of ZIKV identified by the attending physician based on proposed clinical criteria. The samples were also screened for ZIKV, DENV and CHIKV using PCR. RESULTS: Of the 433 patients analyzed, 168 (38.8%) were laboratory-confirmed for arboviruses: 96 were positive for ZIKV, 67 were positive for DENV (56 for DENV-2, 9 for DENV-1, and 2 for DENV-4), four were positive for co-infection with ZIKV/DENV-2, and one was positive for CHIKV. The most common signs or symptoms in the patients with laboratory-confirmed ZIKV were rash (100%), arthralgia (77.1%), fever (74.0%), myalgia (74.0%) and non-purulent conjunctivitis (69.8%). In patients with laboratory-confirmed DENV infections, the most frequently observed symptoms were rash (100%), fever (79.1%), myalgia (74.6%), headache (73.1%) and arthralgia (70.1%). The measure of association between clinical manifestations and laboratory manifestations among patients with ZIKV and DENV detected a statistically significant difference only in abdominal pain (p=0.04), leukopenia (p=0.003), and thrombocytopenia (p=0.01). CONCLUSION: Our data suggests that clinical and epidemiological criteria alone are not a good tool for ZIKV and DENV differentiation, and that laboratory diagnosis should be mandatory.

Evaluation and optimization of SYBR Green real-time reverse transcription polymerase chain reaction as a tool for diagnosis of the Flavivirus genus in Brazil.

INTRODUCTION: The genus Flavivirus includes several pathogenic species that cause severe illness in humans. Therefore, a rapid and accurate molecular method for diagnosis and surveillance of these viruses would be of great importance. Here, we evaluate and optimize a quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) method for the diagnosis of the Flavivirus genus. METHODS: We evaluated different commercial kits that use the SYBR Green system for real-time RT-PCR with a primer set that amplifies a fragment of the NS5 flavivirus gene. The specificity and sensitivity of the assay were tested using twelve flaviviruses and ribonucleic acid (RNA) transcribed from the yellow fever virus. Additionally, this assay was evaluated using the sera of 410 patients from different regions of Brazil with acute febrile illness and a negative diagnosis for the dengue virus. RESULTS: The real-time RT-PCR amplified all flaviviruses tested at a melting temperature of 79.92 to 83.49°C. A detection limit of 100 copies per ml was determined for this assay. Surprisingly, we detected dengue virus in 4.1% (17/410) of samples from patients with febrile illness and a supposedly negative dengue infection diagnosis. The viral load in patients ranged from 2.1×107to 3.4×103copies per ml. CONCLUSIONS: The real-time RT-PCR method may be very useful for preliminary diagnoses in screenings, outbreaks, and other surveillance studies. Moreover, this assay can be easily applied to monitor viral activity and to measure viral load in pathogenesis studies.

Dengue virus surveillance: Detection of DENV-4 in the city of São José do Rio Preto, SP, Brazil.

Dengue viruses are the most common arbovirus infection worldwide and are caused by four distinct serotypes of the dengue virus (DENV). In the present study, we assessed DENV transmission in São José do Rio Preto (SJRP) from 2010 to 2014. We analyzed blood samples from febrile patients who were attended at health care centers in SJRP. DENV detection was performed using multiplex RT-PCR, using flavivirus generic primers, based on the genes of the non-structural protein (NS5), followed by nested-PCR assay with species-specific primers. We analyzed 1549 samples, of which 1389 were positive for NS1 by rapid test. One thousand and eight-seven samples (78%) were confirmed as positive by multiplex RT-PCR: DENV-4, 48.5% (528/1087); DENV-1, 41.5% (449/1087); DENV-2, 9.5% (104/1087); and co-infection (5 DENV-1/DENV-4, 1 DENV-1/DENV-2), 0.5% (6/1087). Phylogenetic analysis of the DENV-4 grouped the isolates identified in this study with the American genotype and the showed a relationship between isolates from SJRP and isolates from the northern region of South America. Taken together, our data shows the detection and emergence of new dengue genotype in a new region and reiterate the importance of surveillance programs to detect and trace the evolution of DENV.

Eficácia do uso do álcool etílico 70% na antissépsia da pele antes da coleta de sangue / Efficacy of 70% ethyl alcohol use o9n skin antissepsia before blood collection

Objetivo ­ Quantificar a presença de agentes da flora microbiana da pele de usuários da sala de coleta de uma unidade básica de saúde ­ UBS, assim como verificar a eficácia da antissepsia da pele com o uso do álcool etílico 70% prévia à coleta de sangue. Métodos ­ A coleta da região antecubital do braço dos 40 pacientes selecionados aleatoriamente, foi realizada antes e após antissepsia do local com álcool etílico 70%, com auxílio de um swab embebido em solução fisiológica estéril para posterior transporte e processamento o qual foi realizado no laboratório escola da Universidade Paulista UNIP, campus JK, São José do Rio Preto. Resultados ­ Das 80 amostras (40 antes da antissepsia e 40 depois da antissepsia) coletadas, foi observada uma redução de 87,01% do número de colônias de micro-organismos identificados como cocos e bacilos Gram-positivos, catalase positivos. Conclusão ­ Os resultados da pesquisa permitem concluir que o álcool etílico a 70% é eficaz na antissepsia da pele e que a antissepsia com álcool etílico 70% reduz a flora microbiana da pele conferindo maior segurança e liberdade de riscos ao procedimento prévio à coleta de sangue.

Objective ­ To quantify the presence of agents of the microbial flora of the skin of patients in the collection room of a hospital (UBS) as well as to verify the efficacy of skin antisepsis with the use of 70% ethyl alcohol prior to blood collection. Methods ­ The collection of the antecubital region of the arm of the 40 randomly selected patients was performed before and after site antisepsis with 70% ethyl alcohol, with the aid of a swab soaked in sterile solution for subsequent transportation and processing, which was performed in the laboratory University of São Paulo UNIP, JK campus, São José do Rio Preto. Results ­ From the 80 samples (40 before antisepsis and 40 after antisepsis) collected, a reduction of 87.01% in the number of colonies of microorganisms identified as cocci and gram positive bacilli, catalase positive, was observed. Conclusion ­ The results of the research allow us to conclude that 70% ethyl alcohol is effective in antisepsis of the skin and that antisepsis with 70% ethyl alcohol reduces the microbial flora of the skin giving greater safety and reduced risk to the procedure prior to blood collection

Monitoramento de fungos anemófilos no ambiente de uma biblioteca no Município de São José do Rio Preto - Sp, Brasil / Anemophilic fungi monitoring in a library environment in the city of São Jose do Rio Preto, Brazil

Os fungos desempenham vários papéis que impactam a humanidade de diversas maneiras. Suas características metabólicas são importantes na biotecnologia, porém, tais microrganismos podem desencadear alguns problemas de saúde pública e até mesmo serem letais. Objetivo: detectar a presença de fungos no acervo de uma biblioteca no município de São José do Rio Preto. Metodologia: foram coletadas quarenta amostras nas superfícies inanimadas (livros, estantes, documentos, mapas, artigos e revistas) das principais salas da biblioteca com o auxílio de swabs umedecidos em solução salina estéril, posteriormente encaminhados ao laboratório de Biomedicina da Universidade Paulista ­ UNIP. As amostras foram semeadas em meio de cultura ágar Sabouraud Dextrose (SDA), tendo adicionado cloranfenicol e incubadas a 30 °C. Foi realizada a colônia gigante em todas as cepas crescidas em SDA para a realização da técnica de microcultivo para a identificação dos fungos, de acordo com o Manual de Detecção e Identificação dos Fungos de Importância Médica da Agência Nacional de Vigilância Sanitária. Resultados: Houve positividade em trinta e uma amostras (78%) e em quatro delas foi observado mais de um tipo de colônia (13%). Das vinte e duas superfícies de livros analisadas, foram isolados e identificados: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. e Nigrospora sp. Nas oito superfícies de estantes: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. e Scopulariopsis sp. e, nos dez documentos: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. e Trichoderma sp. Conclusão: Os fungos encontrados estão amplamente distribuídos no ambiente como solo e ar e, por diversos fatores, instalam-se em locais como bibliotecas. Em condições favoráveis, podem infectar o homem e causar perdas patrimoniais para os acervos.

Fungi play many roles that impact humankind in different ways. Their metabolic characteristics are important in biotechnology; however, these microorganisms can trigger some public health problems or may even be lethal. Objective: detect the presence of fungi in the collection of a public library in the city of São José do Rio Preto, Brazil. Methods: a total of forty samples were collected from inanimate surfaces (books, shelves, documents, maps, articles and magazines) located in the main rooms of the library with swabs soaked in sterile saline solution and sent to the Universidade Paulista ­ UNIP laboratories. The samples were plated in Sabouraud Dextrose Agar (SDA) supplemented with chloramphenicol and incubated at 30 °C. The colonies that grew in SDA were isolated in Potato Dextrose Agar for performing the slide culture technique for the identification of the fungi, performed according to the Manual of Detection and Identification of Fungi of Medical Importance from the Brazilian Health Surveillance Agency (ANVISA). Results: Thirty-one samples (78%) were positive, and in four of them more than one fungus genus was observed (13%). From the twenty-two book surfaces analyzed, the following fungi were isolated and identified: Aspergillus flavus, Aspergillus niger, Cunninghamella sp., Cladosporium sp., Curvularia sp., Mucor sp. and Nigrospora sp. On the eight shelves: Aspergillus flavus, Aspergillus niger, Aspergillus versicolor, Penicillium sp. and Scopulariopsis sp. The ten documents analyzed presented the following fungi: Aspergillus nidulans, Aspergillus sp., Cladosporium sp., Cunninghamella sp. and Trichoderma sp.. Conclusion: These fungi are widely distributed in the environment such as in the soil and air, and due to several factors, they colonize public places, such as libraries. In favorable conditions, they may infect humans and cause diseases.

Detection of Mayaro virus infections during a dengue outbreak in Mato Grosso, Brazil.

Arboviruses are common agents of human febrile illness worldwide. In dengue-endemic areas illness due to other arboviruses have been misdiagnosed as dengue based only on clinical-epidemiological data. In this study we investigated the presence of Brazilian arboviruses in sera of 200 patients presenting acute febrile illness, during a dengue outbreak in Sinop, MT, Brazil. The results showed that 38 samples were positive to Dengue virus (DENV) type 1, two samples to DENV type 4, and six to Mayaro virus. These results indicate that arboviruses others than DENV are circulating in Sinop and the surrounding region, which are going undiagnosed. In addition, molecular and evolutionary analyses indicate that two MAYV genotypes are co-circulating in Mato Grosso, Brazil. Thus, a strong surveillance program must be implemented to evaluate and monitor the distribution and the true importance of non-dengue arboviruses in the etiology of acute febrile illnesses.

Isolamento e identificação de bactérias presentes em superfícies inanimadas de uma unidade básica de saúde no interior de São Paulo / Isolation and identification bacteria present in inanimate surfaces of a basic health units in the state of São Paulo

Objetivo ­ Identificar e isolar bactérias em superfícies inanimadas de uma Unidade Básica de Saúde no interior de São Paulo e verificar a suscetibilidade bacteriana frente aos antibióticos testados. Métodos ­ Foram coletadas 60 amostras de superfícies inanimadas de uma Unidade Básica de Saúde realizado um esfregaço da superfície utilizando um swab umidificado em solução salina estéril e inoculado em caldo BHI. Após foram semeadas em meio de cultura não seletivo e identificadas através de coloração de gram, catalase, crescimento no manitol, coagulase, DNAse, fermentação da lactose, EPM, Mili e Citrato. O teste de suscetibilidade antimicrobiana por difusão de disco foi aplicado. Resultados ­ Das 60 superfícies inanimadas analisadas, 59 (98,3%) estavam contaminadas, sendo 29% (17) contaminadas por Staphylococcus aureus, 71,1% (41) por Staphylococcus coagulase negativa e 72,9% por enterobactérias (20,9% Escherichia coli, 16,3% Citrobacter freundii, 13,9% Serratia marcescens, 13,9% Serratia liquefaciens, 7% Salmonella paratyphi, 7% Klebsiella pneumoniae, 4,6% Shigella sp., 4,6% Yersinia enterocolitica, 2,3% Klebsiella oxytoca, 2,3% Serratia odorifera, 2,3% Providencia sp. e 2,3% Enterobacter sp.). Foi observada uma multirresistência dessas bactérias frente aos antibióticos testados. Conclusão ­ A contaminação em superfícies inanimadas, assim como o perfil de resistência microbiana, foram evidentes na UBS analisada, resultados estes que demonstram a necessidade de uma maior vigilância no que diz respeito ao controle de infecções, assim ao que se refere a reavaliação das estratégias de limpeza e desinfecção dessas superfícies inanimadas

Objective ­ Identify and bacteria isolates on inanimate surfaces of a Basic Health Unit in the interior of São Paulo and to verify the bacterial susceptibility against the tested antibiotics. Methods ­ Sixty samples of inanimate surfaces from a Basic Health Unit were collected by swabbing the surface using a swab humidified in sterile saline and inoculated into BHI broth. After sowing in non-selective culture medium and identified by staining of gram, catalase, growth in mannitol, coagulase, DNAse, fermentation of lactose, EPM, Mili, Citrate. The antimicrobial susceptibility test by disk diffusion were applied. Results ­ Of the 60 inanimate surfaces analyzed, 59 (98.3%) were contaminated, with 28.9% (17) being contaminated by Staphylococcus aureus, 71.1% (42) Staphylococcus coagulase negative and 72.9% by enterobacteria (20.9% Escherichia coli, 16.3% Citrobacter freundii, 13.9% Serratia marcescens, 13.9% Serratia liquefaciens, 7% Salmonella paratyphi, 7% Klebsiella pneumoniae, 4.6% Shigella sp., 4.6% Yersinia enterocolitica, 2.3% Klebsiella oxytoca, 2.3% Serratia odorifera, 2.3% Providencia sp. e 2.3% Enterobacter sp.) A multiresistance of these bacteria was observed against the tested antibiotics. Conclusions ­ The contamination on inanimate surfaces, as well as the microbial resistance profile, were evident in the Basic Health Unit analyzed, which results demonstrate the need for greater vigilance with regard to infection control, as well as the reevaluation of strategies for cleaning and disinfecting these inanimate surfaces.

Bactérias causadoras de infecções urinárias e seu perfil de resistência aos antimicrobianos / Bacteria who causes urinary tract infection and their antimicrobial resistance profile

Objetivo ­ Identificar quais os microrganismos responsáveis por esta infecção e seu perfil de resistência aos antimicrobianos. Metódos ­ A pesquisa foi realizada no Laboratório CDAC-Centro de Diagnósticos de Análises Clínicas, situado no município de Nova Granada - SP. Foram analisadas amostras de uroculturas positivas e seus respectivos antibiogramas, cujos resultados foram obtidos através de laudos laboratoriais digitais no período de janeiro de 2017 a janeiro de 2018 e à partir da realização dessa coleta de dados, foi feito um levantamento dos principais agentes patológicos causadores das infecções do trato urinário que atingem a população do município, assim como foi verificado o perfil de suscetibilidade aos antibioticos testados. Resultados ­ O uropatógeno mais frequente foi Escherichia coli (N = 58/136; 43%), seguido por Enterobacter sp. (N = 30/136; 22%), Klebsiella sp. (N = 15/136; 11%), Shigella sp. (N = 10/136; 7%), Proteus vulgaris. (N = 6/136; 4,5%), Citrobacter sp. (N = 5/136; 3,5%), Providencia sp. (N = 4/136; 3%), Proteus mirabilis. (N = 4/136; 3%), Edwardsiella sp. (N = 2/136; 1,5%) e Proteus sp. (N = 2/136; 1,5%). Oxacilina e ácido nalidixico apresentaram menor poder inibitório contra os uropatógenos encontrados. Conclusão ­ O uropatógeno mais frequente foi Escherichia coli,seguido por Enterobacter sp., Klebsiella sp. e Shigella sp. Os dados aqui relatados demonstram que a etiologia das infecções urinárias é semelhante à encontrada em outros municípios. Porém, o padrão de resistência desses uropatógenos pode possuir características diferenciadas de acordo com o histórico de consumo de antimicrobianos em cada comunidade. Assim, é importante que dados epidemiológicos sejam periodicamente divulgados com a intenção de auxiliar a comunidade médica

Objective ­ To identify the responsible microorganisms for the urine infection and their antimicrobial resistance profile. Methods ­ We carried out the research at the Laboratory-Diagnostic Center for Clinical Analysis (CDAC), located at the Nova Granada city. We analyzed samples of positive urocultures and their respective antibiograms, whose results were taken through laboratory reports from January 2017 to January 2018 and from the data gathering. We performed a data survey of the main pathological agents who causes infections in the urinary tract affecting the city's population, as well as the susceptibility profile of the antibiotics tested. Results ­ Escherichia coli (N = 58/136, 43%) was the most frequent uropathogen identified, followed by Enterobacter sp. (N = 30/136, 22%), Klebsiella sp. (N = 15/136, 11%), Shigella sp. (N = 10/136, 7%), Proteus vulgaris (N = 6/136, 4,5%), Citrobacter sp. (N = 5/136; 3,5%), Providencia sp. (N = 4/136, 3%), Proteus mirabilis (N = 4/136, 3%), Edwardsiella sp. (N = 2/136, 1.5%), and Proteus sp. = 2/136, 1.5%). Oxacillin and nalidixic acid showed lower inhibitory power against the found uropathogens. Conclusion ­ Escherichia coli was the most frequent uropathogen found, followed by Enterobacter sp, Klebsiella sp and Shigella sp. We demonstrate with the data reported here that the etiology of urinary infections of our research is similar to the found in other cities. However, the antimicrobial resistance profile of these uropathogens may have different characteristics according to the history of antibiotics use in each community. It is therefore important that epidemiological data be periodically disclosed with the intention of assisting the medical community

Isolamento e identificação de bactérias presentes em estojos de lentes de contato / Isolation and indentification of bacteria presents of contact lens cases

Objetivo ­ Esta pesquisa teve o intuito de realizar um estudo microbiológico em estojos de lentes de contato, bem como correlacionar com os hábitos de higiene dos usuários. Métodos ­ Após aprovação do comitê de ética, 50 estojos de lente de contato foram cedidos por acadêmicos voluntários, os quais também responderam a um questionário contendo dados pessoais, assim como sobre a forma de manipulação das lentes. As amostras foram coletadas da superfície interna do estojo, isoladas em meios de cultura e identificadas através de provas bioquímicas específicas. Resultados ­ Dos 50 estojos, 49 (98%) apresentaram crescimento de microrganismos, tais como Staphylococcus sp (27), Staphylococcus aureus (9), Serratia liquefaciens (4), Citrobacter freundii (2), Escherichia coli (1) e Hafnia alvei (1), sendo que em um estojo foi identificado ambas as bactérias Staphylococcus aureus e Serratia liquefaciens. As lentes mais utilizadas (96% dos usuários) foram dos tipos gelatinosas descartáveis, seguida do tipo rígida (4%). O uso terapêutico esteve presente em 76% dos usuários, já o uso estético em 24%. O tempo de uso das mesmas lentes foi de 30 dias até quatro anos. Quando questionados sobre higienização, 34% afirmaram lavar as mãos sempre que manipulavam as lentes, 10% armazenavam os estojos em locais fechados, enquanto os outros 84% armazenavam em gabinetes de banheiros ou expostos sobre as pias. Em relação a incômodos na utilização, 32% relataram prurido, ardência ou vermelhidão. Conclusão ­ A falta de cuidado na higienização e no armazenamento do

Objective ­ This research aimed to carry out a microbiological study in contact lens cases, as well as to correlate with users' hygiene habits. Methods ­ After approval by the ethics committee, 50 contact lens cases were provided by volunteer academics, who also answered a questionnaire containing personal data, as well as how to manipulate the lens. Samples were collected from the inner surface of the kit, isolated in culture media and identified by specific biochemical tests. Results ­ Of the 50 kits, 49 (98%) showed growth of microorganisms such as Staphylococcus sp (27), Staphylococcus aureus (9), Serratia liquefaciens (4), Citrobacter freundii (2), Escherichia coli (1) and Hafnia alvei (1), and in one kit both Staphylococcus aureus and Serratia liquefaciens bacteria were identified. The most used lenses (96% of users) were of the disposable soft types, followed by the rigid type (4%). Therapeutic use was present in 76% of users, while aesthetic use in 24%. The wearing time of the same lenses was from 30 days to four years. When asked about sanitation, 34% said they washed their hands whenever they manipulated the lenses, 10% stored the cases indoors, while the other 84% stored in bathroom cabinets or exposed on the sinks. Regarding discomfort in use, 32% reported itching, burning or redness. Conclusions ­ The lack of care in the hygiene and storage of the cases were extremely important for its microbiological contamination, answering the hypothesis raised at the beginning of the study.

Origin, tempo, and mode of the spread of DENV-4 Genotype IIB across the state of São Paulo, Brazil during the 2012-2013 outbreak

BACKGROUND Dengue virus type 4 (DENV-4) was first reported in Brazil in 1982 and since then no more cases were detected again in Brazil until 2010, when the virus was reintroduced. Over the following years, the virus spread to several Brazilian states and resulted in about 1,400,000 dengue cases, in 2013. The largest number of cases were documented in the Southeast macro-region. OBJECTIVES To determine the phylogeography of DENV-4 Genotype IIB strains isolated during the epidemics in 2012-2013 in São Paulo, Brazil, we aimed to contextualise the contribution of viruses sampled in different localities across the overall movement of DENV-4 in Brazil. METHODS Based on the envelope gene sequences retrieved from GenBank, we employed a Bayesian phylogeographic approach to assess the spatiotemporal dynamics of DENV-4 Genotype IIB in São Paulo, Brazil. FINDINGS The dispersal dynamics of DENV-4 Genotype IIB in Brazil indicated Rio de Janeiro and Mato Grosso states as the most likely routes toward São Paulo before the 2012-2013 outbreak. Likewise, Guarujá and São José do Rio Preto facilitated viral spread and transmission to other localities in the South and Southeast macro-regions in Brazil. CONCLUSIONS The spread pattern of DENV-4 Genotype IIB strains across the country supports two independent introductions of the virus in São Paulo in a short period of time. Furthermore, São Paulo appears to have played a pivotal role in the dissemination of DENV-4 to other locations in Brazil.

Frequência dos alelos do sistema antígeno leucocitário humano em doadores e pacientes pré-transplante de medula óssea / Human leukocyte antigen alleles frequency in donors and pre-transplant patients from bone marrow

Introdução: O estudo da frequência dos alelos detectados nos doadores e pacientes previamente selecionados para o transplante de medula óssea permite estimar as reais chances de um paciente em lista de espera encontrar um doador com antígeno leucocitário humano (Human leucocite antigen; HLA) idêntico não relacionado, além de facilitar e direcionar o planejamento do crescimento do Registro Nacional deDoadores de Medula Óssea. Objetivo: Descrever e analisar afrequência dos alelos do sistema HLA de classe I (HLA-A, -B e -C) e classe II (HLA-DRB1 e -DQB1) de doadores e pacientespré-transplante de medula óssea, do Hospital de Câncer deBarretos. Material e Métodos: Um total de 98 amostras dedoadores e 106 amostras de pacientes foi selecionado comtipificações em alta resolução, no período de outubro de 2014a outubro de 2015. As amostras foram tipificadas para os lociHLA-A, -B, -C, -DR e -DQ. Resultados: O predomínio daraça branca reflete a composição étnica do Brasil. As doençasde base mais comuns que levaram o paciente ao transplanteforam a leucemia aguda linfóide (34%) e mieloide (29,2%).Os grupos alélicos mais frequentes nos registros foramA*02, A*24, A*03, A*01, B*35, B*44, C*07, DQB1*03,DQB1*05, DQB1*06, DRB1*01 e DRB1*13. Conclusão: Osresultados encontrados reforçam a importância de conhecero perfil demográfico e imunogenético das regiões do Brasil,contribuindo desta forma na redução do tempo de espera porum doador histocompatível

Introduction: The study of allele frequencies detected in donors and patients previously selected for bone marrow transplantation allows us to estimate the real chances of a patient in the waiting list to find an Human leucocite antigen (HLA) identical unrelated donor. This also facilitates and drives the growth planning of the Brazilian Registry of planning Bone Marrow Transplantation (REDOME). Objective: Describe and analyze the frequency of HLA class I alleles (HLA-A*, -B* and ­C*) and class II alleles, genotypes, and haplotypes(HLA-DRB1* and -DQB1*) from donors and bone marrowpre-transplant patients. Material and Methods: A total of 98donor samples and 106 patient samples were selected withhigh resolution typing, from October 2014 to October 2015.Samples were typed for HLA-A, -B, -C, -DR and -DQ loci.Results: The predominance of the white race reflects theethnic composition of Brazil. The most common underlyingdiseases that led to transplantation patients were acutelymphoid leukemia (34%) and myeloid (29.2%). The mostfrequent allelic groups were A*02, A*24, A*03, A*01, B*35,B*44, C*07, DQB1*03, DQB1*05, DQB1*06, DRB1*01 andDRB1*13. Conclusion: The results reinforce the importanceof understanding the demographic and immunogenic profilefrom Brazilian Regions. This can contribute to the reduction ofwaiting time for a histocompatible donor.