Comparative performance evaluation of hepatitis C virus genotyping based on the 5' untranslated region versus partial sequencing of the NS5B region of brazilian patients with chronic hepatitis C

<p>Abstract</p> <p>Background</p> <p>Genotyping of hepatitis C virus (HCV) has become an essential tool for prognosis and prediction of treatment duration. The aim of this study was to compare two HCV genotyping methods: reverse hybridization line probe assay (LiPA v.1) and partial sequencing of the NS5B region.</p> <p>Methods</p> <p>Plasma of 171 patients with chronic hepatitis C were screened using both a commercial method (LiPA HCV Versant, Siemens, Tarrytown, NY, USA) and different primers targeting the NS5B region for PCR amplification and sequencing analysis.</p> <p>Results</p> <p>Comparison of the HCV genotyping methods showed no difference in the classification at the genotype level. However, a total of 82/171 samples (47.9%) including misclassification, non-subtypable, discrepant and inconclusive results were not classified by LiPA at the subtype level but could be discriminated by NS5B sequencing. Of these samples, 34 samples of genotype 1a and 6 samples of genotype 1b were classified at the subtype level using sequencing of NS5B.</p> <p>Conclusions</p> <p>Sequence analysis of NS5B for genotyping HCV provides precise genotype and subtype identification and an accurate epidemiological representation of circulating viral strains.</p

ePAGe: Sistematização do Plano de Atenção Gerontológica (PAGe)

ABSTRACTThe Gerontological Care Plan, originally “Plano de Atenção Gerontológica(PAGe, in Portuguese)”, is an instrument used to assistGerontology professionals in the multidimensional assessment ofthe elderly. Since its creation, the instrument has undergone a seriesof changes and in this study, we adopted the most currentversion, which has 104 questions. Taking into account the numberof questions present in the instrument, it is possible to infer thatthe manual application of the PAGe on paper demands time andattention from the professional. This paper presents a systematizedversion of the PAGe, whose main objective is to optimize the processof application and analysis of the instrument. The design andevaluation of the systematized version took place in partnershipwith Gerontology professionals, those who created the PAGe orwho frequently apply it. The results point to a positive evaluationby the specialists, especially concerning the application time andthe practicality of generating, visualizing, and analyzing the results

An unsettling explanation for the failure of skatole‐baited ovitraps to capture Culex mosquitoes

Culex mosquitoes are primarily found in temperate and tropical regions worldwide where they play a crucial role as main vectors of filarial worms and arboviruses. In Recife, a northeast city in Brazil, high densities of Culex quinquefasciatus are often found in association with human populated areas. In marked contrast to another part of the city, field tests conducted in the neighborhood of Sítio dos Pintos showed that trapping of mosquitoes in skatole-baited ovitraps did not differ significantly from captures in control (water) traps. Thus, classical and molecular taxonomic approaches were used to analyze the Culex species circulating in Sítio dos Pintos. Results obtained from both approaches agreed on the cocirculation of Culex quinquefasciatus and Culex nigripalpus in three different areas of this neighborhood. What was initially considered as an unexpected failure of this lure turned out to be a more unsettling problem, that is, the first report in Recife of Culex nigripalpus, a vector of Venezuelan equine encephalitis&nbsp;virus and West Nile virus. Unplanned urbanization processes close to remnants of the Atlantic forest, such as observed in Sítio dos Pintos, may have contributed to the introduction of Cx. nigripalpus in urban areas

Correction: Development of Hepatitis C Virus Genotyping by Real-Time PCR Based on the NS5B Region

Background: Hepatitis C virus (HCV) genotyping is the most significant predictor of the response to antiviral therapy. The aim of this study was to develop and evaluate a novel real-time PCR method for HCV genotyping based on the NS5B region. Methodology/Principal Findings: Two triplex reaction sets were designed, one to detect genotypes 1a, 1b and 3a; and another to detect genotypes 2a, 2b, and 2c. This approach had an overall sensitivity of 97.0%, detecting 295 of the 304 tested samples. All samples genotyped by real-time PCR had the same type that was assigned using LiPA version 1 (Line in Probe Assay). Although LiPA v. 1 was not able to subtype 68 of the 295 samples (23.0%) and rendered different subtype results from those assigned by real-time PCR for 12/295 samples (4.0%), NS5B sequencing and real-time PCR results agreed in all 146 tested cases. Analytical sensitivity of the real-time PCR assay was determined by end-point dilution of the 5000 IU/ml member of the OptiQuant HCV RNA panel. The lower limit of detection was estimated to be 125 IU/ml for genotype 3a, 250 IU/ml for genotypes 1b and 2b, and 500 IU/ml for genotype 1a. Conclusions/Significance: The total time required for performing this assay was two hours, compared to four hours required for LiPA v. 1 after PCR-amplification. Furthermore, the estimated reaction cost was nine times lower than that of available commercial methods in Brazil. Thus, we have developed an efficient, feasible, and affordable method for HCV genotype identification.Secretaria de Estado da Saude do ParanaAraucaria FoundationFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Alves de Queiroz Family Fund for Researc

An unsettling explanation for the failure of skatole-baited ovitraps to capture Culex mosquitoes

Submitted by Kamylla Nascimento ([email protected]) on 2018-03-23T13:10:01Z No. of bitstreams: 1 PE-IAM (PREPRINT) - Ovitraps failure to capture mosquitoes .pdf: 1345690 bytes, checksum: 86bdba7a7a6069a3a716974419bdef92 (MD5)Approved for entry into archive by Kamylla Nascimento ([email protected]) on 2018-03-23T14:20:14Z (GMT) No. of bitstreams: 1 PE-IAM (PREPRINT) - Ovitraps failure to capture mosquitoes .pdf: 1345690 bytes, checksum: 86bdba7a7a6069a3a716974419bdef92 (MD5)Made available in DSpace on 2018-03-23T14:20:14Z (GMT). No. of bitstreams: 1 PE-IAM (PREPRINT) - Ovitraps failure to capture mosquitoes .pdf: 1345690 bytes, checksum: 86bdba7a7a6069a3a716974419bdef92 (MD5) Previous issue date: 2018Este trabalho foi apoiado pelo Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), sob o prêmio 407475 / 2012-5; Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco, sob premiaçãoAPQ-2141-4.00 / 12 PPSUS-FACEPE; e o Instituto Nacional de Alergia e Infecciosidade Doenças dos Institutos Nacionais de Saúde, sob o prêmio R01AI095514 e R21AI128931.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia. Recife, PE, Brasil / Universidade Federal de Pernambuco. Centro Acadêmico do Agreste. Caruaru, PE, Brasil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia. Recife, PE, Brasil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia. Recife, PE, Brasil.Universidade Federal de Santa Catarina. Centro de Ciências Biológicas. Departamento de Biologia Celular, Embriologia e Genética. Florianópolis, SC, Brazil.Universidade de São Paulo. Faculdade de Saúde Pública. Departamento de Epidemiologia. São Paulo, SP, Brazil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia. Recife, PE, Brasil.University of California-Davis. Department of Molecular and Cellular Biology. Davis, CA, USA.Culex mosquitoes are primarily found in temperate and tropical regions worldwide where they play a crucial role as main vectors of filarial worms and arboviruses. In Recife, a northeast city in Brazil, high densities of Culex quinquefasciatus are often found in association with human populated areas. In marked contrast to another part of the city, field tests conducted in the neighborhood of Sítio dos Pintos showed that trapping of mosquitoes in skatole-baited ovitraps did not differ significantly from captures in control (water) traps. Thus, classical and molecular taxonomic approaches were used to analyze the Culex species circulating in Sítio dos Pintos. Results obtained from both approaches agreed on the co-circulation of Culex quinquefasciatus and Culex nigripalpus in three different areas of this neighborhood. What was initially considered as an unexpected failure of this lure turned out to be a more unsettling problem, i.e., the first report in Recife of Culex nigripalpus, a vector of Venezuelan equine encephalitis virus and West Nile virus. Unplanned urbanization processes close to remnants of the Atlantic forest, such as observed in Sítio dos Pintos, may have contributed to the introduction of Cx. nigripalpus in urban areas. This article is protected by copyright. All rights reserved

Primers and Probes used in Real-Time PCR genotyping of NS5B region.

1<p>1a_.H77.NC_004102–reference sequence used to calculate positions of NS5B region.</p