ABSTRACT
Abstract Noroviruses (NVs) are responsible for most cases of human nonbacterial gastroenteritis worldwide. Some parameters for the purification of NV virus-like particles (VLPs) such as ease of production and yield were studied for future development of vaccines and diagnostic tools. In this study, VLPs were produced by the expression of the VP1 and VP2 gene cassette of the Brazilian NV isolate, and two purification methods were compared: cesium chloride (CsCl) gradient centrifugation and ion-exchange chromatography (IEC). IEC produced more and purer VLPs of NV compared to CsCl gradient centrifugation.
Subject(s)
Child , Humans , Centrifugation, Density Gradient/methods , Chromatography, Ion Exchange/methods , Norovirus/genetics , Viral Structural Proteins/genetics , Virosomes/isolation & purification , Brazil , Viral Structural Proteins/metabolism , Virosomes/genetics , Virosomes/metabolismABSTRACT
Watermelon (Citrullus lanatus) cultivated in almost all tropical and subtropical regions of the world, has its largest output in China, and then, according to FAO data, Turkey, Iran and Brazil, being one of the main crops cultivated in State of Tocantins, Brazil. In this work was investigated the occurrence and distribution of the watermelon viruses, totaling 752 samples taken in a stratified experimental design in four representative regions of production: Gurupi (150), Lagoa da Confusao (232), Formoso do Araguaia (265) and Porto Nacional (105). The sampling and collecting the leaves of plants with the presence of symptoms were performed once a week during the entire cultivation cycle. As a result, were observed by Dot-ELISA method, different types of viruses, such as Papaya ringspot W (PRSV-W), Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV) (potyvirus), Cucumber mosaic virus ( CMV) (Cucumovirus) and Zucchini lethal chlorosis virus (ZLCV) (Tospovirus). Of these, PRSV-W was predominant (22%), followed by WMV (15%), ZLCV (11%), CMV (5%) and ZYMV (4%). Mixed infections with PRSV-W + WMV and PRSV-W + ZLCV were also observed around 20% frequency (expressed with symptoms differently from a single infection). The results provide important support for the program management viruses.
ABSTRACT
O aumento na ocorrência de begomoviroses (geminiviroses) em tomateiros, Lycopersicon esculentum Mill., em várias regiões brasileiras, vem causando grandes prejuízos para o agronegócio de tomate, devido à ocorrência do inseto vetor, Bemisia argentifolii (Bemisia tabaci biotipo B). A diagnose é realizada em geral por "polymerase chain reaction" (PCR) ou por hibridização com sondas radioativas. A PCR é um método de alta sensibilidade, porém apresenta a desvantagem da possibilidade de obtenção de resultados falso-positivos, devido a contaminações, ou falso-negativos causados por inibidores contaminantes da reação, ou pela extrema especificidade dos iniciadores. A hibridização de ácidos nucléicos com sondas radioativas tem o seu uso limitado devido à necessidade de infra-estrutura especial, treinamento de pessoal, riscos para a saúde do manipulador e demanda constante de radioquímicos. O presente trabalho tem por finalidade demonstrar a viabilidade do uso do método de hibridização com sondas não-radioativas para a detecção de um begomovírus de tomateiro do Brasil. A sensibilidade do teste foi alta, obtendo-se detecção de até 0,1fg do DNA homólogo e em extrato bruto foliar diluído até 100 vezes.
Major outbreaks of tomato begomoviruses (geminiviruses) in several tomato Lycopersicon esculentum Mill growing in many parts of Brazil have been imposing significant losses upon the tomato agribusiness, due to the introduction of the Bemisia argentifolii (Bemisia tabaci biotype B). Polymerase chain reaction (PCR) and hybridization are generally used for diagnosis. The PCR is a detection method with high sensitivity, however it has the disadvantage of producing false-positives, due to contamination, or false-negatives caused by inhibitors or because of the high primer specifity. The use of nucleic acid hybridization with radio-labelled probes is restricted due to the requirement of special infrastructure and handling experience, the risk for the userÆs health and a regular radiochemical element supply. This study is aimed at demonstrating the usefulness of the hybridization method with non-radioactive probes for detection of a tomato begomoviruses in Brazil. The sensitivity of this method was high enabling the detection of 0.1fg of homologous DNA and in crude sap extract diluted up to 100-fold.