O vírus da Artrite-encefalite caprina (CAEV) é um Lentivírus de pequenos ruminantes, que causa uma doença crônica e progressiva, caracterizada por encefalomielite, mastite, pneumonia e artrite. O diagnóstico é baseado na detecção de anticorpos contra o vírus através da Imunodifusão em Gel de Agarose (IDGA), técnica sorológica de referência para CAEV, porém de baixa sensibilidade. O objetivo deste trabalho foi produzir um antígeno a partir da cultura de células de membrana sinovial caprina (MSC) infectadas com a CAEV, para ser utilizado em um teste diagnóstico (ELISA indireto). O antígeno foi obtido de culturas de células de MSC infectadas e posterior tratamento com SDS 0,1%. Amostras de soros caprinos (n=343) foram utilizadas para detectar a presença de anticorpos para CAEV pelo teste ELISA indireto e a técnica IDGA. Nessas amostras, o teste ELISA detectou 72 ( 21%) amostras positivas. Entretanto, o teste IDGA detectou 30 (8%) amostras positivas. O ELISA indireto também detectou precocemente uma soroconversão em 5 animais de um total de 13 controlados periodicamente durante 2 anos. A sensibilidade e a especificidade do teste ELISA com relação a IDGA foi de 73,3% e 84% respectivamente. Esse ELISA com o antígeno viral assim produzido mostrou-se efetivo, de baixo custo e sensível para o diagnóstico sorológico de anticorpos para CAEV.
283Crossing and diagnostic methods of cubiu hybrid plants based on genetic markers Crop Breeding and Applied Biotechnology 8: [283][284][285][286][287][288][289][290] 2008
Chickpea (Cicer arietinum L.) is an essential grain for human consumption owing to its high protein content, nutritional quality and energy-efficient production. The aim of this study was to compare the protein extracts of 24 chickpea genotypes by biochemically characterising the storage proteins. The storage protein content was characterised by protein separation with polyacrylamide gel electrophoresis and visualisation of the banding patterns, which revealed considerable genetic variability within and between genotypes in this chickpea collection. High performance liquid chromatography showed that all genotypes had balanced amino acid content and some were rich in seven amino acids. Two chickpea genotypes, Flip97-171C and Elite, representative of the kabuli and desi types, respectively, were chosen for total proteome analysis. Two-dimensional electrophoresis and subsequent mass spectrometry were used to identify 454 protein spots from the Flip97-171C and Elite proteomes. By using Mascot Server software, 37% of the spots were identified as 47 different proteins involved in a large range of metabolic functions. Most proteins from both proteomes were assigned to nutritional storage activity. Chickpea proteome analysis is essential in reaffirming the quality of this grain protein for human nutrition, and will be important in future nutriproteomics and plant-breeding studies.
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