Picobirnavirus : pesquisa de novos hospedeiros animais, identificação atraves de ensaio de dot-blot e metodos de purificação

AUTOR(ES)

Ismar Rocha Haga

DATA DE PUBLICAÇÃO

1998

RESUMO

A novel virus tentatively named picobirnavirus (PBV) has been described in faeces of several species of vertebrates (humans, rats, guinea pigs, pigs, rabbits and chickens). Examination by polyacrylamide gel electrophoresis (PAGE) of faecal specimens of giant anteaters (Myrmecophaga tridactyla) kept in captivity revealed the presence of the two equimolar bands resembling those of the picobirnaviruses. This is the first report of picobirnaviruses in faeces of a wild animal in captivity, suggesting that there might be other natural PBV hosts. Different methods for virus concentration were tested: addition of calcium chloride (CaCI2), addition of polyethylene glycol (PEG) and sodium chloride (NaCl) and centrifugation through sucrose cushion, the method that provided the most reliable results. Cesium chloride (CsCI) solutions have been widely used for PBV purification protocols. Alternative protocols for density gradient centrifugation (rubidium chloride, sucrose and potassium tartrate) were tested. The results obtained by using rubidium chloride (RbCI) were compatible with those previously reported for CsCI solutions, indicating that RbCI solutions may be used as an alternative protocol for virus purification, not only for picobirnaviruses but for other viruses as well. An enzyme immunoassay using nitrocellulose membranes is described for the detection of picobirnaviruses in faeces from different origins. In this study, positive results were detected in faecal samples from three different animaIs (rats, pigs and giant anteaters), suggesting that viruses identified in these animaIs might share common antigens. The test characteristics indicate that it can be used as a screening test in routine diagnosis, along with a confirmatory polyacrylamide gel electrophoresis (PAGE) test. This is the first report of such an assay used to detect this new, and yet little known, virus in faecal specimens

ASSUNTO(S)

imunoquimica virus microscopia eletronica

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