Abstract

LAMPREA, Nina Paola et al. Production and evaluation of an antiserum for immunohistochemical detection of rabies virus in aldehyde fixed brain tissues. Biomédica [online]. 2010, vol.30, n.1, pp.146-151. ISSN 0120-4157.

Introduction. The standard procedure for rabies diagnosis requires fresh samples of infected brain to be analyzed by two techniques, direct immunofluorescence and inoculation in mice. Rabies-infected, aldehyde-fixed brain tissues can be examined by immunohistochemistry, but the required commercial antibodies are scarce and expensive. Objectives. An anti-rabies antiserum was produced and tested to evaluate the effectiveness of rabies antigen detection in aldehyde preserved brain tissue. Materials and methods. Rabbits were inoculated with a rabies vaccine produced in Vero cells (origin-African green monkey kidney). Anti-rabies antiserum was obtained and tested by immunohistochemistry in aldehyde-fixed brain sections of rabies-infected mice. Several experimental conditions were assayed. The usefulness of the antiserum in human pathology samples was also tested. Results. The specificity of the antiserum was demonstrated for immunohistochemical detection of rabies antigen in fixed aldehydes nervous tissue both from experimental material and pathology archival collection. In addition, the antiserum was successful in detecting rabies virus under conditions that have been considered unfavorable for the preservation of antigens. Conclusions. The inoculation of rabies vaccine in rabbits is an easy and safe procedure for obtaining antiserum useful for the detection of rabies antigen in samples of nervous tissue. Sections obtained on vibratome better preserve the viral antigenicity in comparison with paraffin-embedded tissues. This methods permit less expensive and more rapid immunohistochemical diagnosis of rabies.

Keywords : rabies; rabies virus; rabies vaccines; antibodies; fixatives; tissue fixation; immuno-histochemistry.

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