Desenvolvimento e padronização de método imunoenzimático para a detecção de antígenos de poliovírus 1 e herpesvírus bovino 1

AUTOR(ES)
DATA DE PUBLICAÇÃO

2004

RESUMO

(first article) An indirect solid-phase enzyme-immunoassay (EIA) was developed for the detection of bovine herpes virus 1 (BHV-1) antigen. BHV-1 antigen was obtained in HEp-2 cell culture and used to immunize rabbits and guinea pigs. The BHV-1 antibodies were evaluated by double immunodifusion in agar gel and neutralization test. Optimal concentrations of guinea pig and rabbit immunoglobulins were determined by checkerboard titration. Microtitre plates were coated with 9.0 ug/ml guinea pig anti- BHV-1 immunoglobulin and rabbit anti-BHV-1 immunoglobulin at the concentration of 15.0 ug/ml was used as detecting antibody. The performance of EIA was analysed from standard curve by measuring the coefficient of variation (CV) and the establishment of dose response relationship. The standard curve with eight different concentrations in five replicates resulted in a CV between 3.14% to 15.76%. The dose response relationship was determined by simple linear regression (OD = 0,0149 x [Ag] + 0,4041) with a coefficient of correlation, R2, equal to 96.98%. The assay detected a minimum of 1.6 ug/ml BHV-1 antigen.

ASSUNTO(S)

teste imunoenzimático antigens enzime immunoassay antígenos

ACESSO AO ARTIGO

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