Development of an enzyme-linked immunosorbent assay for the detection of humoral antibody to Pasteurella anatipestifer.

摘要: An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Pasteurella anatipestifer in duck sera is described. As part initial development, micro-titration plates from different manufacturers were assayed their suitability bind P. antigen. The Nunc Immunopiate II was chosen, on account its overall reproducibility (5.5% coefficient variation) and absence an edge effect. Optimum concentrations reagents determined inclusion 1.0M NaCl wash buffer found reduce non-specific binding increase sensitivity. specific that detected only those ducks either exposed or following vaccination with anatipestifer; immunised other heterologous bacterial antigens, having agglutinating them, gave no detectable response ELISA. Between-assay coefficients variation quality control serum pools representing high, medium low levels 6.8%, 8.3% 8.6% respectively. A precision-dose profile derived. graph absorbance versus log(2) end point titre showed a linear relationship (r = 0.99) over range investigated. derived regression line (P<0.001) used transform measurement obtained single 1:100 dilution into value. It demonstrated ELISA much superior method rapid slide agglutination agar gel precipitin tests measuring antibody responses exposure against type 2.