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Detection of antibody by the Ouchterlony double immunodiffusion technique
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Antibodies were detected using double immunodiffusion [11]. Petri plates containing casted agarose with six peripheral wells contained serially diluted serum, and a central well contained recombinant and native PirBvp antigen. Antigen and serum were filled into wells (20 μL per well). The double diffusion plates were stored in a humidified chamber at 4 °C, and the precipitin lines were examined daily for 2 days.
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