ELISAによるブリ血清中の抗Pasteurella piscicidaLPS特異抗体の定量

概要

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Quantitation of antibody (Ig) in yellowtail, Seriola quinqueradiata serum against P. piscicida LPS was carried out by the enzyme-linked immunosorbent assay (ELISA). Pure LPS extracted from P. piscicida was immobilized on Polystyrene bead (LPS-bead). Diluted yellowtail serum was added to the LPS-bead complex. The yellowtail antibody-coupled LPS-bead was incubated with anti-yellowtail Ig rabbit Fab' conjugated with peroxidase and enzyme activity bound on the bead was measured.Using 3 sera differing in agglutination titer from yellowtail immunized with formalin-killed P. piscicida, within assay variation was small showing a mean coefficient of variation of 4.4 to 13.7%.Specific antibody in sera from non-immunized yellowtail was determined. The result showed that the range of specific antibody concentration for 23 non-immunized fish was 6.0 to 30.0μg/ml. In sixty-five immunized by immersion the specific Ig concentration was 5.9 to 112.5μg/ml and in 25 fish intraperitoneally immunized the specific Ig concentration was 97.0 to 1, 030.0μg/ml. There was high correlation (r=0.944) between the concentration of specific antibody and agglutination titer of the sera against formalin-killed P. piscicida cells.