Development of a monoclonal blocking ELISA for the detection of antibodies against fowlpox virus.
To provide a fast and easy method to detect antibodies against fowlpox virus (FWPV) particularly in high numbers of chicken sera we established a monoclonal blocking enzyme-linked immunosorbent assay (ELISA). We chose two different monoclonal antibodies (mAb), anti-FWPV 3D9/2B3 and anti-FWPV 8F3/2E11, which are both directed against the 39-kDa protein of FWPV strain HP-1. The blocking ELISA depends on the blocking of mAb binding to solid-phase antigen in the presence of positive serum. For an epidemiological study a total of 184 serum samples from Gambian chicken flocks were analysed against each of the mAbs. Four of the sera were shown to contain FWPV antibodies. These four sera showed a positive cut-off value of more than 50% inhibition exclusively in the test against the mAb anti-FWPV 8F3/2E11. This phenomenon can be explained by the binding of the mAbs to distinct epitopes on the same protein.
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Bavarian Animal Health Service, Poing, Germany.
Pfister
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Kaaden
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D000818:Animals; D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral / Q000097:blood*; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000592:standards* / Q000662:veterinary; D005586:Fowlpox / Q000175:diagnosis* / Q000821:virology; D005587:Fowlpox virus / Q000276:immunology* / Q000302:isolation & purification; D012680:Sensitivity and Specificity
D019138:Antibodies, Blocking; D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral