Date of Award

Summer 1981

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

James C. Johnson

Committee Member

Lloyd Wolfinbarger, Jr.

Committee Member

Kneeland Nesius

Call Number for Print

Special Collections LD4331.B46 L68

Abstract

Duck plague virus causes a hemorrhagic disease affecting the family Anatidae, resulting in 60-80% mortality in captive flocks. Plaque and neutralization assays for Anatid herpesvirus (AHV) require 5-10 days for completion and are restricted by the seasonal availability of fertile duck eggs. The enzyme linked innnunosorbent assay (ELISA) is rapid and specific and avoids the use of living cells. The Holland strain of AHV was grown on duck embryo fibroblasts and purified by banding on CsCl gradients. The resulting virus was used to raise antisera in prebled New Zealand white male rabbits. Antibody production was confirmed in double diffusion plates and by neutralization. The standard direct ELISA of 0.25 ml used AHV specific IgG, prepared by annnonium sulfate precipitation of the antisera, coupled to horseradish peroxidase and 2,2'- azino di(3-ethyl-benzthiazoline sulfonate) as the enzyme substrate. Reaction parameters including time, pH, antigen and conjugated-antibody concentration, solid phase, as well as interference by host cells, were varied for optimization of the assay. The assay appears to be sensitive to as little as 32 ng of protein. With the preparation of a highly purified virus and antiserum with high specificity, the ELISA system may become a basic procedure for AHV identification.

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DOI

10.25777/fa0g-w451

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