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Update Date: [2013-11-04]

NO.: 8

AHRI report No.08


STUDIES ON PSEUDORABIES DISEASE (AUJESZKY'S DISEASE) IN SWINE: Ⅰ. Preparation and application of fluorescent-labaled antibody of pseudorabies


Department Taiwan Animal Health Researdi Institute
Author J.F. Su;T.C. UnC.M, Shieli

An epiacotic of Auszky disease cccured in a large herd in southern Taiwan from June through Novemit in 1971. Pseudorabies virus was isolaI from the ham of 1by pigs susJ of dying of Auszkys disease. limnunofluoxescence had arivantuges of sJ and &niracy over conventional methods. So that the authors tiled to prepare paeudorabies fluorescent antihdy conjugate emploJ in the diagnosis of thediseaseinthisisland. The menlresults aresummar&Iasfollows:

1 .The sific antiPrV hyperimmune serum could ire obtained and used as the source of preparation of pseudorabies fluorescent antibody by employing three month old SPFpigleis incculatlvAth2mlofPrV Ongtung&Auszkystain 10”(3) TCIDSOfrrJ) for lsic iromunition, and 4 more booster injections of 1O—3C1) ml of PrV (1O TCID5OkD1) were achninistexel following the basic immunization. The sea thus obtained showed a high neutralizing antibody titer, Anti-Pingtung was 1:256, AntiAujszky was 1:51 Z and were considered satisfatory for the preparation of pseudorabies fluorescent antibody.

2.The r-globulin was prthpitatl by adding the saturatrd ammonium sulfate solution to the hyperimmune serum, then the protein concentration of the r-globulin solution was determined and conjugaI with FITC at the fluoxescein-protein ratio of 1/50. The conjugate was passed through Sephadex and DEAE columns to remove the unconjugatl FITC dye and non-spthfic fluorescein. The conjugate thus obtained through three gccedures was very spezific for PrV and showed a stain titer of 1:2 to 1:4.

3.Specific fluorescence was oberved at &-8 hours following incculating Pry on the PK-1 5 cell line and Pr/ appeared to form a plaqne. At 24 hours the cytopathic effect could ire oherveri, and many infeotd cells appeared shrunken without discrenible nuclei. The entire cell fluoresced at this stage, and had a vacancy in center of p1ue.

4.Anti-Pingtung and Anti-Auzky serum showed cross reaction, but never did with HCV. Therefore, the isolate of Pingtung stain had the same antigenicity as Aujs7ky strain and differed from HCV.


pseudoxabies HCVAujszky strain