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Clinical and Diagnostic Laboratory Immunology, March 2001, p. 352-356, Vol. 8, No. 2
Retrovirology Centre of Expertise, Canadian
Food Inspection Agency, Charlottetown, Prince Edward Island, Canada
C1A 5T11; Health of Animals and Food
Laboratory, Canadian Food Inspection Agency, St-Hyacinthe, Quebec,
Canada J2S 8E32; and Atlantic Veterinary
College, University of Prince Edward Island, Charlottetown, Prince
Edward Island, Canada C1A 4P33
Received 19 July 2000/Returned for modification 13 November
2000/Accepted 21 December 2000
Polyethylene glycol (PEG) was used to produce whole-virus antigen
derived from tissue culture cells infected with a Canadian strain of
caprine arthritis-encephalitis virus. PEG antigen batches were obtained
after precipitation and concentration of infected tissue culture
material with PEG 8000 and final treatment with sodium dodecyl sulfate.
The optimum time of harvest of tissue culture extracted material to
produce the maximum amount of viral proteins was determined in roller
bottles, after cocultivation of infected and noninfected fetal lamb
corneal cells. Samples from day 9 to day 25 postculture were collected
and processed. By Western blotting, the optimum time of harvest was
found to be day 25 following the coculture. Two large batches of PEG
antigen were prepared at the optimum time of harvest. Both batches gave similar results when tested by Western blotting and enzyme-linked immunosorbent assay (ELISA), using reference control sera from infected
and noninfected goats. For further testing in ELISA, cutoff values and
ratios were determined for PEG batch 1, using 200 known serum samples
from goats free of the disease. The PEG antigen batch was compared with
an in-house ELISA antigen in a kinetic mode, using 498 serum samples
from field goats. The in-house ELISA antigen was produced following two
rounds of ultracentrifugation and treatment with sodium dodecyl sulfate
(R. A. Heckert, W. B. McNab, S. M. Richardson, and
M. R. Briscoe, Can. J. Vet. Res. 56:237-241, 1992). The PEG
antigen batch was found suitable for ELISA, with a relative specificity
of 100% and a relative sensitivity of 99.4% compared to the in-house
ELISA antigen. This method of antigen production for ELISA was found to
be rapid, inexpensive, and reliable for the diagnosis of
caprine-arthritis encephalitis, without requiring the use of
sophisticated laboratory equipment.
1071-412X/01/$04.00+0 DOI: 10.1128/CDLI.8.2.352-356.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Simple and Rapid Method for Production of Whole-Virus Antigen for
Serodiagnosis of Caprine Arthritis-Encephalitis Virus by
Enzyme-Linked Immunosorbent Assay
*
Corresponding author. Mailing address: Health of
Animals and Food Laboratory, CFIA, 3400 Blvd. Casavant West,
St-Hyacinthe, Quebec, Canada, J2S 8E3. Phone: (450) 773-7730. Fax:
(450) 773-8152. E-mail: simardc{at}em.agr.ca.
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