Double-antibody sandwich enzyme-linked immunosorbent assay and immunoblot analysis used for control of caseous lymphadenitis in goats and sheep

E. A. ter Laak From the Department of Bacteriology, Central Veterinary Institute, PO Box 65, 8200 AB Lelystad, The Netherlands.

Search for other papers by E. A. ter Laak in
Current site
Google Scholar
PubMed
Close
 DVM
,
J. Bosch From the Department of Bacteriology, Central Veterinary Institute, PO Box 65, 8200 AB Lelystad, The Netherlands.

Search for other papers by J. Bosch in
Current site
Google Scholar
PubMed
Close
,
G. C. Bijl From the Department of Bacteriology, Central Veterinary Institute, PO Box 65, 8200 AB Lelystad, The Netherlands.

Search for other papers by G. C. Bijl in
Current site
Google Scholar
PubMed
Close
, and
B. E. C. Schreuder From the Department of Bacteriology, Central Veterinary Institute, PO Box 65, 8200 AB Lelystad, The Netherlands.

Search for other papers by B. E. C. Schreuder in
Current site
Google Scholar
PubMed
Close
 DVM

Click on author name to view affiliation information

Summary

A double-antibody sandwich elisa for detection of antibodies directed against the exotoxin of Corynebacterium pseudotuberculosis, the cause of caseous lymphadenitis (CL) in small ruminants, was developed. A concentrated exotoxin was used. For interpretation of elisa results, these sera were tested: sequentially obtained sera of C pseudotuberculosis-inoculated goats and sheep that were monitored for 68 weeks; sequentially obtained sera from 80 goats of 3 flocks with CL; sera from 652 goats of 7 flocks without CL; sera from 160 sheep of 4 flocks without CL; and 2,265 caprine and 208 ovine sera submitted for diagnostic testing. Data regarding the infection status and history of 10,454 of the 23,302 animals were collected after testing; most of these were goats that had been part of a CL control program. Specificity and sensitivity of the elisa were nearly 100%. Subsequently, 31,978 animals from which no data on infection status of flocks had been collected were then tested. It was concluded that the elisa is a useful diagnostic test for CL eradication programs. Sera with doubtful or inconclusive elisa results were examined by use of immunoblot analysis. Proteins from C pseudotuberculosis culture supernatant were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto nitrocellulose. Six proteins with molecular mass of 68, 65, 39, 38, 31, and 29 kDa reacted with sera from goats and sheep with experimentally induced or naturally acquired infection. Immunoblot analysis was valuable in classifying sera with doubtful or inconclusive results by elisa.

Summary

A double-antibody sandwich elisa for detection of antibodies directed against the exotoxin of Corynebacterium pseudotuberculosis, the cause of caseous lymphadenitis (CL) in small ruminants, was developed. A concentrated exotoxin was used. For interpretation of elisa results, these sera were tested: sequentially obtained sera of C pseudotuberculosis-inoculated goats and sheep that were monitored for 68 weeks; sequentially obtained sera from 80 goats of 3 flocks with CL; sera from 652 goats of 7 flocks without CL; sera from 160 sheep of 4 flocks without CL; and 2,265 caprine and 208 ovine sera submitted for diagnostic testing. Data regarding the infection status and history of 10,454 of the 23,302 animals were collected after testing; most of these were goats that had been part of a CL control program. Specificity and sensitivity of the elisa were nearly 100%. Subsequently, 31,978 animals from which no data on infection status of flocks had been collected were then tested. It was concluded that the elisa is a useful diagnostic test for CL eradication programs. Sera with doubtful or inconclusive elisa results were examined by use of immunoblot analysis. Proteins from C pseudotuberculosis culture supernatant were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto nitrocellulose. Six proteins with molecular mass of 68, 65, 39, 38, 31, and 29 kDa reacted with sera from goats and sheep with experimentally induced or naturally acquired infection. Immunoblot analysis was valuable in classifying sera with doubtful or inconclusive results by elisa.

All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 35 35 6
PDF Downloads 22 22 1
Advertisement