Development of monoclonal antibody ELISA for simultaneous detection of bovine coronavirus, rotavirus serogroup A, and Escherichia coli K99 antigen in feces of calves

Christopher J. Thorns From the Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey KT15 3NB United Kingdom (Thorns, Bell, Chasey, Roeder) and Cambridge Veterinary Sciences, Littleport, Ely, Cambridgeshire CB6 ISE, United Kingdom (Chesham).

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 MSc, MIBiol
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Michael M. Bell From the Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey KT15 3NB United Kingdom (Thorns, Bell, Chasey, Roeder) and Cambridge Veterinary Sciences, Littleport, Ely, Cambridgeshire CB6 ISE, United Kingdom (Chesham).

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 MIBiol
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David Chasey From the Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey KT15 3NB United Kingdom (Thorns, Bell, Chasey, Roeder) and Cambridge Veterinary Sciences, Littleport, Ely, Cambridgeshire CB6 ISE, United Kingdom (Chesham).

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 PhD
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John Chesham From the Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey KT15 3NB United Kingdom (Thorns, Bell, Chasey, Roeder) and Cambridge Veterinary Sciences, Littleport, Ely, Cambridgeshire CB6 ISE, United Kingdom (Chesham).

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 PhD
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Peter L. Roeder From the Ministry of Agriculture, Fisheries, and Food, Central Veterinary Laboratory, Weybridge, Surrey KT15 3NB United Kingdom (Thorns, Bell, Chasey, Roeder) and Cambridge Veterinary Sciences, Littleport, Ely, Cambridgeshire CB6 ISE, United Kingdom (Chesham).

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 BVet Med, PhD

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Summary

A rapid elisa was developed for simultaneous detection of bovine coronavirus (bcv), rotavirus (rv) serogroup A, and Escherichia coli K99 antigen in feces of calves. A mixture of 3 monoclonal antibodies specific for bcv, rv, or K99 was used successfully to capture the antigens; the same antibodies labeled with peroxidase were used to detect bcv, rv, or K99. The triple elisa was compared with standard reference diagnostic methods by examining feces from experimentally and naturally infected and healthy calves. All the components of the test were highly specific (> 90%) and sensitive (bcv, 77%; K99, 93%; rv, 100%) when used in a format requiring short incubation steps at 20 C and visual recording of results.

Summary

A rapid elisa was developed for simultaneous detection of bovine coronavirus (bcv), rotavirus (rv) serogroup A, and Escherichia coli K99 antigen in feces of calves. A mixture of 3 monoclonal antibodies specific for bcv, rv, or K99 was used successfully to capture the antigens; the same antibodies labeled with peroxidase were used to detect bcv, rv, or K99. The triple elisa was compared with standard reference diagnostic methods by examining feces from experimentally and naturally infected and healthy calves. All the components of the test were highly specific (> 90%) and sensitive (bcv, 77%; K99, 93%; rv, 100%) when used in a format requiring short incubation steps at 20 C and visual recording of results.

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