Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies Babesia bovis-Infected Cattle

Will L Goff; Terry F McElwain; Carlos E Suarez; Wendell C Johnson; Wendy C Brown; Junzo Norimine; Donald P Knowles

doi:10.1128/CDLI.10.1.38-43.2003

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Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies Babesia bovis-Infected Cattle

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Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies Babesia bovis-Infected Cattle

Will L Goff, Terry F McElwain, Carlos E Suarez, Wendell C Johnson, Wendy C Brown, Junzo Norimine and Donald P Knowles

Clinical and diagnostic laboratory immunology, Vol.10(1), pp.38-43

01/2003

DOI: https://doi.org/10.1128/CDLI.10.1.38-43.2003

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https://hdl.handle.net/2376/104979

PMCID: PMC145267

PMID: 12522037

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https://doi.org/10.1128/CDLI.10.1.38-43.2003View

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Abstract

Veterinary Immunology

The competitive enzyme-linked immunosorbent assay (cELISA) format has proven to be an accurate, reliable, easily standardized, and high-throughput method for detecting hemoparasite infections. In the present study, a species-specific, broadly conserved, and tandemly repeated B-cell epitope within the C terminus of the rhoptry-associated protein 1 of the hemoparasite Babesia bovis was cloned and expressed as a histidine-tagged thioredoxin fusion peptide and used as antigen in a cELISA. The assay was optimized with defined negative and positive bovine sera, where positive sera inhibited the binding of the epitope-specific monoclonal antibody BABB75A4. The cELISA accurately differentiated animals with B. bovis -specific antibodies from uninfected animals and from animals with antibodies against other tick-borne hemoparasites (98.7% specificity). In addition, B. bovis -specific sera from Australia, Argentina, Bolivia, Puerto Rico, and Morocco inhibited the binding of BABB75A4, confirming conservation of the epitope. The assay first detected experimentally infected animals between 13 and 17 days postinfection, and with sera from naturally infected carrier cattle, was comparable to indirect immunofluorescence (98.3% concordance). The assay appears to have the characteristics necessary for an epidemiologic and disease surveillance tool.

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Title: Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies Babesia bovis-Infected Cattle
Creators: Will L Goff - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Terry F McElwain - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Carlos E Suarez - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Wendell C Johnson - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Wendy C Brown - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Junzo Norimine - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Donald P Knowles - Animal Disease Research Unit, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99164-6630
Publication Details: Clinical and diagnostic laboratory immunology, Vol.10(1), pp.38-43
Academic Unit: Veterinary Microbiology and Pathology, Department of; Paul G. Allen School for Global Animal Health
Publisher: American Society for Microbiology
Identifiers: 99900546967701842
Language: English
Resource Type: Journal article