Journal article
Validation of a Commercially Available Monoclonal Antibody-Based Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to Neospora caninum in Cattle
Journal of clinical microbiology, Vol.39(11), pp.3851-3857
11/2001
Handle:
https://hdl.handle.net/2376/113635
PMCID: PMC88454
PMID: 11682497
Abstract
A previously described monoclonal antibody (MAb)-based competitive-inhibition enzyme-linked immunosorbent assay (cELISA) was modified to optimize performance, and the assay was validated in various defined cattle populations for detection of serum antibody to
Neospora caninum
, a major cause of bovine abortion. Modifications to the cELISA included capturing native
N. caninum
antigen with a parasite-specific MAb (MAb 5B6-25) and directly conjugating the competitor MAb (MAb 4A4-2), with both MAbs binding different epitopes of a conserved, immunodominant 65-kDa tachyzoite surface antigen. The assay was validated using three serum sets, a “gold standard” set of 184 cow sera defined by fetal histopathology and
N. caninum
immunohistochemistry and by maternal
N. caninum
indirect fluorescence assay (IFA) at a 1:200 serum dilution, a relative standard set of 330 cow sera defined by IFA alone, and a set of 4,323 cow sera of unknown
N. caninum
status. A test cutoff of 30% inhibition was identified. The diagnostic sensitivity was 97.6%, and diagnostic specificity was 98.6% for the gold standard abortion-defined sera. The diagnostic sensitivity was 96.4%, and diagnostic specificity was 96.8% for the relative standard IFA-defined sera. Testing of the 4,323 bovine sera of unknown
N. caninum
status revealed a distinct bimodal distribution and steep sigmoid frequency curve with only 1.8% of samples within 5% of the test cutoff, indicating a sharp discrimination between test-positive and test-negative samples. In summary, the modified
N. caninum
cELISA provided a simple, rapid, and versatile method to accurately identify
N. caninum
infection status in cattle using a single cutoff value.
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Details
- Title
- Validation of a Commercially Available Monoclonal Antibody-Based Competitive-Inhibition Enzyme-Linked Immunosorbent Assay for Detection of Serum Antibodies to Neospora caninum in Cattle
- Creators
- Timothy V Baszler - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040Scott Adams - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040John Vander-Schalie - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040Bruce A Mathison - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040Miladin Kostovic - Department of Veterinary Microbiology and Pathology, Washington State University, Pullman, Washington 99164-7040
- Publication Details
- Journal of clinical microbiology, Vol.39(11), pp.3851-3857
- Academic Unit
- Paul G. Allen School for Global Animal Health
- Publisher
- American Society for Microbiology
- Identifiers
- 99900548191101842
- Language
- English
- Resource Type
- Journal article