Evaluation of Enzyme-Linked Immunosorbent Assays for Detection of Mycoplasma bovis-Specific Antibody in Bison Sera

Abstract

Mycoplasma bovis has recently emerged as a significant and costly infectious disease problem in bison. A method for the detection of M. bovis-specific serum antibodies is needed in order to establish prevalence and transmission patterns. Enzyme-linked immunosorbent assays (ELISAs) validated for the detection of M. bovis-specific serum IgG in cattle are commercially available, but their suitability for bison sera has not been determined. A collection of bison sera, most from animals with a known history of infection or vaccination with M. bovis, was tested for M. bovis-specific IgG using commercially available kits as well as an inhouse ELISA in which either cattle or bison M. bovis isolates were used as a source of antigen. Comparison of the results demonstrates that ELISAs optimized for cattle sera may not be optimal for the identification of bison seropositive for M. bovis, particularly those with low to moderate antibody levels. The reagent used for the detection of bison IgG and the source of the antigen affect the sensitivity of the assay. Optimal performance was obtained when the capture antigen was derived from bison isolates rather than cattle isolates and when a protein G conjugate rather than an anti-bovine IgG conjugate was used for the detection of bison IgG.

Association of MicroRNAs with Antibody Response to Mycoplasma bovis in Beef Cattle

“…Cattle sera were tested for antibodies reactive with M . bovis using a direct ELISA, as previously reported [ 16 ], with the following modifications: 0.5 ug of antigen was used per well. Anti-bovine IgG-peroxidase conjugate (KPL, Inc.), was diluted 1:3000 in wash buffer to detect cattle IgG and color development was halted after 45 min.…”

A serological and molecular study on the occurrence of mycoplasmas in European bison (Bison bonasus) from two areas of Eastern Poland

“…In Podlaskie province of Poland where BF is situated, M. bovis seroprevalence in cattle was assessed at 77% . Although the usefulness of the Bio-X M. bovis ELISA in bison testing was demonstrated earlier (Register et al 2013a) the possibility of false positive results in this study cannot be excluded. Nevertheless it has been previously demonstrated that M. bovis does infect bison and can cause serious disease problems (Dyer et al 2008, Janardhan et al 2010, Register et al 2013b.…”

“…Unfortunately, these results were not confirmed by other methods. The Bio-X M. bovis ELISA adopted in this study is intended for use in cattle; however, Register et al (2013a) demonstrated its application for use in bison when they compared it to their in-house ELISA designed for testing bison with agreement at more than 96%. That difference related to two samples which came from bison with an unknown history of exposure to M. bovis.…”

A review of mycoplasma diagnostics in cattle

“…Because of M. bovis currently being considered the most common and important species, much of the assay development and research available is specific to this species. This includes the commercially available indirect ELISA kits for anti‐ M. bovis antibody detection by Bio‐X Diagnostics (Rochefort, Belgium) for use on serum and milk samples, which has been used in several field and assay validation studies . The Bovichek M. bovis antibody ELISA test kit by Biovet Inc (Quebec, Canada) is also commercially available for use on serum samples and has been used in studies evaluating in‐house assays …”

“…69 test kit by Biovet Inc (Quebec, Canada) is also commercially available for use on serum samples and has been used in studies evaluating inhouse assays. 95,96 When developing an ELISA assay, the potential for cross-reactivity of an M. bovis specific ELISA with other bacterial species found in cattle and ruminants is difficult to assess but is important from a diagnostic perspective, particularly if these species are not of clinical importance.…”