Use of blood matrices and alternative biological fluids for antibody detection in animal tuberculosis

Authors: LYASCHCENKO, KONSTANTIN - Chembio Diagnostic Systems, Inc; SIKAR-GANG, ALINA - Chembio Diagnostic Systems, Inc; SRIDHARA, ARCHANA - Chembio Diagnostic Systems, Inc; JOHNATHAN-LEE, ASHLEY - Chembio Diagnostic Systems, Inc; ELAHI, RUBYAT - Chembio Diagnostic Systems, Inc; GREENWALD, RENA - Chembio Diagnostic Systems, Inc; LAMBOTTE, PAUL - Chembio Diagnostic Systems, Inc; ESFANDIARI, JAVAN - Chembio Diagnostic Systems, Inc; ROOS, EDUARD - Stellenbosch University; KERRY, TANYA - Stellenbosch University; MILLER, MICHELE - Stellenbosch University; Thacker, Tyler; Palmer, Mitchell; Waters, Wade

Submitted to: Veterinary Immunology and Immunopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/19/2021
Publication Date: 7/22/2021
Citation: Lyaschcenko, K., Sikar-Gang, A., Sridhara, A., Johnathan-Lee, A., Elahi, R., Greenwald, R., Lambotte, P., Esfandiari, J., Roos, E., Kerry, T., Miller, M., Thacker, T.C., Palmer, M.V., Waters, W.R. 2021. Use of blood matrices and alternative biological fluids for antibody detection in animal tuberculosis. Veterinary Immunology and Immunopathology. 239. Article 110303. https://doi.org/10.1016/j.vetimm.2021.110303.
DOI: https://doi.org/10.1016/j.vetimm.2021.110303

Interpretive Summary: The USDA has been engaged in a bovine tuberculosis eradication effort since 1917. Although great progress has been made, there is a critical need for improved diagnostic tests that are quicker than current skin testing methods. Detection of antibodies to M. bovis in blood is attractive as blood is easily obtained and most labs are equipped with instrumentation for antibody detection. We analyzed various bodily fluids, plasma, serum, saliva, broncho-alveolar lavage, urine, diaphragm extract and bile from experimentally infected cattle for antibodies to M. bovis. A high correlation was observed between all blood-derived specimens. Broncho-alveolar lavage and diaphragm fluid yielded positive results in all infected animals. Antibody titers in other fluids were relatively low. These findings support the use of some non-traditional biological fluids in serological assays for bovine tuberculosis.

Technical Abstract: Bovine tuberculosis (bTB) control programs can be improved by implementation of advanced ante-mortem testing algorithms. Serodiagnostic methods using traditional blood or blood-derived specimens may benefit from the use of less invasive alternative biological fluids, provided those mirror systemic antibody responses. In the present study, we used Dual Path Platform (DPP) and Multiantigen Print Immunoassay (MAPIA) to compare antibody levels in ten sample types including whole blood (fresh and hemolyzed), plasma (fresh and leftover from Bovigam testing), serum, saliva, broncho-alveolar lavage, urine, diaphragm extract, and bile collected from cattle aerosol-infected with Mycobacterium bovis. High correlation (r =0.97-0.99) in measurments of IgG antibodies to MPB70/MPB83 fusion antigen by DPP assay was found between all blood-derived specimens supporting matrix equivalency. Broncho-alveolar lavage and diaphragm extract yielded positive results in all the infected animals tested, showing high correlation with matching serum data (r =0.94 and r =0.95, respectively) and suggesting their potential use in antibody assays. Characterized by MAPIA, the antigen reactivity patterns obtained with paired sera and alternative specimens were nearly identical, with slight differences in intensity. Antibodies were also found by DPP assay in saliva, urine, and bile from some of the infected animals, but the titers were relatively low, thus reducing the diagnostic value of such specimens. The proposed approach was evaluated in a pilot field study on warthogs diagnosed with M. bovis infection. Relative levels of antibody in tissue fluid obtained from lymph nodes or lungs were consistent with those detected in sera and detectable in all infected warthogs. The findings support the diagnostic utility of non-traditional biological fluids and tissue samples when used as alternative test specimens in serologic assays for bTB.