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Development and evaluation of an interferon gamma assay for the diagnosis of tuberculosis in red deer experimentally infected with Mycobacterium bovis

Investigation published in BMC Veterinary Research

November 1st, 2017

BACKGROUND:
Red deer (Cervus elaphus) is regarded as an epidemiologically relevant host for Mycobacterium bovis (M. bovis) and closely related members of the Mycobacterium tuberculosis complex that cause animal tuberculosis (TB). The standard antemortem screening test for the detection of TB in deer is the intradermal tuberculin skin test, but the detection of interferon-gamma (IFNγ) produced by white blood cells exposed to M. bovis antigens can be used as an alternative or supplemental assay in most TB eradication/control programs. This study aims to develop an in-house sandwich ELISA for deer IFNγ, based on the cross-reactivity of the antibodies to both cervid and bovine IFNγ, and to evaluate the potential of this assay to detect M. bovis-infected red deer in response to the in vitro stimulation of whole-blood cells with bovine purified protein derivative (bPPD), p22 protein complex derived from bPPD or using the specific tuberculous mycobacterial proteins ESAT-6/CFP-10, Rv3615c and Rv3020c. The positive control stimulant used in this study was pokeweed mitogen, which resulted in a consistent induction of IFNγ in samples from red deer, thus allowing the interpretation of the assay.
RESULTS:
The percentage of animals correctly classified by this technique as M. bovis non-infected was 100%. The detection of infected animals as positive was high and ranged widely depending upon the antigen and the cut-off value applied, as well as the time after infection. Our findings indicate that this protocol may serve as a reliable assay for the antemortem diagnosis of TB from the initial stage of M. bovis-infection, and may also be adequately sensitive.
CONCLUSIONS:
The suggested optimal antigens and cut-off are bPPD, p22 and the combination of ESAT-6/CFP-10 and Rv3020c with a 0.05 Δ optical density, which yielded a up to 100% correct classification of TB positive and negatve red deer under our experimental conditions. This technique will aid in TB testing of farmed and translocated deer. Future studies should evaluate the ability of this IFNγ assay to detect specific responses under field conditions




Risalde MA., Thomas J., Sevilla I., Serrano M., Ortiz JA., Garrido J., Dominguez M., Dominguez L., Gortazar C. and Ruiz-Fons JF.




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Development and evaluation of an interferon gamma assay for the diagnosis of tuberculosis in red deer experimentally infected with Mycobacterium bovis

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Development and evaluation of an interferon gamma assay for the diagnosis of tuberculosis in red deer experimentally infected with Mycobacterium bovis


Participants:

Gobierno de Castilla-La ManchaSanidad y Biotecnología (SaBio). Instituto de Investigación en Recursos Cinegéticos (IREC). Consejo Superior de Investigaciones Científicas (CSIC). Universidad de Castilla La Mancha (UCLM). Gobierno de Castilla-La Mancha (JCCM).

Instituto Maimonides de Investigación Biomédica de CordobaInstituto Maimonides de Investigación Biomédica de Cordoba.

Indian Council of Agricultural ResearchIndian Council of Agricultural Research (ICAR).

Gobierno VascoNeiker-Tecnalia, Instituto Vasco de Investigación y Desarrollo Agrario. Gobierno Vasco.

Medianilla Red Deer GeneticsMedianilla Red Deer Genetics.

Instituto de Salud Carlos IIIServicio de Inmunología. Centro Nacional de Microbiología (CNM). Instituto de Salud Carlos III (ISCIII).

Universidad ComplutenseServicio de Micobacterias (MYC). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).







BMC Veterinary Research
FACTOR YEAR Q
1.958 2017

NLMID: 101249759

PMID: 29145844

ISSN: 1746-6148



TITLE: Development and evaluation of an interferon gamma assay for the diagnosis of tuberculosis in red deer experimentally infected with Mycobacterium bovis


JOURNAL: BMC Vet. Res


NUMERACIÓN: 16;13(1):341


AÑO: 2017


PUBLISHER: BioMed Central


AUTHORS: Risalde MA., Thomas J., Sevilla I., Serrano M., Ortiz JA., Garrido J., Dominguez M., Dominguez L., Gortazar C. and Ruiz-Fons JF.


VISAVET PARTICIPANTS


Lucas Domínguez Rodríguez

DOI: https://doi.org/ 10.1186/s12917-017-1262-6


CITE THIS PUBLICATION:

Risalde MA., Thomas J., Sevilla I., Serrano M., Ortiz JA., Garrido J., Dominguez M., Dominguez L., Gortazar C. and Ruiz-Fons JF. Development and evaluation of an interferon gamma assay for the diagnosis of tuberculosis in red deer experimentally infected with Mycobacterium bovis. BMC Veterinary Research. 16;13(1):341. 2017. (A). ISSN: 1746-6148. DOI: 10.1186/s12917-017-1262-6


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