J Vet Sci.  2014 Jun;15(2):259-265. 10.4142/jvs.2014.15.2.259.

Extent of Mycobacterium bovis infection in dairy cattle herds subject to partial culling as determined by an interferon-gamma assay

• Je S
• Yeo UC
• Song T
• Kim KC
• Park SY
• Kim MJ
• Cho SN

• KMID: 1784647
• DOI: http://doi.org/10.4142/jvs.2014.15.2.259

Abstract

The interferon-gamma (IFN-gamma) assay is employed as a complementary diagnostic test for bovine tuberculosis (BTB) in many countries. To simplify this assay, we established a 96-well plate format using the ESAT-6 and CFP-10 antigens and then employed it to determine the extent of Mycobacterium (M.) bovis infection in dairy herds with a history of BTB outbreaks in a country where only selective culling is practiced. The sensitivity and specificity of this IFN-gamma assay were 85.9% and 100%, respectively, based on comparison with the conventional single intradermal tuberculin test (SIDT). The IFN-gamma assay was also positive in 30.4% and 36.8% of SIDT-negative animals from herds with recent and remote BTB outbreaks, respectively. Of 14 SIDT-negative, IFN-gamma positive cattle, five (35.7%) were culture positive and an additional six were positive based on a polymerase chain reaction-based test for M. bovis. Therefore, the IFN-gamma assay has the potential to serve as a specific and sensitive test for M. bovis infection in dairy cattle. Further, the results indicated that a substantial portion of SIDT-negative animals in herds with previous BTB outbreaks were actually infected with M. bovis. Accordingly, the present selective-culling strategy may require modifications to include this more sensitive assay.

Keyword

bovine tuberculosis; IFN-gamma assay; Mycobacterium bovis

MeSH Terms

Animals
Antigens, Bacterial/*diagnostic use
Bacterial Proteins/diagnostic use
Cattle
Female
Interferon-gamma Release Tests/*veterinary
Mycobacterium bovis/*isolation & purification
Polymerase Chain Reaction/veterinary
Republic of Korea/epidemiology
Tuberculosis, Bovine/*diagnosis/*epidemiology/microbiology
Antigens, Bacterial
Bacterial Proteins

Figure

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  Fig. 1 Standard curve for IFN-γ enzyme-linked immunosorbent assay showing measurement of recombinant bovine IFN-γ. Linearity was determined at IFN-γ concentrations ranging from 0.078 to 10 ng/mL.

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  Fig. 2 IFN-γ secretion of mycobacterial antigens in SIDT-positive and -negative cattle. ***p < 0.0001.

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  Fig. 3 ROC curve calculated for IFN-γ assay using the ESAT6 and CFP10 antigen cocktail. IFN-γ assay AUC = 0.958, 95% confidence interval (CI) = 0.930 to 0.985.

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  Fig. 4 PCR analysis and visible lesions on hilar lymph nodes of SIDT-negative cattle. (A) Electrophoresis on 1.5% agarose gel showing the 113 bp PCR products after amplification with the IS1081 F/R primers. Lane M, 100 bp DNA size marker; Lane 1, M. bovis DNA; Lanes 2 ~ 7, samples of hilar lymph nodes. (B) Visible lesions of hilar lymph nodes from cattle showing positive response to IFN-γ assay, but negative response to SIDT.

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