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Development of a test for bovine tuberculosis in cattle based on measurement of gamma interferon mRNA by real-time PCR
  1. W. Gan, MSc1,
  2. X. Zhou, PhD1,
  3. H. Yang, PhD2,
  4. H. Chen, MSc3,
  5. J. Qiao, MSc4,
  6. S. H. Khan, MSc1,
  7. L. Yang, PhD1,
  8. X. Yin, PhD1 and
  9. D. Zhao, PhD1
  1. 1The State Key Lab of Agrobio-technology, Key Lab of Animal Epidemiology and Zoonosis, National TES Lab, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 2 Yuanmingyuan West road, Beijing 100193, China
  2. 2Center for Cow Research, Shandong Academy of Agriculture, 159 Gongye North road, Shandong 250183, China
  3. 3Beijing general station of animal husbandry and veterinary service, Beijing Municipal Bureau of Agriculture, A15 Beiyuan road, Beijing 100107, China
  4. 4College of Veterinary Medicine, China Agricultural University, 2 Yuanmingyuan West road, Beijing 100193, China
  1. E-mail for correspondence: zhaodm{at}cau.edu.cn

Abstract

The infection status of cattle for bovine tuberculosis (bTB) was determined by real-time PCR, comparing the levels of IFN-γ mRNA in blood cultures stimulated with either bovine or avian tuberculin with non-stimulated control (phosphate buffer saline, PBS) blood culture. Totally, 137 cattle were tested to validate the assay, in which 54 were IFN-γ real-time quantitative PCR (RT-qPCR) positive, while the remaining 83 were found negative. Meanwhile, the IFN-γ ELISA test was carried out using the Bovigam IFN-γ detection ELISA kit and these results were used as a standard. The results of the single intradermal tuberculin tests (SIDT) and IFN-γ RT-qPCR tests were compared and revealed that the RT-qPCR correlated better with the ELISA and its accuracy was higher than SIDT. This indicates the RT-qPCR is a useful diagnostic method for bTB in cattle. However, several limitations remain for our approach, such as lack of a TB lesions or postmortem test results as a gold standard. Further improvements should be made in the future to increase accuracy of diagnosis of bTB in cattle.

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