The State Key Lab of Agrobio-technology, National TES Lab, Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, 2 Yuanmingyuan West road, Beijing 100193, China.
Vet Rec. 2013 Aug 3;173(5):117. doi: 10.1136/vr.101552. Epub 2013 Jul 6.
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.
本研究采用实时 PCR 方法检测牛结核分枝杆菌（bTB）的感染状态，比较牛结核菌素或禽结核菌素刺激与未刺激（磷酸盐缓冲盐水，PBS）血培养的血液中 IFN-γ mRNA 水平。共检测了 137 头牛以验证该检测方法，其中 54 头 IFN-γ 实时定量 PCR（RT-qPCR）阳性，其余 83 头为阴性。同时，使用 Bovigam IFN-γ 检测 ELISA 试剂盒进行 IFN-γ ELISA 检测，并将这些结果作为标准。比较了单点皮内结核菌素试验（SIDT）和 IFN-γ RT-qPCR 检测结果，结果表明 RT-qPCR 与 ELISA 相关性更好，其准确性高于 SIDT。这表明 RT-qPCR 是一种有用的牛结核病诊断方法。然而，我们的方法仍存在一些局限性，例如缺乏结核病病变或尸检结果作为金标准。未来应进一步改进，以提高牛结核病诊断的准确性。
