title: Development of a test for bovine tuberculosis in cattle based on measurement of gamma interferon mRNA by real-time PCR.
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.
authors.affiliation: 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.
authors.forename: W
authors.initials: W
authors.lastname: Gan
authors.forename: X
authors.initials: X
authors.lastname: Zhou
authors.forename: H
authors.initials: H
authors.lastname: Yang
authors.forename: H
authors.initials: H
authors.lastname: Chen
authors.forename: J
authors.initials: J
authors.lastname: Qiao
authors.forename: S H
authors.initials: SH
authors.lastname: Khan
authors.forename: L
authors.initials: L
authors.lastname: Yang
authors.forename: X
authors.initials: X
authors.lastname: Yin
authors.forename: D
authors.initials: D
authors.lastname: Zhao
mesh_terms: D000818:Animals; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D007371:Interferon-gamma / Q000097:blood*; D007428:Intradermal Tests / Q000662:veterinary; D012333:RNA, Messenger / Q000097:blood*; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary*; D015203:Reproducibility of Results; D014374:Tuberculin Test / Q000662:veterinary; D014380:Tuberculosis, Bovine / Q000097:blood / Q000175:diagnosis*
chemical_list: D012333:RNA, Messenger; D007371:Interferon-gamma
keywords: Bovine tuberculosis; Cattle; Diagnostics; RT-PCR techniques