title: [Development of an interferon-gamma ELISPOT for bovine tuberculosis].
abstract: We established an ELISPOT for bovine interferon-gamma (BoIFN-γ), and applied it in the diagnosis of bovine tuberculosis (bTB). Monoclonal antibodies that can bind with native BoIFN-γ were screened as the coating antibody and detecting antibody. After optimization of detecting conditions including coating antibody concentration, cell number, and detecting antibody concentration, the ELISPOT assay was established. Peripheral mononuclear cells (PBMCs) isolated from 30 cows were co-cultured with PPD, and detected with the ELISPOT assay. The optimal conditions of ELISPOT assay were 2.5 μg/mL coating antibody 2G5, 2.5 x 10(5) cells/well, and 1 μg/mL detecting antibody Bio-5E11. In these 30 cows tested both with the ELISPOT assay and the BOVIGAM kit, 11 cows were proved to be positive in ELISOPT assay with the sensitivity of 78.6%, and 12 cows were proved to be negative in ELISOPT assay with the specificity of 75%. The ELISPOT assay for BoIFN-γ could be used to detect bTB efficiently and it might be an alternative method for the diagnosis of bTB.
authors.forename: Zhengzhong
authors.initials: Z
authors.lastname: Xu
authors.forename: Fa
authors.initials: F
authors.lastname: Shan
authors.forename: Fengli
authors.initials: F
authors.lastname: Shan
authors.forename: Chuang
authors.initials: C
authors.lastname: Meng
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Xie
authors.forename: Jiaying
authors.initials: J
authors.lastname: Liu
authors.forename: Jingjing
authors.initials: J
authors.lastname: Min
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.forename: Xin'an
authors.initials: X
authors.lastname: Jiao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D058501:Enzyme-Linked Immunospot Assay / Q000662:veterinary*; D005260:Female; D007371:Interferon-gamma / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014380:Tuberculosis, Bovine / Q000175:diagnosis*
chemical_list: D000911:Antibodies, Monoclonal; D007371:Interferon-gamma