title: [Establishment of human IFN-gamma in vitro release assay and its application in tuberculosis diagnosis].
abstract: This study aimed to establish human IFN-gamma (hIFN-gamma) in vitro release assay and to apply it in diagnosis of human tuberculosis. Human IFN-gamma gene was cloned and expressed in Escherichia coli. The recombinant hIFN-gamma was purified and used as immunogen to immunize mice and rabbits respectively. Monoclonal and polyclonal antibodies were respectively developed and a sandwich ELISA was established. The heparized whole blood from 111 active tuberculosis patients and 292 clinical healthy controls were collected. The blood was stimulated with tuberculosis specific fused antigen ESAT-6/CFP-10 and the plasma was collected for IFN-gamma detection. The sensitivity for tuberculosis diagnosis was 95.5%, whereas the positive detection rate for the healthy controls was 16.7%. There was a significant difference between the patients and healthy controls (P<0.01) indicating that this assay had a high sensitivity and specificity, and thus could be promising in tuberculosis diagnosis.
authors.affiliation: The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
authors.forename: Yingyu
authors.initials: Y
authors.lastname: Chen
authors.forename: Quantao
authors.initials: Q
authors.lastname: Deng
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Zhan
authors.forename: Aizhen
authors.initials: A
authors.lastname: Guo
authors.forename: Jie
authors.initials: J
authors.lastname: Xiang
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
authors.forename: Jinhai
authors.initials: J
authors.lastname: Zhou
authors.forename: Qinzhi
authors.initials: Q
authors.lastname: Zeng
authors.forename: Wu
authors.initials: W
authors.lastname: Wei
authors.forename: Qingwei
authors.initials: Q
authors.lastname: Tong
authors.forename: Yanjie
authors.initials: Y
authors.lastname: Chao
authors.forename: Youji
authors.initials: Y
authors.lastname: Kuang
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D011817:Rabbits; D013601:T-Lymphocytes / Q000276:immunology*; D014376:Tuberculosis / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007371:Interferon-gamma