[Establishment of human IFN-gamma in vitro release assay and its application in tuberculosis diagnosis].
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.
Chen
Yingyu
Y
The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
Deng
Quantao
Q
Zhan
Zhihua
Z
Guo
Aizhen
A
Xiang
Jie
J
Chen
Jun
J
Zhou
Jinhai
J
Zeng
Qinzhi
Q
Wei
Wu
W
Tong
Qingwei
Q
Chao
Yanjie
Y
Kuang
Youji
Y
Chen
Huanchun
H
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
D000911:Antibodies, Monoclonal; D007371:Interferon-gamma