Establishment of human IFN-gamma in vitro release assay and its application in tuberculosis diagnosis.
- Author:
Yingyu CHEN
1
;
Quantao DENG
;
Zhihua ZHAN
;
Aizhen GUO
;
Jie XIANG
;
Jun CHEN
;
Jinhai ZHOU
;
Qinzhi ZENG
;
Wu WEI
;
Qingwei TONG
;
Yanjie CHAO
;
Youji KUANG
;
Huanchun CHEN
Author Information
1. The State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
Enzyme-Linked Immunosorbent Assay;
Female;
Humans;
Interferon-gamma;
immunology;
secretion;
Mice;
Mice, Inbred BALB C;
Mycobacterium tuberculosis;
immunology;
Rabbits;
T-Lymphocytes;
immunology;
Tuberculosis;
diagnosis;
immunology
- From:
Chinese Journal of Biotechnology
2008;24(9):1653-1657
- CountryChina
- Language:Chinese
-
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.
