1.Establishment of human IFN-gamma in vitro release assay and its application in tuberculosis diagnosis.
Yingyu CHEN ; Quantao DENG ; Zhihua ZHAN ; Aizhen GUO ; Jie XIANG ; Jun CHEN ; Jinhai ZHOU ; Qinzhi ZENG ; Wu WEI ; Qingwei TONG ; Yanjie CHAO ; Youji KUANG ; Huanchun CHEN
Chinese Journal of Biotechnology 2008;24(9):1653-1657
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.
Animals
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Antibodies, Monoclonal
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Enzyme-Linked Immunosorbent Assay
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Female
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Humans
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Interferon-gamma
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immunology
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secretion
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Mice
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Mice, Inbred BALB C
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Mycobacterium tuberculosis
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immunology
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Rabbits
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T-Lymphocytes
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immunology
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Tuberculosis
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diagnosis
;
immunology