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Establishment of Human IFN-g in vitro Release Assay and Its Application in Tuberculosis Diagnosis
人IFN-g体外释放检测法的建立及其在结核病诊断中的应用

Keywords: tuberculosis,IFN-g,diagnosis,ELISA,ESAT-6/CFP-10,monoclonal antibody
结核病
,IFN-g,诊断,ELISA,ESAT-6/CFP-10,单克隆抗体

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Abstract:

This study aimed to establish human IFN-g (hIFN-g) in vitro release assay and to apply it in diagnosis of human tuberculosis. Human IFN-g gene was cloned and expressed in Escerichia coli. The recombinant hIFN-g 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-g 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.

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