Development of Elispot assay for diagnosis of pulmonary tuberculosis

Abstract

Abstract: Objective To develop Elispot assays for detection of Mycobacterium tuberculosis antigen-specific IFN-γin peripheral blood and evaluate its applicability in diagnosis of pulmonary tuberculosis. MethodsRecombinant protein library containing the early secretary antigen target 6 kDa protein (ESAT-6) and peptide library were used as antigens to develop an Elispot assay for measuring antigen-specific IFN-γ. IFN-γresponses in peripheral bloods were determined in 32 patients with pulmonary tuberculosis(TB), 205 healthy controls (HC) and 18 patients with lung diseases other than tuberculosis (disease control, DC). For comparison, 32 patients with TB and 20 healthy controls were also tested with the whole-blood IFN-γ assay（Quantiferon-TB-Gold，QFT-G）in parallel. ResultsThe proportion of positive IFN-γresponse using our in-house Elispot in TB, HC, and DC is 75.0%, 7.3% and 11.1%, respectively. The parallel study using QFT-G showed that 78.1% of TB was positive. There is no significant difference between the Results got by our in-house Elispot and QFT-G（χ²=1.6，P>0.05）. ConclusionsAn Elispot assay for detection of MTB antigen-specific IFN-γ was developed with higher sensitivity and specificity. The Elispot assay can be applied to diagnosis of TB infection.

Key words: pulmonary tuberculosis/diagnosis, Mycobacterium tuberculosis, interferon type Ⅱ, immanoenzyme techiques

Yang Qianting¹,Xu Ping²,Zeng Jianfeng¹，Zhu Xiuyun¹, Huang Hua¹, Deng Qunyi¹,Liu Guohui,Zhou Boping,Chen Xinchun. Development of Elispot assay for diagnosis of pulmonary tuberculosis[J]. Chinese Journal of Antituberculosis, 2009, 31(3): 144-148.

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