目的探讨酶联免疫斑点试验（ELISPOT）检测结核分枝杆菌特异抗原T淋巴细胞的频率，在快速诊断结核病中的临床应用价值。方法利用ELISPOT检测对结核菌特异抗原刺激反应，分泌γ干扰素的效应T淋巴细胞数量方法，测定30名健康体检者，24例丙肝患者，65例非结核呼吸道疾病患者和112例结核病患者外周血单个核细胞中结核菌抗原特异的T淋巴细胞的频率。结果112例结核病患者中，107例结核病患者结核抗原特异ELISPOT阳性，提示方法敏感度为95．5％，30名健康对照中，2名健康体检者阳性，其他健康体检者阴性，24例丙肝患者和65例非结核呼吸道疾病患者均为阴性，方法的特异度为93．3％。结论ELISPOT是较灵敏和特异的快速检测结核菌感染的方法，可用于结核病的快速诊断。

Clinical application of enzyme-linked immunospot assay for rapid diagnosis of active tuberculosis

YUE Jun , LIANG Li, LI Su-hui, ZHANG Yue-fian, XIAO He-ping.

Department of Clinical Laboratory, Shanghai Pulmonary Hospital, Shanghai 200433, China

Abstract:

Objective To evaluate the clinical application of enzyme-linked immunospot （ELISPOT） assay for detection of M. tuberculosis antigen specific T cells in the rapid diagnosis of active tuberculosis. Methods Using the rapid enzyme-linked immunospot assay for detection of T cells with secretion of IFN-γ specific for M. tuberculosis antigens in blood samples from 112 patients with tuberculosis, 24 patients with hepatitis C, 30 healthy persons, and 65 with other respiratory diseases. Results 107 of 112 tuberculosis patients had detectable M. tuberculosis antigen-specific T cells, whereas 2 of 30 healthy subjects and 65 patients with non-tuberculosis illnesses responded. This assay has a sensitivity of 95. 5%, specificity of 93.3%. Conclusions M. tuberculosis specific T cells could serve as accurate markers of M. tuberculosis infection in an area of high tuberculosis prevalence. ELISPOT is a rapid, sensitive and specific method for detecting M. tuberculosis specific T cells. It also gives objective evidence to the diagnosis of active tuberculosis.

Key words:

Enzyme-linked immunosorbent assay; Mycobacterium tuberculosis; Diagnosis