【作者】 杨芳芳； 方毅敏； 董涛； 涂志全； 赖小敏；

【Author】 Yang Fangfang~1,Fang Yimin~2,Dong Tao~1,Tu Zhiquan~1,Lai Xiaomin~1 (1.Department of Microbiology,Ministry of Education Key Laboratory of Tropical Diseases Control, Gangdong Provincial Department of Education Key Laboratory of Functional Molecules from Marine Microorganisms,Gangdong Provincial Research Center for Severe Infectious Disease Prevention and Control Technology,Zhongshan School of Medicine,Sun Yat-sen University,Guangzhou 510080,China.2. Guangzhou Chest Hospital,Guangzhou 510095,China)

【机构】 中山大学中山医学院微生物学教研室热带病防治研究教育部重点实验室海洋微生物功能分子广东省高校重点实验室广东省重大传染病预防和控制技术研究中心； 广州市胸科医院；

【摘要】 目的建立结核分枝杆菌特异性(Mycobacterium tuberculosis,MTB)多肽酶联免疫斑点试验(Enzyme linked immunospot assay,ELISPOT)检测MTB感染情况的方法,评价其临床诊断价值。方法通过比较不同的多肽组确定ELISPOT检测MTB感染方法,建立IFN-γ-ELISPOT方法。以PPD皮肤试验作对照。同时检测部分健康人群做对照。结果用ELISPOT方法检测结核患者标本,E6、E7、C14、E6+E7、E6+E7+C14五个多肽实验组检测阳性率依次为52.32%、76.89%、73.89%、91.23%、91.30%。五个组患者对应的PPD试验检测阳性率都高于95%。用E6+E7+C14混合多肽ELISPOT检测两个社区的健康人群其检测阳性率分别为9%和3%,而PPD试验的阳性检出率为45%和60%。结论应用混合多肽建立的IFN-γ-ELISPOT方法可初步应用于诊断MTB感染、辅助结核病诊断。更多还原

【Abstract】 Objective:To establish IFN-γ-based enzyme-linked immunospot assay(ELISPOT) methods by using peptides from ESAT-6/CFP-10 and evaluate the application value for detection of Mycobacterium tuberculosis(MTB) infection and diagnosis of tuberculosis(TB).Methods:By comparing different peptides groups and detecting the samples from TB patients and healthy control donors to evaluate the application value of IFN-γ-ELISPOT for detection of MTB infection and diagnosis of TB.PPD test was used as control method. Results:Several IFN-γ-ELISPOT methods,by using five peptide groups inculding E6,E7,C14,E6 + E7 and E6 + E7 + C14,were used to detect the samples from TB patients,and the positive detection rates were 52.32%,76.89%,73.89%,91.23%,91.30%,respectively.While the positive rates of PPD test were all higher than 95%.Moreover,E6+E7+C14-ELISPOT was used to test the samples from healthy donors of two communities,and the positive rates were 9%and 3%,respectively,in contrast,the positive rates for PPD test were 45%and 60%.Conclusion:The peptides-IFN-γ-ELISPOT we established could detect MTB infection efficiently,and could be used in the assistant diagnosis of TB.更多还原