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王艳芳, 梁卫华, 郝卫, 潘玉先, 车小燕. 曲霉抗原双抗体夹心ELISA法特异性检测[J]. 中国公共卫生, 2008, 24(4): 476-478. DOI: 10.11847/zgggws2008-24-04-54
引用本文: 王艳芳, 梁卫华, 郝卫, 潘玉先, 车小燕. 曲霉抗原双抗体夹心ELISA法特异性检测[J]. 中国公共卫生, 2008, 24(4): 476-478. DOI: 10.11847/zgggws2008-24-04-54
WANG Yan-fang, LIANG Wei-hua, HAO Wei, . Study of double monoclonal antibody sandwich ELISA in detection of Aspergillus specific antigen[J]. Chinese Journal of Public Health, 2008, 24(4): 476-478. DOI: 10.11847/zgggws2008-24-04-54
Citation: WANG Yan-fang, LIANG Wei-hua, HAO Wei, . Study of double monoclonal antibody sandwich ELISA in detection of Aspergillus specific antigen[J]. Chinese Journal of Public Health, 2008, 24(4): 476-478. DOI: 10.11847/zgggws2008-24-04-54

曲霉抗原双抗体夹心ELISA法特异性检测

Study of double monoclonal antibody sandwich ELISA in detection of Aspergillus specific antigen

  • 摘要: 目的 用2组曲霉单克隆抗体(mAbs)建立特异性识别不同种类曲霉抗原的检测方法。方法 采用天然烟曲霉抗原免疫,获得广谱针对曲霉抗原的单克隆抗体;采用重组烟曲霉抗原获得特异性针对烟曲霉抗原的单克隆抗体,用间接免疫荧光鉴定,并分别建立2种双抗体夹心ELISA法,对19种常见的环境和临床分离曲霉株、马尔尼菲氏青霉菌及念珠菌培养液进行检测。结果 间接免疫荧光显示,用天然烟曲霉抗原免疫获得的单克隆抗体(mAbs-1)可广谱识别多种曲霉分离株,而重组烟曲霉抗原获得的单克降抗体(mAbs-2)仅能特异性结合临床和环境分离的烟曲霉抗原。用mAbs-1建立的双抗体夹心ELISA法可检测19种常见曲霉株培养液;用特异性针对烟曲霉抗原单克降抗体(mAbs-2)建立的双抗体夹心ELISA法可特异性检测临床和环境分离株烟曲霉培养液;与其他曲霉株无交叉反应;2种双抗体夹心ELISA法与马尔尼菲氏青霉菌及念珠菌培养液均无交叉反应。结论 2种曲霉单克降抗体双抗体夹心ELISA法,除可广谱检测环境和临床分离曲霉株,还可以区分烟曲霉与其他曲霉,可作为监测环境、农产品、食品中的曲霉污染和早期诊断曲霉病的新技术手段。

     

    Abstract: Objective To establish immunological methods specific for detecting antigens in different groups of monoclonal antibodies.Methods Indirect immnofluorescence assay was applied to identify specificity of the two groups of monoclonal antibodies prepared with crude antigen and recombinant antigen of Aspergillus fumigatus,respectively.Two different double monoclonal antibody sandwich EL ISA assays established with the two groups of antibodies were performed to detect antigents in the cell culture supermatants of 19 common species of Aspergillus,Penicillium marneffei,and 5 species of Candidas.Results The results of indirect immnofluorescence assay indicated that the monoclonal antibodies prepared with crude antigen of Aspergillus fumigatus were specific for antigens in both clinical isolates and environmental isolates of Aspergillus,whereas the other group of monoclonal antibodies was proved to be specific for Aspergillus fumigatus of both clinical and environmental isolates.The ELISA assay established with the crude antigen-specfic monoclonal antibodies could detect both of the clinical and environmental isolates of Aspergllius,while the other assay could only detect Aspergillus fumigatus of both clinical and environmental isolates.And no cross reaction with the cell culture of Penialllium marneffei and Candidas was observed with the two methods.Conclusion The EL ISA assays can detect both of the clinical and environmental isolates of Aspergillus,and differentiate Aspergillus fumigatus from other species of Aspergillus.The study provides us with new tools for monitoring the contaminmetion of Aspergillus in the environment,agricultural commodities and food,and this is also useful for early diagnosis of patients with aspergillosis.

     

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