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| 双抗原夹心ELISA法检测马尔尼菲青霉Mp1p抗体 | |
| 引用本文： | 王艳芳,曾磊,陈学东,郝卫,杨梅,蔡建飘,王压娣,袁国勇,车小燕. 双抗原夹心ELISA法检测马尔尼菲青霉Mp1p抗体[J]. 南方医科大学学报, 2013, 33(3): 439-443 |
| 作者姓名： | 王艳芳 曾磊 陈学东 郝卫 杨梅 蔡建飘 王压娣 袁国勇 车小燕 |
| 作者单位： | 王艳芳 (南方医科大学珠江医院检验医学部,广东广州,510282)；曾磊 (南方医科大学珠江医院检验医学部,广东广州,510282)； 陈学东 (南方医科大学珠江医院检验医学部,广东广州,510282)； 郝卫 (南方医科大学珠江医院检验医学部,广东广州,510282)；杨梅 (南方医科大学珠江医院检验医学部,广东广州,510282)； 蔡建飘 (南方医科大学珠江医院检验医学部,广东广州,510282)； 王压娣 (南方医科大学珠江医院检验医学部,广东广州,510282)； 袁国勇 (香港大学微生物系,香港)； 车小燕 (南方医科大学珠江医院检验医学部,广东广州,510282)； |
| 基金项目： | 国家自然科学基金(项目编号：81101299)广东省教育部产学研结合项目(项目编号：2010B090400498)广州市科技计划项目(项目编号：2012Y2-00021) |
| 摘 要： | 目的建立一种检测马尔尼菲青霉特异性抗体的双抗原夹心ELISA方法。方法利用毕赤酵母系统表达重组马尔尼菲青 霉特异性甘露糖蛋白Mp1p，并利用改良过碘酸钠法标记Mp1p，经棋盘滴定法建立一种可检测马尔尼菲青霉Mp1p特异性抗体 的双抗原夹心ELISA法，并检测100例健康人群对照血清、21例血培养确诊其他真菌感染病人血清和15例血培养确诊马尔尼 菲青霉病人血清，联合本实验室前期建立的马尔尼菲青霉抗原检测方法评价其临床应用价值。结果成功建立一种检测马尔 尼菲青霉Mp1p特异性抗体的双抗原夹心ELISA法，经健康人群对照及其他真菌感染病人血清评价特异度为100%（121/121）， 检测15例马尔尼菲青霉病人血清，Mp1p特异性抗体2例阳性，Mp1p特异性抗原12例阳性，Mp1p抗体与抗原联合检测可明显 提高灵敏度，达到93.3%（14/15）。结论双抗原夹心ELISA法检测马尔尼菲青霉Mp1p特异性抗体具有高度的特异性，联合抗 原检测可提高马尔尼菲青霉感染诊断率。 |
| 关 键 词： | 马尔尼菲青霉 双抗原夹心ELISA法 真菌 Mp1p 抗体检测 |
A double-antigen sandwich ELISA for detecting Penicillium marneffei Mp1p-specific antibody |
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| WANG Yanfang,ZENG Lei,CHEN Xuedong,HAO Wei,YANG Mei,CAI Jianpiao,WANG Yadi,YUAN Guoyong,CHE Xiaoyan. A double-antigen sandwich ELISA for detecting Penicillium marneffei Mp1p-specific antibody[J]. Journal of Southern Medical University, 2013, 33(3): 439-443 | |
| Authors: | WANG Yanfang ZENG Lei CHEN Xuedong HAO Wei YANG Mei CAI Jianpiao WANG Yadi YUAN Guoyong CHE Xiaoyan |
| Affiliation: | 1 Center for Clinical Laboratory,Zhujiang Hospital,Southern Medical University,Guangzhou 510282,China;2 Department of Microbiology,The University of Hong Kong,Hong Kong Special Administrative Region,China |
| Abstract: | Objective To establish an immunological method for detecting antibodies of Penicillium marneffei. Methods The recombinant Mp1p protein of Penicillium marneffei was expressed in Pichia pastoris and labeled with HRP (Mp1p-HRP) with a modified sodium periodate method. A double-antigen sandwich enzyme-linked immunosorbant assay (ELISA) was established by determining the optimal coating concentration of Mp1p protein and the concentration of the detecting protein Mp1p-HRP. The sensitivity and specificity of the assay was evaluated by detecting Mp1p antibodies in 100 serum samples from healthy donors, 15 samples from culture-confirmed penicilliosis patients, and 21 samples from patients with culture-confirmed other fungal infections. Results A double-antigen sandwich ELISA was successfully established for detecting Mp1p-specific antibody. The specificity of the assay was 100% (121/121) for detecting Mp1p-specific antibody in the sera from healthy donors and patients with other fungal infection. The detection results of the 15 serum samples from patients with culture-confirmed penicilliosis showed positivity for Mp1p antibody in 2 samples and Mp1p antigen positivity in 12 samples; combining the detection results of Mp1p antigen and antibody obviously increased the diagnostic sensitivity to 93.3% (14/15). Conclusion The double-antigen sandwich ELISA shows a high specificity in detecting Mp1p-specific antibody, and simultaneous detection of Mp1p antigen and antibody can increase the diagnostic sensitivity for penicilliosis. |
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