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Ultra-high detection of pathogenic virus on the basis of destruction of a liposome that capsulates nanoparticles

基于纳米粒子脂质体破坏的病原病毒的超高检测

基本信息

批准号：
18510105
负责人：
EGASHIRA Naoyoshi
金额：
$ 2.58万
依托单位：
Prefectural University of Hiroshima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

By using a novel detection method combining electrochemiluminescence (ECL) and liposome, we have searched development of the rapid detection for hemagglutinin protein that exists on the surface of influenza virus.1. We successfully have synthesized the ruthenium complex that shows adequate adsorption to a gold electrode and have clarified the optimal conditions for liposome preparation and measurement procedure.2. Detection of BSA protein: We have determined the optimal conditions for the number of antibody on the liposome, the liposome concentration, and the time of immunoreaction and thereby have realized the rapid and highly sensitive detection of BSA; lower detection limit of 10^-(13) mol/mL and measurement time within 90 min.3. Detection of hemagglutinin peptide: We have prepared the immunoliposome using the antibody to discriminate the peptide of the reservative am no acid combination (20 mer) in hemagglutinin of A type influenza virus. The analytical procedure is as follows: a, binding of the peptide onto the gold electrode; b, additions of peptide solution and then the immunoliposome solution on the electrode; c, immunoreaction for 1h; d, ECL measurement on application of potential. We have detected the peptide lithe concentration 10^<-17> mol/mL within 90 min4. Detection of hemagglutinin protein : The hemagglutinin protein prepared by genetic engineering was bound onto the electrode and then was detected in the same way mentioned above.5. Detection of influenza virus: By the same method, we have accomplished highly sensitive detection for A type of influenza virus (several hundreds of virus particles/mL) and hereafter will improve the precision of ECL data.Conclusively, we have rapidly detected influenza virus by our novel method. The method is promising for ninny fields because of applicability to many proteins as well as other viruses.

利用电化学发光（ECL）和脂质体相结合的新检测方法，探索了流感病毒表面血凝素蛋白的快速检测方法。我们成功地合成了在金电极上具有良好吸附性能的钌配合物，并明确了脂质体制备的最佳条件和测定方法。BSA蛋白检测：确定了脂质体上抗体数目、脂质体浓度、免疫反应时间的最佳条件，实现了BSA的快速、高灵敏度检测；低检出限为10^-(13)mol/mL，检测时间在90 min以内。血凝素肽的检测：利用抗体制备免疫脂质体，对A型流感病毒血凝素中保留的无酸组合（20 mer）的肽进行鉴别。分析步骤如下：a，将肽结合到金电极上；B，先添加肽溶液，再在电极上添加免疫脂质体溶液；C，免疫反应1h；d、电势应用的ECL测量。我们在90分钟内检测到肽脂质浓度10^<-17> mol/mL 4。4 .血凝素蛋白的检测：将基因工程制备的血凝素蛋白结合到电极上，用上述方法进行检测。流感病毒的检测：用同样的方法，我们已经完成了A型流感病毒的高灵敏度检测（几百个病毒颗粒/mL），今后将提高ECL数据的精度。总之，我们已经快速检测流感病毒的新方法。由于该方法适用于许多蛋白质和其他病毒，因此在许多领域都很有前景。