Virus detection using filament-coupled antibodies.
Virus detection using filament-coupled antibodies.
复制标题
使用丝耦合抗体进行病毒检测。
DOI：
10.1002/bit.20537
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
Haselton,FrederickR
中科院分区：
文献类型：
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作者：
Stone,GregoryP;Mernaugh,Ray;Haselton,FrederickR
酶联免疫吸附试验的两个吸引人的特点是抗体-抗原识别的特异性和酶扩增的敏感性。本报告描述了一种非酶分子识别平台的开发，该平台适用于医疗保健临床环境和生物危险物质的现场检测。这种细丝抗体识别分析(FARA)是基于直径为120μm的聚酯长丝上的抗体探针的圆周条带。这种设计的一个优点是，通过一系列25-60μ的L液体填充微毛细管小室顺序定位细丝耦合探针，实现了自动化处理。通过使用与细丝偶联的抗M13KO7 IgG1单抗检测M13KO7细菌病毒的存在，对该方法进行了评估。细丝运动首先将抗体定位在含有M13KO7病毒溶液的微毛细管内，然后将探针移动到随后的小室，在那里清洗细丝偶联的探针，暴露于荧光标记的抗M13K07抗体，然后再次清洗。然后使用平板微阵列扫描仪测量细丝荧光。病毒在溶液中的存在仅在含有偶联抗体探针的区域产生特征性的细丝荧光增加。即使没有典型的ELISA的酶扩增，8.3 × 108病毒颗粒的存在也比固定的阴性对照抗体产生的荧光增加30倍。在ELISA法的比较研究中，基于细丝的方法具有与∼1.7 × 107病毒颗粒类似的灵敏度下限。该平台可能会被证明具有吸引力，适用于医疗设备、生物危险材料检测或其他需要灵敏、快速和自动化分子识别的应用。©2005威利期刊公司。
Two attractive features of ELISA are the specificity of antibody–antigen recognition and the sensitivity achieved by enzymatic amplification. This report describes the development of a non‐enzymatic molecular recognition platform adaptable to point‐of‐care clinical settings and field detection of biohazardous materials. This filament‐antibody recognition assay (FARA) is based on circumferential bands of antibody probes coupled to a 120 μm diameter polyester filament. One advantage of this design is that automated processing is achieved by sequential positioning of filament‐coupled probes through a series of 25–60 μL liquid filled microcapillary chambers. This approach was evaluated by testing for the presence of M13KO7 bacterial virus using anti‐M13KO7 IgG1monoclonal antibody coupled to a filament. Filament motion first positioned the antibodies within a microcapillary tube containing a solution of M13KO7 virus before moving the probes through subsequent chambers, where the filament‐coupled probes were washed, exposed to a fluorescently labeled anti‐M13K07 antibody, and washed again. Filament fluorescence was then measured using a flatbed microarray scanner. The presence of virus in solution produced a characteristic increase in filament fluorescence only in regions containing coupled antibody probes. Even without the enzymatic amplification of a typical ELISA, the presence of 8.3 × 108virus particles produced a 30‐fold increase in fluorescence over an immobilized negative control antibody. In an ELISA comparison study, the filament‐based approach had a similar lower limit of sensitivity of ∼1.7 × 107virus particles. This platform may prove attractive for point‐of‐care settings, the detection of biohazardous materials, or other applications where sensitive, rapid, and automated molecular recognition is desired. © 2005 Wiley Periodicals, Inc.