Zeng Haijuan, Zhang Duoqiang, Zhai Xuzhao, Wang Shujuan, Liu Qing
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai, 200093, China.
Hepatobiliary Surgery Department, Ningxia People's Hospital, Yinchuan, 750002, China.
Anal Bioanal Chem. 2018 Jan;410(1):71-77. doi: 10.1007/s00216-017-0690-2. Epub 2017 Oct 30.
A rapid lateral flow immunochromatographic strip (ICS) using fluorescein isothiocyanate (FITC) labeled antigen and antibody was developed for the detection of Acidovorax citrulli (Ac) in melons and vegetable samples. In the ICS, signal amplification was realized based on antigen Ac and anti-Ac monoclonal antibody (McAb) 4F conjugated with FITC, respectively, which were forming two probes. The control line and the test line were obtained by immobilizing the goat anti-mouse IgG antibody and anti-Ac McAb 6D on both sides of the nitrocellulose membrane. The visual detection limit of the strip was 10 CFU/mL, which was 10-fold sensitive compared to the strip of FITC only labeling antigen or antibody. Signal amplification ICS was successfully applied to the detection of Ac in melon and vegetable samples with less detection time and operation procedures compared to the traditional enzyme-linked immunosorbent assay (ELISA) and PCR methods. This is the first report of using FITC labeled antigen and McAb as dual fluorescent probes to develop a direct-type immunofluorescence strip for the rapid and sensitive detection of Ac, which demonstrates a powerful tool for rapidly screening Ac in plant materials and other samples. Graphical abstract The schematic presentation of the test strip (a) and the positive result (b) or negative result
开发了一种使用异硫氰酸荧光素（FITC）标记抗原和抗体的快速侧向流动免疫层析试纸条（ICS），用于检测甜瓜和蔬菜样品中的西瓜嗜酸菌（Ac）。在该ICS中，基于分别与FITC偶联的抗原Ac和抗Ac单克隆抗体（McAb）4F实现信号放大，二者形成两种探针。通过将山羊抗小鼠IgG抗体和抗Ac McAb 6D固定在硝酸纤维素膜的两侧获得对照线和测试线。该试纸条的目视检测限为10 CFU/mL，比仅用FITC标记抗原或抗体的试纸条灵敏10倍。与传统的酶联免疫吸附测定（ELISA）和聚合酶链反应（PCR）方法相比，信号放大ICS成功应用于甜瓜和蔬菜样品中Ac的检测，检测时间和操作步骤更少。这是首次报道使用FITC标记抗原和McAb作为双荧光探针开发直接型免疫荧光试纸条用于快速灵敏检测Ac，这证明了其是一种用于快速筛选植物材料和其他样品中Ac的强大工具。图形摘要 试纸条示意图（a）以及阳性结果（b）或阴性结果
