Label-free immunoassay for porcine circovirus type 2 based on excessively tilted fiber grating modified with staphylococcal protein A
Label-free immunoassay for porcine circovirus type 2 based on excessively tilted fiber grating modified with staphylococcal protein A
复制标题
基于葡萄球菌蛋白A修饰过度倾斜光纤光栅的猪圆环病毒2型无标记免疫分析
DOI：
10.1016/j.bios.2016.07.100
复制
发表时间：
2016-12-15
影响因子：
12.6
通讯作者：
Zhang, Lin
中科院分区：
文献类型：
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作者：
Luo, Binbin;Wu, Shengxi;Zhang, Lin
利用标准单模光纤内切的过度倾斜光纤光栅（Ex-TFG），建立了一种新型的无标记免疫检测方法，用于猪圆环病毒2型（PCV2）的特异性检测。利用葡萄球菌蛋白A （Staphylococcal protein A， SPA）修饰硅化纤维表面形成SPA层，可大大提高抗PCV2单克隆抗体（MAb）的生物活性比例，从而提高抗PCV2单克隆抗体与PCV2抗原特异性吸附和结合事件的有效性。通过监测该传感器在不同PCV2滴度下的共振波长位移进行免疫分析实验。通过尿素处理将抗PCV2单克隆抗体从SPA层完全分离，并重新结合到传感器表面的SPA层上，进行PCV2的重复免疫分析。先检测猪繁殖与呼吸综合征病毒（PRRSV），后检测PCV2，检验免疫传感器的特异性。结果表明，PCV2免疫传感器的检出限（LOD）为9.371TCID(50)/mL，饱和值为4.801 × 10(3)TCID(50)/mL，重复性好，特异性好。(C) 2016 Elsevier B.V.版权所有
Using excessively tilted fiber grating (Ex-TFG) inscribed in standard single mode fiber, we developed a novel label-free immunoassay for specific detection of porcine circovirus type 2 (PCV2), which is a minim animal virus. Staphylococcal protein A (SPA) was used to modify the silanized fiber surface thus forming a SPA layer, which would greatly enhance the proportion of anti-PCV2 monoclonal antibody (MAb) bioactivity, thus improving the effectiveness of specific adsorption and binding events between anti-PCV2 MAbs and PCV2 antigens. Immunoassay experiments were carried out by monitoring the resonance wavelength shift of the proposed sensor under different PCV2 titer levels. Anti-PCV2 MAbs were thoroughly dissociated from the SPA layer by treatment with urea, and recombined to the SPA layer on the sensor surface for repeated immunoassay of PCV2. The specificity of the immunosensor was inspected by detecting porcine reproductive and respiratory syndrome virus (PRRSV) first, and PCV2 subsequently. The results showed a limit of detection (LOD) for the PCV2 immunosensor of similar to 9.371TCID(50)/mL, for a saturation value of similar to 4.801 x 10(3)TCID(50)/mL, with good repeatability and excellent specificity. (C) 2016 Elsevier B.V. All rights reserved.