Development of colloidal gold-based immunochromatographic assay for rapid detection of Mycoplasma suis in porcine plasma
Development of colloidal gold-based immunochromatographic assay for rapid detection of Mycoplasma suis in porcine plasma
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DOI：
10.1016/j.bios.2013.12.048
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发表时间：
2014-05-15
影响因子：
12.6
通讯作者：
Chai, Tongjie
中科院分区：
文献类型：
--
作者：
Meng, Kai;Sun, Wenjing;Chai, Tongjie
用抗猪支原体多克隆抗体包被金纳米颗粒，建立了一步法免疫层析检测方法。suis）的检测方法。猪血浆中的猪。用柠檬酸钠还原金盐，偶联抗M.猪。用重组M. suis在大肠杆菌中表达。捕获抗体和包被抗体的最佳浓度分别为12 μ g/ml和1.5 mg/ml，封闭缓冲液为1%牛血清白蛋白。在最佳分析条件下目视检测，免疫层析测定试验的检测下限为100 ng/ml。用猪瘟病毒、猪繁殖与呼吸综合征病毒、猪肺炎支原体、猪弓形虫和猪细小病毒对免疫层析试纸条进行特异性评价。该抗体与其它相关猪病原体无交叉反应。该定性试验基于结果的目视评价，不需要任何设备。M.猪流感检测时间小于10 min，适合基层快速检测。所研制的一步法胶体金免疫层析试纸条具有较高的特异性和灵敏度。因此，该方法是一种可行、方便、快速、有效的方法。猪血浆中的猪。(C)2013爱思唯尔有限公司版权所有。
A one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pAb) against Mycoplasma suis (M. suis) was developed in this study for the detection of M. suis in porcine plasma. The colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pAb against M. suis. The pAb was produced by immunizing the BALB/c mice with recombinant MSG1 (rMSG1) protein from M. suis expressed in Escherichia coli. The optimal concentrations of the capture antibody and the coating antibody were 12 mu g/m1 and 1.5 mg/ml, respectively, and that of the blocking buffer was 1% bovine serum albumin. The lower detection limit of the immunochromatographic assay test was 100 ng/ml with visual detection under optimal conditions of analysis. Classical swine fever virus, porcine reproductive and respiratory syndrome virus, swine pneumonia mycoplasma, swine toxoplasma, and porcine parvovirus were used to evaluate the specificity of the immunochromatographic strips. No cross-reaction of the antibodies with other related swine pathogens was observed. This qualitative test based on the visual evaluation of the results did not require any equipment. The assay time for M. suis detection was less than 10 min, suitable for rapid detection at the grassroots level. The one-step colloidal gold immunochromatographic strips that we developed had high specificity and sensitivity. Therefore, this method would be feasible, convenient, rapid, and effective for detecting M. suis in porcine plasma. (C) 2013 Elsevier B.V. All rights reserved.