Development of a sandwich vertical flow immunogold assay for rapid detection of oxytetracycline residue in fish tissues
Development of a sandwich vertical flow immunogold assay for rapid detection of oxytetracycline residue in fish tissues
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DOI：
10.1016/j.foodchem.2018.07.124
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发表时间：
2019-01-01
期刊：
影响因子：
8.8
通讯作者：
Shamasundar, B. A.
中科院分区：
文献类型：
--
作者：
Moumita, M.;Shankar, K. M.;Shamasundar, B. A.
建立了一种基于一次性多孔滤膜的多重银增强夹心垂直流动免疫金分析法（SVIA），用于鱼肉组织中土霉素（OTC）残留的现场快速检测。合成人工抗原，制备选择性抗土霉素的兔抗血清和高特异性单克隆抗体。该测定由三层组成。第一层由固定在膜上的抗OTC兔抗血清（捕获抗体）组成，第二层是目标分子（OTC），第三层用检测剂免疫金标记的mAb精制。银增强SVIA的灵敏度低至2 ng/mL（4 min内），这表明其灵敏度比HRP标记的夹心垂直流测定法提高了125倍。开发的技术的可靠性进行了共同评估与HPLC和验证，使用115个领域的鱼类样品来自印度南部的不同地区。结果表明，SVIA具有很高的潜力，建立一个半定量的常规现场筛选工具。
A rapid multiplex silver enhanced 'sandwich vertical flow immunogold assay (SVIA)' based on disposable porous filter-membrane was developed for on-site detection of oxytetracycline (OTC) residues in fish tissues. Artificial antigens were synthesized to raise selective anti-OTC rabbit antiserum and highly specific monoclonal antibody (mAb). The assay consists of three layers. The first layer was composed of immobilized anti-OTC rabbit antiserum (capture antibody) onto the membrane, the second layer was the molecule of interest (OTC) and the third layer was refined with detector immunogold labeled mAb. The sensitivity of the silver enhanced SVIA was as low as 2 ng mL(-1) (within 4 min) representing a 125-fold increase in sensitivity over the HRP labeled sandwich vertical flow assay. The reliability of developed technique was co-evaluated with HPLC and validated using 115 field fish samples from different regions of southern India. The results demonstrated that SVIA has the high potentiality to be established as a semi-quantitative routine on-site screening tool.