An octuplex lateral flow immunoassay for rapid detection of antibiotic residues, aflatoxin M1 and melamine in milk
An octuplex lateral flow immunoassay for rapid detection of antibiotic residues, aflatoxin M1 and melamine in milk
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DOI：
10.1016/j.snb.2019.04.019
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发表时间：
2019-08-01
影响因子：
8.4
通讯作者：
Chen, Yiqiang
中科院分区：
文献类型：
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作者：
Han, Miaomiao;Cong, Lu;Chen, Yiqiang
抗生素残留、黄曲霉毒素M-1和三聚氰胺是牛奶中的三大化学污染物。然而，目前还没有同时检测这些化学物质的分析方法。在本研究中，开发了八重侧流免疫测定法来解决这个问题。制备特异性单克隆抗体和受体，并将其结合到金纳米颗粒上，形成八种抗体/受体金纳米颗粒缀合物作为检测探针。合成了八种相应的半抗原-蛋白质缀合物，并将其固定在硝酸纤维素膜的不同测试区域上作为捕获抗原。因此，八重免疫反应可以同时发生在单个条带上。定性分析时，肉眼目测即可判断结果，测得喹诺酮类、四环素类、β-内酰胺类、磺胺类、氯霉素、链霉素、黄曲霉毒素M-1、三聚氰胺的截断值为12-80 ng/mL、16-32 ng/mL、2-80 ng/mL、4-80 ng/mL、1.2分别为 ng/mL、50 ng/mL、0.5 ng/mL 和 200 ng/mL。对于半定量分析，可以通过智能手机的内置摄像头拍摄测试线，并通过Image J软件分析线强度。所有化学品的检测限均远低于当局规定的监管限。此外，该方法的检测时间小于20分钟。因此，本研究中开发的免疫测定法是快速筛查牛奶中常见化学污染物的有前途的方法。
Antibiotic residues, aflatoxin M-1 and melamine are three major types of chemical contaminants in milk. However, analytical approach for the simultaneous detection of these chemicals is currently not available. In this study, an octuplex lateral flow immunoassay was developed to address this problem. Specific monoclonal antibodies and receptor were prepared and were bound onto gold nanoparticles to form eight antibody/receptorgold nanoparticle conjugates as detection probes. Eight corresponding hapten-protein conjugates were synthesized and immobilized on distinct test zones of nitrocellulose membrane as capture antigens. Thus octuplex immune-reactions can then simultaneously occur on a single strip. For qualitative analysis, the result can be visually judged by naked eyes and the cut-off values of quinolones, tetracyclines, beta-lactams, sulfonamides, chloramphenicol, streptomycin, aflatoxin M-1 and melamine were measured to be 12-80 ng/mL, 16-32 ng/mL, 2-80 ng/mL, 4-80 ng/mL, 1.2 ng/mL, 50 ng/mL, 0.5 ng/mL and 200 ng/mL, respectively. For semi-quantitative analysis, the test lines can be photographed by a built-in camera of smartphone and the line intensities can be analyzed by Image J software. The limits of detection for all the chemicals were far below the regulatory limits set by authorities. Furthermore, the detection time of this method was less than 20 min. Consequently, the developed immunoassay in this study is a promising approach for rapidly screening common chemical contaminants in milk.