Development of multianalyte flow-through and lateral-flow assays using gold particles and horseradish peroxidase as tracers for the rapid determination of carbaryl and endosulfan in agricultural products
Development of multianalyte flow-through and lateral-flow assays using gold particles and horseradish peroxidase as tracers for the rapid determination of carbaryl and endosulfan in agricultural products
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DOI：
10.1021/jf0531407
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发表时间：
2006-04-05
影响因子：
6.1
通讯作者：
Wang, S
中科院分区：
文献类型：
--
作者：
Zhang, C;Zhang, Y;Wang, S
开发了以金颗粒和辣根过氧化物酶（HRP）为示踪剂的膜基竞争性免疫测定法，用于西维因和硫丹的多重分析。对于基于金的免疫测定，膜条涂有山羊抗兔IgG（对照线）和甲萘威半抗原-卵清蛋白（OVA）和硫丹半抗原-卵清蛋白（测试线）。在基于金的检测中，西维因和硫丹的视觉检测极限分别为100和10微克/升。以HRP为示踪剂的免疫检测方法中，将抗西维因和抗硫丹抗体分别包被在膜上作为检测线，西维因的目视检测限为10 μ g/L，硫丹的目视检测限为1 μ g/L。所开发的测定分别使用金颗粒和HRP作为标记物;在研究中使用HRP标记物获得了10倍的视觉检测限增强。通过用缓冲液适当稀释样品提取物消除基质干扰。为了验证多分析物测定法，通过多分析物金基测定法对样品进行了筛选，并通过高效液相色谱法（西维因测定法）和气相色谱法（硫丹测定法）进行了确认。西维因和硫丹的多分析物金基流动分析法测定结果与仪器分析结果（高效液相色谱-紫外检测器和气相色谱-电子捕获检测器）一致。所开发的多分析物免疫测定法，其结果进行了直观的解释，可作为方便的定性工具，现场快速筛查西维因和硫丹同时在农产品中。
Membrane-based competitive immunoassays using gold particles and horseradish peroxidase (HRP) as tracers in flow-through and lateral-flow formats for multianalysis of carbaryl and endosulfan were developed. For gold-based immunoassay, membrane strips were coated with goat anti-rabbit IgG (control line) and carbaryl hapten-ovalbumin (OVA) and endosulfan hapten-OVA (test lines). The visual detection limits for carbaryl and endosulfan were 100 and 10 mu g/L in gold-based assays, respectively. For immunoassay using HRP as tracer, anti-carbaryl and anti-endosulfan antibodies were separately coated on the membrane as test lines, and the visual detection limits were 10 mu g/L for carbaryl and 1 mu g/L for endosulfan. The developed assays used gold particles and HRP as labels, respectively; 10 times enhancement in the visual detection limit using HRP label was obtained in the study. Matrix interference was eliminated by appropriate dilution of sample extracts with buffer. For the validation of the multianalyte assay, the samples were screened by multianalyte gold-based assay and confirmed by HPLC for carbaryl determination and by GC for endosulfan determination. The results of multianalyte gold-based flow-through assay for the determination of carbaryl and endosulfan were in good agreement with the results of instrumental analysis (HPLC with ultraviolet detection and GC with electron capture detection). The developed multianalyte immunoassays for which the results were interpreted visually can be used as convenient qualitative tools for on-site rapid screening of carbaryl and endosulfan simultaneously in agricultural products.