Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China.
Molecules. 2012 Jun 15;17(6):7401-14. doi: 10.3390/molecules17067401.
Due to the widespread use and potential toxicity of avermectins (AVMs), multi-residue monitoring of AVMs in edible tissues, especially in milk, has become increasingly important. With the aim of developing a broad-selective immunoassay for AVMs, a broad-specific monoclonal antibody (Mab) was raised. Based on this Mab, a homologous indirect enzyme-linked immunosorbent assay (ELISA) for the rapid detection of AVMs in milk was developed. Under the optimized conditions, the IC₅₀ values in assay buffer were estimated to be 3.05 ng/mL for abamectin, 13.10 ng/mL for ivermectin, 38.96 ng/mL for eprinomectin, 61.00 ng/mL for doramectin, 14.38 ng/mL for emamectin benzoate. Detection capability (CCβ) of the ELISA was less than 5 ng/mL and 2 ng/mL in milk samples prepared by simple dilution and solvent extraction, respectively. The optimized ELISA was used to quantify AVMs in milk samples spiked at different amounts. The mean recovery and coefficient of variation (CV) were 95.90% and 15.42%, respectively. The Mab-based ELISA achieved a great improvement in AVMs detection. Results proved this broad-selective ELISA would be useful for the multi-residue determination of AVMs in milk without purification process.
由于阿维菌素(AVMs)的广泛使用和潜在毒性,对食用组织中 AVMs 的多残留监测,特别是在牛奶中,变得越来越重要。本研究旨在开发一种用于 AVMs 的广谱选择性免疫分析方法,制备了一种广谱特异性单克隆抗体(Mab)。基于该 Mab,开发了一种用于快速检测牛奶中 AVMs 的同源间接酶联免疫吸附测定法(ELISA)。在优化条件下,在测定缓冲液中,阿维菌素、伊维菌素、埃普里诺菌素、多拉菌素、苯甲酰氨基阿维菌素的 IC₅₀ 值估计分别为 3.05、13.10、38.96、61.00、14.38 ng/mL。通过简单稀释和溶剂提取制备的牛奶样品中 ELISA 的检测能力(CCβ)分别小于 5 和 2 ng/mL。优化的 ELISA 用于定量分析不同添加量的牛奶样品中的 AVMs。平均回收率和变异系数(CV)分别为 95.90%和 15.42%。基于 Mab 的 ELISA 在 AVMs 检测方面取得了重大进展。结果表明,该广谱选择性 ELISA 无需净化过程即可用于牛奶中 AVMs 的多残留测定。