Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University, Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Integrated Management of Crop Diseases and Pests (Nanjing Agricultural University), Ministry of Education, Nanjing 210095, PR China.
Ecotoxicol Environ Saf. 2011 Sep;74(6):1595-9. doi: 10.1016/j.ecoenv.2011.04.026. Epub 2011 May 4.
An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) has been developed for detection of pretilachlor in water and soil. An immunogen was prepared from haptens of pretilachlor conjugated to bovine serum albumin(BSA). The specific polyclonal antibodies were obtained by immunizing New Zealand white rabbits. The influence of parameters including concentrations of methanol, ionic strength and pH values were optimized to improve the sensitivity of the assay. The optimized ELISA was shown to have a high sensitivity and specificity for pretilachlor. Under optimal conditions, the ELISA has demonstrated an 50% inhibitory concentration (IC(50)) value of 0.0359 mg/L with a limit of detection (LOD, IC(10)) of 6.9 ng/L. The cross-reactivities to some analogs of pretilachlor (acetochlor, butachlor, metazachlor and metalaxyl) were below 1.5%. The average recoveries of pretilachlor from distilled water, tap water, paddy water and soil were in the range of 77.0-115.2% between 0.005 and 5.0mg/L. The results of ELISA for spiked samples were confirmed by GC-ECD with a high correlation coefficient of 0.9950(n=11). Thus, the ELISA proven to be a sensitive, specific, inexpensive and quantitative tool for detection of pretilachlor from four kinds of spiked samples.
一种间接竞争酶联免疫吸附测定法（ic-ELISA）已被开发用于检测水中和土壤中的丙草胺。免疫原由丙草胺与牛血清白蛋白（BSA）偶联的半抗原制备。通过免疫新西兰白兔获得了特异性的多克隆抗体。优化了包括甲醇浓度、离子强度和 pH 值等参数，以提高测定的灵敏度。优化的 ELISA 对丙草胺具有高灵敏度和特异性。在最佳条件下，ELISA 的 50%抑制浓度（IC（50））值为 0.0359mg/L，检测限（IC（10））为 6.9ng/L。对丙草胺的一些类似物（乙草胺、丁草胺、异丙甲草胺和甲霜灵）的交叉反应性低于 1.5%。在 0.005 至 5.0mg/L 之间，从蒸馏水、自来水、稻田水和土壤中提取的丙草胺的平均回收率在 77.0-115.2%之间。用 GC-ECD 对加标样品进行 ELISA 验证，相关系数为 0.9950（n=11）。因此，该 ELISA 被证明是一种灵敏、特异、廉价且可定量检测四种加标样品中丙草胺的工具。
