动物营养学报2019，Vol.31Issue(11)：5230-5238,9.DOI:10.3969/j.issn.1006⁃267x.2019.11.039
脱氧雪腐镰刀菌烯醇间接竞争酶联免疫吸附测定试剂盒的研制及初步应用
Development and Preliminary Application of an Indirect Competitive ELISA Kit for Detecting Deoxynivalenol
摘要
Abstract
This study was conducted to develop a self?assembled blocking indirect competitive ELISA ( icELISA) kit for the detection of deoxynivalenol in feed. Based on the preparation of anti?DON monoclonal antibodies, we established the standard curve of icELISA and optimized the detection conditions, which was used to examine the performance of the kit, then the determination results were compared with the high?per?formance liquid chromatography ( HPLC) method. The results showed that the minimum detectable concentra?tion of the kit for DON was 1.147 ng／mL, with the detection range of 1.50 to 130.31 ng／mL, and the 50%inhibiting concentration ( IC50) of 9.84 ng／mL. The recovery experiment by adding standard samples was car?ried out for four samples of wheat, corn, pig feed and chicken feed. The recovery was in the range of 77.1%to 113.2%, and the relative standard deviation ( RSD) ranged from 4.2% to 12.6%. Besides, the kit had the advantages of high specificity, good stability, long effective life and negligible sample matrix interference. Fi?nally, twenty blind samples were determined by the kit, and the results were in good agreement with the HPLC method. These results indicate that the icELISA kit developed in this experiment has good performance to meet the relevant technical requirements, and with the characteristics of accuracy, reliability, convenience and high throughput screening to detect DON. Therefore, it is suitable for rapid screening of DON for the market.[ Chi?nese Journal of Animal Nutrition, 2019, 31(11) :5229?5237]关键词
脱氧雪腐镰刀菌烯醇/icELISA试剂盒/性能测定/初步应用Key words
deoxynivalenol/icELISA kit/performance measurement/preliminary application分类
农业科技引用本文复制引用
韩丽,吕俊美,周叶,张文举,王自良,李月涛,姜金庆,李任峰,范国英,张海棠,牛琳琳..脱氧雪腐镰刀菌烯醇间接竞争酶联免疫吸附测定试剂盒的研制及初步应用[J].动物营养学报,2019,31(11):5230-5238,9.基金项目
"十二五"国家科技支撑计划项目"激素类药物快速检测技术研究与示范" ( 2014BAD13B05?01) （ 2014BAD13B05?01）
新疆生产建设兵团重点领域科技攻关项目(2018AB041) （2018AB041）
第八师现代农业科技攻关与成果转化项目(2018NY05) （2018NY05）
