酶联适体分析法检测动物源性食品中的卡那霉素残留  

Enzyme-linked aptamer assay for detection of kanamycin residues in animal derived food

作　　者：赵秋伶[1] 史素青[2] 张尤华[1] 于晓艳[1] 

机构地区：[1]辽宁工程技术大学矿业技术学院,辽宁葫芦岛125105 [2]西北大学化学与材料学院,陕西西安710069

出　　处：《食品工业科技》2015年第10期86-89,100,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金(21004047);陕西省青年科技新星人才项目(2014KJXX-62)

摘　　要：建立了基于核酸外切酶Ⅰ的酶联适体分析法,用于测定动物源性食品中的卡那霉素残留。核酸适体和卡那霉素结合后,不再被核酸外切酶Ⅰ剪切,而能进一步联结辣根过氧化物酶（HRP）,催化四甲基二苯胺底物显色,以450nm的吸光度与浓度的线性关系确定卡那霉素的检出限。考察了包被浓度、竞争反应时间、核酸外切酶I用量及封闭液等因素对检测的影响。在优化的实验条件下,建立的方法对卡那霉素检测有高灵敏度,检测限为3.26μg/L,线性范围5~100μg/L;在4种肉类中添加5.0、20.0、50.0μg/kg的卡那霉素时,加标回收率可达75.8%~90.6%,相对标准偏差RSD为4.2%~7.8%。该方法无需大型仪器,操作简单,可用于动物源性食品中卡那霉素残留的快速检测。Exonuclease I-based enzyme-linked aptamer assay for the detection of kanamycin residues in animal derived food was established in this paper. Aptamer binding with kanamycin could no longer be digested by exonuclease I, but could further specifically capture the horseradish peroxidase（HRP） that catalyzed methyenedianiline substrate to produce color change. Determination of the limit of detection was based on linear relationship between absorbance value at 450nm and the concentration of kanamycin. The effects of coated concentration,competitive reaction time,dosage of exonuclease I and blocking solutions on the detection were investigated. The results showed that under the optimal conditions,the established exonuclease I-depended enzyme-linked aptamer analysis of kanamycin have high sensitivity,the detection limit was 3.26μg/L with linear ranging from 5 to 100μg/L. When kanamycin of 5.0,20.0 and 50.0μg/kg were added to four animal derived foods,the recovery rates ranged from 75.8% to 90.6% ,the relative standard deviation for RSD ranged from 4.2% to 7.8%. The method had simple operation without need large instrument and could be used for fast detection of kanamycin residues in animal derived foods.

关 键 词：卡那霉素 核酸适体 酶联适体分析 动物源性食品 

分 类 号：TS201.3[轻工技术与工程—食品科学]