适体荧光分析法检测养殖废水中四环素类抗生素  

Detection of Tetracycline Antibiotics in Aquaculture Wastewater by DNA-based Fluorescence Assay

作　　者：赵秋伶[1] 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

出　　处：《分析测试学报》2016年第7期869-873,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(21506087)

摘　　要：建立了核酸适体识别-荧光探针技术检测养殖废水中土霉素、四环素、金霉素及强力霉素4种四环素类抗生素（TCs）总残留量的新方法。两段DNA对TCs共同识别后折叠成稳定的“发卡型”双链结构,核酸染料4＇-6-二脒基-2-苯基吲哚（DAPI）能插入“发卡”部位产生荧光信号发射,据此可实现对TCs的定量检测。在最优实验条件下,以土霉素为目标分子建立工作曲线,在10～50 nmol/L范围内,荧光强度与对应浓度呈良好线性关系,土霉素的检出限为2.3 nmol/L。用四环素、土霉素、金霉素和强力霉素分别进行加标回收实验,平均回收率为88.5%～102.3%,相对标准偏差（n=5）为3.2%～6.7%。5个养殖场水样中均有TCs检出,检出量在7.6～42.7 nmol/L之间。该方法简单、灵敏、快速,可满足养殖场废水中TCs总残留量的测定要求。A novel analysis method based on specific recognition of aptamer and fluorescence probe technique was established for the detection of total residues of tetracycline antibiotics （TCs）, inclu- ding oxytetraeycline, tetracycline, chlortetracycline and Doxycycline etc. in aquaculture wastewater. Two segments of DNAs work together to identify TCs to fold into a stable ＂hairpin＂ double chain structure that can be inserted by nucleic acid dye DAPI to generate of fluorescence emission. Based on this, TCs can be quantified. Under the optimal experimental conditions, the working curve was established with oxytetracycline as target molecule, the fluorescence intensity showed a good linear re- lationship with concentration of oxytetracycline in the range of 10 -50 nmol/L, and the detection lim- it of oxytetracycline was 2. 3 nmol/L. When having oxytetracycline, tetracycline, chlortetracycline and doxycycline as additives, the average recoveries ranged from 88.5% to 102.3% , with relative standard deviations （ n = 5 ） of 3.2% - 6.7% . The method is simple, sensitive and rapid, and could satisfy the requirements for determination of total TCs residues in the farm wastewater.

关 键 词：核酸适体 荧光分析法 四环素类抗生素 快速检测 废水 

分 类 号：O657.6[理学—分析化学] R978.1[理学—化学]