兽药及非法添加物量子点标记检测试纸条研制

发布时间：2018-03-06 09:03

本文选题：兽药　切入点：非法添加物　出处：《天津科技大学》2017年硕士论文　论文类型：学位论文

【摘要】：本文选择量子点作为荧光标记材料,研制了喹诺酮类兽药量子点标记免疫检测试纸条和非法添加物三聚氰胺量子点标记免疫检测试纸条,该方法可用于实际样品的快速检测。采用活化酯法制备喹诺酮类兽药包被原(NOR-OVA)。喹诺酮类兽药量子点标记免疫检测试纸条最佳工作条件为:量子点标记抗体偶联反应的最佳pH值为8.4;量子点(QDs)与活化剂(EDC)与抗体的摩尔比为1:2000:10;包被原的稀释倍数为1:40;二抗的稀释倍数为1:30;标准品稀释液为PBST (pH 7.4);工作液添加量为10 μL;硝酸纤维素膜型号为Milipore HF 135s; QDs-Ab偶联物添加量为0.5 μL。该试纸条方法检出限为2 ng/mL,检测时间为10 min。与5种喹诺酮类兽药环丙沙星、恩诺沙星、培氟沙星、氧氟沙星和洛美沙星,有明显的交叉反应;与其它的10种喹诺酮类兽药和5种常见的其它不同种类兽药没有明显的交叉反应,特异性良好。实际样品中检测限为 10 ng/g。采用活化酯法合成三聚氰胺包被原(MEL-BSA)。三聚氰胺量子点标记免疫检测试纸条最佳工作条件为:量子点标记抗体偶联反应的最佳pH值为8.4;量子点(QDs)与活化剂(EDC)与抗体的摩尔比为1:1500:15;包被原的稀释倍数为1:80;二抗的稀释倍数为1:140;标准品稀释液为PBS (pH 7.4);工作液添加量为5μL;硝酸纤维素膜型号为Milipore HF 135s,用1%乳粉溶液封闭处理;QDs-Ab偶联物添加量为1μL。该试纸条方法检出限为100 ng/mL,检测时间为10 min。结构类似物中除了环丙氨嗪以外,都不存在交叉反应;与5种常见的其它不同种类兽药也没有明显的交叉反应,特异性良好。实际样品中检测限为400 ng/g。本研究当中所建立的喹诺酮类兽药和三聚氰胺量子点标记免疫检测试纸条样品前处理简单、操作简便、检测时间短、特异性好,与传统的胶体金试纸条相比灵敏度更高,能够适用于现场大量样品的快速检测。
[Abstract]:In this paper, Quantum Dots (QDs) were selected as fluorescent labeling materials, and the QDs immunoassay strips for quinolones and the illegal additives, melamine QDs, were developed. This method can be used for rapid detection of real samples. Quinolones coated with NOR-OVAX were prepared by activation ester method. The best working conditions of QDIA strips for quinolones were as follows: QDD-labeled antibody coupling reaction. The optimum pH value is 8.4; the molar ratio of QDs to activator is 1: 2000: 10; the dilution factor of the coating is 1: 40; the dilution multiple of the second antibody is 1: 30; the dilution of the standard sample is PBST / pH 7.4; the working solution is 10 渭 L; the nitrocellulose membrane is coated with nitrocellulose. Model Milipore HF 135s, QDs-Ab conjugate amount 0.5 渭 L. the detection limit of the test strip is 2 ng / mL, the detection time is 10 min. with 5 kinds of quinolones, ciprofloxacin, Enrofloxacin, pefloxacin, ofloxacin and lomefloxacin had significant cross reactions, but not with 10 other quinolones and 5 other common different veterinary drugs. The detection limit in real samples is 10 ng / g. Melamine coated with MEL-BSAA is synthesized by activated ester method. The best working conditions of the test strip for immunological detection of melamine quantum dots are: QD labeled antibody coupling reaction. The optimum pH value is 8.4; the molar ratio of QDs to activator EDC) and antibody is 1: 1500: 15; the dilution factor of the coating is 1: 80; the dilution multiple of the second antibody is 1: 140; the dilution of the standard product is PBS pH 7.4; the working solution is 5 渭 L; the nitrocellulose membrane type is 1: 140; Milipore HF 135s, the addition amount of QDs-Ab conjugate was 1 渭 L. the detection limit of the method was 100ng / mL and the detection time was 10min. There was no cross reaction; there was no significant cross reaction with five other common veterinary drugs. The detection limit in the actual sample is 400 ng / g. The sample of quinolones and melamine quantum dot labeled immunoassay strip established in this study has the advantages of simple pretreatment, simple operation, short detection time and good specificity. Compared with the traditional colloidal gold test strip, it is more sensitive and can be used for rapid detection of a large number of samples in the field.
【学位授予单位】：天津科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S859.84

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