侧流胶体金免疫试纸快速测定牛奶中的泰乐菌素

Lateral flow colloidal gold immunoassay paper for rapid determination of tylosin in milk

大环内酯类抗生素是非常重要的一类抗菌化合物,而泰乐菌素(tylosin,TYL)是最常用的预混大环内酯类抗生素。但是,这些药物的残留物会在人体中引起过敏反应,并对抗生素产生抗药性。为了评估TYL的残基,需要灵敏的分析方法以确保低浓度下的测定。该文开发了一种快速免疫层析侧流试纸条,用于专门测定牛奶样品中的TYL残留。该文筛选了抗TYL单克隆抗体。抗体滴度为2.56×10^(-5),IC_(50)为4.6091 ng/mL。该测试条由样品垫、共轭垫、包含质控线和线的硝酸纤维素(nitrocellulose membrane,NC)膜以及吸收垫组成。用试纸条测定TYL的不同标准样品(0、2、4、8、16、32、64 ng/mL)。用扫描仪计算出试纸的检出限(limit of detection,LOD)为1.5434 ng/mL,肉眼计算为10 ng/mL,计算其IC_(50)为7.8362 ng/mL,无需特殊设备,测试结果将在20 min内获得。牛奶的平行分析显示从试纸条和HPLC获得的结果相当,试纸与其他兽药无交叉反应。因此,该试纸条非常简便快捷,可作为定量、半定量或定性检测牛奶中TYL残留的筛选方法。

Macrolide antibiotics are a very important class of antibacterial compounds,and tylosin(TYL)is the most commonly used premixed macrolide antibiotics.However,the residues of these drugs can cause allergic reactions in the human body and become resistant to antibiotics.In order to evaluate the residues of TYL,sensitive analytical methods are required to ensure the determination at low concentrations.In this work,we developed a rapid immunochromatographic lateral flow test strip for the determination of TYL residues in milk samples.We screened anti-TYL monoclonal antibodies.The antibody titer was 2.56×10^(-5),and the IC_(50) was 4.6091 ng/mL.The test strip was composed of a sample pad,a conjugate pad,a control line and a test line,and a NC membrane and an absorbent pad.Different standard samples(0,2,4,8,16,32,and 64 ng/mL)of TYL were measured with test strips.The limit of detection(LOD)of the test paper calculated with the scanner was 1.5434 ng/mL,while LOD calculated by naked eye was 10 ng/mL,and the IC_(50) was calculated to be 7.8362 ng/mL.No special equipment was required.The test result could be obtained within 20 minutes.The parallel analysis of milk showed that the results obtained from the test strips and HPLC were comparable,and the test strips did not cross-react with other veterinary drugs.Therefore,this test strip is very simple and fast,and can be used as a screening method for quantitative,semi-quantitative or qualitative detection of TYL residues in milk.