鸡蛋中泰妙菌素化学发光酶免疫分析方法研究

李露; 杨金易; 王宇; 张政; 苑婷婷; 肖毅美; 徐振林; 沈玉栋

Development of a Chemiluminescence Enzyme Immunoassay Method for Detection of Tiamulin Residues in Eggs

DOI：

KeyWord:tiamulin egg monoclonal antibody chemiluminescene enzyme immunoassay（CLEIA）

Author	Institution
LI Lu,YANG Jin?yi,WANG Yu,ZHANG Zheng,YUAN Ting?ting,XIAO Yi?mei,XU Zhen?lin,SHEN Yu?dong	1.Guangdong Provincial Key Laboratory of Food Quality and Safety，College of Food Science，South China Agricultural University，Guangzhou ， China; 2. Guangzhou Institute for Food Inspection， Guangzhou ，China; 3. Shenzhen Care - green Agriculture Products Testing & Certification Co. Ltd.，Shenzhen ，China

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Abstract:

A chemiluminescence enzyme immunoassay（CLEIA） method was established for the monitoring of tiamulin residues in egg samples，based on a monoclonal antibody with excellent specificity and sensitivity obtained by synthesizing antigen to immunize BALB/c mice and cell experiments.The optimal conditions were finalized as follows：the coating antigen concentration：0.26 μg/mL，the antibody concentration：0.94 μg/mL，the reaction buffer system：PBS（0.02 mol/L，pH 7.4），and the dilution buffer，the concentration and the reaction time for the enzyme-labeled secondary antibody were PBST buffer，2 μg/mL and 40 min，respectively.Under the optimal conditions，a CLEIA method for tiamulin detection was established，with an IC50 of 0.54 ng/mL，a limit of detection（LOD，IC10） of 0.03 ng/mL，and a linear detection range （IC20-IC80） of 0.17－1.74 ng/mL.There was no obvious cross-reaction between the method and the other structural and functional analogs of tiamulin.The CLEIA method was used for the detection of real egg samples.The spiked recoveries ranged from 100% to 118%，with the relative standard deviations（RSDs） less than 10%. The test results of the developed CLEIA had a good correlation with those of HPLC－MS/MS（r2 = 0.998 7），indicating that the method was suitable for the detection of tiamulin residues in egg samples.