Development of an enzyme-linked immunoassay for the detection of gentamicin in swine tissues
Development of an enzyme-linked immunoassay for the detection of gentamicin in swine tissues
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DOI：
10.1016/j.foodchem.2007.10.022
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发表时间：
2008-05-01
期刊：
影响因子：
8.8
通讯作者：
Xiao, Xilong
中科院分区：
文献类型：
--
作者：
Chen, Yiqiang;Shang, Yanhong;Xiao, Xilong
我们开发了一种酶联免疫测定法，可快速、灵敏地检测猪组织中的庆大霉素。用庆大霉素-牛血清白蛋白偶联物免疫家兔，第5次免疫后收集抗血清。在优化免疫试剂浓度后，竞争性间接ELISA（ciELISA）的IC_（50）值为0.98 ng/ml，而竞争性直接ELISA（cdELISA）的IC_（50）值较低，仅为0.92 ng/ml。优化后的酶联免疫吸附试验可在45 min内完成，与其他氨基糖苷类抗生素的交叉反应性可忽略不计。猪组织中庆大霉素的回收率为64.7%~ 101.2%，变异系数为4.5- 12.1%，肌肉、肝脏和肾脏中庆大霉素的检出限分别为6.2 μ g/kg、3.6 μ g/kg和2.7 μ g/kg。(C)2007爱思唯尔有限公司保留所有权利。
We developed an enzyme-linked immunoassay that provides rapid and sensitive detection of gentamicin in swine tissues. Rabbit was immunized with gentamicin-BSA conjugate and antiserum was collected after the fifth immunization. After optimizing the concentration of immunoreagents, competitive indirect ELISA (ciELISA) gave an IC50 value of 0.98 ng/ml, while competitive direct ELISA (cdELISA) exhibited lower IC50 value of 0.92 ng/ml, thus cdELISA was further optimized under various pH values and ionic strengths of assay buffer, different coating methods and incubation time. The optimized ELISA can be completed within 45 min and it showed negligible cross-reactivity with other aminoglycosides. The recoveries of gentamicin from spiked swine tissues at levels of 25-200 mu g/kg ranged from 64.7% to 101.2% with CVs of 4.5-12.1%, and the detection limits were 6.2 mu g/kg in muscle, 3.6 mu g/kg in liver and 2.7 mu g/kg in kidney, respectively. (C) 2007 Elsevier Ltd. All rights reserved.