Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (16): 3470-3477.doi: 10.3864/j.issn.0578-1752.2013.16.018

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Development and Identification of ELISA Kit for Norfloxacin Determination

 XING  Guang-Xu-12, WANG  Guo-Dong-3, HU  Xiao-Fei-2, WANG  Fang-Yu-2, DENG  Rui-Guang-2, ZHANG  Gai-Ping-2   

  1. 1.College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450000
    2. Key Lab for Animal Immunology, Henan Academy of Agricultural Science, Zhengzhou 450002
    3.Anyang Institute of Technology, Anyang 455000, Henan
  • Received:2013-03-13 Online:2013-08-15 Published:2013-06-25

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Abstract: 【Objective】The objective of this experiment is to develop an ELISA kit for norfloxacin measurement. 【Method】Based on the prepared norfloxacin monoclonal antibody, the best work concentration of monoclonal antibody and HRP labeled antibody were selected by matrix method to develop an indirect competition ELISA kit for norfloxacin detection, and the properties of which also were examined and conformed. 【Result】The kit has average IC50 of 5.18 μg•L-1, a sensitivity of 0.31 μg•L-1, and the detection limit of 1.00 μg•L-1. The kit has different cross-reactivity with peifloxacin, ofloxacin, ciprofloxacin, lomefloxacin, oxilinic acid. The recoveries of norfloxacin in milk was 103.23%, and the CV was 9.33%. Among the detection results by three different batch kits, the inter-assay and intra-assay coefficient variation were all below 10%. The kit could remain effective for more than six months when stored at 4℃. There was no significant difference in the tested value of norfloxacin in milk between with NEO-kit and with LC-MS/ MS (P≥0.05). Furthermore, the tested value of norfloxacin in practice milk was no difference between with NEO-kit and with LC-MS/MS (P≥0.05). 【Conclusion】 A Nor-kit with high sensitivity, specificity and accuracy was developed in this experiment.

Key words: Norfloxacin , monoclonal antibody , ELISA , kit

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