食品科学 ›› 2011, Vol. 32 ›› Issue (10): 110-114.doi: 10.7506/spkx1002-6630-201110025

• 分析检测 • 上一篇    下一篇

恩诺沙星时间分辨荧光免疫分析方法的建立

赵莉莉,黄 飚,王晓岚,*,宓晓黎,金 坚,陆茂林   

  1. 1.江南大学医药学院 2.江苏省微生物研究所有限责任公司 3.江苏省原子医学研究所
  • 出版日期:2011-05-25 发布日期:2011-04-08
  • 基金资助:
    国家“863”计划项目(2008AA10Z425)

Establishment of a Time-resolved Fluoroimmunoassay for Measuring Enrofloxacin

ZHAO Li-li1,2,HUANG Biao3,WANG Xiao-lan1,*,MI Xiao-li2,JIN Jian1,LU Mao-lin2   

  1. 1. School of Medicine and Pharmaceutics, Jiangnan University, Wuxi 214122, China;2. Jiangsu Institute of Microbiology Co. Ltd., Wuxi 214063, China;3. Jiangsu Institute of Nuclear Medicine, Wuxi 214063, China
  • Online:2011-05-25 Published:2011-04-08

摘要: 目的:采用时间分辨荧光免疫分析(time-resolved fluoroimmunoassay,TRFIA)技术建立高灵敏的恩诺沙星(enrofloxacin,ENR)快速检测方法。方法:以包被抗原(ENR-OVA)包被微孔板,与游离的恩诺沙星共同竞争抗恩诺沙星抗体,以铕(Eu3+)标记的羊抗兔抗体进行示踪。结果:方法的批内变异系数小于10%、批间变异系数小于15%,平均回收率为91.62%,灵敏度为5ng/L,可测范围为0.01~100μg/L,ED20、ED50和ED80分别为0.088、3.40μg/L和32.81μg/L。结论:ENR-TRFIA方法稳定性好、可测范围宽,具有很好的应用前景。

关键词: 恩诺沙星, 时间分辨荧光免疫分析, 兽药残留

Abstract: A time-resolved fluoroimmunoassay (TRFIA) was developed for the rapid and sensitive detection of enrofloxacin (ENR). In indirect TRFIA format, the envelope antigen composed of ENR and ovalbumin (OVA) was coated onto the microplate, and followed by the incubation with free ENR and anti-ENR polyclonal antibody. A goat anti-rabbit IgG conjugated with europium (Eu3+) was used for the detection. The intra-assay CV (coefficient of variation) of the developed method was less than 10% and the inter-assay CV less than 15%. The average recovery rate was 91.62% and the sensitivity was 5 ng/L. The antibody had a linear response over the range from 0.01μg/L to 100μg/L. The ED20, ED50 and ED80 (80%, 50% and 20% effective doses) were 0.088, 3.40μg/ L and 32.81μg/ L, respectively. The high stability and broad detection range allows the TRFIA method to have promising application prospects.

Key words: enrofloxacin, time-resolved fluoroimmunoassay, veterinary drug residues

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