›› 2007, Vol. 34 ›› Issue (6): 79-81.

• 疾病防治 • 上一篇    下一篇

达氟沙星间接竞争ELISA检测方法的建立

李振华1,2, 斯坎达尔·买合木提1, 魏东2,3, 张乃生2

  

  1. 1.新疆农业大学动医学院,乌鲁木齐 830052;2.吉林大学畜牧兽医学院,长春 130062;3.河北北方学院动物医学系,张家口 075000
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2007-06-20 发布日期:2007-06-20

Establishment of the Method of Detecting the Residues of DFLX by Indirect Competitive Enzyme-linked Immunosorbent Assay

LI Zhenhua1,2, SIKANDAER Maihemuti1,WEI Dong2,3,ZHANG Naisheng2
  

  1. 1.College of Animal Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052,China; 2. College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062,China; 3.Department of Veterinary Medicine, North Hebei University, Zhangjiakou 075000,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2007-06-20 Published:2007-06-20

摘要: 将达氟沙星(danofloxacin, DFLX)与牛血清白蛋白(BSA)、卵清白蛋白(OVA)偶联分别作为免疫原与包被原, 达氟沙星为竞争的半抗原,建立间接竞争ELISA检测方法。试验结果表明,理想的包被抗原浓度为1.25 μg/L, 多抗(DFLX-PcAb)工作浓度为1∶10000,酶标二抗的工作浓度为1∶4000,最适检测范围为0.1~10 ng/L,最低检测限为0.2 ng/L, 批内和批间变异系数分别为3.81%和6.25%。得到回归方程y=0.7975-0.125x(R2=0.989)和标准曲线,从而建立了DFLX的快速检测残留的间接竞争酶联免疫吸附试验(ci-ELISA)。

关键词: 达氟沙星; 残留检测; 间接竞争ELISA

Abstract: DFLX hapten was coupled to BSA and OVA, DFLX-OVA was coating antigen, DFLX-BSA was immunity antigen. The indirect competitive ELISA for DFLX was developed. The result of experiment that the optimal concentration of the coating antigen, polyclonal antibody against DFLX and sheep anti-rabbit IgG were 1.25 μg/L, 1∶10000 and 1∶4000 respectively. Quantization of the DFLX was linear from 0.1~10 ng/L. with the detection limit being 0.2 ng/l. The coefficients of variation of intra-assay and inter-assay were 3.81%and 6.25% respectively. Then, the regression equation was y=0.7975-0.125x(R2=0.989)and there was a standard curve. So indirect competitive enzyme-linked immunosorbent assay was established to detect rapidly the residues of DFLX in food of animal origin.

Key words: danofloxacin(DFLX); residue detection; indirect competitive ELISA

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