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CLC number: TS207.3

On-line Access: 2012-07-05

Received: 2011-12-31

Revision Accepted: 2012-02-29

Crosschecked: 2012-06-19

Cited: 12

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Citations:  Bibtex RefMan EndNote GB/T7714

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Journal of Zhejiang University SCIENCE B 2012 Vol.13 No.7 P.545-554

http://doi.org/10.1631/jzus.B1200001


Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk


Author(s):  Guo-ying Fan, Ruo-song Yang, Jin-qing Jiang, Xin-yao Chang, Jun-jie Chen, Yong-hua Qi, Shi-xiu Wu, Xue-feng Yang

Affiliation(s):  College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China; more

Corresponding email(s):   jjq5678@126.com

Key Words:  Norfloxacin, Fluoroquinolones, Indirect competitive ELISA, Class-specificity, Milk


Guo-ying Fan, Ruo-song Yang, Jin-qing Jiang, Xin-yao Chang, Jun-jie Chen, Yong-hua Qi, Shi-xiu Wu, Xue-feng Yang. Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk[J]. Journal of Zhejiang University Science B, 2012, 13(7): 545-554.

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author="Guo-ying Fan, Ruo-song Yang, Jin-qing Jiang, Xin-yao Chang, Jun-jie Chen, Yong-hua Qi, Shi-xiu Wu, Xue-feng Yang",
journal="Journal of Zhejiang University Science B",
volume="13",
number="7",
pages="545-554",
year="2012",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1200001"
}

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%A Guo-ying Fan
%A Ruo-song Yang
%A Jin-qing Jiang
%A Xin-yao Chang
%A Jun-jie Chen
%A Yong-hua Qi
%A Shi-xiu Wu
%A Xue-feng Yang
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%P 545-554
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T1 - Development of a class-specific polyclonal antibody-based indirect competitive ELISA for detecting fluoroquinolone residues in milk
A1 - Guo-ying Fan
A1 - Ruo-song Yang
A1 - Jin-qing Jiang
A1 - Xin-yao Chang
A1 - Jun-jie Chen
A1 - Yong-hua Qi
A1 - Shi-xiu Wu
A1 - Xue-feng Yang
J0 - Journal of Zhejiang University Science B
VL - 13
IS - 7
SP - 545
EP - 554
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PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1200001


Abstract: 
Modified 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) method was employed to synthesize the artificial antigen of norfloxacin (NOR), and New Zealand rabbits were used to produce anti-NOR polyclonal antibody (pAb). Based on the checkerboard titration, an indirect competitive enzyme-linked immunosorbent assay (icELISA) standard curve was established. This assay was sensitive and had a working range from 0.12 to 68.40 ng/ml, with the half maximal inhibitory concentration (IC50) and limit of detection (LOD) values of 2.7 ng/ml and 0.06 ng/ml, respectively. The produced pAb exhibited high cross-reactivity to fluoroquinolones (FQs) tested, and the IC50 values to enoxacin, ciprofloxacin, and pefloxacin were 3.1, 3.4, and 4.1 ng/ml, respectively. It also indicated that the concentrations of NaOH and methanol in assay buffer should not be higher than 10% and 30%. When spiked in milk at 5, 20, and 50 ng/ml, the recoveries for NOR, enoxacin, ciprofloxacin, and pefloxacin ranged 90.5%–98.0%, 84.0%–95.2%, 94.0%–106.0%, and 89.5%–100.0%, respectively. The results suggest that this class-specific pAb-based icELISA could be utilized for the primary screening of FQ residues in animal-original products.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

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