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On-line Access: 2010-01-01

Received: 2009-07-21

Revision Accepted: 2009-10-19

Crosschecked: 2009-12-16

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Journal of Zhejiang University SCIENCE B 2010 Vol.11 No.1 P.52-60

10.1631/jzus.B0900215


Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine


Author(s):  Jian-xiang WU, Shao-en ZHANG, Xue-ping ZHOU

Affiliation(s):  Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China; more

Corresponding email(s):   wujx@zju.edu.cn

Key Words:  Streptomycin, Monoclonal antibody, Antibiotic residue, Enzyme-linked immunosorbent assay (ELISA), Colloidal gold-based immunochromatographic assay (CGIA)


Jian-xiang WU, Shao-en ZHANG, Xue-ping ZHOU. Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine[J]. Journal of Zhejiang University Science B, 2010, 11(1): 52-60.

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author="Jian-xiang WU, Shao-en ZHANG, Xue-ping ZHOU",
journal="Journal of Zhejiang University Science B",
volume="11",
number="1",
pages="52-60",
year="2010",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B0900215"
}

%0 Journal Article
%T Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine
%A Jian-xiang WU
%A Shao-en ZHANG
%A Xue-ping ZHOU
%J Journal of Zhejiang University SCIENCE B
%V 11
%N 1
%P 52-60
%@ 1673-1581
%D 2010
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B0900215

TY - JOUR
T1 - Monoclonal antibody-based ELISA and colloidal gold-based immunochromatographic assay for streptomycin residue detection in milk and swine urine
A1 - Jian-xiang WU
A1 - Shao-en ZHANG
A1 - Xue-ping ZHOU
J0 - Journal of Zhejiang University Science B
VL - 11
IS - 1
SP - 52
EP - 60
%@ 1673-1581
Y1 - 2010
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B0900215


Abstract: 
A protein conjugate of streptomycin (streptomycin-bovine serum albumin (BSA) conjugate) was prepared and used as immunogen to produce monoclonal antibodies (MAb). One hybridoma secreting anti-streptomycin MAb was obtained and then used to produce MAb. The MAb named 13H5 showed the 50% maximal inhibitory concentration (IC50) value of 4.65 ng/ml and the IC20 value of 0.21 ng/ml in phosphate buffered saline (PBS). At optimum conditions, an indirect competitive enzyme-linked immunosorbent assay (ELISA) and a colloidal gold-based immunochromatographic assay (CGIA) were developed and applied to detect streptomycin residues in milk and swine urine samples. The developed ELISA showed that the minimum detection limit was 2.0 and 1.9 ng/ml for milk and swine urine samples, respectively, without obvious cross-reactivity to other tested antibiotics except dihydrostreptomycin which gave a 118.32% cross reaction value. Milk and swine urine samples spiked with streptomycin at 10, 50, 100 and 200 ng/ml were analyzed by the established ELISA. The mean recovery of streptomycin was from 81.9% to 105.5% and from 84.3% to 92.2% for milk and swine urine, respectively. The optimized CGIA showed that the minimum detection limit was 20.0 ng/ml for milk and swine urine samples. The results of spiked analysis and specific analysis demonstrate that the CGIA could be applicable for screening milk and swine urine samples for the presence of streptomycin residues on-site. The established ELISA and CGIA allow the rapid, low-cost, and sensitive determination of streptomycin residues in food samples.

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