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Development of a lateral flow fluorescent microsphere immunoassay for the determination of sulfamethazine in milk

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Abstract

The fluorescent microsphere has been increasingly used as detecting label in immunoassay because of its stable configuration, high fluorescence intensity, and photostability. In this paper, we developed a novel lateral flow fluorescent microsphere immunoassay (FMIA) for the determination of sulfamethazine (SMZ) in milk in a quantitative manner with high sensitivity, selectivity, and rapidity. A monoclonal antibody to SMZ was covalently conjugated with the carboxylate-modified fluorescent microsphere, which is polystyrene with a diameter of 200 nm. Quantitative detection of SMZ in milk was accomplished by recording the fluorescence intensity of microspheres captured on the test line after the milk samples were diluted five times. Under optimal conditions, the FMIA displays a rapid response for SMZ with a limit of detection of as low as 0.025 ng mL−1 in buffer and 0.11 μg L−1 in milk samples. The FMIA was then successfully applied on spiked milk samples and the recoveries ranged from 101.1 to 113.6 % in the inter-batch assay with coefficient of variations of 6.0 to 14.3 %. We demonstrate here that the fluorescent microsphere-based lateral flow immunoassay (LFIA) is capable of rapid, sensitive, and quantitative detection of SMZ in milk.

Schematic illustration of the strategy for sulfamethazine detection using a lateral flow fluorescent microsphere immunoassay

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Acknowledgments

This work is supported by grants from Trans-Century Training Programme Foundation for the Talents by the Ministry of Education (NCET-12-0529), Key Projects in the National Science and Technology Pillar Program during the 12th Five-Year Plan Period (2011BAK10B01, 2012BAK17B00), and Sino-Russian International Scientific and Technological Cooperation (2011DFR30470).

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Authors and Affiliations

  1. College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, Beijing, 100193, China

    Rui Chen, Heng Li, Han Zhang, Suxia Zhang, Weimin Shi, Jianzhong Shen & Zhanhui Wang

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  1. Rui Chen
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  2. Heng Li
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  3. Han Zhang
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  4. Suxia Zhang
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  5. Weimin Shi
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  6. Jianzhong Shen
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  7. Zhanhui Wang
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Corresponding author

Correspondence to Zhanhui Wang.

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Chen, R., Li, H., Zhang, H. et al. Development of a lateral flow fluorescent microsphere immunoassay for the determination of sulfamethazine in milk. Anal Bioanal Chem 405, 6783–6789 (2013). https://doi.org/10.1007/s00216-013-7150-4

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  • DOI: https://doi.org/10.1007/s00216-013-7150-4

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