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Aptamer-functionalized magnetic nanoparticles for simultaneous fluorometric determination of oxytetracycline and kanamycin

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Abstract

This work describes a method for the simultaneous detection of oxytetracycline (OTC) and kanamycin (KMY) using aptamers acting as both recognition and separation elements, and complementary oligonucleotides labeled with a green emitting fluorophore (carboxyfluorescein, FAM) and a yellow emitting fluorophore (carboxy-X-rhodamine, ROX), respectively, as signal labels. An OTC aptamer and a KMY aptamer were immobilized on the surface of magnetic nanoparticles (MNPs) via avidin-biotin chemistry. The aptamers preferentially bind their respective targets and thereby cause the upconcentration of analytes. However, in their absence they bind fluorescently-tagged complementary oligonucleotide later added to the reaction system. This cause the NPs to become fluorescent, with emission peaks located at 520 and 608 nm, respectively. The effects of the concentration of avidin, aptamer, complementary oligonucleotide, incubation temperature and incubation time were optimized. Under the optimal conditions, linear relationships were obtained in the range of 1–50 ng∙mL−1 for OTC and KMY, with limits of detection of 0.85 ng∙mL−1 and 0.92 ng∙mL−1, respectively. The method was applied to the analysis of pork, milk, and honey samples spiked with OTC and MKY. Recoveries ranged from 76.5 to 94.7 % and 77.8 to 93.1 %, respectively, and the relative standard deviation was <10.0 %.

This work describes an assay for the simultaneous detection of oxytetracycline and kanamycin using aptamer-modified as both recognition and separation elements, and complementary oligonucleotide labeled with FAM and ROX, respectively, as signal labels. The developed method possesses high sensitivity and selectivity, and short analysis time.

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Acknowledgments

This work is financially supported by the Science & Technology to guide project of Xinjiang Academy of Agriculture and Reclamation Science (62YYD201309).

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Authors and Affiliations

  1. Institute of Animal Husbandry and Veterinary Science, Xinjiang Academy of Agriculture and Reclamation Science, Wuyi Road No. 221, Shihezi, 832000, People’s Republic of China

    Changbin Liu

  2. Analysis and Testing Center, Xinjiang Academy of Agriculture and Reclamation Science, Wuyi Road No. 221, Shihezi, 832000, People’s Republic of China

    Chunxia Lu, Zonggui Tang, Xia Chen, Guohong Wang & Fengxia Sun

Authors
  1. Changbin Liu
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  2. Chunxia Lu
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  3. Zonggui Tang
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  4. Xia Chen
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  5. Guohong Wang
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  6. Fengxia Sun
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Corresponding authors

Correspondence to Chunxia Lu or Fengxia Sun.

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Changbin Liu and Chunxia Lu contributed equally to this work.

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Liu, C., Lu, C., Tang, Z. et al. Aptamer-functionalized magnetic nanoparticles for simultaneous fluorometric determination of oxytetracycline and kanamycin. Microchim Acta 182, 2567–2575 (2015). https://doi.org/10.1007/s00604-015-1628-z

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  • DOI: https://doi.org/10.1007/s00604-015-1628-z

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