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Visual detection and microplate assay for Staphylococcus aureus based on aptamer recognition coupled to tyramine signal amplification

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Abstract

We have developed a specific method for the visual detection of Staphylococcus aureus based on aptamer recognition coupled to tyramine signal amplification technology. A biotinylated aptamer specific for S. aureus was immobilized on the surface of the wells of a microplate via biotin-avidin binding. Then, the target bacteria (S. aureus), the biotinylated-aptamer-streptavidin-HRP conjugates, biotinylated tyramine, hydrogen peroxide and streptavidin-HRP were successively placed in the wells of the microplate. After adding TMB reagent and stop solution, the intensity of the yellow reaction product can be visually inspected or measured with a plate reader. Under optimized conditions, there is a linear relationship between absorbance at 450 nm and the concentration of S. aureus in the 10 to 107 cfu mL−1 concentration range (with an R2 of 0.9976). The limit of detection is 8 cfu mL−1.

A visual detection method for Staphylococcus aureus was based on aptamer recognition coupled to tyramine signal amplification. The linear range was from 10 to 107 cfu mL-1 and the limit of detection was 8 cfu mL-1.

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Acknowledgments

This work was partly supported by the S&T Supporting Project of Jiangsu (BE2011621, BE2010679, BE2012614), the National S&T Support Program of China (2012BAK08B01, 2011YQ170067), NSFC (21375049) NCET-11-0663 and JUSRP51309A.

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Synergetic Innovation Center of Food Safety and Nutrition, School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Jinglei Yuan, Can Li, Xiaoyuan Ma, Yu Xia, Jie Chen & Zhouping Wang

  2. Zhangjiagang Entry-Exit Inspection and Quarantine Bureau, Zhangjiangang, 215600, China

    Ye Yu

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  1. Jinglei Yuan
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  2. Ye Yu
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  3. Can Li
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  4. Xiaoyuan Ma
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  5. Yu Xia
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  6. Jie Chen
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  7. Zhouping Wang
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Correspondence to Xiaoyuan Ma or Zhouping Wang.

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Yuan, J., Yu, Y., Li, C. et al. Visual detection and microplate assay for Staphylococcus aureus based on aptamer recognition coupled to tyramine signal amplification. Microchim Acta 181, 321–327 (2014). https://doi.org/10.1007/s00604-013-1120-6

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  • DOI: https://doi.org/10.1007/s00604-013-1120-6

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