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Silver/copper bimetallic nanoclusters integrating with cryonase-assisted target recycling amplification detection of Salmonella typhimurium

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Abstract

An unsophisticated fluorescence-enabled strategy is brought forward to process the highly sensitive fluorescence detection of Salmonella typhimurium (S. typhimurium) which based on polyethyleneimine (PEI)-templated silver/copper nanoclusters (Ag/CuNCs) (λ excitation = 334 nm and λ emission = 466 nm) with cryonase-assisted target recycling amplification. The Ag/CuNCs nanoclusters are synthesized as fluorescent materials due to their strong and stable fluorescence characteristics and are modified with S. typhimurium aptamers to form aptamer-Ag/CuNCs probes. The probes can be adsorbed on the surface of quenching agents-polydopamine nanospheres (PDANSs), thereby inducing fluorescence quenching of the probes. Once the aptamers are bound to the target, the aptamers/targets complexes are separated from the PDANSs surface, and the Ag/CuNCs recover the fluorescence signal. The released complexes will immediately be transformed into a substrate digested by cryonase (an enzyme that can digest all types of nucleic acids), and the released targets are bound to another aptamers to initiate the next round of cleavage. This reaction will be repeated continuously until all relevant aptamers are consumed and all Ag/CuNCs are released, resulting in a significant amplification of the fluorescence signal and improved sensitivity. Using Ag/CuNCs as fluorescent probes combined with cryonase-assisted amplification strategy, the fluorescence aptasensor is constructed with detection limits as low as 3.8 CFU mL−1, which is tenfold better than without the cryonase assistance. The method developed has been applied to milk, orange juice, chicken, and egg white samples with excellent selectivity and accuracy providing an approach for the early and rapid detection of S. typhimurium in food.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 31872909, No.31772068).

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Authors and Affiliations

  1. College of Agricultural Engineering and Food Science, Shandong University of Technology, No. 266 Xincun Xilu, Zibo, 255049, Shandong, China

    Shouyi Dou, Mengyue Liu, Fengjuan Zhang, Baoxin Li, Yuhao Zhang, Falan Li, Yemin Guo & Xia Sun

  2. Shandong Provincial Engineering Research Center of Vegetable Safety and Quality Traceability, No. 266 Xincun Xilu, Zibo, 255049, Shandong, China

    Shouyi Dou, Mengyue Liu, Fengjuan Zhang, Baoxin Li, Yuhao Zhang, Falan Li, Yemin Guo & Xia Sun

  3. Zibo City Key Laboratory of Agricultural Product Safety Traceability, No. 266 Xincun Xilu, Zibo, 255049, Shandong, China

    Shouyi Dou, Mengyue Liu, Fengjuan Zhang, Baoxin Li, Yuhao Zhang, Falan Li, Yemin Guo & Xia Sun

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  1. Shouyi Dou
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  2. Mengyue Liu
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  6. Falan Li
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  7. Yemin Guo
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  8. Xia Sun
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Contributions

Shouyi Dou, Mengyue Liu, and Xia Sun designed the experiments. Shouyi Dou wrote the manuscript and performed the experiments. Falan Li and Yemin Guo provided writing assistance for the manuscript. Fengjuan Zhang, Baoxin Li, and Yuhao Zhang analyzed the data and drafted the manuscript. All authors have approved and read the final manuscript.

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Correspondence to Yemin Guo or Xia Sun.

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Highlights

1. The silver/copper bimetal nanoclusters combined with cryonase-assisted amplification are adopted to accomplish the highly sensitive detection of Salmonella typhimurium.

2. The aptamer-Ag/CuNCs fluorescent probes are successfully constructed for fluorescence resonance energy transfer to achieve Salmonella typhimurium rapid and highly sensitive detection.

3. The detection limit is as low as 3.8 CFU mL−1, which is 10 times lower than without the cryonase assistance.

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Supplementary file1 (DOCX 12838 KB)

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Dou, S., Liu, M., Zhang, F. et al. Silver/copper bimetallic nanoclusters integrating with cryonase-assisted target recycling amplification detection of Salmonella typhimurium. Microchim Acta 190, 403 (2023). https://doi.org/10.1007/s00604-023-05973-y

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