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A detection method of Escherichia coli O157:H7 based on immunomagnetic separation and aptamers-gold nanoparticle probe quenching Rhodamine B’s fluorescence

Escherichia coli O157:H7 detection method based on IMS and Apt-AuNPs probe quenching Rho B’ s fluorescence

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Abstract

This research aimed to detect Escherichia coli O157:H7 in milk based on immunomagnetic probe separation technology and quenching effect of gold nanoparticles to Rhodamine B. Streptavidin-modified magnetic beads (MBs) were combined with biotin-modified antibodies to capture E. coli O157:H7 specifically. Gold nanoparticle (AuNPs) was incubated with sulfhydryl-modified aptamers (SH-Aptamers) to obtain the Aptamers-AuNPs probe. After magnetic beads captured target bacteria and formed a sandwich structure with the gold nanoprobe, Rhodamine B was added into complex to obtain fluorescent signal changes. Our results demonstrated that the established method could detect E. coli O157:H7 in the range of 101–107 CFU/mL, and the limit of detection (LOD) was 0.35 CFU/mL in TBST buffer (pH = 7.4). In milk simulation samples, the LOD of this method was 1.03 CFU/mL. Our research provides a promising approach on the detection of E. coli O157:H7.

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Acknowledgements

This work was financially supported by the Jilin Province Development and Reform Commission (the Grant Number: 2020C038-7), the Development of Science and Technology, Jilin Province, China (Grant Number: 2018010195JC), National Natural Science Foundation of China (Grant Number: 81401721), the Education Department of Jilin Province, China (Grant Number: JJKH20180239KJ) and the Health and Family Planning Commission of Jilin Province (Grant Number: 2017J074)

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Authors and Affiliations

  1. School of Public Health, Jilin University, 130021, Changchun, China

    Fengnan Lian, Dan Wang, Shuo Yao, Lirui Ge, Yue Wang, Yuyi Zhao, Jinbin Zhao, Xiuling Song, Chao Zhao, Jinhua Li, Yajuan Liu, Minghua Jin & Kun Xu

  2. Jilin Engineering Research Center of Public Health Detection, 130021, Changchun, China

    Fengnan Lian, Dan Wang, Shuo Yao, Lirui Ge, Yue Wang, Yuyi Zhao, Jinbin Zhao, Xiuling Song, Chao Zhao, Jinhua Li, Yajuan Liu, Minghua Jin & Kun Xu

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  1. Fengnan Lian
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  2. Dan Wang
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  3. Shuo Yao
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  4. Lirui Ge
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  5. Yue Wang
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  6. Yuyi Zhao
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  7. Jinbin Zhao
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  8. Xiuling Song
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  9. Chao Zhao
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  10. Jinhua Li
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  11. Yajuan Liu
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  12. Minghua Jin
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  13. Kun Xu
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Contributions

FL, KX, CZ conceived and designed the study. FL, DW, SY, lirui Ge performed the assay of experimental detection. JZ, YZ and YW analyzed the data. FL drafted the manuscript. CZ and XS provided constructive opinions and suggestions. KX, CZ, XS, YL, JL and MJ reviewed and made improvements in the manuscript. All authors read and approved the final version of the paper.

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Correspondence to Kun Xu.

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Lian, F., Wang, D., Yao, S. et al. A detection method of Escherichia coli O157:H7 based on immunomagnetic separation and aptamers-gold nanoparticle probe quenching Rhodamine B’s fluorescence. Food Sci Biotechnol 30, 1129–1138 (2021). https://doi.org/10.1007/s10068-021-00947-3

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