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Saltatory rolling circle amplification assay coupled with photosensitization method for rapid and sensitive detection of Salmonella in food

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Abstract

Colorimetry has emerged as a promising option for pathogenic detection. Herein, a novel method based on saltatory rolling circle amplification (SRCA) combined with photosensitization colorimetric assay (SRCA-C) was developed for rapid and sensitive detection of Salmonella in food. The target identification and signal amplification are realized by SRCA to produce a large amount of double-stranded DNA. Subsequently, SYBR Green I as photosensitizer and 3,3',5,5"-tetramethylbenzidine (TMB) as colorimetric oxidase substrate are added to the colorimetric reaction system. The photosensitization reaction occurs under cyan LED irradiation, and TMB is oxidized to convert the double-stranded DNA signal into absorbance signal, thus realizing the detection of Salmonella. This colorimetric method benefits from convenience (label-free and no requirement of costly and complex equipment), short detection time (DNA amplification for 50 min and color development for 15 min) and high sensitivity (detected as low as 13 CFU/mL of Salmonella in pure culture). Five target strains and 10 nontarget strains were used to validate the specificity of the method. The recovery of Salmonella in the spiked milk samples ranged from 98.4% to 118.4%. Compared with the standard culture method, this method was 100% sensitivity, 98.2% specificity, and 98.3% accuracy for the detection of the real samples. Overall, this method highlights the importance for convenient, rapid, and sensitive detection of Salmonella in food.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (32172288 and 31371772), the Natural Science Foundation of Hebei Province (C2019204342), the Scientific Research Program of Hebei Education Department (QN2022073), the Advanced Talents Incubation Program of the Hebei University (521100222024), Medical Science Foundation of Hebei University (2022A06), the Central Government Guided Local Science and Technology Development Fund Projects (Basic Research Projects) (216Z5501G), the Talent Introduction Project of Hebei Province (360-0803-JSN-3YGS), and the Food Processing Discipline Group of Hebei Agricultural University (No. 2021-06).

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  1. Huibin Wang and Qian Yang have contributed equally to this work.

Authors and Affiliations

  1. College of Food Science and Technology, Hebei Agricultural University, Baoding, 071001, China

    Huibin Wang, Yunzhe Zhang & Wei Zhang

  2. School of Public Health, Hebei University, Baoding, 071002, China

    Qian Yang

  3. College of Science and Technology, Hebei Agricultural University, Cangzhou, 061100, China

    Hui Xu & Xin Lu

  4. College of Life Sciences, Hebei Agricultural University, Baoding, 071001, China

    Wei Zhang

  5. Hebei Key Laboratory of Analysis and Control of Zoonotic Pathogenic Microorganism, Baoding, 071001, China

    Wei Zhang

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  1. Huibin Wang
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  3. Hui Xu
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  4. Yunzhe Zhang
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  5. Xin Lu
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Contributions

HW: Writing—Original Draft, Conceptualization, Methodology, Investigation. QY: Validation, Funding acquisition, Data curation, Writing—Review & Editing. HX: Formal analysis, Software. YZ: Resources, Visualization. XL: Data curation, Software. WZ: Conceptualization, Methodology, Supervision, Funding acquisition, Project administration.

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Correspondence to Wei Zhang.

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Wang, H., Yang, Q., Xu, H. et al. Saltatory rolling circle amplification assay coupled with photosensitization method for rapid and sensitive detection of Salmonella in food. Eur Food Res Technol 249, 2067–2075 (2023). https://doi.org/10.1007/s00217-023-04278-x

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