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A Sensitive Nucleic Acid Detection Platform for Foodborne Pathogens Based on CRISPR-Cas13a System Combined with Polymerase Chain Reaction

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Abstract

Foodborne diseases attributable to foodborne pathogens directly impact food safety and public health. This study aimed to establish a nucleic acid detection platform for sensitive detection of foodborne pathogens. Staphylococcus aureus (ATCC® 29213™) as a model pathogen was used to evaluate the detection performances of the developed method. The advantages of combining rapid acquisition of amplification products, crRNA programmability, and collateral cleavage effects of Cas13a were applied to achieve the detection goal. The findings implied that the target gene as low as 2.4 copies µL−1 could be detected by the method. Furthermore, compared with standard real-time PCR, the PCR-Cas13a-based nucleic acid detection method achieved satisfactory accuracy (99.44% for 180 water samples and 99.00% for 200 milk samples). With the desired simplicity, specificity, sensitivity, and accuracy, this detection strategy could be extended for the detection of other pathogens in food safety, environmental analysis, and medical diagnostics.

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Data Availability

All data generated or analyzed during this study are available from the corresponding author on reasonable request.

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Acknowledgements

The authors would like to thank Suleiman Abdulrahman and Waqas Ahmad for the language correction of the manuscript.

Funding

This work has been financially supported by Key R&D Program of Jiangsu Province (BE2020379), Jiangsu Provincial Agricultural S&T Innovation Foundation (CX(20)2015), Project of Faculty of Agricultural Equipment of Jiangsu University, Postgraduate Research and Practice Innovation Program of Jiangsu Province (KYCX19_1630), and Interdisciplinary Sciences Project of Nanyang Institute of Technology.

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Authors and Affiliations

  1. School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013, People’s Republic of China

    Rui Liu, Yunlian Zhang & Quansheng Chen

  2. College of Electrical and Electronic Engineering, Wenzhou University, Wenzhou, 325035, People’s Republic of China

    Shujat Ali

  3. School of Life Sciences, Jiangsu University, Zhenjiang, 212013, People’s Republic of China

    Dong Huang & Peng Lü

  4. Zhang Zhongjing School of Chinese Medicine, Nanyang Institute of Technology, Nanyang, 473004, People’s Republic of China

    Rui Liu

Authors
  1. Rui Liu
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  2. Shujat Ali
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  3. Dong Huang
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  4. Yunlian Zhang
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  5. Peng Lü
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Contributions

Rui Liu: Conceptualization; methodology; software; writing, original draft preparation; writing, review and editing.

Shujat Ali: Data curation, writing—review and editing.

Dong Huang: Investigation, resources, data curation.

Yunlian Zhang: Software, validation, formal analysis.

Peng Lü: Conceptualization, investigation, supervision, validation.

Quansheng Chen: Supervision, project administration, funding acquisition.

Corresponding authors

Correspondence to Peng Lü or Quansheng Chen.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Conflict of Interest

Rui Liu declares that she has no conflict of interest. Shujat Ali declares that he has no conflict of interest. Dong Huang declares that he has no conflict of interest. Yunlian Zhang declares that she has no conflict of interest. Peng Lü declares that he has no conflict of interest. Quansheng Chen declares that he has no conflict of interest.

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Liu, R., Ali, S., Huang, D. et al. A Sensitive Nucleic Acid Detection Platform for Foodborne Pathogens Based on CRISPR-Cas13a System Combined with Polymerase Chain Reaction. Food Anal. Methods 16, 356–366 (2023). https://doi.org/10.1007/s12161-022-02419-8

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  • DOI: https://doi.org/10.1007/s12161-022-02419-8

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