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Novel Single Primer Isothermal Amplification Method for the Visual Detection of Vibrio parahaemolyticus

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Abstract

As a typical halophilic bacterium, Vibrio parahaemolyticus (V. parahaemolyticus) is regarded as one of the marine seafood-borne pathogens causing severe illnesses in humans and aquatic animals. Herein, a novel single primer isothermal amplification (SPIA) technique which was modified with SYBR Green II was successfully developed for the visual detection of V. parahaemolyticus. Interestingly, the visual superiority of SPIA can be achieved by fluorescence or precipitate to analyze the target pathogens in the seafood samples. The experimental results showed that all of V. parahaemolyticus strains were positive by using the novel SPIA assay, whereas the non-V. parahaemolyticus counterparts were negative. Furthermore, the limit of detection (LOD) of the SPIA assay was 5.7 CFU/mL in pure culture and 42 CFU/g in raw oysters by the visual fluorescence, respectively. Compared with the standard method, the developed SPIA approach achieved the satisfactory results with the characteristics of 100% sensitivity, 98.21% specificity, and 99.17% accuracy in the detection of 241 seafood samples. The developed SPIA method has been demonstrated as a rapid, specific, sensitive, and visual tool for the detection of V. parahaemolyticus, which could be adopted for routine surveillance of food-borne pathogens in the seafood samples.

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Funding

This study was supported by the National Natural Science Foundation of China (31371772), the Natural Science Foundation of Hebei Province (C2019204342), the Scientific Research Program of Hebei Education Department (ZD2017237), the Key Research and Development Project of Hebei Province of China (18275501D), the Food Processing Discipline Group of Hebei Agricultural University (No. 2021-06) and the Program of Introduced Doctor of Hebei Agricultural University (ZD201623).

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Author notes

  1. Qian Yang, Wei Guo and Yi Liu contributed equally to this work.

Authors and Affiliations

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

    Qian Yang, Wei Guo, Yunzhe Zhang, Ruoyang Ming, Yaowu Yuan, Jianxin Tan & Wei Zhang

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

    Qian Yang & Wei Zhang

  3. Baoding Product Quality Supervision and Inspection Institute, Baoding, 071000, Hebei, China

    Yi Liu

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

    Wei Zhang

Authors
  1. Qian Yang
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  2. Wei Guo
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  3. Yi Liu
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  4. Yunzhe Zhang
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  7. Jianxin Tan
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  8. Wei Zhang
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Contributions

Qian Yang, Wei Guo, and Yi Liu contributed equally to this work. Qian Yang and Ruoyang Ming developed methodology and performed data acquisition. Wei Guo contributed to the conception, revision, and editing. Yi Liu and Yunzhe Zhang contributed to the methodology and analysis. Wei Zhang, Yaowu Yuan, and Jianxin Tan contributed to the study conception and design. The draft of the manuscript was written by Qian Yang. Wei Zhang made critical corrections. All authors read and approved the final manuscript.

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Correspondence to Wei Zhang.

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Yang, Q., Guo, W., Liu, Y. et al. Novel Single Primer Isothermal Amplification Method for the Visual Detection of Vibrio parahaemolyticus. Food Anal. Methods 14, 1995–2002 (2021). https://doi.org/10.1007/s12161-021-02033-0

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  • DOI: https://doi.org/10.1007/s12161-021-02033-0

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