Abstract
Vibrio parahaemolyticus has become an important public threat to human health. Rapid and robust pathogen diagnostics are necessary for monitoring its outbreak and spreading. Herein, we report an assay for the detection of V. parahaemolyticus based on recombinase aided amplification (RAA) combined with lateral flow dipstick (LFD), namely RAA-LFD. The RAA-LFD took 20 min at 36~38 ℃, and showed excellent specificity. It detected as low as 6.4 fg/µL of V. parahaemolyticus in genomic DNA, or 7.4 CFU/g spiked food samples with 4 h of enrichment. The limit of detection in shrimp (Litopenaeus Vannamei), fish (Carassius auratus), clam (Ruditapes philippinarum) evidenced that sensitivity was considerably affected by the food matrix. The presence of food matrix reduced the sensitivity of spiked food samples by 10 ~ 100 times. In the filed samples detection, RAA-LFD method showed good coincidence with GB4789.7-2013 method and PCR method at rates of 90.6% and 94.1%, respectively. RAA-LFD has high accuracy and sensitivity for the detection of V. parahaemolyticus, which can serve as a model tool to meet the growing need for point-of-care diagnosis of V. parahaemolyticus.
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Data Availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
This study was financed by Science & Technology Commission of Shanghai Municipality (Grant No. 21N31900300) and Shanghai Agriculture Applied Technology Development Program, China (Grant No. 2019-02-08-00-10-F01149).
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Conceptualization: Xiaohong Sun; Methodology: Darong Li; Writing - original draft preparation: Darong Li, Jiayi Zhao; Writing - review and editing: Darong Li, Jiayi Zhao, Weiqing Lan; Funding acquisition: Xiaohong Sun; Supervision: Xiaohong Sun, Yong Zhao.All authors reviewed the manuscript.
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Li, D., Zhao, J., Lan, W. et al. Effect of food matrix on rapid detection of Vibrio parahaemolyticus in aquatic products based on toxR gene. World J Microbiol Biotechnol 39, 188 (2023). https://doi.org/10.1007/s11274-023-03640-1
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DOI: https://doi.org/10.1007/s11274-023-03640-1