Rapid and Visualized Detection of Virulence-Related Genes of Vibrio cholerae in Water and Aquatic Products by Loop-Mediated Isothermal Amplification

What is it about?

Vibrio cholerae can cause pandemic cholera in humans. The bacterium resides in aquatic environments worldwide. Continuous monitoring of V. cholerae contamination in water and aquatic products is imperative for food safety control and human health. We have developed a rapid and visualized method based on loop-mediated isothermal amplification (LAMP) for diagnosis of virulence-related genes ace, zot, cri, and nanH of V. cholerae. Three pairs of molecular probes targeting each of these genes were designed and synthesized. The one-step LAMP reaction was conducted at 65oC for 40 min. Positive results were simply inspected by the production of light green color under visible light or green fluorescence under UV light (302 nm). Limit of detection (LOD) of the LAMP method was 1.85 pg/reaction of genomic DNA or 2.50 CFU/reaction of cell culture of V. cholerae. The LAMP method showed high efficiency in rapid testing of spiked or collected specimens of water and aquatic products.

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