Development and evaluation of a sensitive recombinase aided amplification assay for rapid detection of Vibrio parahaemolyticus

J Microbiol Methods. 2022 Feb:193:106404. doi: 10.1016/j.mimet.2021.106404. Epub 2022 Jan 3.

Abstract

Vibrio parahaemolyticus (V. parahaemolyticus) is a widely distributed pathogen in the coastal areas, which causes food poisoning and leads to gastroenteritis and sepsis. Therefore, developing a simple, sensitive, and rapid detection method for V. parahaemolyticus is a major concern globally. This study established a sensitive and rapid technique based on recombinase aided amplification (RAA) to detect V. parahaemolyticus. The RAA reaction was carried out successfully at 39 °C within 30 min. The sensitivity of the RAA assay was 101 copies/μL using the recombinant plasmid and 10-3 ng/μL using the V. parahaemolyticus strain. In addition, RAA directly detected 7 × 103 CFU/mL of simulated fecal samples and 0.1 CFU/mL after enrichment for 4 h. The sensitivity and specificity of the RAA assay using fecal and fish samples were 100% similar to that of the real-time PCR. We conclude that the RAA assay is an ideal screening method for detecting V. parahaemolyticus due to its rapidity, high accuracy, and simplicity in operation.

Keywords: Rapid detection; Recombinase aided amplification; Sensitivity; Specificity; ToxR; Vibrio parahaemolyticus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Primers
  • Nucleic Acid Amplification Techniques / methods
  • Real-Time Polymerase Chain Reaction / methods
  • Recombinases
  • Sensitivity and Specificity
  • Vibrio parahaemolyticus* / genetics

Substances

  • DNA Primers
  • Recombinases