ABSTRACT Surveillance of pathogenic bacteria in aquatic products is vital to public health. Vibrio parahaemolyticus, Vibrio vulnificus, Shigella spp. , and Staphylococcus aureus represent important and complementary bacterial hazards associated with aquatic products, highlighting the need for rapid and simultaneous detection methods. This study aimed to develop a novel quadruplex real‐time PCR system for detecting these four pathogens in aquatic products. Four sets of primers and TaqMan probes were newly designed, specific to the pR72H, vvhA, ipaH, and kipA₂ genes, respectively. The assay exhibited high specificity against other common pathogens and no cross‐reaction among these different bacteria. Analytical sensitivity ranged from 10 −6 ng μL −1 for Shigella spp. to 10 −5 ng μL −1 for Vibrio species and Staphylococcus aureus. The limits of detection in pure culture and spiked samples were 10 1 CFU mL −1 for Shigella spp. , 10 2 CFU mL −1 for Vibrio species, and 10 3 CFU mL −1 for Staphylococcus aureus, respectively. To enhance the sensitivity to 10 0 CFU mL −1 directly from real samples, a 9‐h pre‐enrichment step was incorporated. The effectiveness of the method was validated using 365 diverse aquatic products from multiple coastal and inland regions in China, demonstrating diagnostic accuracy above 97. 3% compared to traditional culture‐based methods while offering a significant time reduction (3–7 days vs. 11 h). Overall pathogen prevalence across samples ranged from 0% to 21. 4%. This study offers a practical tool for large‐scale surveillance, enabling timely detection of key pathogens and supporting proactive food‐safety interventions.
Zou et al. (Mon,) studied this question.
Synapse has enriched 5 closely related papers on similar clinical questions. Consider them for comparative context: