A targeted mass spectrometry-based method for the simultaneous detection of four pathogenic Vibrio species in aquatic products

The widespread occurrence of pathogenic Vibrios highlights the need for efficient and accurate screening techniques. In this study, we established a novel targeted mass spectrometry (MS)-based method that enables the rapid and simultaneous detection of four pathogenic Vibrio species—Vibrio vulnificus, Vibrio parahaemolyticus, Vibrio alginolyticus, and Vibrio cholerae—in aquatic products. Eleven peptides were identified as Vibrio biomarkers, allowing for accurate differentiation of these bacterial species from other conventional food-borne pathogens. By integrating the targeted MS method with buoyant density centrifugation (BDC), we efficiently enriched and separated Vibrio from seafood matrices, thereby reducing interference from complex matrix components. The sensitivity for V. vulnificus, V. parahaemolyticus, V. alginolyticus, and V. cholerae was 81.8 %, 92.3 %, 87.5 %, and 88.2 %, respectively. The level of detection (LOD₅₀) for this newly developed method ranging from 0.462 to 1.15 CFU/25 g across all targets, which was 1.15–1.33 times higher than that of the reference method. Both inclusivity and exclusivity precision were 100 %, attributable to the high specificity of the peptide biomarkers used. As a proof-of-concept, this method was employed to screen commercial aquatic products for the four pathogenic Vibrio species, and the detection results were consistent with the culture method. These results demonstrate its potential to enhance food safety and public health initiatives by providing a reliable detection strategy.