Rapid subtyping of pathogenic and nonpathogenic Vibrio parahaemolyticus by fourier transform infrared spectroscopy with chemometric analysis.

Vibrio parahaemolyticus which naturally inhabits marine and estuarine environment represents pathogenic strains (virulence genes tdh or trh positive) and non-pathogenic strains (virulence genes negative). In this study, a rapid method for subtyping pathogenic and non-pathogenic V. parahaemolyticus was established using fourier transform infrared (FTIR) spectroscopy with chemometric analysis. This method targeted three strains of genotypes of V. parahaemolyticus including tdh positive, trh positive and virulence gene-negative (nonpathogenic) V. parahaemolyticus. The FTIR absorption spectra between 1800 and 900 cm-1 highlighted the most distinctive variations and were the most useful for characterizing the three bacteria. The successful differentiation and identification of the three bacteria could be accomplished in less than 1 h by FTIR using principal component analysis (PCA), or another cluster model of hierarchical cluster analysis (HCA). The method was verified by analyzing spiked V. parahaemolyticus fish samples. Furthermore, all of ten clinical isolates of V. parahaemolyticus were identified as tdh-positive, none of the clinical isolates were trh-positive, and all of ten environmental isolates were identified as non-pathogenic by the subtyping method, which were confirmed by PCR assays. All data demonstrated that the newly established subtyping method by FTIR is practical, time-saving, labor-saving, specific and cost-effective, especially suitable for the basic laboratories of CDC and port quarantine departments to perform suiveillance and epidemiological traceability of pathogenic V. parahaemolyticus.