Detection of Virulence Factors of Vibrio parahaemolyticus by Real-Time PCR / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To develop a real-time fluorescence PCR assay to detect the genes encoding thermolabile(hemolysin)(TLH),thermostable direct hemolysin(TDH) and TDH-related hemolysin(TRH) of Vibrio(parahaemolyticus).METHODS The genes of TDH and TRH were selected as target ones of thermostable direct and TDH-related hemolysin,and TLH gene as a specific genomic marker for V.parahaemolyticus.Designed and synthesized the primers and Taqman probes,we investigated 487 stool samples of doubt foodborne illness patients by real-time fluorescence PCR.RESULTS The sensitivity of the assay for TLH and TDH was 1.0?10~2copies,but the sensitivity of TRH was 1.0?10~3copies. Among 487 samples,112 V.parahaemolyticus strains were found;101 samples of these strains showed the production of TDH;none of them was positive for TRH.CONCLUSIONS The Taqman PCR is a rapid and sensitive method to detect the TLH,TDH and TRH of V.parahaemolyticus,it is well suited for screening large numbers of samples at the same time;and TDH is one of the primary virulence factors in clinical isolated V. parahaemolyticus.