Abstract:
Objective To detect the thermostable direct hemolysin(tdh)gene of Vibrio parahaemolyticus using molecular beacon PCR.
Methods The tdh gene of 2 strains of Vibrio parahaemolyticus and 13 strains of other enteric pathogens was respectively amplified in the PCR reaction system containing a molecular beacon probe, and record the fluorescent value in the terminal of PCR and during the real time PCR.
Results The fluorescent value of 2 strains of Vibrio parahaemolyticus and the positive plasmid were 114.9, 95.2, 90.0 when reading in the terminal of PCR, others were 47.0~69.1;and the Ct value of 2 strains of V. parahaemolyticus and the positive plasmid were 26.2, 26.8, 32.0 using molecular beacon real time PCR, and others were over 32.The results using PCR were identical with the agarose gel electrophrosis.
Conclusion Molecular beacon PCR is a rapid, special, sensitive, convenient technique to detect the V. parahaemolyticus possessing tdh gene.