Detection of tlh and tdh expression differences in Vibrio parahaemolyticus by real-time quantitative PCR

Abstract:

Vibrio parahaemolyticus is a foodborne pathogenic bacteria which could be widely found in coastal areas, salt lakes and seafoods, it could cause large-scale food poisoning. tlh (Themolabile hemolysin) and tdh (Thermostable direct hemolyticus) are two of the most important virulence genes of Vp, we can compare the virulence of strains which in different stress conditions, as well as differences strains by comparing the expression levels of tlh and tdh indirectly. In this paper, three kinds of Vp cultured under different conditions, total RNA were extracted from them, 16S rRNA as an internal standard gene, using real-time quantitative PCR (RT-PCR) detect Vp’s tlh and tdh gene expression under different conditions. The results showed that gene tlh and tdh expression difference were significant in different strains and in same strains that under different condition, tlh optimal expression conditions were 5% salinity and 20 °C; tdh optimum expression conditions were 1% salinity and 25 °C. The use of SPSS software for statistical analysis of experimental results show that: factor that affect the expression of tlh is strain greater than salinity greater than temperature; factor that affect the expression of tdh is strain greater than temperature greater than salinity.