Optimize of Loop-mediated isothermal amplification technology in detection of Vibrio parahaemolyticus

In this study,two pairs of primers were designed according to the heat-sensitive hemolysin gene tlh of V. parahaemolyticus and the LAMP amplification reaction conditions were optimized to develop the rapid detection method of V. parahaemolyticus,and verified the specificity and sensitivity. These results suggested that the LAMP mixture containing concentration ratio for outer primer and inner primer was 1 to 8,4 mmol/L Mg2+,3.5 mmol/L d NTPs,performed at 65 ℃ for 1 h. No crossreaction was found with other common bacteria which showed a good specificity. The detection limits of LAMP assay and pure cultures were 5.45 ×101fg/μL LAMP mixture of genomic DNA and 140 cfu/m L respectively. The detection limit of V. parahaemolyticus in food samples were 2.8 ×102cfu/m L. The result indicated that LAMP had the advantage of rapidity,specificity,and sensitivity.