Objective: This study aimed to establish a loop-mediated isothermal amplification (LAMP) assay for detection of Vibrio parahaemolyticus (V. parahaemolyticus).
Methods: The specificity of this assay was evaluated by using a panel of 33 strains of V. parahaemolyticus and 22 strains of other species bacteria. The sensitivity was determined by using serial dilutions of V. parahaemolyticus (ATCC 17802) chromosomal DNA (5×10(0) - 5×10(5) copies/µl). The samples were also tested by using qualification PCR assay and Taqman real-time PCR assay in parallel for comparison with LAMP.
Results: Both sensitivity and specificity of LAMP assay, PCR assay and Taqman real-time PCR assay were 100% (22/22, 33/33, respectively). The detection limits of above three methods assay were 5×10(1) copies/µl, 5×10(3) copies/µl and 5×10(2) copies/µl, respectively. The reaction period of time needed of the above three assays was 22 min, 3 h, 50 min, respectively.
Conclusion: Compared to qualification PCR assay and Taqman real-time PCR assay, the established LAMP assay was better in low detection limit and less reaction time, which made it an ideal method for quick detection of V. parahaemolyticus.
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