LB performed the growth study, determined the susceptibility
to whole blood and helped to draft the manuscript. MCDP performed the animal study. JS constructed the Tn917 library. MG participated in the design of the study and helped to draft the manuscript. DG conceived the study and drafted the manuscript. All selleck chemical authors read and approved the final manuscript.”
“Background The Gram-negative, halophilic marine bacterium Vibrio parahaemolyticus has emerged as a major cause of seafood-associated outbreaks throughout the world and become a significant concern of seafood safety [1–3]. Shellfish, particularly oysters, has been frequently implicated in V. parahaemolyticus infections [4, 5]. Typically within 24 h after eating contaminated seafood, V. parahaemolyticus causes acute, Anlotinib concentration self-limiting gastroenteritis characterized by diarrhea, abdominal cramps, nausea, MLN2238 chemical structure vomiting, fever, and chills, which lasts for 1-3 days . Two hemolysins, the thermostable direct hemolysin (TDH) and the TDH-related hemolysin (TRH) are well-characterized virulence factors for pathogenic V. parahaemolyticus strains . However, the majority of V. parahaemolyticus strains in the environment
and seafood samples lack these two hemolysin genes [8–10], thus the number of total V. parahaemolyticus has been used as an indicator for preventing V. parahaemolyticus infections from seafood consumption [11, 12]. Traditional culture-based methods for isolating and enumerating V. parahaemolyticus from seafood samples involve the most probable number (MPN) technique . Although widely used, such methods are labor-intensive and time-consuming (4-7 days). Molecular-based methods such as DNA probe hybridization and PCR assays have been developed for V. Etofibrate parahaemolyticus and yielded rapid and specific results [14–18]. However, the probe hybridization
procedure and the gel electrophoresis technique used to analyze PCR amplicons are tedious and time-consuming. Recently, several real-time PCR assays have been developed for the detection of V. parahaemolyticus with increased speed and sensitivity [12, 19–21]. Nonetheless, these assays require a dedicated real-time PCR machine, which is rather expensive and not yet widely available. Loop-mediated isothermal amplification (LAMP), a novel DNA amplification technique invented in 2000 , has since been applied in detecting many bacterial and viral agents [23–26]. Because the LAMP assay was carried out under isothermal conditions, a simple heater that maintains a constant temperature (60-65°C) is sufficient. LAMP assays were reported to be highly specific, sensitive, rapid, and cost-effective [23–26]. Very recently, LAMP was adopted to detect V. parahaemolyticus and yielded promising results . However, in this LAMP assay, primers were designed to target the V.