The Establishment And Applification Of Multiplex Real-Time PCR For Detection Of Vibrio Parahaemolyticus And Vibrio Vulnificus

Virio parahaemolyticus and Vibrio vulnificus are two of pathogenic vibrio species primarily involved in causing gastrointestinal illnesses. V. parahaemolyticus is considered to be the causative agent in 50% to 70% of all cases of diarrhea associated with the consumption of fishery products in the summer months. Vibrio vulnificus produces a rapidly fatal septicemia, with a mortality rate of up to 60%, which is primarily associated with the ingestion of raw oysters. Contact of wounds with seawater or shellfish can also lead to serious infections that can progress to septicemia. Therefore, the detection of the presence and the number of V.parahaemolyticus and V. vulnificus were essential.The multiplex Real-time PCR system uses fluorogenic probes to detect PCR products as they form; the exonuclease activity of Taq polymerase releases a labeled reporter dye at the 5' end of the probe from the quencher dye at the 3' end with each cycle of amplification. Thus, increased fluorescence is directly proportional to the formation of PCR products. Plotting the increase in fluorescence versus cycle number gives a comprehensive picture of the PCR process, and quantification of initial template concentration can be calculated from data on the exponential phase of amplification. Multiplex Real-time PCR offers rapid, sensitive, and specific detection V. parahaemolyticus and V. vulnificus.In present paper, a new technique based on real-time PCR for quantitative detection V. parahaemolyticus and a multiplex real-time PCR for simultaneous and quantitative detection of Vibrio parahaemolyticus and Vibrio vulnificus were established. Then a screening of seafood in Eastern China for the presence of Vibrio parahaemolyticus and Vibiro vulnificus was carried out base on the mutiplex real-time PCR and Culture Method. 1 Application of real-time PCR for quantitative detection of Vibrio parahaemolyticus Vibrio parahaemolyticus is one of important human food pathogens. Traditional diagnostic tests for V. parahaemolyticus are laborious and aways present flase negative...