title: Comparison of molecular detection methods for Vibrio parahaemolyticus and Vibrio vulnificus.
abstract: Pathogenic vibrios are a global concern for seafood safety and many molecular methods have been developed for their detection. This study compares several molecular methods for detection of total and pathogenic Vibrio parahaemolyticus and Vibrio vulnificus, in MPN enrichments from oysters and fish intestine samples. This study employed the DuPont Qualicon BAX® System Real-Time PCR assay for detection of V. parahaemolyticus and V. vulnificus. Multiplex real-time PCR detection of total (tlh+), tdh+, and trh+V. parahaemolyticus was conducted on the Cepheid SmartCycler II. Total (rpoD) and tdh+V. parahaemolyticus were also detected using LAMP. V. vulnificus detection was performed using real-time PCR methods developed for the SmartCycler and the AB 7500 Fast. Recommended template preparations were compared to BAX® lysis samples for suitability. There was no significant difference in detection of V. parahaemolyticus and V. vulnificus using the BAX® or SmartCycler assays. The AB assay showed no difference from other methods in detection of V. vulnificus unless boiled templates were utilized. There was a significant difference in detection of tdh+V. parahaemolyticus between SmartCycler and LAMP assays unless the total (tlh+) V. parahaemolyticus gene target was omitted from the SmartCycler assay; a similar trend was observed for trh+V. parahaemolyticus.
authors.affiliation: Gulf Coast Seafood Laboratory, Division of Seafood Science and Technology, FDA, 1 Iberville Drive, Dauphin Island, AL 36528, USA. Jessica.Jones@fda.hhs.gov
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Jones
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Hara-Kudo
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Krantz
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Benner
authors.forename: Amy B
authors.initials: AB
authors.lastname: Smith
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Dambaugh
authors.forename: John C
authors.initials: JC
authors.lastname: Bowers
authors.forename: Angelo
authors.initials: A
authors.lastname: Depaola
mesh_terms: D015169:Colony Count, Microbial; D003257:Consumer Product Safety; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005506:Food Contamination / Q000032:analysis*; D005516:Food Microbiology / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D017747:Seafood / Q000382:microbiology*; D012680:Sensitivity and Specificity; D014736:Vibrio parahaemolyticus / Q000235:genetics / Q000302:isolation & purification*; D041261:Vibrio vulnificus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial