Open AccessDetection of pandemic Vibrio parahaemolyticus O3:K6 serovar in Gulf of Mexico water and shellfish using real‐time PCR with Taqman ® fluorescent probes
Author(s) -
Rizvi Amy V.,
Panicker Gitika,
Myers Michael L.,
Bej Asim K.
Publication year - 2006
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2006.00387.x
Subject(s) - vibrio parahaemolyticus , taqman , shellfish , serotype , vibrio , microbiology and biotechnology , vibrionaceae , fluorescence , seawater , virology , biology , real time polymerase chain reaction , fishery , bacteria , fish <actinopterygii> , aquatic animal , ecology , gene , physics , biochemistry , genetics , quantum mechanics
We describe a real‐time multiplexed PCR method using Taqman ® probes for the detection of total and pandemic Vibrio parahaemolyticus O3:K6 serovar in oysters and Gulf of Mexico water (gulf water). The specificity of these primers and probes was tested for amplification of a 450 bp thermolabile hemolysin ( tlh ) and a 369 bp ORF8 amplicon representing all V. parahaemolyticus and post‐1996 clinical isolates of pandemic serovar O3:K6, respectively. The sensitivity of detection was 10 pg purified DNA or 10 3 CFU in 1 mL pure culture. Enrichment of this pathogen in oyster tissue homogenate or gulf water for 5 or 8 h resulted in the detection of an initial inoculum of 1 CFU in 1 mL or 1 g of samples. Application of the Taqman ® PCR assay on natural oysters exhibited a positive detection of V. parahaemolyticus , ranging from 16% to 100% of the samples collected primarily during the summer months. None of the samples exhibited a positive detection of O3:K6 serovar. Rapid and sensitive detection of this pathogen will help shellfish industry and Interstate Shellfish Sanitation Conference (ISSC) undertake appropriate measures to monitor this pathogen in oysters and oyster‐growing waters, thereby preventing disease outbreaks and consequently protecting consumer health.