Development of a quantitative real-time PCR method for estimation of the total number of Vibrio parahaemolyticus in contaminated shellfish and seawater.
J Food Prot
; 68(5): 1083-8, 2005 May.
Article
de En
| MEDLINE
| ID: mdl-15895747
ABSTRACT
A real-time PCR method targeting the toxR gene of Vibrio parahaemolyticus was developed to quantify the number of V. parahaemolyticus cells, including those of both the hemolysin-producing and nonproducing strains. The specificity of the primer and probe set was confirmed using 25 strains of V. parahaemolyticus and 30 strains of other microbial species. We determined the threshold cycle number using the real-time PCR and the number of V. parahaemolyticus cells by plate count using serially diluted pure culture and developed a standard curve for quantification. Standard curves for V. parahaemolyticus in seawater and seafood were established using artificially inoculated samples. The threshold cycle number and the number of V. parahaemolyticus cells were correlated with 10(1) to 10(7) CFU/ml in pure culture, seawater, and shellfish homogenate. The real-time PCR method developed in this study was compared with the most-probable-number method in seafood samples that were naturally contaminated. The differences in the number of V. parahaemolyticus cells as determined by the culture method and the PCR method were less than 10-fold.
Recherche sur Google
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Eau de mer
/
Fruits de mer
/
Vibrio parahaemolyticus
/
Contamination des aliments
/
Réaction de polymérisation en chaîne
Type d'étude:
Diagnostic_studies
Limites:
Animals
Langue:
En
Journal:
J Food Prot
Année:
2005
Type de document:
Article
Pays d'affiliation:
Japon