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Detection of pandemic Vibrio parahaemolyticus O3:K6 serovar in Gulf of Mexico water and shellfish using real-time PCR with Taqman fluorescent probes.
Rizvi, Amy V; Panicker, Gitika; Myers, Michael L; Bej, Asim K.
Afiliação
  • Rizvi AV; Department of Biology, University of Alabama at Birmingham, Birmingham, AL 35294-1170, USA.
FEMS Microbiol Lett ; 262(2): 185-92, 2006 Sep.
Article em En | MEDLINE | ID: mdl-16923074
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.
Assuntos
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ostreidae / Água do Mar / Frutos do Mar / Vibrioses / Vibrio parahaemolyticus / Reação em Cadeia da Polimerase Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Animals / Humans País/Região como assunto: Mexico Idioma: En Revista: FEMS Microbiol Lett Ano de publicação: 2006 Tipo de documento: Article País de afiliação: Estados Unidos País de publicação: Reino Unido
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Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ostreidae / Água do Mar / Frutos do Mar / Vibrioses / Vibrio parahaemolyticus / Reação em Cadeia da Polimerase Tipo de estudo: Diagnostic_studies / Evaluation_studies Limite: Animals / Humans País/Região como assunto: Mexico Idioma: En Revista: FEMS Microbiol Lett Ano de publicação: 2006 Tipo de documento: Article País de afiliação: Estados Unidos País de publicação: Reino Unido