Identification of
            <i>Vibrio parahaemolyticus</i>
            Strains at the Species Level by PCR Targeted to the
            <i>toxR</i>
            Gene

Kim, Yung Bu; Okuda, Jun; Matsumoto, Chiho; Takahashi, Naoki; Hashimoto, Satoru; Nishibuchi, Mitsuaki

doi:10.1128/jcm.37.4.1173-1177.1999

Cited by 445 publications

(245 citation statements)

References 19 publications

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“…Suspected V. parahaemolyticus isolates were then ¢nally iden-ti¢ed using ID TEST EB-20 kit (Nissui, Tokyo, Japan) following published criteria [17]. For PCR-aided identi¢cation of V. parahaemolyticus, we used species-speci¢c primers targeting the toxR gene as described by Kim et al [18].…”

Section: Methodsmentioning

confidence: 99%

Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan

Alam

2002

FEMS Microbiology Letters

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Seawater and organic material (live and/or dead matter deposited on any substratum submersed in seawater) were collected during the cool weather season from a coast of the Seto-Inland Sea, Japan, and analyzed to determine Vibrio parahaemolyticus densities and the occurrence of pathogenic strains, defined as those possessing tdh and/or trh genes by the polymerase chain reaction (PCR), using isolated DNA from enrichment culture of the samples. About 95% of the samples were positive for V. parahaemolyticus (with densities of 3 to s 1400 cells per 100 ml water or 10 g organic samples) by the most-probable-number (MPN)-PCR technique with species-specific toxR primers, but only 40% were positive by the conventional MPN-culture technique (with densities ranging from 3 to 240 cells per 100 ml water or 10 g organics). Furthermore, the tdh and trh genes were positive in 55% and 20% of samples, respectively, by the MPN-PCR technique. No tdh and trh gene-positive strains were isolated by the conventional MPN-culture procedure. The difference in detection between the MPN-culture and the MPN-PCR techniques appeared to be significant and may be attributed to different detection sensitivities and other factors. ß

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Section: Methodsmentioning

confidence: 99%

Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan

Alam

2002

FEMS Microbiology Letters

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“…Among the six blocks of reactions, only one was selected for species identification by using the following preliminary tests: arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase (Ottaviani et al, 2003). The strains were also tested by a PCR assay targeting a species-specific sequence of the toxR gene usually used for identification of Vibrio parahaemolyticus strains (Kim et al, 1999).…”

Section: Identification Of Vibrio Strainsmentioning

confidence: 99%

An extensive investigation into the prevalence and the genetic and serological diversity of toxigenic Vibrio parahaemolyticus in Italian marine coastal waters

Ottaviani

Leoni

Rocchegiani

et al. 2012

Environmental Microbiology

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Summary The relationship between Vibrio parahaemolyticus strains isolated from the aquatic environment and those isolated from cases of infection in humans is poorly understood due to the low prevalence of tdh‐ and/or trh‐positive strains in the environment. To address this concern, it would be useful to analyse the genetic relationships among environmental and food strains and with reference to clinical isolates, also applying molecular typing methods. The aim of this study was to evaluate the prevalence of toxigenic V. parahaemolyticus in Italian coastal waters and seafood, to examine intra‐species variability and to identify, using serotyping and pulsed‐field gel electrophoresis (PFGE), relationships among strains from different sources, geographical origin and period of isolation. Of the 192 V. parahaemolyticus strains isolated in different Italian areas and examined in this study, 25 (13.0%) proved to carry the trh gene while none of the strains proved positive to the search by PCR for tdh and Group‐Specific‐toxRS genes. The prevalence of toxigenic strains in the Tyrrhenian Sea was significantly lower than that calculated for the Ligurian coasts. Regarding the sources of isolation, the higher prevalence of trh‐positive V. parahaemolyticus was revealed in fish, followed by clams, plankton, oysters, mussels and lastly seawater. Within the toxigenic strains, 16 serotypes and 20 distinct PFGE patterns were identified. Two clusters, which included a total of 8 V. parahaemolyticus strains, were specifically associated with the North Adriatic Sea area and were stable over time. Our results demonstrate that trh‐positive V. parahaemolyticus strains circulated in Italy in the period 2002–2009 with a prevalence higher than that reported from other European and extra‐European countries, confirming that toxigenic V. parahaemolyticus is an emerging public health concern in Italy, regardless of its pandemic potential.

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“…PCR assays were carried out to test for the presence of V. parahaemolyticus-specific toxR gene, tdh and trh, the virulence genes of pathogenic V. parahaemolyticus as previously described (Tada et al, 1992;Kim et al, 1999). A 535 bp region of ureR gene of V. parahaemolyticus was amplified using primers ure-F (5′-TCTGAGAAAAGGTATAGT TGG-3′) and ure-R (5′-AGTCGTGTATCCTGCACTCTA-3′) designed in this study.…”

Section: Genotyping Of V Parahaemolyticusmentioning

confidence: 99%

Molecular characterization of thermostable direct haemolysin‐related haemolysin (TRH)‐positive Vibrio parahaemolyticus from oysters in Mangalore, India

Parvathi

Kumar

Bhanumathi

et al. 2006

Environmental Microbiology

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Pathogenic Vibrio parahaemolyticus strains producing either or both of a thermostable direct haemolysin (TDH) and a TDH-related haemolysin (TRH) encoded by tdh and trh genes, respectively, are isolated at a low rate from the environment. However, recently we observed that a considerable percentage of APW (alkaline peptone water) enrichment broths of oysters collected off Mangalore India, were trh(+), rather than tdh(+) by PCR. In order to further investigate the prevalence and genetic diversity of trh bearing V. parahaemolyticus in our coast, we attempted to isolate and characterize trh(+)V. parahaemolyticus from oysters. A total of 27 trh(+) strains were isolated during the period between March 2002 and February 2004, of which nine were also tdh(+). All the trh(+) isolates were positive for urease phenotype. The isolates belonged to diverse phenotypes. In order to explore the possible presence of heterogeneity in the trh gene region among trh(+)V. parahaemolyticus, a 1.5 kb region around trh gene was PCR amplified and restriction digested using selected restriction enzymes. The whole genome comparison of strains was performed by randomly amplified polymorphic DNA PCR (RAPD PCR). The PCR-RFLP results revealed fairly well conserved nature of the trh gene region studied in different serotypes. Though 11 strains were positive by PCR for a genomic fragment that has been reported to be amplified in pandemic strains, all strains were negative by group-specific PCR (GS-PCR), orf8 PCR and showed a different RAPD pattern compared with pandemic strains. The results suggest that genetically diverse V. parahaemolyticus carrying virulence genes are associated with the aquatic environment in this region.

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Identification of Vibrio parahaemolyticus Strains at the Species Level by PCR Targeted to the toxR Gene

Cited by 445 publications

References 19 publications

Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan

Environmental investigation of potentially pathogenic Vibrio parahaemolyticus in the Seto-Inland Sea, Japan

An extensive investigation into the prevalence and the genetic and serological diversity of toxigenic Vibrio parahaemolyticus in Italian marine coastal waters

Molecular characterization of thermostable direct haemolysin‐related haemolysin (TRH)‐positive Vibrio parahaemolyticus from oysters in Mangalore, India

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