Detection of the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus by polymerase chain reaction

Tada, J; Ohashi, Tetsuo; Nishimura, Naoyuki; Shirasaki, Yoshinari; Ozaki, Hiroko; Fukushima, Shigeru; Takano, Junpei; Nishibuchi, Mitsuaki; Takeda, Yoshifumi

doi:10.1016/0890-8508(92)90044-x

Cited by 260 publications

(177 citation statements)

References 24 publications

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“…UtoxR/VvtoxR fragments were amplified under the following PCR conditions: 4 min at 94°C, 30 cycles of 94°for 30 s, 61°C for 30 s, 68°C for 30 s with a final 68°C extension of 7 min. All V. parahaemolyticus strains were additionally screened for the hemolysin genes tdh and trh [43,45]. Parameters used for all V. parahaemolyticus PCRs (VptoxR/tdh/trh) were the same as for the identification of toxR genes in V. vulnificus with two exceptions: annealing was performed at 62°C for 1 min and elongation at 68°C for 1 min.…”

Section: Pcr Detection Of Species-specific and Virulence-associated Gmentioning

confidence: 99%

Temporal and Spatial Distribution Patterns of Potentially Pathogenic Vibrio spp. at Recreational Beaches of the German North Sea

Böer

Heinemeyer²,

Luden³

et al. 2013

Microb Ecol

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The number of reported Vibrio-related wound infections associated with recreational bathing in Northern Europe has increased within the last decades. In order to study the health risk from potentially pathogenic Vibrio spp. in the central Wadden Sea, the seasonal and spatial distribution of Vibrio vulnificus, Vibrio parahaemolyticus, Vibrio alginolyticus and Vibrio cholerae were investigated at ten recreational beaches in this area over a 2-year period. V. alginolyticus and V. parahaemolyticus were found to be omnipresent all year round in the study area, while V. vulnificus occurrence was restricted to summer months in the estuaries of the rivers Ems and Weser. Multiple linear regression models revealed that water temperature is the most important determinant of Vibrio spp. occurrence in the area. Differentiated regression models showed a species-specific response to water temperature and revealed a particularly strong effect of even minor temperature increases on the probability of detecting V. vulnificus in summer. In sediments, Vibrio spp. concentrations were up to three orders of magnitude higher than in water. Also, V. alginolyticus and V. parahaemolyticus were found to be less susceptible towards winter temperatures in the benthic environment than in the water, indicating an important role of sediments for Vibrio ecology. While only a very small percentage of tested V. parahaemolyticus proved to be potentially pathogenic, the presence of V. vulnificus during the summer months should be regarded with care.

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Section: Pcr Detection Of Species-specific and Virulence-associated Gmentioning

confidence: 99%

Temporal and Spatial Distribution Patterns of Potentially Pathogenic Vibrio spp. at Recreational Beaches of the German North Sea

Böer

Heinemeyer²,

Luden³

et al. 2013

Microb Ecol

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“…For PCR amplification, 3 ul of the bacterial culture was heated initially to 100 'C for 5 min to disrupt bacteria and denature DNA, and was put into a total volume of 30 ,tl reaction mixture composed of 10 mM-Tris-HCI (pH 9 0), 50 mMKCl, 1-5 mM-MgCl2, 0 01 % gelatin, 0-6 ,tM each of the primers [8] electrophoresed on a 1P5 % agarose gel. After electrophoresis, the gel was stained in 0 5 gug/ml ethidium bromide solution and photographed under UV light.…”

Section: Pcr Amplificationmentioning

confidence: 99%

“…The sequence structure of the genetic locus for producing hemolysin in V. mimicus is homologous to that in TDH-gene of V. parahaemolyticus [7]. The primers to detect the TDH-gene of V. parahaemolyticus by polymerase chain reaction (PCR) have been established [8].…”

Section: Introductionmentioning

confidence: 99%

High prevalence of thermostable direct hemolysin (TDH)-like toxin inVibrio mimicusstrains isolated from diarrhoeal patients

et al. 1993

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SUMMARYA total of 17 isolates of Vibrio mimicus from patients, 29 from environment and 2 from food was examined for toxigenicity. Sixteen (94 %) clinical isolates and one (50%) from food produced TDH-like toxin, whereas none of the environmental isolates did so. The food from which V. mimicus with TDH-like toxin production was isolated, was one which had caused food poisoning. Only one environmental strain produced CT-like toxin, whilst ST-like toxin was not detected from any strains tested.

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“…The test adopted for Kanagawa phenomenon using Wagatsuma agar consumes time and labor. Moreover, the test may not be determinative as many environmental strains are Kanagawa negative and do not produce TDH (Tada et al 1992). Presently there is no in-vitro test for the detection of TRH in V. parahaemolyticus.…”

Section: Introductionmentioning

confidence: 99%

“…Nasu et al (2000) isolated a filamentous phage possessing orf8 in another recently emerging serovar (O4:K68), and reported the orf8 gene fragment as a useful genetic marker for the pandemic group. A toxin regulatory gene (toxR) sequence specific to V. parahaemolyticus, which is present in all the strains irrespective of their ability to produce TDH or TRH, has been applied for definitive identification of the bacterial isolates by several workers (Jacksic et al 2002;Tada et al 1992;Kim et al 1999). …”

Section: Introductionmentioning

confidence: 99%

Occurrence and distribution of virulent strains of Vibrio parahaemolyticus in seafoods marketed from Cochin (India)

Chakraborty

Surendran

2008

World J Microbiol Biotechnol

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This study was aimed for the detection of Vibrio parahaemolyticus by biochemical and molecular methods in seafood samples collected from the markets of Cochin located at the southwest coast of India. A total of seventy-two V. parahaemolyticus cultures were isolated by selecting sucrose and cellobiose non-fermenting colonies. All the biochemically confirmed strains were found to have 368-bp toxR gene fragment, while an additional 24% of the samples were confirmed as V. parahaemolyticus by toxR based polymerase chain reaction (PCR) from enrichment broths. PCR based methods are used to detect tdh, trh, and orf8 genes for the identification of pathogenic and pandemic V. parahaemolyticus. Only one out of two urease positive isolates amplified the trh (500bp) gene. About 10% of the isolates showed weak haemolysis and none were found to amplify tdh (269 bp) and orf8 (746 bp) genes, thus indicating the meager incidence of pandemic strains from this area. The incidence of trh positive isolates from market samples signals towards the adoption of stringent seafood safety measures for the products meant for human consumption.

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Detection of the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus by polymerase chain reaction

Cited by 260 publications

References 24 publications

Temporal and Spatial Distribution Patterns of Potentially Pathogenic Vibrio spp. at Recreational Beaches of the German North Sea

Temporal and Spatial Distribution Patterns of Potentially Pathogenic Vibrio spp. at Recreational Beaches of the German North Sea

High prevalence of thermostable direct hemolysin (TDH)-like toxin inVibrio mimicusstrains isolated from diarrhoeal patients

Occurrence and distribution of virulent strains of Vibrio parahaemolyticus in seafoods marketed from Cochin (India)

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