Molecular epidemiologic evidence for association of thermostable direct hemolysin (TDH) and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis

Shirai, Hiroyuki; Itô, Hisashi; Hirayama, Toshiya; Nakamoto, Yuya; Nakabayashi, Naomi; Kumagai, Kazuaki; Takeda, Yoshifumi; Nishibuchi, Mitsuaki

doi:10.1128/iai.58.11.3568-3573.1990

Cited by 256 publications

(117 citation statements)

References 27 publications

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“…A strong correlation was reported between clinically sig-ni¢cant strains and the presence of the tdh (thermostable direct hemolysin) or trh (thermostable direct-related hemo- lysin) genes, suggesting that the TDH and TRH are virulence factors in V. parahaemolyticus [21]. The V. parahaemolyticus outbreak isolates used in this study were TDHpositive based on hybridization with the tdh gene probe [22] (Table 1); however, V. parahaemolyticus O3:K6 and non-03 :K6 isolates cannot be discriminated on the basis of the tdh gene.…”

Section: Resultsmentioning

confidence: 99%

Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates

Khan

2002

FEMS Microbiology Letters

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Outbreaks of Vibrio parahaemolyticus gastroenteritis in the United States (Texas, New York and Pacific Northwest) in 1997^98 emphasized the need to develop molecular methods for identification and differentiation of these organisms. When outbreak isolates were analyzed for the enterobacterial repetitive intergenic consensus sequences, the Texas and New York outbreak isolates had a specific 850-bp DNA fragment that was absent in Pacific Northwest isolates. The 850-bp polymerase chain reaction (PCR) product was found in isolates of serovar O3:K6, which have an unusual potential to spread and cause infections. To develop a specific molecular detection method for serovar O3:K6, the nucleotide sequence of the 850-bp product was determined. The GenBank blast analysis did not show homology with any known Vibrio spp. gene sequences. Two PCR primers were designed to specifically amplify the unique sequences from serovar O3:K6 isolates. Genomic DNA from 10 Texas, eight New York, and seven Pacific Northwest outbreak isolates of V. parahaemolyticus was assayed by PCR. Texas and New York isolates were positive in the PCR assay, giving a 327-bp PCR product as predicted ; however, Pacific Northwest isolates were negative, indicating the absence of the target gene. Texas and New York isolates were all serovar O3:K6; the Pacific Northwest isolates were not. The primers were tested with other Vibrio spp. and other closely related species and no amplification of the 327-bp PCR product was found. The PCR method can be used to specifically identify O3:K6 V. parahaemolyticus isolates in less than 6 h. ß 2002 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.

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Section: Resultsmentioning

confidence: 99%

Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates

Khan

2002

FEMS Microbiology Letters

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“…This study represents a rare look at the distribution of a poorly understood community of naturally occurring bacteria from which human pathogens can arise. Although the great majority of infections come from V. parahaemolyticus isolates containing tdh and ⁄ or trh and epidemiological evidence has correlated the two gene products (haemolysins) with pathogenicity (Shirai et al 1990), almost all naturally occurring (environmental) isolates lack these two haemolysins (DePaola et al , 2003. Therefore, human infections most commonly arise from rare, naturally occurring tdh + and ⁄ or trh + V. parahaemolyticus strains found in finfish, shellfish, coastal waters and sediment.…”

Section: Discussionmentioning

confidence: 99%

Distribution of type III secretion systems in Vibrio parahaemolyticus from the northern Gulf of Mexico

Noriea

Johnson

Griffitt

et al. 2010

Journal of Applied Microbiology

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Aims: Two well‐characterized Vibrio parahaemolyticus pathogenicity factors – thermostable direct haemolysin (TDH) and TDH‐related haemolysin – are produced by strains containing the tdh and trh genes, respectively. Most strains of V. parahaemolyticus contain two nonredundant type III secretion systems (T3SS), T3SS1 and T3SS2, both of which contribute to pathogenicity. Furthermore, a recent study has revealed two distinct lineages of the V. parahaemolyticus T3SS2: T3SS2α and T3SS2β. The aim of this study was to determine the incidence of these pathogenicity factors in environmental isolates of V. parahaemolyticus. Methods and Results: We collected 130 V. parahaemolyticus isolates (TCBS agar) containing tdh and/or trh (determined by colony hybridization) from sediment, oyster and water in the northern Gulf of Mexico and screened them and 12 clinical isolates (PCR and agarose gel electrophoresis) for pathogenicity factors tdh, trh, T3SS1, T3SS2α and T3SS2β. The majority of potential pathogens were detected in the sediment, including all tdh−/trh+ isolates. T3SS2α components were detected in all tdh+/trh− isolates and zero of 109 trh+ isolates. One T3SS2α gene, vopB2, was found in all tdh+/trh− clinical strains but not in any of the 130 environmental strains. Fluorescence in situ hybridization adapted for individual gene recognition (RING‐FISH) was used to confirm the presence/absence of vopB2. T3SS2β was found in all tdh−/trh+ isolates and in no tdh+/trh− isolates. Conclusions: The combination of haemolysins found in each isolate consistently corresponded to the presence and type of T3SS detected. The vopB2 gene may represent a novel marker for identifying increased virulence among strains. Significance and Impact of the Study: This is the first study to confirm the presence of T3SS2β genes in V. parahaemolyticus strains isolated from the Gulf of Mexico and one of the few that examines the distribution and co‐existence of tdh, trh, T3SS1, T3SS2α and T3SS2β in a large collection of environmental strains.

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“…The TRH was first found to be produced by a V. parahaemolyticus O3:K6 strain implicated in an outbreak of gastro-enteritis (Honda et al, 1987;1988). Subsequent molecular epidemiological study with gene probes demonstrated that the trh gene encoding TRH was strongly associated with clinical strains of V. parahaemolyticus (Shirai et al, 1990). Some of the recent studies on clinical and environmental V. parahaemolyticus have recorded increasing prevalence of trh + V. parahaemolyticus.…”

Section: Discussionmentioning

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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Molecular epidemiologic evidence for association of thermostable direct hemolysin (TDH) and TDH-related hemolysin of Vibrio parahaemolyticus with gastroenteritis

Cited by 256 publications

References 27 publications

Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates

Characterization of United States outbreak isolates of Vibrio parahaemolyticus using enterobacterial repetitive intergenic consensus (ERIC) PCR and development of a rapid PCR method for detection of O3:K6 isolates

Distribution of type III secretion systems in Vibrio parahaemolyticus from the northern Gulf of Mexico

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

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