Phenotypic and Genotypic Characterization of Canadian Clinical Isolates of Vibrio parahaemolyticus Collected from 2000 to 2009

Banerjee, Swapan; Kearney, Ashley; Nadon, Céline; Peterson, Christy-Lynn; Tyler, Kevin; Bakouche, Laurene; Clark, Clifford G.; Hoang, Linda; Gilmour, Matthew W.; Farber, Jeffrey M.

doi:10.1128/jcm.03047-13

Cited by 54 publications

(62 citation statements)

References 33 publications

(34 reference statements)

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“…Confirmation could be done by additional genotyping, such as for the flp locus ( Fig. 2 and Table 1) (16), by application of one of the other primer sets for prp (Table 1), through other typing methods, including PFGE and serotyping (15), by four-or seven-gene MLST (17,18,21,30) (Fig. 1), or, when resources are available, by whole-genome sequencing and phylogeny (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…Only a few virulence markers are known and routinely applied to pathogen identification, including the tdh and trh hemolysin genes, but these do not detect all pathogens. Indeed, Ͼ10% of infections in North America are apparently caused by strains that lack these genes, whose prevalence among nonpathogens and other Vibrio species is not known (10)(11)(12)(13)(14)(15). The detection of a combination of traits or markers that can identify pathogen lineages of most concern, along with quickly and affordably identifying virulence traits, could improve the reliability of pathogen discrimination, identification, and surveillance.…”

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confidence: 99%

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Use of Whole-Genome Phylogeny and Comparisons for Development of a Multiplex PCR Assay To Identify Sequence Type 36 Vibrio parahaemolyticus

Whistler

Hall

et al. 2015

J Clin Microbiol

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dVibrio parahaemolyticus sequence type 36 (ST36) strains that are native to the Pacific Ocean have recently caused multistate outbreaks of gastroenteritis linked to shellfish harvested from the Atlantic Ocean. Whole-genome comparisons of 295 genomes of V. parahaemolyticus, including several traced to northeastern U.S. sources, were used to identify diagnostic loci, one putatively encoding an endonuclease (prp), and two others potentially conferring O-antigenic properties (cps and flp). The combination of all three loci was present in only one clade of closely related strains of ST36, ST59, and one additional unknown sequence type. However, each locus was also identified outside this clade, with prp and flp occurring in only two nonclade isolates and cps in four. Based on the distribution of these loci in sequenced genomes, prp identified clade strains with >99% accuracy, but the addition of one more locus increased accuracy to 100%. Oligonucleotide primers targeting prp and cps were combined in a multiplex PCR method that defines species using the tlh locus and determines the presence of both the tdh and trh hemolysin-encoding genes, which are also present in ST36. Application of the method in vitro to a collection of 94 clinical isolates collected over a 4-year period in three northeastern U.S. states and 87 environmental isolates revealed that the prp and cps amplicons were detected only in clinical isolates identified as belonging to the ST36 clade and in no environmental isolates from the region. The assay should improve detection and surveillance, thereby reducing infections. Vibrio parahaemolyticus is typically harmless, but pathogenic strains can cause severe inflammatory gastroenteritis infections that rarely progress to lethal sepsis (1). It is the leading cause of bacterial seafood-borne illness worldwide, with raw or improperly handled seafood as a major vector. In the United States, it has been of greatest concern for shellfish harvested in the Gulf of Mexico and the Pacific Northwest (2-5). Infections linked to shellfish from the northeastern United States have been rare, but a steep rise in infections occurred in 2012 to 2013 that was concurrent with the probable ecological invasion of a serotype O4:K12 sequence type 36 (ST36) strain. This strain type has been linked to recurrent infections in the Pacific Northwest for more than a decade, suggesting that it may have expanded its geographic range (6, 7). Furthermore, unlike native strains present in the northern Atlantic that cause infrequent infections, the ST36 strain is responsible for an ongoing multistate outbreak (7) (Fig. 1). Rapid identification of this strain complex in clinical samples might aid in the prevention of more widespread infections. Additionally, accurate quantification of this strain in shellfish growing areas could inform harvest strategies that maintain a safe product.Identifying rare pathogenic strains among mostly nonpathogenic populations of V. parahaemolyticus has been a long-standing challenge. A few strains, such as tho...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Use of Whole-Genome Phylogeny and Comparisons for Development of a Multiplex PCR Assay To Identify Sequence Type 36 Vibrio parahaemolyticus

Whistler

Hall

et al. 2015

J Clin Microbiol

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“…Recent studies have reported the isolation of strains that lack both tdh and trh (tdh-and trh-negative strains) from clinical samples (22)(23)(24)(25)(26)(27). However, the ability of tdh-and trh-negative strains to independently cause clinical illness is still controversial.…”

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confidence: 99%

Genomic Features of Environmental and Clinical Vibrio parahaemolyticus Isolates Lacking Recognized Virulence Factors Are Dissimilar

Ronholm

Petronella

Leung

et al. 2016

Appl Environ Microbiol

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Vibrio parahaemolyticus is a bacterial pathogen that can cause illness after the consumption or handling of contaminated seafood. The primary virulence factors associated with V. parahaemolyticus illness are thermostable direct hemolysin (TDH) and Tdh-related hemolysin (TRH). However, clinical strains lacking tdh and trh have recently been isolated, and these clinical isolates are poorly understood. To help understand the emergence of clinical tdh-and trh-negative isolates, a genomic approach was used to comprehensively compare 4 clinical tdh-and trh-negative isolates with 16 environmental tdh-and trh-negative isolates and 34 clinical isolates positive for tdh or trh, or both, with the objective of identifying genomic features that are unique to clinical tdh-and trh-negative isolates. The prevalence of pathogenicity islands (PAIs) common to clinical isolates was thoroughly examined in each of the clinical tdh-and trh-negative isolates. The tdh PAI was not present in any clinical or environmental tdhand trh-negative isolates. The trh PAI was not present in any environmental isolates; however, in clinical tdh-and trh-negative isolate 10-4238, the majority of the trh PAI including a partial trh1 gene was present, which resulted in reclassification of this isolate as a tdh-negative and trh-positive isolate. In the other clinical tdh-and trh-negative isolates, neither the trh gene nor the trh PAI was present. We identified 862 genes in clinical tdh-and trh-negative isolates but not in environmental tdh-and trh-negative isolates. Many of these genes are highly homologous to genes found in common enteric bacteria and included genes encoding a number of chemotaxis proteins and a novel putative type VI secretion system (T6SS) effector and immunity protein (T6SS1). The availability of genome sequences from clinical V. parahaemolyticus tdh-and trh-negative isolates and the comparative analysis may help provide an understanding of how this pathotype is able to survive in vivo during clinical illness. Vibrio parahaemolyticus is a Gram-negative, halophilic bacterium that is ubiquitous in marine and estuarine environments and is often found colonizing shellfish or shrimp. While most strains are nonpathogenic, many have acquired virulence factors that result in illness when individuals are exposed to V. parahaemolyticus strains carrying these virulence factors (1). V. parahaemolyticus is recognized to be a leading cause of foodborne illness worldwide and is transmitted via the handling and consumption of raw or undercooked contaminated seafood (1). Infections occur both sporadically and in very large outbreaks. The most common manifestation of V. parahaemolyticus infection is acute, watery diarrhea accompanied by abdominal pain and nausea, although symptoms can also be severe and include a dysenterylike illness or septicemia (2). Since most cases of illness caused by V. parahaemolyticus are self-limiting, rates of infection are probably underestimated due to underreporting.Clinical V. parahaemolyticus isolates generally have at...

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“…Among the infections associated with the genus Vibrio, the majority was caused by V. parahaemolyticus (Crim et al, 2014). While most foodborne pathogens (i.e., Listeria, Campylobacter, and Escherichia coli O157) have shown a decrease in incidence from 1998 to 2008, V. parahaemolyticus has increased 47% within this time period (Banerjee et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…Underreporting can be attributed by the lack of awareness of the disease, the self-limiting nature of the infection, and the lack of urgency to visit physicians (Banerjee et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Virulence gene expression of Vibrio parahaemolyticus in the viable but nonculturable state

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Phenotypic and Genotypic Characterization of Canadian Clinical Isolates of Vibrio parahaemolyticus Collected from 2000 to 2009

Cited by 54 publications

References 33 publications

Use of Whole-Genome Phylogeny and Comparisons for Development of a Multiplex PCR Assay To Identify Sequence Type 36 Vibrio parahaemolyticus

Use of Whole-Genome Phylogeny and Comparisons for Development of a Multiplex PCR Assay To Identify Sequence Type 36 Vibrio parahaemolyticus

Genomic Features of Environmental and Clinical Vibrio parahaemolyticus Isolates Lacking Recognized Virulence Factors Are Dissimilar

Virulence gene expression of Vibrio parahaemolyticus in the viable but nonculturable state

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