Vibrio parahaemolyticus O serotypes from O1 to O13 all produce R-type lipopolysaccharide: SDS-PAGE and compositional sugar analysis

Iguchi, Takehiro

doi:10.1016/0378-1097(95)00220-y

Cited by 21 publications

(32 citation statements)

References 11 publications

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“…The structural genes for the hemolysins, tdh and trh, respectively, are encoded chromosomally; and PCR-based methods for the detection of the genes have been successfully developed (16,19). V. parahaemolyticus can be classified into 13 O serotypes and 71 K serotypes (9). Although various serovars of the bacterium can cause infections, O3:K6 has been recognized as the predominant serovar responsible for most outbreaks worldwide since 1996 (5,13,17).…”

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confidence: 99%

Genotypic Analyses of Vibrio parahaemolyticus and Development of a Pandemic Group-Specific Multiplex PCR Assay

et al. 2003

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A total of 54 Vibrio parahaemolyticus strains including pandemic O3:K6 strains and newly emerged O4:K68, O1:K25, O1:K26, and O1:K untypeable strains (collectively referred to as the "pandemic group") were examined for their pulsed-field gel electrophoresis (PFGE) and arbitrarily primed PCR (AP-PCR) profiles and for the presence or absence of genetic marker DNA sequences, toxRS/new or orf8, that had been reported elsewhere to be specific for the pandemic group. Both PFGE and AP-PCR analyses indicated that all strains of the pandemic group formed a distinct genotypic cluster, suggesting that they originated from the same clone. In addition to the pandemic group, four O3:K6 strains that did not possess the thermostable direct hemolysin (tdh) gene also belonged to this cluster and possessed the toxRS/new sequence. However, three O3:K6 strains that clearly belonged to the pandemic group by PFGE and AP-PCR did not possess the orf8 sequence. The evidence suggests that neither the toxRS/new nor the orf8 sequence is a reliable gene marker for definite identification of the pandemic group. We therefore developed a novel multiplex PCR assay specific for the pandemic group. The assay successfully distinguished pandemic group strains from other V. parahaemolyticus strains by yielding two distinct PCR products for tdh (263 bp) and the toxRS/new sequence (651 bp).Vibrio parahaemolyticus causes one of the major forms of seafood-borne gastroenteritis, often associated with the consumption of raw or undercooked seafood (4). Past epidemiological studies (7,8,14) revealed a strong association between the thermostable direct hemolysin (TDH) and another hemolysin termed TDH-related hemolysin (TRH), which are produced by members of V. parahaemolyticus, and its pathogenicity. Both hemolysins are thus considered major virulence factors of V. parahaemolyticus. The structural genes for the hemolysins, tdh and trh, respectively, are encoded chromosomally; and PCR-based methods for the detection of the genes have been successfully developed (16,19).

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Genotypic Analyses of Vibrio parahaemolyticus and Development of a Pandemic Group-Specific Multiplex PCR Assay

et al. 2003

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“…Isolates of V. parahaemolyticus can be differentiated by serotyping, and 13 O groups and 71 K types have been identified (7). Serotyping is generally unable to differentiate all isolates originating from different regions or sources.…”

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Evaluation of Typing of Vibrio parahaemolyticus by Three PCR Methods Using Specific Primers

Wong

Lin

2001

J Clin Microbiol

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Vibrio parahaemolyticus is a halophilic bacterium frequently involved in human outbreaks of seafoodassociated gastroenteritis. For epidemiological purposes, different molecular typing methods, such as pulsedfield gel electrophoresis (PFGE) or ribotyping, have been developed for this pathogen; however, these methods are mostly labor-intensive and time-consuming. In this work, we designed and evaluated three rapid PCR typing methods for this pathogen using primers designed on the basis of the following specific sequences: conserved ribosomal gene spacer sequence (RS), repetitive extragenic palindromic sequence (REP), and enterobacterial repetitive intergenic consensus sequence (ERIC). Typing patterns and clustering analysis indicated that these methods apparently differentiated V. parahaemolyticus strains from reference strains of interspecific Escherichia coli, V. cholerae, and V. vulnificus and were also valuable in subspecies typing of this pathogen. Forty domestic strains of V. parahaemolyticus, representing a wide range of PFGE patterns, were grouped into 15, 27, and 27 patterns, with discrimination indexes of 0.91, 0.97, and 0.98, by RS-, REP-, and ERIC-PCR, respectively. The discriminative abilities of these PCR methods closely approached or even exceeded those of PFGE and ribotyping. REP-PCR is preferable to ERIC-PCR because of the greater reproducibility of its fingerprints, while RS-PCR may be a practical method because it generates fewer amplification bands and patterns than the alternatives.Vibrio parahaemolyticus is a halophilic gram-negative bacterium that causes acute gastroenteritis in humans. Food poisoning caused by this pathogen is generally associated with the consumption of contaminated seafood; this organism is a crucial food-borne pathogen in Taiwan, Japan, and other coastal countries with high rates of seafood consumption (17). Clinical manifestations include diarrhea, abdominal cramps, nausea, vomiting, headache, fever, and chills, with incubation periods ranging from 4 to 96 h (4, 9).Isolates of V. parahaemolyticus can be differentiated by serotyping, and 13 O groups and 71 K types have been identified (7). Serotyping is generally unable to differentiate all isolates originating from different regions or sources. Reliable molecular methods for strain typing would significantly aid epidemiological investigations. Recently, several molecular methods were developed for the subspecies typing of V. parahaemolyticus, namely, pulsed-field gel electrophoresis (PFGE) (33), ribotyping (29), and random amplified polymorphic DNA (RAPD) analysis (30). The PFGE method using SfiI digestion is reliable, achieves high discrimination efficiency, and has been applied to typing of V. parahaemolyticus strains in many situations, such as the first pandemic O3:K6 strains (32), food poisoning outbreaks (28), environmental strains from seafood (31), and nosocomial outbreaks (12). However, the whole process takes several days to complete. Compared with PFGE, RAPD analysis has the merits of being less labor-intensi...

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“…Thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) are considered major virulence factors for the organism (7). V. parahaemolyticus can be classified into 13 O serotypes and 71 K serotypes (3). Although various serovars of the bacterium can cause infections, O3:K6 has been recognized as the predominant serovar responsible for most outbreaks worldwide since 1996 (5).…”

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confidence: 99%

Genotyping of Pandemic Vibrio parahaemolyticus O3:K6Still Open to Question

et al. 2002

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Vibrio parahaemolyticus O serotypes from O1 to O13 all produce R-type lipopolysaccharide: SDS-PAGE and compositional sugar analysis

Cited by 21 publications

References 11 publications

Genotypic Analyses of Vibrio parahaemolyticus and Development of a Pandemic Group-Specific Multiplex PCR Assay

Genotypic Analyses of Vibrio parahaemolyticus and Development of a Pandemic Group-Specific Multiplex PCR Assay

Evaluation of Typing of Vibrio parahaemolyticus by Three PCR Methods Using Specific Primers

Genotyping of Pandemic Vibrio parahaemolyticus O3:K6Still Open to Question

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