Genetic and serological identification of three Vibrio parahaemolyticus strains as candidates for novel provisional O serotypes

Guo, Xi; Liu, Bin; Chen, Min; Wang, Yuanyuan; Wang, Lu; Chen, Hongyou; Wang, Yao; Tu, Lihong; Zhang, Xi; Feng, Lu

doi:10.1016/j.ijfoodmicro.2017.01.010

Cited by 15 publications

(33 citation statements)

References 28 publications

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“…Traditionally, 13 kinds of O antigens and 71 kinds of K antigens have been identified [33]. However, V. parahaemolyticus with untypeable and new O or K antigens have been frequently isolated [10,34,35]. Therefore, further new O and K antigens could be expected in V. parahaemolyticus in the future.…”

Section: Discussionmentioning

confidence: 99%

Co-existence of multiple distinct lineages in Vibrio parahaemolyticus serotype O4:K12

et al. 2020

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Section: Discussionmentioning

confidence: 99%

Co-existence of multiple distinct lineages in Vibrio parahaemolyticus serotype O4:K12

et al. 2020

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“…To our knowledge, only four studies have explored the use of molecular methods for serogroup or serotype identification of V. parahaemolyticus to-date. Conventional PCR-based assays have been developed for the identification of O-serogroups (Chen et al, 2012; Guo et al, 2017) and serotype O3:K6 (Yeung et al, 2003), whereas the utility of MALDI-ToF has been explored for the identification for serotype O4:K8 (Li et al, 2018). However, the practical application of these methods has significant limitations, such as the need for cumbersome procedures associated with gel electrophoresis and the ability to only identify limited or single serogroups and serotypes simultaneously.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, a better understanding of the OGD through genetic analyses would further improve the current O-antigenic scheme. A recent study has shown that the frequent occurrence of untypeable O-serogroups (OUTs) may have resulted from intra-species recombination events that occurred within the OGD region and evolved from the current O-serogroups, where several provisional and rare O-serogroup candidates were proposed (Guo et al, 2017). Whereas, for K- serogroups, the genetic locations for encoding K-antigen synthesis in V. parahaemolyticus (Guvener and Mccarter, 2003; Okura et al, 2003) have been a subject of debate, but was subsequently confirmed to be between gmhD and rjg using an O3:K6 isolate by construction of gene deletions (Chen et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Despite these drawbacks, the development of molecular methods for the serotyping of V. parahaemolyticus has been scarce, with only a few methods described to-date. A conventional PCR-based method has been described for the detection and identification of V. parahaemolyticus O-serogroups based on O-serogroup genetic determinants (OGDs) (Chen et al, 2012), which was subsequently extended to include three additional provisional O-serogroups, but without any K- serogroups (Guo et al, 2017). Another conventional PCR was developed to detect an arbitrary marker for the identification of a single serotype O3:K6 (Yeung et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous Identification of Clinically Common Vibrio parahaemolyticus Serotypes Using Probe Melting Curve Analysis

Jiang

Shi

et al. 2019

Front. Cell. Infect. Microbiol.

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The dynamic nature of Vibrio parahaemolyticus epidemiology has presented a unique challenge for disease intervention strategies. Despite the continued rise of disease incidence and outbreaks of vibriosis, as well as the global emergence of pandemic clones and serovariants with enhanced virulence, there is a paucity of molecular methods for the serotyping of V. parahaemolyticus strains to improve disease surveillance and outbreak investigations. We describe the development of a multiplex ligation reaction based on probe melting curve analysis (MLMA) for the simultaneous identification of 11 clinically most common V. parahaemolyticus serotypes spanning a 10-year period. Through extensive sequence analyses using 418 genomes, specific primers and probes were designed for a total of 22 antigen gene targets for the O- and K- serogroups. Additionally, the toxR gene was incorporated into the assay for the confirmation of V. parahaemolyticus. All gene targets were detected by the assay and gave expected Tm values, without any cross reactions between the 11 clinically common serotypes or with 38 other serotypes. The limit of identification for all gene targets ranged from 0.1 to 1 ng/μL. The intra- and inter-assay standard deviations and the coefficients of variation were no more than 1°C and <1% respectively, indicating a highly reproducible assay. A multicenter double-blind clinical study was conducted using the traditional V. parahaemolyticus identification workflow and the MLMA assay workflow in parallel. From consecutive diarrheal stool specimens (n = 6118) collected over a year at 10 sentinel hospitals, a total of 153 V. parahaemolyticus isolates (2.5%) were identified by both workflows. A total agreement (kappa = 1.0) between the serotypes identified by the MLMA assay and conventional serological method was demonstrated. This is the first molecular assay to simultaneously identify multiple clinically important V. parahaemolyticus serotypes, which satisfies the acute need for a practical, rapid and robust identification of V. parahaemolyticus serotypes to facilitate the timely detection of vibriosis outbreaks and surveillance.

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“…Second, serotyping is labor-intensive, and only a few reference laboratories have the capacity to perform the analyses necessary to identify serotypes ( Ballmer et al, 2007 ; Lacher et al, 2014 ). In addition, conventional serological assays that are based on agglutination reactions are also limited by the risks of the high prevalence of non-typeable isolates due to the expression of novel polysaccharide forms, which is common in clinical isolates ( Jenney et al, 2006 ; Thrane et al, 2016 ; Guo et al, 2017 ). The development of NGS technologies and automated data analysis pipelines has led to a number of proof-of-concept studies.…”

Section: Discussionmentioning

confidence: 99%

Establishment of a Molecular Serotyping Scheme and a Multiplexed Luminex-Based Array for Enterobacter aerogenes

Guo

Wang

et al. 2018

Front. Microbiol.

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Serotyping based on surface polysaccharide antigens is important for the clinical detection and epidemiological surveillance of pathogens. Polysaccharide gene clusters (PSgcs) are typically responsible for the diversity of bacterial surface polysaccharides. Through whole-genome sequencing and analysis, eight putative PSgc types were identified in 23 Enterobacter aerogenes strains from several geographic areas, allowing us to present the first molecular serotyping system for E. aerogenes. A conventional antigenic scheme was also established and correlated well with the molecular serotyping system that was based on PSgc genetic variation, indicating that PSgc-based molecular typing and immunological serology provide equally valid results. Further, a multiplex Luminex-based array was developed, and a double-blind test was conducted with 97 clinical specimens from Shanghai, China, to validate our array. The results of these analyses indicated that strains containing PSgc4 and PSgc7 comprised the predominant groups. We then examined 86 publicly available E. aerogenes strain genomes and identified an additional seven novel PSgc types, with PSgc10 being the most abundant type. In total, our study identified 15 PSgc types in E. aerogenes, providing the basis for a molecular serotyping scheme. From these results, differing epidemic patterns were identified between strains that were predominant in different regions. Our study highlights the feasibility and reliability of a serotyping system based on PSgc diversity, and for the first time, presents a molecular serotyping system, as well as an antigenic scheme for E. aerogenes, providing the basis for molecular diagnostics and epidemiological surveillance of this important emerging pathogen.

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Genetic and serological identification of three Vibrio parahaemolyticus strains as candidates for novel provisional O serotypes

Cited by 15 publications

References 28 publications

Co-existence of multiple distinct lineages in Vibrio parahaemolyticus serotype O4:K12

Co-existence of multiple distinct lineages in Vibrio parahaemolyticus serotype O4:K12

Simultaneous Identification of Clinically Common Vibrio parahaemolyticus Serotypes Using Probe Melting Curve Analysis

Establishment of a Molecular Serotyping Scheme and a Multiplexed Luminex-Based Array for Enterobacter aerogenes

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