Septaplex PCR assay for rapid identification of<i>Vibrio cholerae</i>including detection of virulence and<i>int</i>SXT genes

Mantri, Chinmay K.; Mohapatra, Saswat S.; Ramamurthy, Thandavarayan; Ghosh, Raikamal; Colwell, Rita R.; Singh, D. V.

doi:10.1111/j.1574-6968.2006.00491.x

Cited by 22 publications

(12 citation statements)

References 25 publications

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“…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. For V. cholerae, a multiplex-PCR was performed with the primer sets UtoxF/VctoxR, O139F/O139R, ctxA1/ctxA2 and O1F/O1R [43,46,47]. Half a micromole of each O1 Primer and 0.125 μmol of every other primer were used.…”

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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“…Specific PCRs targeting thermostable direct haemolysin (tdh) (Nishibuchi and Kaper, 1985) and the tdh related haemolysin (trh) (Honda et al, 1991;Honda and Iida, 1993) genes were performed with the primer sets tdhD3F/ tdhD1R and trhFR2/trhRR6 to strains assigned to V. parahaemolyticus (Bauer and Rorvik, 2007;Tada et al, 1992). To test V. cholerae strains for the presence of a unique chromosomal region indicating the serotypes O139 (Albert et al, 1997) and O1 (Hoshino et al, 1998) and the cholera toxin gene ctxA (Singh et al, 2002) a multiplex PCR was performed with the primer sets O139F/O139R, O1F/O1R and ctxA1/ctxA2 (Bauer and Rorvik, 2007;Mantri et al, 2006;Nandi et al, 2000). All reactions were performed in duplicate.…”

Section: Pcr Of Regulatory and Virulence-related Genesmentioning

confidence: 99%

Dangerous hitchhikers? Evidence for potentially pathogenic Vibrio spp. on microplastic particles

Kirstein

Kirmizi

Wichels

et al. 2016

Marine Environmental Research

704

372

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The taxonomic composition of biofilms on marine microplastics is widely unknown. Recent sequencing results indicate that potentially pathogenic Vibrio spp. might be present on floating microplastics. Hence, these particles might function as vectors for the dispersal of pathogens. Microplastics and water samples collected in the North and Baltic Sea were subjected to selective enrichment for pathogenic Vibrio species. Bacterial colonies were isolated from CHROMagar™Vibrio and assigned to Vibrio spp. on the species level by MALDI-TOF MS (Matrix Assisted Laser Desorption/IonisationTime of Flight Mass Spectrometry). Respective polymers were identified by ATR FT-IR (Attenuated Total Reflectance Fourier Transform -Infrared Spectroscopy). We discovered potentially pathogenic Vibrio parahaemolyticus on a number of microplastic particles, e.g. polyethylene, polypropylene and polystyrene from North/Baltic Sea. This study confirms the indicated occurrence of potentially pathogenic bacteria on marine microplastics and highlights the urgent need for detailed biogeographical analyses of marine microplastics.

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“…Mit den Primern aus der rfb-Region, die für die O-Antigen-Biosynthese verantwortlich ist, kann man Vibrio cholerae O1 oder O139 detektieren und differenzieren (Tab. 1; Mantri et al, 2006). Aus diagnostischer Sicht ist aber alleinig der Nachweis des CTXφ Phagen durch eine ctxA-PCR (Tab.…”

Section: Klassischer Pcr-nachweisunclassified

Der PCR-gestützte Nachweis von Vibrio cholerae in klinischen und nicht klinischen Proben

Prager

Tschäpe

2007

J. Verbr. Lebensm.

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Zusammenfassung: Obwohl es gute und verlässliche klassischmikrobiologische Verfahren zur Differenzierung und Diagnostik von pathogenen Vibrio cholerae Stämmen gibt, sind in den letzten Jahren immer mehr und sehr erfolgreich PCR Verfahren und real-time PCR-Verfahren zur Spezies-Diagnostik, zur Bestimmung der relevanten Serotypen sowie zur Erfassung von Pathovaren bzw. von virulenten sowie nicht-virulenten Varianten entwickelt und in der praktischen Arbeit erprobt worden. Die z. Z. gängigen und etablierten PCR Verfahren werden in der vorliegenden Arbeit tabellarisch zusammengefasst und diskutiert.Abstract: Although classical microbiological methods for the detection and identifi cation of pathogenic Vibrio cholerae are suffi ciently and reliably available today, the successful development and application of PCR-based assays for species diagnostics, determination of relevant serotypes, discrimination of pathovars and virulent variants were carried out in the last couple of years. The currently available PCR-based assays for V. cholerae are summarised and discussed in the present paper.

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Septaplex PCR assay for rapid identification ofVibrio choleraeincluding detection of virulence andintSXT genes

Cited by 22 publications

References 25 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

Dangerous hitchhikers? Evidence for potentially pathogenic Vibrio spp. on microplastic particles

Der PCR-gestützte Nachweis von Vibrio cholerae in klinischen und nicht klinischen Proben

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