Comparison of a commercial biochemical kit and an oligonucleotide probe for identification of environmental isolates of Vibrio vulnificus

Dalsgaard, Anders; Dalsgaard, Inger; Høi, Lise; Larsen, J. L.

doi:10.1111/j.1472-765x.1996.tb01138.x

Cited by 26 publications

(21 citation statements)

References 23 publications

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“…Given the fact that previous reports have already concluded that the use of API 20 E is inadequate for the identification of V. vulnificus (6), only the API 20 NE system was used in this study. API 20 NE strips were inoculated and processed following the manufacturer's recommendations, and the results were analyzed by using Apilab software, version 3.2.2.…”

Section: Methodsmentioning

confidence: 99%

Identification of the Emerging Pathogen Vibrio vulnificus Biotype 3 by Commercially Available Phenotypic Methods

Colodner¹,

Raz²,

Meir

et al. 2004

J Clin Microbiol

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Identification of the emerging pathogen Vibrio vulnificus biotype 3 has become a challenge for clinical laboratories in the last few years. In this study, the abilities of five commercial systems to identify this new species have been evaluated for the first time, using a unique collection of strains. Fifty-one well-documented wild strains of V. vulnificus biotype 3 were processed using API 20 NE, GNI؉ Vitek 1 cards, ID-GNB Vitek 2 cards, Neg Combo 20 Microscan panels, and NMIC/ID-5 BD Phoenix panels. The numbers of strains identified as V. vulnificus by ID-GNB, NMIC/ID-5, and GNI؉ were 50 (98.0%), 46 (90.2%), and 7 (13.7%), respectively. Neg Combo 20 Microscan panels and API 20 NE were unable to identify any of the strains of this emerging pathogen to the species level and mostly misidentifies them as other species of the Vibrionaceae family. Data on the phenotypic pattern of V. vulnificus biotype 3 when processed in all five systems as presented here could help clinical laboratories in identifying this new pathogen.Since it was first described by Reichelt et al. in 1976 (9), for many years only two biotypes or serovars of the human pathogen Vibrio vulnificus have been recognized (1).In the midsummer of 1996, we reported the first isolation of a new pathogen, V. vulnificus biotype 3, which causes septicemia and severe soft-tissue infections following contact with fish from artificial freshwater ponds (2). Since then, the identification of this emerging pathogen has become a challenge for clinical laboratories. With the phenotypic behavior of this new biotype being different from that of the V. vulnificus biotypes known so far, most of the commercial systems in use do not include the relevant data in their software, making correct identification of the new bacteria problematic.In this study, the phenotypic pattern of this new biotype when tested with five commercial systems was defined, and the abilities of the current software of those systems to correctly identify V. vulnificus biotype 3 to the species level have been evaluated for the first time with a unique collection of strains. MATERIALS AND METHODSIdentification systems. Fifty-one well-documented V. vulnificus biotype 3 strains isolated during the years 1996 to 1997 were processed by five commercial identification systems: API 20 NE, GNIϩ Vitek 1 cards, ID-GNB Vitek 2 cards (BioMerieux, Marcy l-Etoile, France), Neg Combo 20 Microscan panels (Dade Behring Inc.), and NMIC/ID-5 BD Phoenix panels (BD Biosciences).Given the fact that previous reports have already concluded that the use of API 20 E is inadequate for the identification of V. vulnificus (6), only the API 20 NE system was used in this study. API 20 NE strips were inoculated and processed following the manufacturer's recommendations, and the results were analyzed by using Apilab software, version 3.2.2.Inoculated Vitek GNIϩ cards were processed with a Vitek 1 system, version VTK-R 07.02.Vitek ID-GNB cards were used with a Vitek 2 system, version VT2-R 02.03. Microscan Neg Combo 20 panels were inoc...

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Section: Methodsmentioning

confidence: 99%

Identification of the Emerging Pathogen Vibrio vulnificus Biotype 3 by Commercially Available Phenotypic Methods

Colodner¹,

Raz²,

Meir

et al. 2004

J Clin Microbiol

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“…The few studies conducted have, however, revealed the presence of V. parahaemolyticus and V. vulnificus (3,9,14).…”

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confidence: 99%

Occurrence of Vibrio parahaemolyticus , V. cholerae , and V. vulnificus in Norwegian Blue Mussels ( Mytilus edulis )

Bauer

Østensvik

Florvåg

et al. 2006

Appl Environ Microbiol

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“…Later, CPC agar was modified (mCPC) by reducing the concentration of colistin (33), which improved the recovery and isolation of V. vulnificus from environmental sources. Recently, a new modification of CPC agar has been described (16).To identify presumptive V. vulnificus recovered from these media, probes based on the cytolysin gene of V. vulnificus have been developed (11,25,37). The possible loss or rearrangement of nonessential genes, such as the cytotoxin-hemolysin gene, can lead to a false-negative result (12).…”

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A Selective Medium and a Specific Probe for Detection of Vibrio vulnificus

Cerdà‐Cuéllar

Jofre

Blanch

2000

Appl Environ Microbiol

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A selective medium (VVM) and a specific 16S rRNA gene (rDNA) probe (V3VV) for the detection of Vibrio vulnificus were developed. The medium contains D-(؉)-cellobiose as the main carbon source and electrolytes (MgCl 2 -6H 2 O and KCl), which stimulate bacterial growth. Polymyxin B, colistin, and moderate alkalinity and salinity provide selectivity properties. V. vulnificus grows on VVM as flat, bright yellow colonies. Other Vibrio species tested either did not grow or showed green-bluish colonies, with the exception of V. campbelli, V. carchariae, and V. navarrensis. There is a higher colony count on VVM agar than on cellobiose-colistin agar or on modified cellobiose-polymyxin B-colistin agar. The specific probe was evaluated by colony hybridization and dot blot hybridization with PCR-amplified 16S rDNA using collection strains and environmental isolates. No strain studied other than V. vulnificus showed positive hybridization with this oligonucleotide. The combined use of VVM agar and the V3VV probe provided the recovery of V. vulnificus from mixed bacterial suspensions and spiked mussels.Vibrio vulnificus is a ubiquitous bacterium found in estuarine and marine environments. It comprises two biotypes: biotype 1 strains are pathogenic in humans, while biotype 2 strains are virulent in eels and opportunistic in humans, although they have been thought to be pathogenic only for eels until recently (2). It has been isolated from a wide variety of marine organisms, such as oysters, crabs, fish, mussels, shellfish, and plankton (10,17,18,36), and from water and sediment (34). It is well known that raw-shellfish consumption increases the risk of human vibrio diseases. Particularly, V. vulnificus can cause fulminant and severe systemic human illness after the consumption of infected raw oysters (30). Mortality rates of up to 60% have been reported in such infections (18). It is remarkably virulent in individuals who are immunocompromised or have liver dysfunction, which results in increased levels of iron in serum (18,28). Thus, its occurrence in aquatic environments is of concern to shellfish industries and public health agencies. Because a great number and variety of indigenous bacteria are present in the environment together with some pathogenic Vibrio species, the isolation of V. vulnificus is usually performed by enrichment in alkaline peptone water (APW) or in APW supplemented with polymyxin B. However, this enrichment also increases the number of other bacteria, which requires further inoculation of the enriched sample on selective media (10, 16) such as V. vulnificus agar (7), sodium dodecyl sulfate (SDS)-polymyxin B-sucrose (SPS) agar (21), cellobiose-polymyxin B-colistin (CPC) agar (23), V. vulnificus enumeration agar (24), modified CPC (mCPC) agar (33), and cellobiose-colistin (CC) agar (16). The recovery of V. vulnificus was higher with CPC agar than with thiosulfate-citrate-bile salts-sucrose (TCBS), V. vulnificus enumeration, or SDS-polymyxin-sucrose agar (20,23,29,31,32). Later, CPC agar was modified (mCPC...

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Comparison of a commercial biochemical kit and an oligonucleotide probe for identification of environmental isolates of Vibrio vulnificus

Cited by 26 publications

References 23 publications

Identification of the Emerging Pathogen Vibrio vulnificus Biotype 3 by Commercially Available Phenotypic Methods

Identification of the Emerging Pathogen Vibrio vulnificus Biotype 3 by Commercially Available Phenotypic Methods

Occurrence of Vibrio parahaemolyticus , V. cholerae , and V. vulnificus in Norwegian Blue Mussels ( Mytilus edulis )

A Selective Medium and a Specific Probe for Detection of Vibrio vulnificus

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