Rapid detection of Vibrio cholerae O139 Bengal from stool specimens by PCR

Albert, M. John; Islam, Dilara; Nahar, Shamsun; Qadri, Firdausi; Falklind, S; Weintraub, Andrej

doi:10.1128/jcm.35.6.1633-1635.1997

Cited by 35 publications

(19 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Probes for O139 serogroup were designed and evaluated for quick determination of V. cholerae Nair et al, 1995). Primers for V. cholerae O139 detection by PCR were designed by Falklind et al (1996) and Albert et al (1997). However, because of the close similarity of the two rfb genes to the O1 serogroup, a tenuous cross-reaction was detected.…”

Section: Discussionmentioning

confidence: 99%

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Rivera

Lipp

Gil

et al. 2003

Environmental Microbiology

View full text Add to dashboard Cite

Vibrio cholerae is a free-living bacterium found in water and in association with plankton. V. cholerae non-O1/non-O139 strains are frequently isolated from aquatic ecosystems worldwide. Less frequently isolated are V. cholerae O1 and V. cholerae O139, the aetiological agents of cholera. These strains have two main virulence-associated factors, cholera toxin (CT) and toxin co-regulated pilus (TCP). By extracting total DNA from aquatic samples, the presence of pathogenic strains can be determined quickly and used to improve a microbiological risk assessment for cholera in coastal areas. Some methods suggested for DNA extraction from water samples are not applicable to all water types. We describe here a method for DNA extraction from coastal water and a multiplex polymerase chain reaction (PCR) for O1 and O139 serogroups. DNA extraction was successfully accomplished from 117 sea water samples collected from coastal areas of Perú, Brazil and the USA. DNA concentration in all samples varied from 20 ng to 480 micro g micro l-1. The sensitivity of the DNA extraction method was 100 V. cholerae cells in 250 ml of water. The specificity of multiplex O1/O139 PCR was investigated by analysing 120 strains of V. cholerae, Vibrio and other Bacteria species. All V. cholerae O1 and O139 tested were positive. For cholera surveillance of aquatic environments and ballast water, total DNA extraction, followed by V. cholerae PCR, and O1/O139 serogroup and tcpA/ctxA genes by multiplex PCR offers an efficient system, permitting risk analysis for cholera in coastal areas.

show abstract

Section: Discussionmentioning

confidence: 99%

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Rivera

Lipp

Gil

et al. 2003

Environmental Microbiology

View full text Add to dashboard Cite

show abstract

“…Realizing the need for quick diagnosis, a variety of rapid tests including co-agglutination tests [11^13], enzyme-linked immunosorbent assays [14,15] and membrane based immunoassays [16] have been developed in the recent past. Recently, Albert et al [17] have developed a polymerase chain reaction (PCR) assay using a primer pair corresponding to a chromosomal region of V. cholerae O139 for rapid detection of the serogroup from stool samples. However, these tests have limitations in that they are directed to detect either the O1 or the O139 serogroup and further these tests do not provide information on the toxigenic potential of the strains.…”

Section: Introductionmentioning

confidence: 99%

Development and evaluation of a multiplex PCR assay for rapid detection of toxigenic Vibrio cholerae O1 and O139

Hoshino

1998

FEMS Immunology and Medical Microbiology

131

171

View full text Add to dashboard Cite

A multiplex polymerase chain reaction assay was developed for concurrent detection of rfb sequences specific for the O1 and the O139 serogroups of Vibrio cholerae and for ctxA specific sequences. The multiplex PCR assay was found to be highly specific and sensitive and was capable of detecting 65 cfu and 200 cfu per assay of V. cholerae O1 and O139, respectively. Evaluation of the multiplex PCR assay using 121 stool samples from patients admitted to the Infectious Diseases Hospital, Calcutta, showed the assay to be 100% sensitive and 95.2% specific when the culture method was taken as the standard. In addition to the 38 PCR positive stool samples, an additional four samples which were negative by culture method but positive by PCR assay belonged to the O139 serogroup. All the 38 stool samples positive for either O1 or O139 serogroup by PCR assay were also positive for the ctxA amplicon indicating that the O1 and O139 V. cholerae strains have the genetic potential of producing cholera toxin.

show abstract

“…Traditional PCR has emerged as a useful molecular based detection technique and there have been a number of reports describing the application of PCR for the detection of V. cholerae. These assays were mostly directed against single gene targets involved in V. cholerae virulence, regulation of virulence, outer membrane proteins or genes involved with O-antigen synthesis (Shirai et al, 1991;Fields et al, 1992;Chowdhury et al, 1994;Albert et al, 1997;Nandi et al, 2000;Chow et al, 2001;Lipp et al, 2003).…”

Section: Introductionmentioning

confidence: 99%

The use of a high resolution melt real-time polymerase chain reaction (PCR) assay for the environmental monitoring of Vibrio cholerae

Roux¹

2011

Afr. J. Microbiol. Res.

View full text Add to dashboard Cite

A real-time polymerase chain reaction (PCR) assay utilizing high resolution melt (HRM) curve analysis was developed and tested for the monitoring of Vibrio cholerae in water samples. The assay utilized previously published primers that are specific to regions of the V. cholerae ompW and ctxAB genes, allowing it to differentiate between toxigenic and non-toxigenic strains. The ompW and ctxAB primers amplify target regions of 588 and 564 bp in length (respectively) and the amplicons could be accurately identified using HRM curve analysis. High resolution melt curve analysis provided additional accuracy for the determination of amplicon melting temperatures, and allowed amplification of the two targets in a multiplex reaction. Two laboratories employed the assay to analyse 178 water samples obtained from diverse environmental water sources, for the presence of V. cholerae. The assay was found to be a rapid, highly accurate, sensitive and cost effective method for the detection and distinction between toxigenic and non-toxigenic V. cholerae strains in water.

show abstract

Rapid detection of Vibrio cholerae O139 Bengal from stool specimens by PCR

Cited by 35 publications

References 15 publications

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Method of DNA extraction and application of multiplex polymerase chain reaction to detect toxigenic Vibrio cholerae O1 and O139 from aquatic ecosystems

Development and evaluation of a multiplex PCR assay for rapid detection of toxigenic Vibrio cholerae O1 and O139

The use of a high resolution melt real-time polymerase chain reaction (PCR) assay for the environmental monitoring of Vibrio cholerae

Contact Info

Product

Resources

About