Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent-monoclonal antibody and culture methods

Huq, A.; Colwell, Rita R.; Rahman, Rosliza Abdul; Ali, Afsar; Chowdhury, M. A. R.; Parveen, Salina; Sack, David A.; Russek‐Cohen, Estelle

doi:10.1128/aem.56.8.2370-2373.1990

Cited by 228 publications

(113 citation statements)

References 11 publications

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“…By culture methods none of the samples were positive, a result that was not surprising since V. cholerae isolation requires that samples be subjected to enrichment in alkaline peptone water prior to plating onto TCBS agar. Therefore, all samples were also tested by DFA (see Section 2); this technique was previously shown to be very sensitive in detecting both culturable and VBNC V. cholerae O1 in the aquatic environment [13,20,22]. In this case too, none of the examined samples were positive, a result not unexpected since V. cholerae are present in low numbers (ca.…”

Section: Resultsmentioning

confidence: 96%

Vibrios associated with plankton in a coastal zone of the Adriatic Sea (Italy)

Montanari¹,

Pruzzo²,

Pane³

et al. 1999

FEMS Microbiology Ecology

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From April 1996 through March 1997, culturable vibrios associated with both large (>200 μm) and small (0.45–200 μm) size plankton collected in the Ancona, Italy, coastal zone of the Adriatic Sea were studied. Vibrio alginolyticus was readily cultured from large size plankton from July to September and represented the only Vibrio species that could be cultured under the conditions employed in the study. Crustaceans (mainly copepods) and tunicates (Oikopleura spp.) comprised the larger sized plankton, with copepods being 90% of the total plankton population in the summer months. Diatoms (Chaetoceros spp.) were also present but in lower numbers. Vibrios associated with smaller sized plankton comprised mainly V. alginolyticus during June–September and in March, whereas Vibrio vulnificus was isolated from samples collected in April, May, and March. The smaller size plankton comprised predominantly tintinnids, nauplii of crustaceans and exuvia of copepods. These results confirm the role of zooplankton as hosts of vibrios in the environment, and suggest that smaller sized plankton may serve as an important reservoir of these bacteria.

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Section: Resultsmentioning

confidence: 96%

Vibrios associated with plankton in a coastal zone of the Adriatic Sea (Italy)

Montanari¹,

Pruzzo²,

Pane³

et al. 1999

FEMS Microbiology Ecology

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“…The fluorescent monoclonal antibody and PCR methods allowed detection of V. cholerae O1 whether cells were culturable or not. Thus, the DFA and PCR methods offer useful and reliable approaches for environmental studies of V. cholerae and related bacteria and, for these reasons, are preferable to culture (Huq et al, 1990;Lipp et al, 2003).…”

Section: Resultsmentioning

confidence: 99%

Occurrence and distribution of Vibrio cholerae in the coastal environment of Peru

Gil

Louis

Rivera

et al. 2004

Environmental Microbiology

118

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The occurrence and distribution of Vibrio cholerae in sea water and plankton along the coast of Peru were studied from October 1997 to June 2000, and included the 1997-98 El Niño event. Samples were collected at four sites in coastal waters off Peru at monthly intervals. Of 178 samples collected and tested, V. cholerae O1 was cultured from 10 (5.6%) samples, and V. cholerae O1 was detected by direct fluorescent antibody assay in 26 out of 159 samples tested (16.4%). Based on the number of cholera cases reported in Peru from 1997 to 2000, a significant correlation was observed between cholera incidence and elevated sea surface temperature (SST) along the coast of Peru (P < 0.001). From the results of this study, coastal sea water and zooplankton are concluded to be a reservoir for V. cholerae in Peru. The climate-cholera relationship observed for the 1997-98 El Niño year suggests that an early warning system for cholera risk can be established for Peru and neighbouring Latin American countries.

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“…In recent years, a concomitant effort has been made to reduce the time required for detection of V. cholerae O1 in clinical and environmental sampies. To this end, several investigators have employed the fluorescent antibody (FA) technique for detection of V. cholerae [1,3,4]. Polyclonal anti-01 serum was originally used in direct fluorescent antibody (DFA) procedures [6] to detect V. cholerae O1 cells in smears prepared directly from specimens.…”

Section: Introductionmentioning

confidence: 99%

“…Despite the more complicated process involved in production of monoclonal antibodies (mAbs), this technology provides a means for producing specific antibodies without the necessity for highly purified antigen or complicated and time-consuming absorption procedures. A monoclonal antibody, COLTA (University of Maryland Biotechnology Institute, MD), specific for the 'A' factor of V. cholerae O1 lipopolysaccharide that reacts with both Inaba and Ogawa serotypes [7], has been shown to maintain high specificity for V. cholerae O1, when used in the IFA procedure [1,3]. The COLTA MAb has also been incorporated, with good success, in two rapid test kits: CholeraScreen TM, a coagglutination test; and Cholera SMART TM, a colloidal gold-based colorimetric test (New Horizons Diagnostics Corporation, Columbia, MD), both of which are commercially available for detection of V. cholerae O1 directly from stool specimens [2,8].…”

Section: Introductionmentioning

confidence: 99%

Cholera DFA: An improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of Vibrio cholerae O1

Hasan

1994

FEMS Microbiology Letters

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An improved fluorescent monoclonal antibody staining kit, Cholera DFA, for direct detection and enumeration of Vibrio cholerae O1 has been developed, employing a highly specific anti-A antigen monoclonal antibody, COLTA, labeled with fluorescein isothiocyanate (FITC). An optimized quantity of anti-photobleaching agent is used in a glycerol mounting medium to retard the rapid fading of immunofluorescent stained cells during fluorescent microscopy, thus enabling prolonged inspection of individual fields, as well as improved photographic recording of results without loss of fluorescence intensity. When tested for specificity, all 30 strains of V. cholerae O1 reacted with Cholera DFA, whereas 100 heterologous species examined did not, yielding 100% specificity for all strains examined in this study. A field trial was conducted in Bangladesh, employing Cholera DFA and the results were compared with those obtained by conventional culture methods. Of 44 diarrheal stool specimens tested, Cholera DFA was positive for V. cholerae O1 in all culture-positive stool specimens and negative for all culture-negative stool specimens. The procedure is sensitive and highly specific, as well as simple, i.e., less complex than the indirect fluorescent assay, requiring only one reagent and less than 30 min to complete the staining process, while retarding rapid fading that often occurs with fluorescent microscopy.

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Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent-monoclonal antibody and culture methods

Cited by 228 publications

References 11 publications

Vibrios associated with plankton in a coastal zone of the Adriatic Sea (Italy)

Vibrios associated with plankton in a coastal zone of the Adriatic Sea (Italy)

Occurrence and distribution of Vibrio cholerae in the coastal environment of Peru

Cholera DFA: An improved direct fluorescent monoclonal antibody staining kit for rapid detection and enumeration of Vibrio cholerae O1

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