Rapid detection of V. cholerae 01

Chaicumpa, Wanpen; Thattiyaphong, Aree; Supawat, Krongkaew; Chongsa-nguan, Manas; Kalambaheti, Thareerat; Eampokalap, Boonchuay; Ruangkunaporn, Yuwaporn; S, Sricharmorn; Tapchaisri, Pramuan

doi:10.1016/0888-0786(96)87295-7

Cited by 8 publications

(8 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2 Monitoring of the bacterium in water sources is traditionally performed by a series of biochemical tests. 3,4 In addition, several immunoassay procedures employed for the detection of V. cholerae are: polymerase chain reaction (PCR), [5][6][7][8] enzyme linked immunosorbent assay (ELISA), 9 coaglutination, 10 immunofluorescence [11][12][13] and quartz crystal microbalance (QCM). 14 Most of these methods somehow require highly qualified personnel (PCR), long time (ELISA) and sophisticated instrumentation (QCM, PCR).…”

Section: Introductionmentioning

confidence: 99%

Amperometric Immunosensor for the Detection of Vibrio cholerae O1 Using Disposable Screen-printed Electrodes

et al. 2006

View full text Add to dashboard Cite

A disposable amperometric immunosensor was studied for the rapid detection of Vibrio cholerae (V. cholerae), the causative agent of cholera, employing an indirect sandwich enzyme linked immunosorbent assay (ELISA) principle. Screen-printed electrodes (SPEs) were fabricated (by using commercial and homemade carbon inks), electrochemically characterized and the assay conditions were optimized for capturing antibodies and antigen. Whole cell lysate (WCL) of V. cholerae was used to raise antibodies in rabbits and mice. The antibodies raised against WCL of V. cholerae were found to be specific, and no cross reactivity was observed with other enteric bacteria. 1-Naphthyl phosphate was used as a substrate with the amperometric detection of its enzymatic hydrolysis product 1-naphthol at a potential of +400 mV vs. Ag/AgCl reference electrode. A comparison between the amperometric detection technique and the standard ELISA was made in terms of the total assay time, the amount of biological materials used and the sensitivity of detection. The minimum detection limit of the amperometric immunosensor for V. cholerae was found to be 10 5 cells/ml in 55 min, while ELISA detected 10 6 cells/ml in 4 h.

show abstract

Section: Introductionmentioning

confidence: 99%

Amperometric Immunosensor for the Detection of Vibrio cholerae O1 Using Disposable Screen-printed Electrodes

et al. 2006

View full text Add to dashboard Cite

show abstract

“…The animal was then sacrificed by cervical dislocation. Spleen cells were fused with P3x-63-Ag8 myeloma cells by using polyethylene glycol 4000 as a fusogen at a spleen cell/ myeloma cell ratio of 10:1 for production of hybridomas as previously described (4,5). Culture fluids from culture wells containing growing hybrids were collected and screened for antibodies against the homologous antigen.…”

Section: Methodsmentioning

confidence: 99%

“…Dot ELISA was used for detecting V. cholerae O139 antigen in the rectal swab specimens and for testing the cross-reactivity of the antibodies produced by the growing hybridomas. It was performed as described previously (4). A 3-l aliquot of the sample (boiled V. cholerae whole cells, Ly, or specimen) was dotted onto the nitrocellulose (NC) strip (nitrocellulose membrane; Bio-Rad, Richmond, Calif.), and the strip was air dried.…”

Section: Methodsmentioning

confidence: 99%

“…In this report, a rapid, simple, specific, sensitive, and inexpensive dot ELISA similar in principle and concept to that for V. cholerae O1 previously reported by our group (4) has been developed to detect the V. cholerae O139 antigen in stools of patients with watery diarrhea by using the specific monoclonal antibody (MAb) directed against O139 lipopolysaccharide (LPS).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Rapid Diagnosis of Cholera Caused by Vibrio cholerae O139

et al. 1998

View full text Add to dashboard Cite

Hybridomas secreting specific monoclonal antibodies (MAbs) toVibrio cholerae serogroup O139 were produced. Six monoclones (hybridomas) secreting MAbs specific only to lipopolysaccharide of V. cholerae O139 strains and which did not cross-react to 137 strains of other enteric microorganisms were obtained. These clones were designated 12F5-G11, 12F5-G2, 15F5-H5, 5B9-F8, 14C9-D2, and 6D2-D8. The immunoglobulin (Ig) heavy chain isotypes secreted by these clones were IgG2b, IgG2b, IgG2b, IgM, IgG2b, and IgG3, respectively. Clone 12F5-G11 was selected for mass production of MAb, which was used as a detection reagent in the antigen detection assay for diagnosis of cholera caused by V. cholerae O139, and this assay was compared to the conventional bacterial isolation method. Five batches of rectal swab cultures in alkaline-peptone water were collected from 6,497 patients with watery diarrhea. These were 6,310 patients admitted to Bamrasnaradura Infectious Diseases Hospital, 16 patients from Krung Thon Hospital, 78 patients from Bangkok Children’s Hospital, 19 patients from Karen refugee camps, and 74 Indian patients from the National Institute of Cholera and Enteric Diseases, Calcutta, India. The V. cholerae O139 isolations from the rectal swab cultures and the antigen detection assays (i.e., the MAb-based dot-blot ELISA) were performed by different persons of different laboratories, and the results were revealed after all specimens had been tested. Of the 6,497 samples tested, the dot-blot ELISA correctly identified 42 of 42 V. choleraeO139-positive samples and gave a result of positive for three samples which were culture negative for V. cholerae O139. The diagnostic sensitivity, specificity, and efficacy of the dot-blot ELISA were 100, 99.95, and 99.26%, respectively. The ELISA is easy to perform and relatively inexpensive. It can test multiple samples at a single time, does not require special equipment, and does not produce great quantities of contaminated waste. Most of all, it reduces the diagnostic time from at least 2 days for the bacterial isolation to less than 90 min. The assay is recommended as a rapid screening test of cholera cases caused by V. cholerae O139.

show abstract

“…In addition to the above assays for detecting the phenotypic expression of CT, a number of DNA probes and PCR methods have been developed to detect the ctx genes. (12)(13)(14)(15)(16)(17) In this study, polyclonal and monoclonal anti-CT antibodies were produced, characterized, and purified. These antibodies will be used for application in the development of microbial toxin immunosensors.…”

Section: Introductionmentioning

confidence: 99%

Production and Purification of Monoclonal and Polyclonal Antibodies Against Cholera Toxin

Chou

2004

Hybridoma and Hybridomics

View full text Add to dashboard Cite

The aim of this study was to produce monoclonal and polyclonal antibodies against cholera toxin (CT). Hyperimmune ICR mice produced polyclonal antibodies (PAbs) after injection with 0.5 mL of pristane and were injected with NS-1 myeloma cells 2 weeks later. Hyperimmune Balb/c mice were used for the production of monoclonal antibodies (MAbs). After these mice were immunized four times and given a final boost, their spleen cells were collected and fused with NS-1 myeloma cells under the presence of PEG 1500. The fused cells were then selected in the hypoxanthine, aminopterin, and thymidine (HAT)-RPMIX medium. Anti-CT antibody-secreting hybridoma cell lines with high titer were cloned by enzyme-linked immunosorbent assay (ELISA) and then subcloned by limiting dilution in 15% fetal bovine serum (FBS) HT-RPMIX medium. Eleven murine hybridoma producing anti-CT MAbs were obtained and designated CT-A2, CT-B4, CT-B11, CT-C7, CT-D7, CT-E8, CT-F4, CT-F2, CT-F8, CT-E3, CT-E6. Isotypes of MAbs were identified as IgM heavy chain and all were lambda light chain. Hitrap rProtein A and Hitrap IgM purification columns were used for the purification of PAbs and MAbs, respectively.

show abstract

Rapid detection of V. cholerae 01

Cited by 8 publications

References 15 publications

Amperometric Immunosensor for the Detection of Vibrio cholerae O1 Using Disposable Screen-printed Electrodes

Amperometric Immunosensor for the Detection of Vibrio cholerae O1 Using Disposable Screen-printed Electrodes

Rapid Diagnosis of Cholera Caused by Vibrio cholerae O139

Production and Purification of Monoclonal and Polyclonal Antibodies Against Cholera Toxin

Contact Info

Product

Resources

About