Polymerase chain reaction for detection of the cholera enterotoxin operon of Vibrio cholerae.

1991 ◽  
Vol 29 (11) ◽  
pp. 2517-2521 ◽  
Author(s):  
H Shirai ◽  
M Nishibuchi ◽  
T Ramamurthy ◽  
S K Bhattacharya ◽  
S C Pal ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Chain Reaction ◽  
Cholera Enterotoxin ◽  
Polymerase Chain

scholarly journals Detection of Toxigenic Vibrio cholerae Ol Using Polymerase Chain Reaction for Amplifying the Cholera Enterotoxin Gene

1990 ◽  
Vol 64 (10) ◽  
pp. 1323-1329 ◽  
Author(s):  
Kazuhiro KOBAYASHI ◽  
Kazuko SETO ◽  
Susumu AKASAKA ◽  
Masanao MAKINO
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Enterotoxin Gene ◽  
Chain Reaction ◽  
Cholera Enterotoxin ◽  
Polymerase Chain

scholarly journals Rapid Detection of the Vibrio cholerae ctx Gene in Food Enrichments Using Real-Time Polymerase Chain Reaction

2007 ◽  
Vol 90 (5) ◽  
pp. 1278-1283 ◽  
Author(s):  
Willis Fedio ◽  
George M Blackstone ◽  
Lynne Kikuta-Oshima ◽  
Chitra Wendakoon ◽  
Timothy H McGrath ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Real Time ◽  
Bottled Water ◽  
Qpcr Assay ◽  
Second Phase ◽  
Chain Reaction ◽  
Isolation Frequency ◽  
Polymerase Chain ◽  
Static Conditions

Abstract A real-time polymerase chain reaction (qPCR) assay for the detection of the ctxA gene of toxigenic Vibrio cholerae (Vc) was validated against standard culture techniques. The first experimental phase determined optimal enrichment conditions for detection by culture and qPCR of Vc in shrimp, bottled water, milk, and potato salad. The conditions tested included temperature (35 and 42C), time (6 and 18 h), and effect of shaking (0 and 100 rpm). No definitive trends were found with enrichment temperature or shaking on Vc isolation frequency or detection by qPCR. Generally, Vc was detected by qPCR more frequently than Vc was isolated, but this difference was significant only in the 35C 6 h enrichment without shaking. In the second phase of experiments, shrimp, bottled water, milk, potato salad, and oysters were inoculated with each of 3 toxigenic Vc strains (Latin American O1 strain, an O139 strain, and an O1 strain from the U.S. Gulf Coast) and enriched under static conditions at 42C for 6 and 18 h. Overall, detection frequency of ctx by qPCR was 98 (88/90) and 100 (90/90) after 6 and 18 h enrichments, respectively, while Vc isolation frequency was 87 (78/90) and 83 (75/90) after 6 and 18 h, respectively. Toxigenic Vc can be detected by qPCR within an 8 h work day using the 6 h enrichment procedure, assuming an initial level of at least 12 colony-forming units/g; however, overnight enrichment may be necessary to detect lower levels. These data indicate that the qPCR assay for ctx is a more reliable, sensitive, and rapid alternative to standard Vc culture methods and is applicable to diverse food products.

scholarly journals Etiology of Diarrhea Requiring Hospitalization in Bangladesh by Quantitative Polymerase Chain Reaction, 2014–2018

2020 ◽  
Author(s):  
Mami Taniuchi ◽  
Kamrul Islam ◽  
Md Abu Sayeed ◽  
James A Platts-Mills ◽  
Md Taufiqul Islam ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Coefficient Of Variation ◽  
Quantitative Polymerase Chain Reaction ◽  
Age Groups ◽  
Public Health Problem ◽  
Major Public Health Problem ◽  
Surveillance Systems ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Background Diarrhea remains a major public health problem and characterization of its etiology is needed to prioritize interventions. However, most data are from single-site studies of children. We tested samples from participants of any age from 11 geographically diverse hospitals in Bangladesh to describe pathogen-specific burdens of diarrhea. Methods We utilized 2 existing diarrhea surveillance systems: a Nationwide network at 10 sentinel hospitals and at the icddr,b hospital. We tested stools from enrolled participants and nondiarrheal controls for enteropathogens using quantitative polymerase chain reaction and calculated pathogen-specific attributable fractions (AFs) of diarrhea. Results We analyzed 5516 patients with diarrhea and 735 controls. Overall, rotavirus had the highest attributable burden of diarrhea (Nationwide AF, 17.7%; 95% confidence interval [CI], 14.3–20.9%; icddr,b AF, 39.9%; 38.0–41.8%), followed by adenovirus 40/41 (Nationwide AF, 17.9%; 95% CI: 13.9–21.9%; icddr,b AF, 16.6%; 95% CI, 14.4–19.4%) and Vibrio cholerae (Nationwide AF, 10.2%; 95% CI, 9.1–11.3%; icddr,b AF, 13.3%; 95% CI: 11.9–15.1%). Rotavirus was the leading pathogen in children <5 years and was consistent across the sites (coefficient of variation = 56.3%). Adenovirus 40/41 was the second leading pathogen in both children and adults. Vibrio cholerae was the leading pathogen in individuals >5 years old, but was more geographically variable (coefficient of variation = 71.5%). Other attributable pathogens included astrovirus, norovirus, Shigella, Salmonella, ETEC, sapovirus, and typical EPEC. Conclusions Rotavirus, adenovirus 40/41, and V. cholerae were the leading etiologies of infectious diarrhea requiring hospitalization in Bangladesh. Other pathogens were important in certain age groups or sites.

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

2003 ◽  
Vol 5 (7) ◽  
pp. 599-606 ◽  
Author(s):  
Irma N. G. Rivera ◽  
Erin K. Lipp ◽  
Ana Gil ◽  
Nipa Choopun ◽  
Anwar Huq ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Dna Extraction ◽  
Aquatic Ecosystems ◽  
Multiplex Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Vibrio Cholerae O1 ◽  
Polymerase Chain

Multiplex polymerase chain reaction-based assay for the specific detection of toxin-producing Vibrio cholerae in fish and fishery products

2011 ◽  
Vol 90 (3) ◽  
pp. 1111-1118 ◽  
Author(s):  
Geevaretnam Jeyasekaran ◽  
Kannan Thirumalai Raj ◽  
Robinson Jeya Shakila ◽  
Albin Jemila Thangarani ◽  
Durairaj Sukumar
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Multiplex Polymerase Chain Reaction ◽  
Specific Detection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Fishery Products ◽  
Fish And Fishery Products

scholarly journals Semi-nested polymerase chain reaction for detection of toxigenic Vibrio cholerae from environmental water samples

2007 ◽  
Vol 47 (3) ◽  
pp. 207-211 ◽  
Author(s):  
Ajay Kumar Goel ◽  
Shweta Bhadauria ◽  
Pramod Kumar ◽  
Dev V. Kamboj ◽  
Lokendra Singh
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Water Samples ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

scholarly journals Comparison between Biotype of Vibrio cholerae O1 and Genotype using Polymerase Chain Reaction

2005 ◽  
Vol 79 (5) ◽  
pp. 307-313
Author(s):  
Eiji YOKOYAMA ◽  
Kenji KOIWAI ◽  
Masako UCHIMURA
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Chain Reaction ◽  
Vibrio Cholerae O1 ◽  
Polymerase Chain

Evaluation of a rapid polymerase chain reaction based identification technique for Vibrio cholerae isolates

2004 ◽  
Vol 50 (1) ◽  
pp. 229-232 ◽  
Author(s):  
W.J. le Roux ◽  
D. Masoabi ◽  
C.M.E. de Wet ◽  
S.N. Venter
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Cholerae ◽  
Molecular Techniques ◽  
Chain Reaction ◽  
Accurate Identification ◽  
Rapid Pcr ◽  
Rapid Polymerase Chain Reaction ◽  
Identification Technique ◽  
Polymerase Chain ◽  
Public Health Protection

Rapid and accurate identification of waterborne pathogens, such as Vibrio cholerae, in drinking-water sources is important to enable effective resource management and public health protection. Phenotypic systems currently being used for the identification of Vibrio cholerae isolates are time-consuming and the need exists for the development of suitable molecular techniques that can offer both fast and reliable identification. During this study, isolates identified as Vibrio cholerae by means of two different biochemical test systems (API 20E and VITEK 32) were analysed with the polymerase chain reaction (PCR) to compare the reliability of the various identification systems. The selected PCR technique amplified a sequence within the outer membrane protein of Vibrio cholerae, a gene specific for V. cholerae. It was found that out of 243 isolates biochemically identified as V. cholerae with either the API or VITEK system, 21 isolates did not give a positive result with the PCR detection method. Sequencing the 16S rDNA of more than half of these isolates and comparison of the sequences with Internet databases indicated that most of the isolates belonged to the genus Aeromonas. The results indicated that the rapid PCR procedure was more accurate than the API or VITEK systems currently being used for the phenotypic identification of Vibrio cholerae isolates.

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