Related Experiment Video
Updated: Jun 20, 2026
07:58
Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates
Published on: May 30, 2017
A multiplatform real-time polymerase chain reaction detection assay for Vibrio cholerae
Katja A Koskela1, Pirjo Matero, Janet M Blatny
1Centres for Military Medicine and for Biological Threat Preparedness, FI-00290 Helsinki, Finland.
Diagnostic Microbiology and Infectious Disease
|September 5, 2009
Summary
A new real-time polymerase chain reaction (PCR) method accurately identifies enterotoxigenic Vibrio cholerae. This rapid assay targets specific genes for reliable cholera detection and differentiation from other Vibrio species.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Vibrio cholerae causes cholera, a significant public health concern.
- Accurate identification of enterotoxigenic strains is crucial for disease control.
- Distinguishing V. cholerae from other Vibrio species requires reliable diagnostic tools.
Purpose of the Study:
- To develop and validate a multiplatform real-time polymerase chain reaction (PCR) assay for detecting enterotoxigenic Vibrio cholerae.
- To differentiate V. cholerae from other Vibrio species.
- To provide a rapid and accurate tool for cholera diagnostics.
Main Methods:
- Development of a real-time PCR assay targeting genes for the toxR activator and enterotoxin A protein.
- Validation of the assay using a collection of 87 bacterial strains.
- Inclusion of 63 V. cholerae strains and 8 noncholera vibrio strains in the validation set.
Main Results:
- The developed real-time PCR methodology demonstrated successful detection and identification of enterotoxigenic V. cholerae.
- The assay effectively differentiated V. cholerae from other Vibrio species.
- Validation confirmed the assay's reliability with a diverse bacterial strain collection.
Conclusions:
- The multiplatform real-time PCR assay provides a rapid and accurate tool for the detection and identification of cholera.
- This methodology aids in the timely diagnosis and management of cholera outbreaks.
- The assay contributes to improved surveillance and control of V. cholerae infections.
Related Concept Videos
01:33Enzyme-Linked Immunosorbent Assay
In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
01:25Cholera
Cholera is an acute gastrointestinal disease caused by the Gram-negative bacterium Vibrio cholerae. It is transmitted primarily via the fecal-oral route through the ingestion of contaminated water or food.Vibrio cholerae is a motile, Gram-negative bacterium of the family Vibrionaceae, primarily associated with waterborne outbreaks in areas with inadequate sanitation. Although over 200 serogroups of V. cholerae exist, only O1 and O139 are responsible for epidemic cholera. The O1 serogroup,...
01:24Automated Microbial Diagnostics
Automated diagnostic analyzers have transformed clinical microbiology by providing rapid and reliable methods for pathogen identification and antibiotic susceptibility testing. Among these systems, the Vitek 2 is widely used because it automates the traditionally labor-intensive processes of microbial identification (ID) and antibiotic susceptibility testing (AST), delivering standardized and timely results that are essential for effective patient care.Microbial Identification with ID CardsThe...
01:25Rapid Identification of Pathogens
MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...
02:57Real Time RT-PCR
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...