Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Detection of toxigenic Vibrio cholerae with new multiplex PCR

(2012) Detection of toxigenic Vibrio cholerae with new multiplex PCR. Journal of Infection and Public Health. pp. 263-267. ISSN 18760341 (ISSN)

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Abstract

Objective: Vibrio cholerae (V. cholerae) is the etiological agent of epidemic cholera. In recent years, cholera has become endemic in different regions of the world. Traditional culture and microscopic methods are considered the standard for the diagnosis of V. cholerae. However, these methods require days for confirmation. Therefore, molecular methods that may be used for the sensitive, accurate, and rapid analysis of V. cholerae are urgently needed. Materials and methods: In the present investigation, a multiplex PCR assay was developed for the detection of virulence- and toxigenic-associated (VTA) genes (ctxA, tcpA and ompW). To evaluate PCR specificity, other bacteria from the Enterobacteriaceae family (Salmonella typhi, Shigella dysenteriae, and enterotoxigenic Escherichia coli) and Aeromonas hydrophila were examined. The assay sensitivity was evaluated using colony counting and genome dilution methods. Result: Our results showed that this PCR-based method represents an ideal tool for the rapid detection of VTA genes due to its simplicity, cost effectiveness, and accuracy. This multiplex PCR method can be used to determine the presence of VTA genes and can therefore distinguish V. cholerae bacteria from other Vibrio species and bacteria. This method can detect 10-100. CFU V. cholerae and 8.5-85. pg genomic DNA. Discussion: This multiplex PCR method has higher sensitivity and specificity than other methods. The proposed test provides an appropriate and sensitive tool for detecting the presence of toxigenic and pathogenic V. cholerae. © 2012 King Saud Bin Abdulaziz University for Health Sciences.

Item Type: Article
Keywords: Multiplex PCR Vibrio cholerae bacterial protein ctxA protein ompW protein tcpA protein unclassified drug accuracy Aeromonas hydrophila article bacterial gene bacterial virulence bacterium colony bacterium detection controlled study cost effectiveness analysis Escherichia coli multiplex polymerase chain reaction nonhuman outcome assessment priority journal Salmonella typhi sensitivity and specificity Shigella dysenteriae Bacterial Proteins Cholera Cholera Toxin Colony Count, Microbial Food Microbiology Humans Virulence Water Microbiology
Divisions:
Page Range: pp. 263-267
Journal or Publication Title: Journal of Infection and Public Health
Journal Index: Scopus
Volume: 5
Number: 3
Identification Number: https://doi.org/10.1016/j.jiph.2012.02.004
ISSN: 18760341 (ISSN)
Depositing User: مهندس مهدی شریفی
URI: http://eprints.bmsu.ac.ir/id/eprint/1148

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