Simultaneous detection of Escherichia coli O157:H7, toxigenic Vibrio cholera, and Salmonella typhimurium by multiplex PCR
Iranian Journal of Clinical Infectious Diseases. 2009; 4 (2): 97-103
in English
| IMEMR
| ID: emr-100222
ABSTRACT
Escherichia coli O157H7, Vibrio cholerae, and Salmonella typhimurium are pathogenic bacteria found inn contaminated water and food. No assay method is currently available on simultaneous detection or identification of all the three pathogens. Our aim was to develop a rapid and reliable method for this purpose. A protocol for sample collection, and a PCR procedure was designed specifically for the assay. Selected fragments of 239 bp, 432 bp, and 360 bp for E. coli O157 lipopolysaccharide [LPS] gene [rfbE], V. cholerae toxin gene [ctx], and Salmonella typhimurium putative cytoplasmic protein gene [STM4497], respectively, were amplified from the extracted bacterial DNA samples in a single tube by multiplex PCR. The multiplex PCR products were analyzed by gel electrophoresis. All unknown samples were verifiably identified. The assay was sensitive enough to detect and identify as few as 100 cells of E. coli O157H7, V. cholerae and Salmonella typhimurium. The presence of other bacteria did not interfere with the analysis. This assay is a specific and reliable tool which allows cost-effective detection o all three bacterial pathogens in one reaction tube
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Index:
IMEMR (Eastern Mediterranean)
Main subject:
Salmonella typhimurium
/
Vibrio cholerae
/
Polymerase Chain Reaction
/
Electrophoresis, Agar Gel
Language:
English
Journal:
Iran. J. Clin. Infect. Dis.
Year:
2009
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