Development of a groEL gene–based species-specific multiplex polymerase chain reaction assay for simultaneous detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus
Article first published online: 6 DEC 2012
© 2012 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 114, Issue 2, pages 448–456, February 2013
How to Cite
Hossain, M.T., Kim, E.-Y., Kim, Y.-R., Kim, D.-G. and Kong, I.-S. (2013), Development of a groEL gene–based species-specific multiplex polymerase chain reaction assay for simultaneous detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus. Journal of Applied Microbiology, 114: 448–456. doi: 10.1111/jam.12056
- Issue published online: 14 JAN 2013
- Article first published online: 6 DEC 2012
- Accepted manuscript online: 2 NOV 2012 02:14AM EST
- Manuscript Accepted: 26 OCT 2012
- Manuscript Revised: 17 OCT 2012
- Manuscript Received: 18 SEP 2012
- Yeongnam Sea Grant
- groEL gene;
- Multiplex PCR;
- Sea water;
- Vibrio cholerae ;
- V. parahaemolyticus ;
- V. vulnificus
To develop an effective multiplex polymerase chain reaction (PCR) for the simultaneous detection of three important Vibrio species, Vibrio cholerae (Vc), V. parahaemolyticus (Vp) and V. vulnificus (Vv) using the groEL gene, a potential phylogenetic marker.
Methods and Results
Three species-specific primer sets were designed to target Vc, Vp and Vv. A total of 131 Vibrio and non-Vibrio strains were used to determine the specificity and sensitivity of primers. The primers produced specific PCR fragments from all target species strains and did not cross-react with other Vibrio and non-Vibrio species. This PCR method showed good efficiency in detecting coexisting target species in the same sample with a detection limit of 100 pg of Vc, Vp and Vv from mixed purified DNA. Detection of three target species was also possible from artificially inoculated shellfish, flounder and sea water.
The groEL gene is a potential marker for accurate simultaneous detection of Vc, Vp and Vv and could be used to detect these species in environmental and clinical samples.
Significance and Impact of the Study
This newly developed multiplex PCR is a useful and cost-effective method that is applicable in a disease-outbreak prediction system and may provide an effective tool for both the epidemiologist and ecologist.