- ORIGINAL ARTICLE - Rapid Molecular Approach for Simultaneous Detection of Salmonella spp., Shigella spp., and Vibrio cholera Reza Ranjbar a, *, Ali Naghoni a , Davoud Afshar b , Farhad Nikkhahi c , Mohsen Mohammadi d a Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. b Department of Microbiology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran. c Department of Pathobiology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. d Department of Pharmaceutical Biotechnology, Faculty of Pharmacy, Lorestan University of Medical Sciences, Khorramabad, Iran. Received: August 21, 2016 Revised: September 22, 2016 Accepted: October 10, 2016 KEYWORDS: multiplex PCR, Salmonella spp., Shigella spp., Vibrio cholerae Abstract Objectives: Gastrointestinal tract infection is still one of the serious public health problems in many geographic areas and is endemic in most countries including Iran. Early detection of the gastrointestinal tract pathogens can be extremely important. The aim of the current study was to apply a shortened time-multiplex polymerase chain reaction (PCR) for rapid and simultaneous detection of Salmonella spp., Shigella spp., and Vibrio cholera. Methods: The standard and clinical strains of Salmonella spp., Shigella spp., and V. cholerae were used in the assay. Multiplex PCR was performed and optimized based on amplification of invA, putative integrase, and ompW genes for detecting Salmonella spp., Shigella spp., and V. cholerae, respectively. The specificity of the assay was evaluated by testing 12 different bacterial species. Results: Only Salmonella spp., Shigella spp., and V. cholerae strains had positive results when subjected to the assay using multiplex PCR. The assay showed a high sensitivity, and no amplification products were observed in multiplex PCR with any of the other microorganisms. Conclusion: Our study indicated that the invA, putative integrase, and ompW- based multiplex PCR assay appears to be an efficient method for rapid and simultaneous detection of Salmonella spp., Shigella spp., and V. cholerae. 1. Introduction Worldwide, gastrointestinal tract infections are the second most important cause of death; about 25 million enteric infections occur each year. These infections cause significant morbidity and death in children younger than 5 years in particular and in elderly people. It has been estimated that 4e6 million children die each year because *Corresponding author. E-mail: ranjbarre@gmail.com (R. Ranjbar). Osong Public Health Res Perspect 2016 7(6), 373e377 http://dx.doi.org/10.1016/j.phrp.2016.10.002 pISSN 2210-9099 eISSN 2233-6052 Copyright ª 2016 Korea Centers for Disease Control and Prevention. Published by Elsevier Korea LLC. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).