Multiplex PCR for the concurrent detection and differentiation of Salmonella spp., Salmonella Typhi and Salmonella Typhimurium

DOI HANDLE 33 References Open Access
  • Pui Chai Fung
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia
  • Wong Woan Chwen
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia
  • Chai Lay Ching
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia
  • Lee Hai Yen
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia
  • Noorlis Ahmad
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia Universiti Teknologi MARA Pahang
  • Zainazor Tuan Chilek Tuan
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia National Public Health Laboratory, Ministry of Health Malaysia
  • Tang John Yew Huat
    Faculty of Food Technology, Universiti Sultan Zainal Abidin
  • Ghazali Farinazleen Mohamad
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia
  • Cheah Yoke Kqueen
    Department of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia
  • Nakaguchi Yoshitsugu
    Centre for Southeast Asian Studies, Kyoto University
  • Nishibuchi Mitsuaki
    Centre for Southeast Asian Studies, Kyoto University
  • Radu Son
    Center of Excellence for Food Safety Research, Department of Food Science, Faculty of Food Science and Technology, Universiti Putra Malaysia

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Abstract

Salmonellosis outbreaks involving typhoid fever and human gastroenteritis are important diseases in tropical countries where hygienic conditions are often not maintained. A rapid and sensitive method to detect Salmonella spp., Salmonella Typhi and Salmonella Typhimurium is needed to improve control and surveillance of typhoid fever and Salmonella gastroenteritis. Our objective was the concurrent detection and differentiation of these food-borne pathogens using a multiplex PCR. We therefore designed and optimized a multiplex PCR using three specific PCR primer pairs for the simultaneous detection of these pathogens. The concentration of each of the primer pairs, magnesium chloride concentration, and primer annealing temperature were optimized before verification of the specificity of the primer pairs. The target genes produced amplicons at 429 bp, 300 bp and 620 bp which were shown to be 100% specific to each target bacterium, Salmonella spp., Salmonella Typhi and Salmonella Typhimurium, respectively.

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