title: Detection of major diarrheagenic bacterial pathogens by multiplex PCR panels.
abstract: Diarrheal diseases remain a major threat to the youngest population in low- and middle-income countries. The main bacterial pathogens causing diarrhea are diarrheagenic Escherichia coli (DEC) that consists of enteroaggregative (EAEC), enteropathogenic (EPEC), enterotoxigenic (ETEC), enterohemorrhagic EHEC and enteroinvasive E. coli (EIEC), Salmonella, Shigella spp. (S. dysenteria, S. sonnei, S. flexneri) Campylobacter (C. coli, C. jejuni), Vibrio (V. vulnificus, V. parahaemolyticusm, V. cholerae), Yersinia enterocolitica and Aeromonas hydrophila. The aim of this study was to set up rapid multiplex PCR (mPCR) panels to identify these diarrheagenic pathogens based on their specific virulence genes. Primers against specific target genes were combined into three mPCR panels: one for diarrheal E. coli, one for pathogens causing mainly bloody diarrhea, and the third for the remaining pathogens. The panels were tested against a set of stool samples from Swedish children with diarrhea and controls and the analysis identified bacterial pathogens in 14/54 (26%) of the samples. These results show that our three developed mPCR panels can detect main bacterial diarrheagenic pathogens in clinical samples.
authors.affiliation: University of Gothenburg Vaccine Research Institute (GUVAX), Department of Microbiology and Immunology, Institute of Biomedicine, The Sahlgrenska Academy of University of Gothenburg, Gothenburg, Sweden; Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Åsa
authors.initials: Å
authors.lastname: Sjöling
authors.affiliation: University of Gothenburg Vaccine Research Institute (GUVAX), Department of Microbiology and Immunology, Institute of Biomedicine, The Sahlgrenska Academy of University of Gothenburg, Gothenburg, Sweden.
authors.forename: Leila
authors.initials: L
authors.lastname: Sadeghipoorjahromi
authors.affiliation: Department of Pediatrics, Queen Silvia Children's Hospital Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Novak
authors.affiliation: University of Gothenburg Vaccine Research Institute (GUVAX), Department of Microbiology and Immunology, Institute of Biomedicine, The Sahlgrenska Academy of University of Gothenburg, Gothenburg, Sweden. Electronic address: Joshua.Tobias@microbio.gu.se.
authors.forename: Joshua
authors.initials: J
authors.lastname: Tobias
mesh_terms: D001419:Bacteria / Q000235:genetics; D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D003967:Diarrhea / Q000175:diagnosis* / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers
keywords: Diarrheagenic pathogens; Human; Multiplex PCR