title: Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients.
abstract: We evaluated a multiplex PCR assay, the Seeplex Diarrhoea ACE detection, that simultaneously detects 15 enteric pathogens, including Salmonella spp., Shigella spp., Vibrio spp., toxin B producer Clostridium difficile, Campylobacter spp., Clostridium perfringens, Yersinia enterocolitica, Aeromonas spp., Escherichia coli O157:H7, verocytotoxin-producing Escherichia coli, adenovirus, Group A rotavirus, norovirus GI and GII, and astrovirus. We compared this assay with clinical methods routinely used in our laboratory, for detecting enteropathogens in stool samples collected from 245 paediatric patients with suspected infectious gastroenteritis. We recovered 61 bacterial pathogens and 121 enteric viruses with our laboratory assays, while we detected 78 bacteria and 167 viruses with the molecular assay. We calculated specificity and sensitivity for both methods after analysis of discordant results and demonstrated greater sensitivity for multiplex PCR than for our routine methods, with the exception of Salmonella spp. and toxigenic C. difficile detection. The multiplex PCR assay proved to be a reliable tool to directly detect the most common enteropathogens in stool samples but with some limitations.
authors.affiliation: Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy. Electronic address: manuela.onori@opbg.net.
authors.forename: Manuela
authors.initials: M
authors.lastname: Onori
authors.affiliation: Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Luana
authors.initials: L
authors.lastname: Coltella
authors.affiliation: Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Livia
authors.initials: L
authors.lastname: Mancinelli
authors.affiliation: Department of Laboratory Medicine, Bacteriology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: Argentieri
authors.affiliation: Medical Direction, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Donato
authors.initials: D
authors.lastname: Menichella
authors.affiliation: Paediatric and Infectious Disease Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Alberto
authors.initials: A
authors.lastname: Villani
authors.affiliation: Paediatric and Infectious Disease Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Grandin
authors.affiliation: Paediatric and Infectious Disease Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Diletta
authors.initials: D
authors.lastname: Valentini
authors.affiliation: Medical Direction, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Raponi
authors.affiliation: Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children's Hospital, IRCCS, Rome, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Russo
mesh_terms: D000293:Adolescent; D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D002648:Child; D002675:Child, Preschool; D005243:Feces / Q000382:microbiology / Q000821:virology; D005260:Female; D005759:Gastroenteritis / Q000175:diagnosis* / Q000382:microbiology / Q000821:virology; D006801:Humans; D007223:Infant; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
keywords: Enteric pathogens; Multiplex PCR; Paediatric patients