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Evaluation of a multiplex PCR assay for simultaneous detection of bacterial and viral enteropathogens in stool samples of paediatric patients
Authors:Manuela Onori  Luana Coltella  Livia Mancinelli  Marta Argentieri  Donato Menichella  Alberto Villani  Annalisa Grandin  Diletta Valentini  Massimiliano Raponi  Cristina Russo
Affiliation:1. Department of Laboratory Medicine, Virology Unit, Bambino Gesù Children''s Hospital, IRCCS, Rome, Italy;2. Department of Laboratory Medicine, Bacteriology Unit, Bambino Gesù Children''s Hospital, IRCCS, Rome, Italy;3. Medical Direction, Bambino Gesù Children''s Hospital, IRCCS, Rome, Italy;4. Paediatric and Infectious Disease Unit, Bambino Gesù Children''s Hospital, IRCCS, Rome, Italy
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.
Keywords:Multiplex PCR   Enteric pathogens   Paediatric patients
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