title: Five commercial DNA extraction systems tested and compared on a stool sample collection.
abstract: In this study, 5 different commercial DNA extraction systems were tested on a stool sample collection containing 81 clinical stool specimens that were culture-positive for diarrheagenic Escherichia coli, Campylobacter jejuni, Salmonella enterica, or Clostridium difficile. The purified DNAs were analyzed by polymerase chain reaction (PCR) directed toward the relevant organisms. The results showed that conventional PCR combined with the extraction systems BioRobot EZ1 (Qiagen, Hilden, Germany), Bugs'n Beads (Genpoint, Oslo, Norway), ChargeSwitch (Invitrogen, Paisley, UK), QIAamp Stool Mini Kit (Qiagen), and 2 protocols (generic and Specific A) for EasyMag (BioMérieux, Marcy I'Etoile, France) were able to identify 89%, 62%, 85%, 88%, 85%, and 91%, respectively, of the pathogens originally identified by conventional culture-based methods. When TaqMan PCR was combined with the EasyMag Specific A protocol, 99% of the samples were correctly identified. The results demonstrate that the extraction efficiencies can vary significantly among different extraction systems, careful optimization may have a significant positive effect, and the use of sensitive and specific detection methods like TaqMan PCR is an ideal choice for this type of analysis.
authors.affiliation: Department of Microbiological Diagnostics, Unit of Gastrointestinal Infection, Statens Serum Institut, 2300 S Copenhagen, Denmark. SPN@ssi.dk
authors.forename: Søren
authors.initials: S
authors.lastname: Persson
authors.forename: Richard F
authors.initials: RF
authors.lastname: de Boer
authors.forename: Anna M D
authors.initials: AM
authors.lastname: Kooistra-Smid
authors.forename: Katharina E P
authors.initials: KE
authors.lastname: Olsen
mesh_terms: D016123:Campylobacter jejuni / Q000235:genetics / Q000302:isolation & purification; D016360:Clostridioides difficile / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification; D005243:Feces / Q000382:microbiology*; D006801:Humans; D058889:Molecular Typing / Q000295:instrumentation* / Q000379:methods; D016133:Polymerase Chain Reaction; D019779:Salmonella enterica / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial