title: Evaluation of the Xpert vanA/vanB assay using enriched inoculated broths for direct detection of vanB vancomycin-resistant Enterococci.
abstract: Rapid and accurate detection of VRE (vancomycin-resistant enterococci) is required for adequate antimicrobial treatment and infection prevention measures. Previous studies using PCR for the detection of VRE, including Cepheid's Xpert vanA/vanB assay, reported accurate detection of vanA VRE; however, many false-positive results were found for vanB VRE. This is mainly due to nonenterococcal vanB genes, which can be found in the gut flora. Our goal was to optimize the rapid and accurate detection of vanB VRE and to improve the positive predictive value (PPV) by limiting false-positive results. We evaluated the use of the Xpert vanA/vanB assay on rectal swabs and on enriched inoculated broths for the detection of vanB VRE. By adjusting the cycle threshold (CT) cutoff value to ≤ 25 for positivity by PCR on enriched broths, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were 96.9%, 100%, 100%, and 99.5% for vanB VRE, respectively. As shown in this study, CT values of ≤ 25 acquired from enriched broths can be considered true positive. For broths with CT values between 25 and 30, we recommend confirming the results by culture. CT values of >30 appeared to be true negative. In conclusion, this study shows that the Cepheid's Xpert vanA/vanB assay performed on enriched inoculated broths with an adjusted cutoff CT value is a useful and rapid tool for the detection of vanB VRE.
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands x.w.zhou@umcg.nl.
authors.forename: X
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: J P
authors.initials: JP
authors.lastname: Arends
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: G A
authors.initials: GA
authors.lastname: Kampinga
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: H M
authors.initials: HM
authors.lastname: Ahmad
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Dijkhuizen
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: van Barneveld
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: J W A
authors.initials: JW
authors.lastname: Rossen
authors.affiliation: Department of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands.
authors.forename: A W
authors.initials: AW
authors.lastname: Friedrich
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis / Q000382:microbiology*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011237:Predictive Value of Tests; D012007:Rectum / Q000382:microbiology; D012680:Sensitivity and Specificity; D065507:Vancomycin-Resistant Enterococci / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; C079913:VanB protein, Enterococcus