title: Screening for vancomycin-resistant enterococci: an efficient and economical laboratory-developed test.
abstract: A laboratory-developed test (Lab Assay), combining enrichment broth and real-time polymerase chain reaction (PCR) for vancomycin-resistant enterococci (VRE) screening, was developed and evaluated in this study. A total of 1,765 faecal or rectal swabs sent to the laboratory for VRE screening were investigated in parallel by Lab Assay and the Roche LightCycler VRE detection kit-based method. The diagnostic values for Lab Assay were as follows: 100% sensitivity, 79.92% specificity, 1.94% positive predictive value and 100% negative predictive value, which were comparable to the results from the LightCycler kit-based assay. The detection limit of Lab Assay was 10(0) to 10(1) colony-forming units (CFU)/ml of inoculum in broth for both VanA-type and VanB-type VRE. The PCR method developed in this study was approved to be applicable on both the Applied Biosystems 7500 Fast Real-Time PCR System and the LightCycler(®) 480 Real-Time PCR System. The flexibility in choosing PCR systems makes it possible that the PCR assay could be fully compatible with the DNA extraction's platform, providing an integrated workflow. Furthermore, the material cost is saved at 7EUR per sample when Lab Assay replaces the commercial kit-based method in our routine screening for VRE. Therefore, the laboratory-developed broth-PCR method is an efficient and economical assay for VRE screening.
authors.affiliation: Division of Clinical Microbiology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge, Karolinska Institute, 141 86, Stockholm, Sweden. hong.fang@karolinska.se
authors.forename: H
authors.initials: H
authors.lastname: Fang
authors.forename: A-K
authors.initials: AK
authors.lastname: Ohlsson
authors.forename: G-X
authors.initials: GX
authors.lastname: Jiang
authors.forename: M
authors.initials: M
authors.lastname: Ullberg
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D019729:Carbon-Oxygen Ligases / Q000235:genetics; D019411:Clinical Laboratory Techniques; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D016983:Enterococcus / Q000187:drug effects / Q000235:genetics*; D005243:Feces / Q000382:microbiology; D016908:Gram-Positive Bacterial Infections / Q000175:diagnosis*; D006801:Humans; D057230:Limit of Detection; D008826:Microbial Sensitivity Tests / Q000379:methods*; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D014640:Vancomycin; D020713:Vancomycin Resistance / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C071437:VanA ligase, Bacteria; C079913:VanB protein, Enterococcus; D014640:Vancomycin; D019729:Carbon-Oxygen Ligases