title: Evaluation of PCR-reverse blot hybridization assay, REBA Sepsis-ID test, for simultaneous identification of bacterial pathogens and mecA and van genes from blood culture bottles.
abstract: BACKGROUND
  The aim of this study was to evaluate a newly developed PCR-based reverse blot hybridization assay (PCR-REBA), REBA Sepsis-ID (M&D, Wonju, Korea), to rapidly detect the presence of bacteremia and antimicrobial resistance gene in blood culture samples.
  
  
  METHODS
  One thousand four hundred consecutive blood culture samples from patients with a delta neutrophil index greater than 2.7% were selected from March to July in 2013. Three hundred positive and 1,100 negative for bacterial growth in blood culture bottles samples were tested by conventional and real-time PCR-REBA, respectively.
  
  
  RESULTS
  The overall agreement between the conventional identification test and the REBA Sepsis-ID test was 95.3% (286/300). Agreement for gram-positive bacteria, gram-negative bacteria, fungi, and polymicrobials was 94.5% (190/201), 97.3% (71/73), 100% (14/14), and 91.7% (11/12), respectively. The detection rate of the mecA gene from methicillin-resistant Staphylococcus isolates was 97.8% (90/92). The vanA gene was detected in one blood culture sample from which vancomycin-resistant Enterococcus was isolated. When the cycle threshold for real-time PCR was defined as 30.0, 2.4% (26/1,100) of negative blood culture samples tested positive by real-time PCR.
  
  
  CONCLUSIONS
  The REBA Sepsis-ID test is capable of simultaneously and quickly detecting both causative agents and antimicrobial resistance genes, such as mecA and van, in blood culture positive samples.
authors.affiliation: Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
authors.forename: Soon Deok
authors.initials: SD
authors.lastname: Park
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Gyusang
authors.initials: G
authors.lastname: Lee
authors.affiliation: M&D, Inc., Wonju Eco Environmental Technology Center, Wonju, Korea.
authors.forename: Hye-Young
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Min
authors.initials: M
authors.lastname: Park
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Sunghyun
authors.initials: S
authors.lastname: Kim
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Hyunjung
authors.initials: H
authors.lastname: Kim
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Jungho
authors.initials: J
authors.lastname: Kim
authors.affiliation: Department of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
authors.forename: Young Keun
authors.initials: YK
authors.lastname: Kim
authors.affiliation: Department of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
authors.forename: Hyo Youl
authors.initials: HY
authors.lastname: Kim
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Hyeyoung
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
authors.forename: Young
authors.initials: Y
authors.lastname: Uh
authors.affiliation: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Korea.
authors.forename: Jong Bae
authors.initials: JB
authors.lastname: Kim
mesh_terms: D016470:Bacteremia / Q000382:microbiology; D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques / Q000379:methods*; D019729:Carbon-Oxygen Ligases / Q000235:genetics*; D024881:Drug Resistance, Bacterial / Q000235:genetics; D016983:Enterococcus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D046915:Penicillin-Binding Proteins; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D001426:Bacterial Proteins; D046915:Penicillin-Binding Proteins; D012336:RNA, Ribosomal, 16S; D011933:Reagent Kits, Diagnostic; C071437:VanA ligase, Bacteria; C487789:mecA protein, Staphylococcus aureus; D019729:Carbon-Oxygen Ligases
keywords: Bacteremia; Blood Culture; Blot; Hybridization; Real-time PCR; Vancomycin resistance; mecA