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

• Published in: Annals of laboratory medicine 2014, 34(6), , pp.446-455
• Main Authors: Park, Soon Deok, Lee, Gyusang, Wang, Hye-Young, Park, Min, Kim, Sunghyun, Kim, Hyunjung, Kim, Jungho, Kim, Young Keun, Kim, Hyo Youl, Lee, Hyeyoung, Uh, Young, Kim, Jong Bae
• Format: Article
• Language: English
• Subjects: Bacteremia - microbiology; Bacterial Proteins - genetics; Bacteriological Techniques - methods; Carbon-Oxygen Ligases - genetics; Drug Resistance, Bacterial - genetics; Enterococcus - genetics; Enterococcus - isolation & purification; Humans; Methicillin-Resistant Staphylococcus aureus - genetics; Methicillin-Resistant Staphylococcus aureus - isolation & purification; Nucleic Acid Hybridization; Original; Penicillin-Binding Proteins; Reagent Kits, Diagnostic; Real-Time Polymerase Chain Reaction; RNA, Ribosomal, 16S - analysis; 병리학
• ISSN: 2234-3806; 2234-3814
• DOI: 10.3343/alm.2014.34.6.446

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. 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. 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. 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.