title: PCR-reverse blot hybridization assay for screening and identification of pathogens in sepsis.
abstract: Rapid and accurate identification of the pathogens involved in bloodstream infections is crucial for the prompt initiation of appropriate therapy, as this can decrease morbidity and mortality rates. A PCR-reverse blot hybridization assay for sepsis, the reverse blot hybridization assay (REBA) Sepsis-ID test, was developed; it uses pan-probes to distinguish Gram-positive and -negative bacteria and fungi. In addition, the assay was designed to identify bacteria and fungi using six genus-specific and 13 species-specific probes; it uses additional probes for antibiotic resistance genes, i.e., the mecA gene of methicillin-resistant Staphylococcus aureus (MRSA) and the vanA and vanB genes of vancomycin-resistant enterococci (VRE). The REBA Sepsis-ID test successfully identified clinical isolates and blood culture samples as containing Gram-positive bacteria, Gram-negative bacteria, or fungi. The results matched those obtained with conventional microbiological methods. For the REBA Sepsis-ID test, of the 115 blood culture samples tested, 47 (40.8%) and 49 (42.6%) samples were identified to the species and genus levels, respectively, and the remaining 19 samples (16.5%), which included five Gram-positive rods, were identified as Gram-positive bacteria, Gram-negative bacteria, or fungi. The antibiotic resistances of the MRSA and VRE strains were identified using both conventional microbiological methods and the REBA Sepsis-ID test. In conclusion, the REBA Sepsis-ID test developed for this study is a fast and reliable test for the identification of Gram-positive bacteria, Gram-negative bacteria, fungi, and antibiotic resistance genes (including mecA for MRSA and the vanA and vanB genes for VRE) in bloodstream infections.
authors: Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Republic of Korea.
authors: Yeonim
authors: Y
authors: Choi
authors: Hye-Young
authors: HY
authors: Wang
authors: Gyusang
authors: G
authors: Lee
authors: Soon-Deok
authors: SD
authors: Park
authors: Bo-Young
authors: BY
authors: Jeon
authors: Young
authors: Y
authors: Uh
authors: Jong Bae
authors: JB
authors: Kim
authors: Hyeyoung
authors: H
authors: Lee
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D001424:Bacterial Infections / Q000175:diagnosis*; D001426:Bacterial Proteins / Q000235:genetics; D019729:Carbon-Oxygen Ligases / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008826:Microbial Sensitivity Tests; D009181:Mycoses / Q000175:diagnosis*; D046915:Penicillin-Binding Proteins; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D018805:Sepsis / Q000175:diagnosis / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins; D046915:Penicillin-Binding Proteins; D011933:Reagent Kits, Diagnostic; C071437:VanA ligase, Bacteria; C079913:VanB protein, Enterococcus; C487789:mecA protein, Staphylococcus aureus; D019729:Carbon-Oxygen Ligases