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.
10.3343/alm.2014.34.6.446
- Author:
Soon Deok PARK
1
;
Gyusang LEE
;
Hye Young WANG
;
Min PARK
;
Sunghyun KIM
;
Hyunjung KIM
;
Jungho KIM
;
Young Keun KIM
;
Hyo Youl KIM
;
Hyeyoung LEE
;
Young UH
;
Jong Bae KIM
Author Information
1. Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea. u931018@yonsei.ac.kr
- Publication Type:Original Article ; Evaluation Studies ; Research Support, Non-U.S. Gov't
- Keywords:
Real-time PCR;
Blot;
Hybridization;
Bacteremia;
mecA;
Vancomycin resistance;
Blood Culture
- MeSH:
Bacteremia/microbiology;
Bacterial Proteins/*genetics;
Bacteriological Techniques/*methods;
Carbon-Oxygen Ligases/*genetics;
Drug Resistance, Bacterial/genetics;
Enterococcus/*genetics/isolation & purification;
Humans;
Methicillin-Resistant Staphylococcus aureus/*genetics/isolation & purification;
*Nucleic Acid Hybridization;
RNA, Ribosomal, 16S/analysis;
Reagent Kits, Diagnostic;
*Real-Time Polymerase Chain Reaction
- From:Annals of Laboratory Medicine
2014;34(6):446-455
- CountryRepublic of Korea
- Language:English
-
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.
