Ann Lab Med.  2014 Nov;34(6):446-455. 10.3343/alm.2014.34.6.446.

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

• Park SD
• Lee G
• Wang HY
• Park M
• Kim S
• Kim H
• Kim J
• Kim YK
• Kim HY
• Lee H
• Uh Y
• Kim JB

• KMID: 2129570
• DOI: http://doi.org/10.3343/alm.2014.34.6.446

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.

Keyword

Real-time PCR; Blot; Hybridization; Bacteremia; mecA; Vancomycin resistance; Blood Culture

MeSH Terms

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
Bacterial Proteins
Carbon-Oxygen Ligases
RNA, Ribosomal, 16S
Reagent Kits, Diagnostic

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