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

Soon Deok Park, Gyusang Lee, Hye Young Wang, Min Park, Sunghyun Kim, Hyunjung Kim, Jungho Kim, YoungKeun Kim, Hyo Youl Kim, Hyeyoung Lee, Young Uh, Jong Bae Kim

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)446-455
Number of pages10
JournalAnnals of laboratory medicine
Volume34
Issue number6
DOIs
Publication statusPublished - 2014 Jan 1

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Bottles
Pathogens
Cell culture
Assays
Sepsis
Blood
Genes
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Bacteria
Methicillin Resistance
Methicillin
Gram-Positive Bacteria
Vancomycin
Korea
Bacteremia
Anti-Infective Agents
Fungi
Gram-Negative Bacteria
Staphylococcus

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Park, Soon Deok ; Lee, Gyusang ; Wang, Hye Young ; Park, Min ; Kim, Sunghyun ; Kim, Hyunjung ; Kim, Jungho ; Kim, YoungKeun ; Kim, Hyo Youl ; Lee, Hyeyoung ; Uh, Young ; Kim, Jong Bae. / 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. In: Annals of laboratory medicine. 2014 ; Vol. 34, No. 6. pp. 446-455.
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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.",
author = "Park, {Soon Deok} and Gyusang Lee and Wang, {Hye Young} and Min Park and Sunghyun Kim and Hyunjung Kim and Jungho Kim and YoungKeun Kim and Kim, {Hyo Youl} and Hyeyoung Lee and Young Uh and Kim, {Jong Bae}",
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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, Soon Deok; Lee, Gyusang; Wang, Hye Young; Park, Min; Kim, Sunghyun; Kim, Hyunjung; Kim, Jungho; Kim, YoungKeun; Kim, Hyo Youl; Lee, Hyeyoung; Uh, Young; Kim, Jong Bae.

In: Annals of laboratory medicine, Vol. 34, No. 6, 01.01.2014, p. 446-455.

Research output: Contribution to journalArticle

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T1 - 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

AU - Park, Soon Deok

AU - Lee, Gyusang

AU - Wang, Hye Young

AU - Park, Min

AU - Kim, Sunghyun

AU - Kim, Hyunjung

AU - Kim, Jungho

AU - Kim, YoungKeun

AU - Kim, Hyo Youl

AU - Lee, Hyeyoung

AU - Uh, Young

AU - Kim, Jong Bae

PY - 2014/1/1

Y1 - 2014/1/1

N2 - 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.

AB - 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.

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