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Sepsis pp 97-108 | Cite as

Molecular Detection of Antibiotic Resistance Genes from Positive Blood Cultures

  • Musa Y. HindiyehEmail author
  • Gill Smollan
  • Shiraz Gefen-Halevi
  • Ella Mendelson
  • Nathan Keller
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1237)

Abstract

Rapid detection of the bacterial causative agent causing sepsis must be coupled with rapid identification of the antibiotic resistant mechanism that the pathogen might possess. Real-time PCR (qPCR)-based assays have been extensively utilized in the clinical microbiology field as diagnostic tools for the rapid detection of specific nucleic acid (NA) targets. In this chapter, we will discuss the technical aspects of using an internally controlled qPCR assay for the rapid detection of Klebsiella pneumoniae carbapenemase gene (bla KPC) in positive Bactec blood culture bottles. The multiplex qPCR (bla KPC/RNase P) utilizes specific primers and probes for the detection of the bacterial carbapenem resistance mechanism, bla KPC gene, and the internal control RNase P. The internal control of the qPCR assay is vital for detecting any inhibitors that are well known to be present in the blood culture bottles. Rapid detection of the antibiotic resistant mechanism present in the bacterial pathogen causing sepsis can help in better managing patients’ infection.

Key words

Blood culture bottles blaKPC Carbapenem resistance Multidrug resistance 

Notes

Acknowledgment

The authors would like to acknowledge the Chaim Sheba Medical Center laboratory team in particular the bacteriology and virology staff members. Without their help, this assay would have not been validated.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Musa Y. Hindiyeh
    • 2
    • 5
    Email author
  • Gill Smollan
    • 3
  • Shiraz Gefen-Halevi
    • 3
  • Ella Mendelson
    • 1
    • 4
  • Nathan Keller
    • 3
  1. 1.Central Virology Laboratory, Public Health ServicesMinistry of Health, Chaim Sheba Medical CenterTel-HashomerIsrael
  2. 2.Real Time Molecular Diagnosis Unit, Israel Central Virology LaboratoryChaim Sheba Medical CenterTel HashomerIsrael
  3. 3.Infectious Disease Unit/Microbiology LaboratorySheba Medical CenterTel HashomerIsrael
  4. 4.Department of Epidemiology and Preventive Medicine, School of Public Health, Sackler Faculty of MedicineTel-Aviv UniversityTel-AvivIsrael
  5. 5.Central Virology Laboratory, Public Health Services, Department of Epidemiology and Preventive Medicine, School of Public Health, Sackler Faculty of MedicineMinistry of Health, Chaim Sheba Medical CenterTel-HashomerIsrael

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