TY - JOUR
T1 - Molecular detection of antibiotic resistance genes from positive blood cultures
AU - Hindiyeh, Musa Y.
AU - Smollan, Gill
AU - Gefen-Halevi, Shiraz
AU - Mendelson, Ella
AU - Keller, Nathan
N1 - Publisher Copyright:
© Springer Science+Business Media New York 2015.
PY - 2014
Y1 - 2014
N2 - Rapid detection of the bacterial causative agent causing sepsis must be coupled with rapid identifi cation of the antibiotic resistant mechanism that the pathogen might possess. Real-time PCR (qPCR)-based assays have been extensively utilized in the clinical microbiology fi eld as diagnostic tools for the rapid detection of specifi c 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 (blaKPC) in positive Bactec blood culture bottles. The multiplex qPCR (blaKPC /RNase P) utilizes specifi c primers and probes for the detection of the bacterial carbapenem resistance mechanism, blaKPC 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.
AB - Rapid detection of the bacterial causative agent causing sepsis must be coupled with rapid identifi cation of the antibiotic resistant mechanism that the pathogen might possess. Real-time PCR (qPCR)-based assays have been extensively utilized in the clinical microbiology fi eld as diagnostic tools for the rapid detection of specifi c 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 (blaKPC) in positive Bactec blood culture bottles. The multiplex qPCR (blaKPC /RNase P) utilizes specifi c primers and probes for the detection of the bacterial carbapenem resistance mechanism, blaKPC 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.
KW - Blood culture bottles
KW - Carbapenem resistance
KW - Multidrug resistance
KW - bla KPC
UR - http://www.scopus.com/inward/record.url?scp=84921768356&partnerID=8YFLogxK
U2 - 10.1007/978-1-4939-1776-1_10
DO - 10.1007/978-1-4939-1776-1_10
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C2 - 25319783
AN - SCOPUS:84921768356
SN - 1064-3745
VL - 1237
SP - 97
EP - 108
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -