A locked nucleic acid (LNA)-based real-time PCR assay for the rapid detection of multiple bacterial antibiotic resistance genes directly from positive blood culture

PloS One
Lingxiang ZhuQuan-Zhen Li

Abstract

Bacterial strains resistant to various antibiotic drugs are frequently encountered in clinical infections, and the rapid identification of drug-resistant strains is highly essential for clinical treatment. We developed a locked nucleic acid (LNA)-based quantitative real-time PCR (LNA-qPCR) method for the rapid detection of 13 antibiotic resistance genes and successfully used it to distinguish drug-resistant bacterial strains from positive blood culture samples. A sequence-specific primer-probe set was designed, and the specificity of the assays was assessed using 27 ATCC bacterial strains and 77 negative blood culture samples. No cross-reaction was identified among bacterial strains and in negative samples, indicating 100% specificity. The sensitivity of the assays was determined by spiking each bacterial strain into negative blood samples, and the detection limit was 1-10 colony forming units (CFU) per reaction. The LNA-qPCR assays were first applied to 72 clinical bacterial isolates for the identification of known drug resistance genes, and the results were verified by the direct sequencing of PCR products. Finally, the LNA-qPCR assays were used for the detection in 47 positive blood culture samples, 19 of which (40.4%) were ...Continue Reading

References

Jan 1, 1986·Antimicrobial Agents and Chemotherapy·B J Hartman, A Tomasz
Feb 2, 2002·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·David M Livermore
Nov 1, 2002·Journal of Clinical Microbiology·C L C WieldersF J Schmitz
Jul 17, 2007·Clinical Microbiology Reviews·Anne Marie Queenan, Karen Bush
Dec 25, 2007·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·G M RossoliniC Mugnaioli
Jan 13, 2009·Clinical Microbiology Reviews·George A Jacoby
Sep 12, 2009·The Journal of Antimicrobial Chemotherapy·Cynthia OxacelayPatrice Nordmann
Jan 15, 2010·The Journal of Antimicrobial Chemotherapy·Caroline DallenneGuillaume Arlet
Jul 6, 2010·Journal of Microbiological Methods·Alma BrolundChristian G Giske
Dec 3, 2010·Journal of Clinical Microbiology·Patrice NordmannTimothy R Walsh
Dec 7, 2010·International Journal of Antimicrobial Agents·Timothy R Walsh
May 14, 2011·The Journal of Antimicrobial Chemotherapy·Danny C T OngThean Yen Tan
Jun 22, 2011·Antimicrobial Agents and Chemotherapy·Thierry NaasPatrice Nordmann
Jul 12, 2013·Clinical Microbiology and Infection : the Official Publication of the European Society of Clinical Microbiology and Infectious Diseases·J YangL Han
Oct 15, 2013·World Journal of Microbiology & Biotechnology·Lingxiang ZhuQuan-Zhen Li
Jan 1, 2014·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·S JiY Yu
Jan 25, 2014·Emerging Infectious Diseases·Guang ZhouJiyong Yang
Apr 4, 2014·Journal of Laboratory Physicians·Gunjan GuptaPurva Mathur
May 28, 2014·Journal of Microbiology, Immunology, and Infection = Wei Mian Yu Gan Ran Za Zhi·Ying LiMinggui Wang
Aug 12, 2014·Infection, Genetics and Evolution : Journal of Molecular Epidemiology and Evolutionary Genetics in Infectious Diseases·Chuanming ZhangLiping Zhang

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Methods Mentioned

BETA
PCR
Assay

Software Mentioned

CLUSTAL W
ProbeFinder Assay Design
BLAST
ABI SDS2

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