A simplified and cost-effective method combining real-time PCR and pyrosequencing for detection of aac(6')-Ib-cr gene

J Microbiol Methods. 2013 Nov;95(2):268-71. doi: 10.1016/j.mimet.2013.09.015. Epub 2013 Sep 26.

Abstract

We previously described a pyrosequencing-based method able to guarantee detection of aac(6')-Ib-cr by characterizing precisely the single nucleotide polymorphisms leading to Trp102Arg and Asp179Tyr. By gaining in simplicity and cost-effectiveness, through the use of real-time PCR, we propose here a cutting-edge evolution of our existing pyrosequencing method.

Keywords: Plasmid-mediated quinolone resistance; Pyrosequencing; Real-time PCR; SYBR Green; aac(6′)-Ib-cr.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases / genetics
  • Acetyltransferases / isolation & purification
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Cost-Benefit Analysis
  • DNA, Bacterial / isolation & purification*
  • Drug Resistance, Bacterial / genetics
  • Escherichia coli / genetics
  • Escherichia coli / isolation & purification
  • Klebsiella pneumoniae / genetics
  • Klebsiella pneumoniae / isolation & purification
  • Polymorphism, Single Nucleotide
  • Real-Time Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Acetyltransferases
  • aminoglycoside acetyltransferase