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

• We developed a cutting-edge evolution of our method for aac(6′)-Ib-cr detection.
• This method combined SYBR Green real-time PCR and pyrosequencing.
• Simplicity and rapidity increased cost-effectiveness compared to our previous method.

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.