Optimization of five qPCR protocols toward the detection and the quantification of antimicrobial resistance genes in environmental samples
Roberta Tolosi, Lisa Carraro, Andrea Laconi, Alessandra Piccirillo
Abstract
Here, we describe the optimization and validation of five quantitati ve PCR (qPCR) assays by employing the SYBRGreen chemistry paired with melting curve analysis to detect and quantify clinically relevant antimicrobial resistance genes (ARGs) (i.e. ermB, bla CTXM1-like , bla CMY-2 , qnrA and qnrS ) from environmental samples (i.e. soil and manure) . These five protocols accurately detected and quantified the aforementioned ARGs in complex environmental matrices and represent useful tools for both diagnostic and monitoring activities of resistant bacteria and ARGs into the environment.
Topics & Concepts
BacteriaAntimicrobialAntibiotic resistanceMelting curve analysisComputational biologyEnvironmental DNAReal-time polymerase chain reactionGeneBiologyMicrobiologyChemistryGeneticsEcologyBiodiversityPharmaceutical and Antibiotic Environmental ImpactsMolecular Biology Techniques and ApplicationsAntibiotic Resistance in Bacteria