Development of a Multiplex Real-Time PCR Assay for Rapid Detection of Tigecycline Resistance Gene <i>tet</i> (X) Variants from Bacterial, Fecal, and Environmental Samples
Yulin Fu, Dejun Liu, Huangwei Song, Zhihai Liu, Haiyang Jiang, Yang Wang
Abstract
We developed a multiplex real-time SYBR green-based PCR assay for rapid detection of tet (X) and its variants, including tet (X1) and tet (X2) and high-level tigecycline resistance genes tet (X3), tet (X4), and tet (X5). We showed that the real-time PCR assay developed had high linearity ( R 2 ≥ 0.996), sensitivity (low detection limit), and specificity (only the target gene could be amplified significantly) and further evaluated it using bacterial, fecal, and environmental samples.
Topics & Concepts
TigecyclineMultiplexSYBR Green IBiologyDetection limitReal-time polymerase chain reactionMicrobiologyMultiplex polymerase chain reactionMolecular biologyGeneFecesPolymerase chain reactionAntibioticsChemistryGeneticsChromatographyPharmaceutical and Antibiotic Environmental ImpactsAntibiotic Resistance in BacteriaBacteriophages and microbial interactions