Establishment of real-time PCR assay to detect resistance gene armA

Objective To establish a TaqMan real-time PCR assay for the detection of resistance gene armA. Methods We designed the primers and probe according to armA gene and evaluated its specificity by using common pathogens. The sensitivity of the assay was evaluated by using different concentrations of recombinant standard plasmid. The simulated fecal specimens made by mixing different concentrations of armA-positive Klebsiella pneumonia with feces were used to evaluate the applicability of the assay in clinical laboratories. Results The results demonstrated that this assay had good specificity. The sensitivity of the assay was found to be 4.07×101copy/ml by standard curve analysis. The lower threshold of real-time PCR assay of the simulated fecal specimens was 1.3×104 cfu/ml. Conclusion The real-time PCR assay established in our study can be used in the detection of armA in resistance gene surveillance.