Pseudomonas aeruginosa

Abstract

This protocol describes a duplex real-time PCR assay for the identification of P. aeruginosa using two different gene sequences, comprising the ecf∈X and gyrB genes. The ecf∈X gene encodes an extra-cytoplasmic function sigma factor, which may be involved in haem uptake or virulence, whereas the gyrB gene encodes the DNA gyrase subunit B [5]. Notably, both these genes offer reliable targets for the detection of P. aeruginosa by PCR [5, 7]. The benefit of using a duplex assay for a single organism is that it provides simultaneous confirmation of isolate identity, in addition to reducing the potential for false negative misidentification caused by sequence variation in primer or probe targets [7, 12]. © 2010 Springer Science+Business Media B.V.