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].
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Anuj, S., Whiley, D.M. (2010). Pseudomonas aeruginosa. In: Schuller, M., Sloots, T., James, G., Halliday, C., Carter, I. (eds) PCR for Clinical Microbiology. Springer, Dordrecht. https://doi.org/10.1007/978-90-481-9039-3_24
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DOI: https://doi.org/10.1007/978-90-481-9039-3_24
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