A diagnostic PCR assay for the detection of an Australian epidemic strain of Pseudomonas aeruginosa.

Annals of Clinical Microbiology and Antimicrobials
Heidi L WilliamsCynthia B Whitchurch

Abstract

Chronic lung infection with the bacterium Pseudomonas aeruginosa is one of the hallmarks of cystic fibrosis (CF) and is associated with worsening lung function, increased hospitalisation and reduced life expectancy. A virulent clonal strain of P. aeruginosa (Australian epidemic strain I; AES-I) has been found to be widespread in CF patients in eastern Australia. Suppression subtractive hybridization (SSH) was employed to identify genetic sequences that are present in the AES-I strain but absent from the sequenced reference strain PAO1. We used PCR to evaluate the distribution of several of the AES-I loci amongst a collection of 188 P. aeruginosa isolates which was comprised of 35 AES-I isolates (as determined by PFGE), 78 non-AES-I CF isolates including other epidemic CF strains as well as 69 P. aeruginosa isolates from other clinical and environmental sources. We have identified a unique AES-I genetic locus that is present in all 35 AES-I isolates tested and not present in any of the other 153 P. aeruginosa strains examined. We have used this unique AES-I locus to develop a diagnostic PCR and a real-time PCR assay to detect the presence of P. aeruginosa and AES-I in patient sputum samples. We have developed diagnostic PCR assa...Continue Reading

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Citations

Jul 31, 2012·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·V WatersD S Guttman
Feb 24, 2011·IEEE Transactions on Bio-medical Engineering·Jane P BearingerDonald P King
Jun 21, 2011·Journal of Cystic Fibrosis : Official Journal of the European Cystic Fibrosis Society·Pieter DeschaghtMario Vaneechoutte
Aug 31, 2018·Clinical Microbiology Reviews·Michael D ParkinsValerie J Waters

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Methods Mentioned

BETA
electrophoresis
PCR
PCRs

Software Mentioned

BLASTN
BLASTX
Primer Express

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