A diagnostic PCR assay for the detection of an Australian epidemic strain of <it>Pseudomonas aeruginosa</it>
<p>Abstract</p> <p>Background</p> <p>Chronic lung infection with the bacterium <it>Pseudomonas aeruginosa </it>is one of the hallmarks of cystic fibrosis (CF) and is associated with worsening lung function, increased hospitalisation and reduced life expectan...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-07-01
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| Series: | Annals of Clinical Microbiology and Antimicrobials |
| Online Access: | http://www.ann-clinmicrob.com/content/9/1/18 |
| Summary: | <p>Abstract</p> <p>Background</p> <p>Chronic lung infection with the bacterium <it>Pseudomonas aeruginosa </it>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 <it>P. aeruginosa </it>(Australian epidemic strain I; AES-I) has been found to be widespread in CF patients in eastern Australia.</p> <p>Methods</p> <p>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 <it>P. aeruginosa </it>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 <it>P. aeruginosa </it>isolates from other clinical and environmental sources.</p> <p>Results</p> <p>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 <it>P. aeruginosa </it>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 <it>P. aeruginosa </it>and AES-I in patient sputum samples.</p> <p>Conclusions</p> <p>We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the <it>P. aeruginosa </it>strain AES-I. We have also shown that Whatman FTA<sup>® </sup>Elute cards may be used with PCR-based assays to rapidly detect the presence of <it>P. aeruginosa </it>strains in CF sputum.</p> |
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| ISSN: | 1476-0711 |