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

Williams, HL; Turnbull, L; Thomas, SJ; Murphy, A; Stinear, T; Armstrong, DS; Whitchurch, CB

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

Williams, HL Turnbull, L

Thomas, SJ Murphy, A Stinear, T Armstrong, DS Whitchurch, CB

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Publication Type:: Journal Article
Citation:: Annals of Clinical Microbiology and Antimicrobials, 2010, 9
Issue Date:: 2010-07-16

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Field	Value	Language
dc.contributor.author	Williams, HL	en_US
dc.contributor.author	Turnbull, L https://orcid.org/0000-0002-9255-9033	en_US
dc.contributor.author	Thomas, SJ	en_US
dc.contributor.author	Murphy, A	en_US
dc.contributor.author	Stinear, T	en_US
dc.contributor.author	Armstrong, DS	en_US
dc.contributor.author	Whitchurch, CB https://orcid.org/0000-0003-2296-3791	en_US
dc.date.available	2010-07-16	en_US
dc.date.issued	2010-07-16	en_US
dc.identifier.citation	Annals of Clinical Microbiology and Antimicrobials, 2010, 9	en_US
dc.identifier.uri	http://hdl.handle.net/10453/37916
dc.description.abstract	Background: 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.Methods: 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.Results: 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.Conclusions: We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the P. aeruginosa strain AES-I. We have also shown that Whatman FTA® Elute cards may be used with PCR-based assays to rapidly detect the presence of P. aeruginosa strains in CF sputum. © 2010 Williams et al; licensee BioMed Central Ltd.	en_US
dc.relation.ispartof	Annals of Clinical Microbiology and Antimicrobials	en_US
dc.relation.isbasedon	10.1186/1476-0711-9-18	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Sputum	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Pseudomonas Infections	en_US
dc.subject.mesh	Pneumonia, Bacterial	en_US
dc.subject.mesh	Cystic Fibrosis	en_US
dc.subject.mesh	DNA, Bacterial	en_US
dc.subject.mesh	DNA Primers	en_US
dc.subject.mesh	Bacteriological Techniques	en_US
dc.subject.mesh	Sensitivity and Specificity	en_US
dc.subject.mesh	Polymerase Chain Reaction	en_US
dc.subject.mesh	Nucleic Acid Hybridization	en_US
dc.subject.mesh	Child	en_US
dc.subject.mesh	Australia	en_US
dc.title	A diagnostic PCR assay for the detection of an Australian epidemic strain of Pseudomonas aeruginosa	en_US
dc.type	Journal Article
utslib.citation.volume	9	en_US
utslib.for	1103 Clinical Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.publication-status	Published	en_US
pubs.volume	9	en_US

Abstract:

Background: 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.Methods: 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.Results: 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.Conclusions: We have developed diagnostic PCR assays that are 100% sensitive and 100% specific for the P. aeruginosa strain AES-I. We have also shown that Whatman FTA® Elute cards may be used with PCR-based assays to rapidly detect the presence of P. aeruginosa strains in CF sputum. © 2010 Williams et al; licensee BioMed Central Ltd.

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