Comparison of real time diagnostic chemistries to detect Pseudomonas aeruginosa in respiratory samples from cystic fibrosis patients |
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| Authors: | J.L. Fothergill M.J. Ledson M.J. Walshaw P.S. McNamara K.W. Southern C. Winstanley |
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| Affiliation: | 1. NIHR Biomedical Research Centre in Microbial Disease, Royal Liverpool Hospital, Prescott Rd, Liverpool L7 8XP, UK;2. Institute of Infection and Global Health, University of Liverpool, Ronald Ross Building, West Derby St, Liverpool L69 7BE, UK;3. Amanda Unit, Liverpool Heart and Chest Hospital, Thomas Drive, Liverpool L14 3PE, UK;4. Department for Women''s and Children''s Health, University of Liverpool, Alder Hey Children''s Hospital, Eaton Rd, West Derby, Liverpool L12 2AP, UK |
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| Abstract: | BackgroundEarly eradication therapy is key to keeping the airways Pseudomonas aeruginosa infection-free and rapid identification is essential.MethodsWe used rapid DNA extraction and qPCR assays to detect bacterial, P. aeruginosa and strain-specific targets in samples using two qPCR chemistries. Using 459 respiratory samples from adult and children CF patients, we compared two qPCR methods to culture-based methods in terms of sensitivity and time to result.ResultsFor adult samples, there was 100% concordance between methods. There was no clear pattern in fluctuations in P. aeruginosa number during exacerbation. In child samples, qPCR methods identified additional P. aeruginosa positive samples. The time-to-result was reduced by over 24 h and copy number and colony forming unit could differ dramatically in some samples.ConclusionIf adopted, these methods could significantly improve early P. aeruginosa detection in diagnostic laboratories and therefore play a pivotal role in prolonging infection-free airways in CF patients. |
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