Comparison of real time diagnostic chemistries to detect Pseudomonas aeruginosa in respiratory samples from cystic fibrosis patients

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• 作者：J.L. Fothergill ; M.J. Ledson ; M.J. Walshaw ; P.S. McNamara ; K.W. Southern ; C. Winstanley
• 关键词：Pseudomonas aeruginosa ; Cystic fibrosis ; Diagnostic testing ; qPCR ; Early colonisation ; Chronic infection
• 刊名：Journal of Cystic Fibrosis
• 出版年：December, 2013
• 年：2013
• 卷：12
• 期：6
• 页码：675-681
• 全文大小：324 K

文摘

Background

Early eradication therapy is key to keeping the airways Pseudomonas aeruginosa infection-free and rapid identification is essential.

Methods

We 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.

Results

For 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.

Conclusion

If 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.