aeruginosa culture and qPCR positive but the follow-up samples were culture and qPCR negative. This may indicate that qPCR still detected DNA of already killed bacteria. Another 10 samples (1%) were P. aeruginosa
qPCR negative but culture positive. False negativity of the qPCR was not the reason for the negative qPCR result, because CH5424802 in vitro qPCR inhibition and primer mismatch could be excluded. Interestingly, for 5 of these 10 patients, there was discordance between both culture techniques, suggestive for borderline detection by culture and thus a low inoculum of the pathogen. Such discordance between culture results was observed in only 11 out of 89 qPCR positive samples. For many samples with discordant qPCR and culture results, a low bacterial inoculum may be the explanation. Based on our results in this study
and a previous study , both approaches have comparable sensitivity, and at low inocula both may be at the border of their detection limit. In addition, at low inocula the distribution of the bacteria in the sample may be more uneven and because we used different parts of each sample to perform qPCR respectively culture, randomization may have influenced the qPCR and/or https://www.selleckchem.com/products/ly3039478.html culture result negatively. The presence of a low inoculum can be concluded from the significantly higher Cq values of qPCR positive/culture negative samples, compared to the qPCR positive/culture positive samples and from the fact that cultures were positive for only one of both media used in 5 out of 10 qCPR negative/culture positive samples. Possibly other factors, such as sample type, the presence of other bacterial species or the genotype of the P. aeruginosa isolate might differentially influence the ease with which P. aeruginosa can be detected by culture versus qPCR. Further research is warranted on a larger set of samples with discordant qPCR – bacterial culture results to determine the Immune system influence of some of these factors. Conclusions The NVP-AUY922 concentration present study indicates that the currently used routine culture techniques perform equally well as DNA amplification
techniques for detection of P. aeruginosa in respiratory samples of CF patients, not chronically infected with P. aeruginosa. Looking at it from a different angle, qPCR was both sensitive and specific compared with a gold standard of culture. These data, gathered on clinical samples, confirm the results of our previous laboratory study in which culture methods were equally sensitive to the combination of the most sensitive DNA extraction method and the most sensitive amplification assay, i.e. probe based qPCR . Therefore, we may conclude that for this study, based on a large amount of patients and samples, qPCR for P. aeruginosa may have a predictive value for impending P. aeruginosa infection in only a limited number of cases. Acknowledgements Pieter Deschaght is indebted to the IWT for PhD research grant IWT-SB/71184.