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dc.contributor.authorPathak, Sharad
dc.contributor.authorAwuh, Jane Atesoh
dc.contributor.authorLeversen, Nils Anders
dc.contributor.authorFlo, Trude Helen
dc.contributor.authorÅsjø, Birgitta
dc.identifier.citationPLoS ONE 2012, 7(4)nb_NO
dc.description.abstractDue to the slow growth rate and pathogenicity of mycobacteria, enumeration by traditional reference methods like colony counting is notoriously time-consuming, inconvenient and biohazardous. Thus, novel methods that rapidly and reliably quantify mycobacteria are warranted in experimental models to facilitate basic research, development of vaccines and anti-mycobacterial drugs. In this study we have developed quantitative polymerase chain reaction (qPCR) assays for simultaneous quantification of mycobacterial and host DNA in infected human macrophage cultures and in mouse tissues. The qPCR method cannot discriminate live from dead bacteria and found a 10- to 100-fold excess of mycobacterial genomes, relative to colony formation. However, good linear correlations were observed between viable colony counts and qPCR results from infected macrophage cultures (Pearson correlation coefficient [r] for M. tuberculosis = 0.82; M. a. avium = 0.95; M. a. paratuberculosis = 0.91). Regression models that predict colony counts from qPCR data in infected macrophages were validated empirically and showed a high degree of agreement with observed counts. Similar correlation results were also obtained in liver and spleen homogenates of M. a. avium infected mice, although the correlations were distinct for the early phase (nb_NO
dc.publisherPublic Library of Sciencenb_NO
dc.rightsNavngivelse 3.0 Norge*
dc.titleCounting mycobacteria in infected human cells and mouse tissue: a comparison between qPCR and CFUnb_NO
dc.typeJournal articlenb_NO
dc.typePeer reviewednb_NO
dc.subject.nsiVDP::Medisinske fag: 700::Basale medisinske, odontologiske og veterinærmedisinske fag: 710::Medisinsk mikrobiologi : 715nb_NO
dc.subject.nsiVDP::Midical sciences: 700::Basic medical, dental and veterinary sciences: 710::Medical microbiology: 715nb_NO
dc.source.journalPLoS ONEnb_NO
dc.description.localcode© 2012 Pathak et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.nb_NO

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Navngivelse 3.0 Norge
Except where otherwise noted, this item's license is described as Navngivelse 3.0 Norge