The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis

Simple item page
dc.contributor.authorDevonshire, Alison S.en_ZA
dc.contributor.authorO’Sullivan, Denise M.en_ZA
dc.contributor.authorHoneyborne, Isobellaen_ZA
dc.contributor.authorJones, Gerwynen_ZA
dc.contributor.authorKarczmarczyk, Mariaen_ZA
dc.contributor.authorPavsic, Jernejen_ZA
dc.contributor.authorGutteridge, Aliceen_ZA
dc.contributor.authorMilavec, Mojcaen_ZA
dc.contributor.authorMendoza, Pabloen_ZA
dc.contributor.authorSchimmel, Heinzen_ZA
dc.contributor.authorVan Heuverswyn, Franen_ZA
dc.contributor.authorGorton, Rebeccaen_ZA
dc.contributor.authorCirillo, Daniela Mariaen_ZA
dc.contributor.authorBorroni, Emanueleen_ZA
dc.contributor.authorHarris, Kathrynen_ZA
dc.contributor.authorBarnard, Marinusen_ZA
dc.contributor.authorHeydenrych, Anthenetteen_ZA
dc.contributor.authorNdusilo, Norahen_ZA
dc.contributor.authorWallis, Carole L.en_ZA
dc.contributor.authorPillay, Keshreeen_ZA
dc.contributor.authorBarry, Thomasen_ZA
dc.contributor.authorReddington, Kateen_ZA
dc.contributor.authorRichter, Elviraen_ZA
dc.contributor.authorMozioglu, Erkanen_ZA
dc.contributor.authorAkyurek, Semaen_ZA
dc.contributor.authorYalcınkaya, Burhanettinen_ZA
dc.contributor.authorAkgoz, Muslumen_ZA
dc.contributor.authorZel, Janaen_ZA
dc.contributor.authorFoy, Carole A.en_ZA
dc.contributor.authorMcHugh, Timothy D.en_ZA
dc.contributor.authorHuggett, Jim F.en_ZA
dc.date.accessioned2017-07-28T10:37:29Z
dc.date.available2017-07-28T10:37:29Z
dc.date.issued2016
dc.descriptionCITATION: Devonshire, A. S., et al. 2016. The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis. BMC Infectious Diseases, 16:366, doi:10.1186/s12879-016-1696-7.
dc.descriptionThe original publication is available at https://bmcinfectdis.biomedcentral.com
dc.description.abstractBackground: Real-time PCR (qPCR) based methods, such as the Xpert MTB/RIF, are increasingly being used to diagnose tuberculosis (TB). While qualitative methods are adequate for diagnosis, the therapeutic monitoring of TB patients requires quantitative methods currently performed using smear microscopy. The potential use of quantitative molecular measurements for therapeutic monitoring has been investigated but findings have been variable and inconclusive. The lack of an adequate reference method and reference materials is a barrier to understanding the source of such disagreement. Digital PCR (dPCR) offers the potential for an accurate method for quantification of specific DNA sequences in reference materials which can be used to evaluate quantitative molecular methods for TB treatment monitoring. Methods: To assess a novel approach for the development of quality assurance materials we used dPCR to quantify specific DNA sequences in a range of prototype reference materials and evaluated accuracy between different laboratories and instruments. The materials were then also used to evaluate the quantitative performance of qPCR and Xpert MTB/RIF in eight clinical testing laboratories. Results: dPCR was found to provide results in good agreement with the other methods tested and to be highly reproducible between laboratories without calibration even when using different instruments. When the reference materials were analysed with qPCR and Xpert MTB/RIF by clinical laboratories, all laboratories were able to correctly rank the reference materials according to concentration, however there was a marked difference in the measured magnitude. Conclusions: TB is a disease where the quantification of the pathogen could lead to better patient management and qPCR methods offer the potential to rapidly perform such analysis. However, our findings suggest that when precisely characterised materials are used to evaluate qPCR methods, the measurement result variation is too high to determine whether molecular quantification of Mycobacterium tuberculosis would provide a clinically useful readout. The methods described in this study provide a means by which the technical performance of quantitative molecular methods can be evaluated independently of clinical variability to improve accuracy of measurement results. These will assist in ultimately increasing the likelihood that such approaches could be used to improve patient management of TB.en_ZA
dc.description.urihttps://bmcinfectdis.biomedcentral.com/articles/10.1186/s12879-016-1696-7
dc.description.versionPublisher's version
dc.format.extent10 pages
dc.identifier.citationDevonshire, A. S., et al. 2016. The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis. BMC Infectious Diseases, 16:366, doi:10.1186/s12879-016-1696-7
dc.identifier.issn1471-2334 (Online)
dc.identifier.otherdoi:10.1186/s12879-016-1696-7
dc.identifier.urihttp://hdl.handle.net/10019.1/102022
dc.language.isoen_ZAen_ZA
dc.publisherBioMed Central
dc.rights.holderAuthors retain copyright
dc.subjectMycobacterium tuberculosis -- Molecular diagnosisen_ZA
dc.subjectPolymerase chain reactoren_ZA
dc.subjectPMRen_ZA
dc.subjectMolecular biologyen_ZA
dc.subjectQuantitative molecular diagnostic methodsen_ZA
dc.titleThe use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosisen_ZA
dc.typeArticleen_ZA
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
devonshire_use_2016.pdf
Size:
684.49 KB
Format:
Adobe Portable Document Format
Description:
Download article
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
1.95 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Research Articles (Molecular Biology and Human Genetics)