Show simple item record

dc.contributor.authorMcMullan, BJ
dc.contributor.authorDesmarini, D
dc.contributor.authorDjordjevic, JT
dc.contributor.authorChen, SC-A
dc.contributor.authorRoper, M
dc.contributor.authorSorrell, TC
dc.date.accessioned2020-06-30T03:50:46Z
dc.date.available2020-06-30T03:50:46Z
dc.date.issued2015-01-27
dc.identifierpii: PONE-D-14-38843
dc.identifier.citationMcMullan, B. J., Desmarini, D., Djordjevic, J. T., Chen, S. C. -A., Roper, M. & Sorrell, T. C. (2015). Rapid Microscopy and Use of Vital Dyes: Potential to Determine Viability of Cryptococcus neoformans in the Clinical Laboratory. PLOS ONE, 10 (1), https://doi.org/10.1371/journal.pone.0117186.
dc.identifier.issn1932-6203
dc.identifier.urihttp://hdl.handle.net/11343/241013
dc.description.abstractBACKGROUND: Cryptococcus neoformans is the commonest cause of fungal meningitis, with a substantial mortality despite appropriate therapy. Quantitative culture of cryptococci in cerebrospinal fluid (CSF) during antifungal therapy is of prognostic value and has therapeutic implications, but is slow and not practicable in many resource-poor countries. METHODS: We piloted two rapid techniques for quantifying viable cryptococci using mixtures of live and heat-killed cryptococci cultured in vitro: (i) quantitative microscopy with exclusion staining using trypan blue dye, and (ii) flow cytometry, using the fluorescent dye 2'-7'-Bis-(2-carboxyethyl)-5-(6)-carboxyfluorescein, acetoxymethyl ester (BCECF-AM). Results were compared with standard quantitative cryptococcal cultures. Quantitative microscopy was also performed on cerebrospinal fluid (CSF) samples. RESULTS: Both microscopy and flow cytometry distinguished between viable and non-viable cryptococci. Cell counting (on log scale) by microscopy and by quantitative culture were significantly linearly associated (p<0.0001) and Bland-Altman analysis showed a high level of agreement. Proportions of viable cells (on logit scale), as detected by flow cytometry were significantly linearly associated with proportions detected by microscopy (p<0.0001) and Bland-Altman analysis showed a high level of agreement. CONCLUSIONS: Direct microscopic examination of trypan blue-stained cryptococci and flow-cytometric assessment of BCECF-AM-stained cryptococci were in good agreement with quantitative cultures. These are promising strategies for rapid determination of the viability of cryptococci, and should be investigated in clinical practice.
dc.languageEnglish
dc.publisherPUBLIC LIBRARY SCIENCE
dc.titleRapid Microscopy and Use of Vital Dyes: Potential to Determine Viability of Cryptococcus neoformans in the Clinical Laboratory
dc.typeJournal Article
dc.identifier.doi10.1371/journal.pone.0117186
melbourne.source.titlePLOS ONE
melbourne.source.volume10
melbourne.source.issue1
melbourne.source.pagese0117186-
dc.rights.licenseCC BY
melbourne.elementsid1454864
melbourne.openaccess.pmchttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4308066
melbourne.contributor.authorMcMullan, Brendan
dc.identifier.eissn1932-6203
pubs.acceptance.date2014-12-19
melbourne.accessrightsOpen Access


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record