Detection and identification of yeasts from formalin-fixed, paraffin-embedded tissue by use of Pcr-Electrospray ionization mass spectrometry

Patricia Simner, Seanne P. Buckwalter, James R. Uhl, Nancy L. Wengenack, Bobbi S. Pritt

Research output: Contribution to journalArticle

Abstract

Diagnosis of yeast infection is typically accomplished by fungal smear and culture, histopathologic examination, and/or serologic studies. Newer assays based on mass spectrometry may be useful for yeast identification when histologic examination is inconclusive, fungal cultures are not ordered, or cultures fail to yield a causative agent. The purpose of this study was to evaluate the ability of the PLEX-ID broad fungal assay to accurately detect and identify yeasts in formalin-fixed paraffin-embedded (FFPE) tissues. Tissue samples from 78 FFPE specimens with both histopathology and corresponding culture results for a variety of yeasts were tested using the PLEX-ID broad fungal assay. A 40-mFFPE tissue section from each specimen was digested with proteinase K followed by nucleic acid extraction and PCR using broad-range fungal primers. Yeast DNA in amplified products was identified using electrospray ionization mass spectrometry. Discordant results were resolved by D2 rRNA gene sequencing. PLEX-ID analysis detected yeast DNA in 78.2% (61/78) of the cases, of which 91.8% (56/61) were concordant with culture results. Of the 5 discordant positive results, 4 PLEX-ID results were considered to result from environmental contaminants, while 1 clinically important discrepancy was observed (Blastomyces dermatitidis by culture and Cryptococcus neoformans by PLEX-ID). Sequencing of the discordant sample was unsuccessful. The majority of histopathology results (89.7% [70/78]) correlated with culture results. The PLEX-ID broad fungal assay identifies fungi directly from FFPE tissues and can be a useful adjunct to traditional culture and histopathology tests.

Original languageEnglish (US)
Pages (from-to)3731-3734
Number of pages4
JournalJournal of Clinical Microbiology
Volume51
Issue number11
DOIs
StatePublished - Nov 2013
Externally publishedYes

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Electrospray Ionization Mass Spectrometry
Paraffin
Formaldehyde
Yeasts
Blastomyces
Endopeptidase K
Cryptococcus neoformans
DNA
rRNA Genes
Nucleic Acids
Mass Spectrometry
Fungi
Polymerase Chain Reaction
Infection

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

Detection and identification of yeasts from formalin-fixed, paraffin-embedded tissue by use of Pcr-Electrospray ionization mass spectrometry. / Simner, Patricia; Buckwalter, Seanne P.; Uhl, James R.; Wengenack, Nancy L.; Pritt, Bobbi S.

In: Journal of Clinical Microbiology, Vol. 51, No. 11, 11.2013, p. 3731-3734.

Research output: Contribution to journalArticle

Simner, Patricia ; Buckwalter, Seanne P. ; Uhl, James R. ; Wengenack, Nancy L. ; Pritt, Bobbi S. / Detection and identification of yeasts from formalin-fixed, paraffin-embedded tissue by use of Pcr-Electrospray ionization mass spectrometry. In: Journal of Clinical Microbiology. 2013 ; Vol. 51, No. 11. pp. 3731-3734.
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