Abstract | INTRODUCTION: The aim of the study was the evaluation of panfungal PCR protocols with subsequent sequence analysis for the diagnostic identification of invasive mycoses in formalin-fixed, paraffin-embedded tissue samples with rare tropical mycoses. MATERIALS AND METHODS: RESULTS: The primers of the panfungal PCRs readily and predominantly reacted with contaminating environmental fungi that had deposited on the paraffin blocks. Altogether three sequence results of histoplasmosis and mycetoma samples that matched the histological assessment were associated with sample age <10 years and virtually without PCR inhibition. CONCLUSIONS: The high risk of amplifying environmental contaminants severely reduces the usefulness of the assessed panfungal PCR/sequencing protocols for the identification of rare and/or tropical mycoses in stored formalin-fixed, paraffin-embedded tissues. Histological assessment remains valuable for such indications if cultural differentiation is impossible from inactivated sample material.
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Authors | Hagen Frickmann, Ulrike Loderstaedt, Paul Racz, Klara Tenner-Racz, Petra Eggert, Alexandra Haeupler, Ralf Bialek, Ralf Matthias Hagen |
Journal | BioMed research international
(Biomed Res Int)
Vol. 2015
Pg. 938721
( 2015)
ISSN: 2314-6141 [Electronic] United States |
PMID | 25961048
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- DNA, Fungal
- RNA, Ribosomal, 18S
- RNA, Ribosomal, 28S
- Formaldehyde
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Topics |
- DNA, Fungal
(genetics, isolation & purification)
- Formaldehyde
- Fungi
(genetics, isolation & purification, pathogenicity)
- Humans
- Mycoses
(diagnosis, genetics, microbiology)
- Paraffin Embedding
- RNA, Ribosomal, 18S
(genetics, isolation & purification)
- RNA, Ribosomal, 28S
(genetics, isolation & purification)
- Tissue Fixation
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