Detection of Tropical Fungi in Formalin-Fixed, Paraffin-Embedded Tissue: Still an Indication for Microscopy in Times of Sequence-Based Diagnosis?

被引:21
作者
Frickmann, Hagen [1 ,2 ]
Loderstaedt, Ulrike [3 ]
Racz, Paul [4 ]
Tenner-Racz, Klara [4 ]
Eggert, Petra [4 ]
Haeupler, Alexandra [4 ]
Bialek, Ralf [5 ]
Hagen, Ralf Matthias [1 ]
机构
[1] German Armed Forces Hosp Hamburg, Bernhard Nocht Inst, Dept Trop Med, D-20359 Hamburg, Germany
[2] Univ Med Rostock, Inst Microbiol Virol & Hyg, D-18057 Rostock, Germany
[3] Univ Med Goettingen, Dept Clin Chem, Cent Lab Dept, D-37075 Gottingen, Germany
[4] Bernhard Nocht Inst Trop Med, Dept Infect Dis Pathol, D-20359 Hamburg, Germany
[5] LADR GmbH MVZ Dr Kramer & Kollegen, D-21502 Geesthacht, Germany
关键词
INVASIVE ASPERGILLOSIS; MOLECULAR-DETECTION; IDENTIFICATION; PCR; DNA; INFECTIONS; CULTURE; AGENTS; BLOOD;
D O I
10.1155/2015/938721
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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. Five different previously described panfungal PCR/sequencing protocols targeting 18S and 28S ribosomal RNA gene fragments as well as internal transcribed spacer 1 and 2 fragments were evaluated with a collection of 17 formalinfixed, paraffin-embedded tissue samples of patients with rare and/or tropical invasive mycoses, comprising chromoblastomycosis, coccidioidomycosis, cryptococcosis, histoplasmosis, mucormycosis, mycetoma/maduromycosis, and rhinosporidiosis, in a proof-of- principle analysis. 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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页数:11
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