Detection and identification of fungi in bronchoalveolar lavage fluid from immunocompromised patients using panfungal PCR
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články
Grantová podpora
65269705
Ministerstvo Zdravotnictv? Cesk? Republiky
TE02000058
Technologick? Agentura ?esk? Republiky
MUNI/A/1105/2018
Masarykova Univerzita
PubMed
30535753
DOI
10.1007/s12223-018-00669-w
PII: 10.1007/s12223-018-00669-w
Knihovny.cz E-zdroje
- MeSH
- bronchoalveolární lavážní tekutina mikrobiologie MeSH
- diagnostické techniky molekulární MeSH
- DNA fungální genetika MeSH
- DNA primery genetika MeSH
- houby genetika izolace a purifikace patogenita MeSH
- imunokompromitovaný pacient * MeSH
- intergenová DNA genetika MeSH
- invazivní mykotické infekce diagnóza MeSH
- lidé MeSH
- polymerázová řetězová reakce MeSH
- sekvenční analýza DNA MeSH
- senzitivita a specificita MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- DNA fungální MeSH
- DNA primery MeSH
- intergenová DNA MeSH
Rapid diagnostics of fungal pneumonia and initiation of appropriate therapy are still challenging. In this study, we used two panfungal assays to test bronchoalveolar lavage fluid (BALF) samples to prove their ability to confirm invasive fungal disease diagnosis and identify causative agents. Two methods targeting different fungal rDNA regions were used, and the obtained PCR products were sequenced directly or after cloning. In total, 106 BALF samples from 104 patients were tested. After sequencing, we obtained 578 sequences. Four hundred thirty-seven sequences were excluded from further analysis due to duplication (n = 335) or similarity with sequences detected in the extraction control sample (n = 102); 141 unique sequences were analyzed. Altogether, 23/141 (16%) of the fungi detected belonged to pathogenic species, and 63/141 (45%) were identified as various yeasts; a variety of environmental or very rare fungal human pathogens represented 29/141 (21%) of the total and 26/141 (18%) were described as uncultured fungus. Panfungal PCR detected fungal species that would be missed by specific methods in only one case (probable cryptococcosis). Panfungal PCR followed by sequencing has limited use for testing BALF samples due to frequent commensal or environmental fungal species pickup.
Department of Clinical Microbiology University Hospital Brno Brno Czech Republic
Department of Internal Medicine Hematology and Oncology University Hospital Brno Brno Czech Republic
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