Emergence of Aureobasidium pullulans as human fungal pathogen and molecular assay for future medical diagnosis
Jazyk angličtina Země Spojené státy americké Médium print-electronic
Typ dokumentu časopisecké články, práce podpořená grantem
- MeSH
- Ascomycota genetika izolace a purifikace MeSH
- biotest * MeSH
- časná diagnóza MeSH
- DNA fungální analýza MeSH
- DNA sondy chemie genetika MeSH
- fenotyp MeSH
- fylogeneze MeSH
- imunokompromitovaný pacient MeSH
- kvantitativní polymerázová řetězová reakce metody MeSH
- lidé MeSH
- mikroskopie MeSH
- molekulární sekvence - údaje MeSH
- mykózy diagnóza imunologie mikrobiologie MeSH
- oportunní infekce diagnóza imunologie mikrobiologie MeSH
- rhodaminy analýza MeSH
- RNA ribozomální 18S analýza MeSH
- sekvence nukleotidů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 5-carboxytetramethylrhodamine succinimidyl ester MeSH Prohlížeč
- DNA fungální MeSH
- DNA sondy MeSH
- rhodaminy MeSH
- RNA ribozomální 18S MeSH
Despite the great importance of Aureobasidium pullulans in biotechnology, the fungus had emerged as an opportunistic human pathogen, especially among immunocompromised patients. Clinical detection of this rare human fungal pathogen presently relies on morphology diagnosis which may be misleading. Thus, a sensitive and accurate quantitative molecular assay for A. pullulans remains lacking. In this study, we presented the microscopy observations of A. pullulans that reveals the phenotypic plasticity of the fungus. A. pullulans-specific primers and molecular beacon probes were designed based on the fungal 18S ribosomal RNA (rRNA) gene. Comparison of two probes with varied quencher chemistry, namely BHQ-1 and Tamra, revealed high amplification efficiency of 104% and 108%, respectively. The optimized quantitative real-time PCR (qPCR) assays could detect and quantify up to 1 pg concentration of A. pullulans DNA. Both assays displayed satisfactory performance parameters at fast thermal cycling mode. The molecular assay has great potential as a molecular diagnosis tool for early detection of fungal infection caused by A. pullulans, which merits future study in clinical diagnosis.
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The current status of Aureobasidium pullulans in biotechnology
