Emergence of Aureobasidium pullulans as human fungal pathogen and molecular assay for future medical diagnosis
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Ascomycota genetics isolation & purification MeSH
- Biological Assay * MeSH
- Early Diagnosis MeSH
- DNA, Fungal analysis MeSH
- DNA Probes chemistry genetics MeSH
- Phenotype MeSH
- Phylogeny MeSH
- Immunocompromised Host MeSH
- Real-Time Polymerase Chain Reaction methods MeSH
- Humans MeSH
- Microscopy MeSH
- Molecular Sequence Data MeSH
- Mycoses diagnosis immunology microbiology MeSH
- Opportunistic Infections diagnosis immunology microbiology MeSH
- Rhodamines analysis MeSH
- RNA, Ribosomal, 18S analysis MeSH
- Base Sequence MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 5-carboxytetramethylrhodamine succinimidyl ester MeSH Browser
- DNA, Fungal MeSH
- DNA Probes MeSH
- Rhodamines MeSH
- RNA, Ribosomal, 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
