PCR-RFLP on β-tubulin gene for rapid identification of the most clinically important species of Aspergillus
| Autor: | Mojtaba Nabili, Mojtaba Taghizadeh Armaki, Tuba Nasri, Mahdi Abastabar, Akbar Hoseinnejad, Mohammad Taghi Hedayati, Alessandro C. Pasqualotto |
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| Rok vydání: | 2015 |
| Předmět: |
Microbiology (medical)
Aspergillus flavus Microbiology Polymerase Chain Reaction law.invention Aspergillus fumigatus Aspergillus nidulans law Tubulin Environmental Microbiology Aspergillosis Humans Aspergillus terreus skin and connective tissue diseases DNA Fungal Molecular Biology Polymerase chain reaction Electrophoresis Agar Gel Aspergillus biology biology.organism_classification Molecular Typing Restriction fragment length polymorphism Aspergillus clavatus Polymorphism Restriction Fragment Length |
| Zdroj: | Journal of microbiological methods. 117 |
| ISSN: | 1872-8359 |
| Popis: | Aspergillus species are important agents of life-threatening infections in immunosuppressed patients. Proper speciation in the Aspergilli has been justified based on varied fungal virulence, clinical presentations, and antifungal resistance. Accurate identification of Aspergillus species usually relies on fungal DNA sequencing but this requires expensive equipment that is not available in most clinical laboratories. We developed and validated a discriminative low-cost PCR-based test to discriminate Aspergillus isolates at the species level. The Beta tubulin gene of various reference strains of Aspergillus species was amplified using the universal fungal primers Bt2a and Bt2b. The PCR products were subjected to digestion with a single restriction enzyme AlwI. All Aspergillus isolates were subjected to DNA sequencing for final species characterization. The PCR-RFLP test generated unique patterns for six clinically important Aspergillus species, including Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus terreus, Aspergillus clavatus and Aspergillus nidulans. The one-enzyme PCR-RFLP on Beta tubulin gene designed in this study is a low-cost tool for the reliable and rapid differentiation of the clinically important Aspergillus species. |
| Databáze: | OpenAIRE |
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