Aspergillus tubingensis and Aspergillus niger as the dominant black Aspergillus, use of simple PCR-RFLP for preliminary differentiation
| Autor: | Marjan Motamedi, Hossein Mirhendi, Sadegh Nouripour-Sisakht, Fereshteh Zarei |
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| Rok vydání: | 2015 |
| Předmět: |
0301 basic medicine
Sequence analysis 030106 microbiology Restriction Mapping Aspergillus acidus Air Microbiology Polymerase Chain Reaction law.invention Microbiology 03 medical and health sciences law Tubulin Environmental Microbiology Aspergillosis Humans DNA Fungal Polymerase chain reaction Aspergillus awamori Soil Microbiology Electrophoresis Agar Gel Aspergillus biology Aspergillus niger biology.organism_classification 030104 developmental biology Infectious Diseases Aspergillus tubingensis Food Microbiology Restriction fragment length polymorphism Genome Fungal Polymorphism Restriction Fragment Length |
| Zdroj: | Journal de mycologie medicale. 26(1) |
| ISSN: | 1773-0449 |
| Popis: | This work aimed to identify the species distribution of common clinical and environmental isolates of black Aspergilli based on simple restriction fragment length polymorphism (RFLP) analysis of the β-tubulin gene. A total of 149 clinical and environmental strains of black Aspergilli were collected and subjected to preliminary morphological examination. Total genomic DNAs were extracted, and PCR was performed to amplify part of the β-tubulin gene. At first, 52 randomly selected samples were species-delineated by sequence analysis. In order to distinguish the most common species, PCR amplicons of 117 black Aspergillus strains were identified by simple PCR-RFLP analysis using the enzyme TasI. Among 52 sequenced isolates, 28 were Aspergillus tubingensis, 21 Aspergillus niger, and the three remaining isolates included Aspergillus uvarum, Aspergillus awamori, and Aspergillus acidus. All 100 environmental and 17 BAL samples subjected to TasI-RFLP analysis of the β-tubulin gene, fell into two groups, consisting of about 59% (n=69) A. tubingensis and 41% (n=48) A. niger. Therefore, the method successfully and rapidly distinguished A. tubingensis and A. niger as the most common species among the clinical and environmental isolates. Although tardy, the Ehrlich test was also able to differentiate A. tubingensis and A. niger according to the yellow color reaction specific to A. niger. A. tubingensis and A. niger are the most common black Aspergillus in both clinical and environmental isolates in Iran. PCR-RFLP using TasI digestion of β-tubulin DNA enables rapid screening for these common species. |
| Databáze: | OpenAIRE |
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