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Identification of Aspergillus Sections Flavi, Nigri, and Fumigati and Their Differentiation Using Specific Primers Pubmed



Ashtiani NM1 ; Kachuei R2 ; Yalfani R3 ; Harchegani AB4 ; Nosratabadi M5
Authors
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Authors Affiliations
  1. 1. Department of Biology, School of Basic Science, Islamic Azad University-Pishva Varamin Branch, Tehran, Iran
  2. 2. Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
  3. 3. Department of Nursery, Faculty of Nursery and Midwifery, Islamic Azad University-Pishva Varamin Branch, Tehran, Iran
  4. 4. Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran
  5. 5. Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran

Source: Infezioni in Medicina Published:2017


Abstract

Aspergillus species are important in medicine, agriculture and various industries. The sections Fumigati, Flavi, and Nigri are the most important members of the Aspergillus genus. This study intended to identify and separate these three Aspergillus sections and to differentiate among them using specific primers. A bioinformatics study was initially performed to analyse the sequences of five genes, namely, beta-tubulin, calmodulin, the pre-rRNA processing protein Tsr1, the DNA-replication licensing factor Mcm7, and RNA polymerase II second largest subunit (RPB2) in the three Aspergillus sections using MEGA6 software and the NCBI database. Primers were designed to select genes for each of the Aspergillus sections being analysed. A total of 134 environmental and clinical Aspergillus species were isolated, purified and ini- tially identified by colony morphology..Subsequently, DNA was extracted using the phenol-chloroform method, specific primers were synthesized, PCR was performed for DNA from all isolates, and the results were compared to morphological characteristics. Of the 134 isolates tested, 56 were Nigri, 32 were Fumigati, 32 were Flavi, and the rest (14 isolates) belonged to other sections. The beta-tubulin and calmodulin genes were found to be the most suitable for differentiating among these three groups; the beta-tubulin gene was used for molecular identification of Aspergillus section Fumigati, and the calmodulin gene for identifying sections Flavi and Nigri. © 2017, EDIMES Edizioni Medico Scientifiche. All rights reserved.