title: Development of RFLP-PCR method for the identification of medically important Aspergillus species using single restriction enzyme MwoI.
abstract: In this study we attempted to modify the PCR-RFLP method using restriction enzyme MwoI for the identification of medically important Aspergillus species. Our subjects included nine standard Aspergillus species and 205 Aspergillus isolates of approved hospital acquired infections and hospital indoor sources. First of all, Aspergillus isolates were identified in the level of species by using morphologic method. A twenty four hours culture was performed for each isolates to harvest Aspergillus mycelia and then genomic DNA was extracted using Phenol-Chloroform method. PCR-RFLP using single restriction enzyme MwoI was performed in ITS regions of rDNA gene. The electrophoresis data were analyzed and compared with those of morphologic identifications. Total of 205 Aspergillus isolates included 153 (75%) environmental and 52 (25%) clinical isolates. A. flavus was the most frequently isolate in our study (55%), followed by A. niger 65(31.7%), A. fumigatus 18(8.7%), A. nidulans and A. parasiticus 2(1% each). MwoI enabled us to discriminate eight medically important Aspergillus species including A. fumigatus, A. niger, A. flavus as the most common isolated species. PCR-RFLP method using the restriction enzyme MwoI is a rapid and reliable test for identification of at least the most medically important Aspergillus species.
authors.affiliation: Cellular and Molecular Research Center School of Medicine Urmia University of Medical Sciences UrmiaWest Azarbayejan Iran Cellular and Molecular Research Center, School of Medicine, Urmia University of Medical Sciences, Urmia, West Azarbayejan, Iran.
authors.forename: K
authors.initials: K
authors.lastname: Diba
authors.affiliation: Department of Parasitology and Mycology School of Public Health and Institute of Public Health Research Tehran University of Medical Sciences Tehran Iran Department of Parasitology and Mycology, School of Public Health and Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: H
authors.initials: H
authors.lastname: Mirhendi
authors.affiliation: Department of Parasitology and Mycology School of Public Health and Institute of Public Health Research Tehran University of medical sciences Tehran Iran Department of Parasitology and Mycology, School of Public Health and Institute of Public Health Research, Tehran University of medical sciences, Tehran, Iran.
authors.forename: P
authors.initials: P
authors.lastname: Kordbacheh
authors.affiliation: Department of Parasitology and Mycology School of Public Health and Institute of Public Health Research Tehran University of medical sciences Tehran Iran Department of Parasitology and Mycology, School of Public Health and Institute of Public Health Research, Tehran University of medical sciences, Tehran, Iran.
authors.forename: S
authors.initials: S
authors.lastname: Rezaie
mesh_terms: D001228:Aspergillosis / Q000382:microbiology*; D001230:Aspergillus / Q000145:classification* / Q000235:genetics*; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D013997:Time Factors
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; C060413:endodeoxyribonuclease MwoI; D015252:Deoxyribonucleases, Type II Site-Specific
keywords: Aspergillus; MwoI; identification