Identification of Aspergillus species using a single restriction enzyme MwoI in RFLP method based on the amplification of rDNA gene.

Diba , kambiz; Mirhendi , seyed hosein; Kordbacheh , parivash; Khorshidvand , zohre

Volume 15, Issue 4 (1-2014) yafte 2014, 15(4): 80-88 | Back to browse issues page

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Diba K, Mirhendi S H, Kordbacheh P, Khorshidvand Z. Identification of Aspergillus species using a single restriction enzyme MwoI in RFLP method based on the amplification of rDNA gene.. yafte 2014; 15 (4) :80-88
URL: http://yafte.lums.ac.ir/article-1-1454-en.html

Identification of Aspergillus species using a single restriction enzyme MwoI in RFLP method based on the amplification of rDNA gene.

Kambiz Diba ^*

, Seyed hosein Mirhendi

, Parivash Kordbacheh

, Zohre Khorshidvand

Abstract: (8615 Views)

Background: Aspergillus species are occasionally associated with human infections. It is essential to find a rapid and reliable method to identify these fungi and can be useful for screening the disease and also epidemiological aims. Materials and Methods: Our subjects included Aspergillus species isolated from clinical and environmental sources of four Iranian educational hospitals. Aspergilli were primarily identified using morphological methods in the level of species. Aspergillus mycelia were harvested from sabouraud broth media and the used for the DNA extraction by Glass beads-Phenol Chloroform method. Ribosomal DNA gene was amplified using the universal primers of ITS. The resulted PCR products were subjected to a digestion with a novel restriction enzyme, MwoI in RFLP method. Results: We could identify eight important species of Aspergillus including: A. flavus, A. fumigatus, A. niger, A. terreus, A. clavatus, A. nidulans, A. ochraceus, A. amsteloidami using tested molecular method. The identification of Aspergillus species was carried out using a protocol including, extraction, PCR and RFLP in a short time 36 hour culture . Conclusion: The method of the PCR-RFLP based on ribosomal DNA gene with the novel restriction enzyme, MwoI enabled us to identify the most medically important Aspergillus species in a short time.

Keywords: Aspergillus, Amplification, Restriction enzyme.

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Type of Study: Research |
Received: 2014/01/21 | Accepted: 2014/01/21 | Published: 2014/01/21