Document Type : Original Article
Authors
1 University of Basrah, Department of Biology, Science College
2 University of Basrah
Abstract
Morphological characterization has gradually been replaced by molecular DNA methods to specifically identify the species of fungi. Therefore, an efficient, fast, and inexpensive procedure to get the fungal genomic DNA is very desirable in a variety of applications including DNA barcoding and genetic epidemiology. This study was aimed at designing a quick ribosomal DNA (rDNA) extraction from fungal pure cultures of ascomycetes and also validate the specificity of monoclonal antibody JF5 in the recognition of antigens of the genus Aspergillus. The development of DNA extraction protocol was tested on 60 isolates of ascomycete fungi and the conventional CTAB methodology was used as the control. Universal ITS primers were used to sequence the amplified ITS regions. At the same time, direct ELISA of surface wash antigens of the tested fungi was carried out. ITS sequencing established the great efficiency (100%) of the rapid protocol and resulted in amplifiable DNA of the appropriate size of the all tested fungal species. This molecular recognition was confirmed immunologically for Aspergillus. JF5 was able to show high specificity toward antigens of the genus Aspergillus. The developed protocol for Aspergillus identification is simple, harmless and can process several samples simultaneously. JF5 exhibited high specificity for Aspergillus antigen detection.
Keywords
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