title: Development of a PCR-based assay for rapid and reliable identification of pathogenic Fusaria.
abstract: Identification of Fusarium species has always been difficult due to confusing phenotypic classification systems. We have developed a fluorescent-based polymerase chain reaction assay that allows for rapid and reliable identification of five toxigenic and pathogenic Fusarium species. The species includes Fusarium avenaceum, F. culmorum, F. equiseti, F. oxysporum and F. sambucinum. The method is based on the PCR amplification of species-specific DNA fragments using fluorescent oligonucleotide primers, which were designed based on sequence divergence within the internal transcribed spacer region of nuclear ribosomal DNA. Besides providing an accurate, reliable, and quick diagnosis of these Fusaria, another advantage with this method is that it reduces the potential for exposure to carcinogenic chemicals as it substitutes the use of fluorescent dyes in place of ethidium bromide. Apart from its multidisciplinary importance and usefulness, it also obviates the need for gel electrophoresis.
authors.affiliation: School of Plant Sciences, The University of Reading, Whiteknights, Berks RG6 6AS, Reading, UK. pmishra@afhe.ualberta.ca
authors.forename: Prashant K
authors.initials: PK
authors.lastname: Mishra
authors.forename: Roland T V
authors.initials: RT
authors.lastname: Fox
authors.forename: Alastair
authors.initials: A
authors.lastname: Culham
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis; D004275:DNA, Ribosomal / Q000235:genetics; D005670:Fusarium / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009181:Mycoses / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D013997:Time Factors
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal