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The use of a CYP51C gene based PCR-RFLP assay for simultaneous detection and identification of Fusarium avenaceum and F. tricinctum in wheat
- Fernández-Ortuño, Dolores, Atkins, Sarah L., Fraaije, Bart A.
- International journal of food microbiology 2011 v.145 no.1 pp. 370-374
- restriction fragment length polymorphism, diagnostic techniques, enzymes, DNA primers, DNA, polymerase chain reaction, wheat, Fusarium avenaceum, genes, Fusarium head blight
- Contamination of cereals with mycotoxins such as beauvericin (BEA), enniatins (Ens) and moniliformin (MON) is mainly caused by Fusarium avenaceum and F. tricinctum. This is a world-wide problem which requires rapid and sensitive detection methods. To allow for high throughput screening of large numbers of samples, a diagnostic PCR method was developed for the simultaneous detection of F. avenaceum and F. tricinctum. The interspecific divergence found in the Fusarium-specific CYP51C gene was used to design species-specific PCR primers. The specificity of the assay was demonstrated for DNA samples extracted from a wide range of Fusarium species belonging to the Fusarium head blight (FHB) complex, as well as for naturally-infected grain samples. The PCR-amplified products were digested with the restriction enzyme XbaI to enable differentiation between F. avenaceum and F. tricinctum. This PCR- restriction fragment length polymorphism (RFLP) assay proved to be a simple and relatively inexpensive method highly suited for routine detection and identification of F. avenaceum and F. tricinctum in wheat samples.