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A PCR-ELISA for the detection of potential fumonisin producing Fusarium species has been developed, using the ribosomal ITS1 sequence as target. For this purpose, the sequences of the ITS1 regions of different fumonisin producing Fusarium species have been determined and compared to the sequences of fumonisin non-producing species. In general, the ITS1 sequences were highly homologous. However, some minor sequence polymorphisms were detected, which differentiates potential fumonisin producingFusarium species from non-producing species. By using these sequence differences, a PCR-ELISA for potential fumonisin producing Fusarium species was developed. All other ubiquitously occurring food-borne fungi tested showed negative results with this test.