The type A trichothecenes T-2 toxin (T-2) and HT-2 toxin (HT-2) are hazardous Fusarium products that contaminate many field crops growing in cold to temperate regions across the world. Toxicity studies in laboratory and farm animals have been used to derive a temporary tolerable daily intake (t-TDI) for the sum of T-2 and HT-2 of no more than 60 ng/kg body weight. To protect the consumers, it is now necessary to screen a large number of food samples for the presence of these poisonous fungal metabolites. Towards that goal, we discovered that the transcriptional apparatus of a human carcinoma cell line (MCF7) provides a sensitive biological sensor of type A trichothecenes. In fact, exposure of this easy-to-culture cell line to T-2 or HT-2 results in the regulation of >2,000 different transcripts with expression changes ranging from >5,000-fold gene inductions to >40-fold gene repressions. These transcriptional responses have been exploited to develop practical microchip and reverse transcription-polymerase chain reaction (RT-PCR) assays for the detection of type A trichothecenes at parts per billion levels.
Type A trichothecenes (primarily T-2 and HT-2 toxins) are common fungal metabolites found in a wide range of grains and other field crops grown in temperate climatic zones. By acting as potent inhibitors of protein synthesis, T-2 and HT-2 exert adverse effects particularly against rapidly proliferating tissues, including the bone marrow, the immune system and epithelial cells. Based on toxicity studies in laboratory and farm animals, a temporary tolerable daily intake for the sum of T-2 and HT-2 has been issued in the European Union. However, exposure assessments suggest that the combined intake of these natural compounds exceeds in many cases the proposed threshold. To further protect the consumers, it is therefore necessary to screen a large number of food samples for parts per billion levels of both T-2 and HT-2. Towards that goal, we are the first to report that these two type A trichothecenes induce fast and high-amplitude transcriptional changes in cultured human breast cancer cells. This specific response involving marker gene inductions by more than 1000-fold has been exploited to develop a real-time PCR-based screening method that displays a limit of detection of 5 ng g(-1) for T-2 and 10 ng g(-1) for HT-2. The practicability of this bioassay is demonstrated by its application to the detection of type A trichothecenes in different food matrices.
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