ศูนย์นวัตกรรมเทคโนโลยีหลังการเก็บเกี่ยว

Highly sensitive protein translation assay for trichothecene toxicity in airborne particulates. Comparison with cytotoxicity assays.

YIKE, L., ALLAN, T., SORENSON, W.G. and DEARBORN, D.G.

Applied and Environmental Microbiology 65: 88-94.

1999

บทคัดย่อ

Screening assays for environmental mycotoxins in bulk samples currently use cytotoxicity in cell cultures, but their application to air particulate samples often lacks sensitivity and specificity for fungal spores. An assay based on inhibition of protein synthesis using translation of firefly luciferase in a rabbit reticulocyte system has been developed for the detection of trichothecenes found in  the spores of toxigenic fungi. Ethanol extracts of air particulates trapped on polycarbonate filters are ultrafiltered and applied at several dilutions to a  translation reaction mixture. The activity of translated luciferase is measured directly in a luminometer, eliminating the need for radioisotopes and time consuming sample processing. Comparison to the 3-(4,5- dimethylthiazol-2yl)-2,5- diphenyltetrazolium bromide cytotoxicity assay indicates a 400 fold increase in sensitivity of trichothecene detection in addition to a much higher specificity for these toxins. Initial field testing indicates a strong correlation between the measured level of toxicity and the presence of toxigenic tungi detected with microbiological methods.