University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal
Gewählte Master / Diploma Thesis:
Raphaela Weber
(2017):
Novel Molecular Methods for the Detection of Food Safety Related Viruses in Frozen Fruit Processing.
Master / Diploma Thesis - Institut für Lebensmittelwissenschaften,
BOKU-Universität für Bodenkultur,
pp 167.
UB BOKU
obvsg
- Abstract:
- In recent years, large outbreaks by food-borne enteroviruses, especially noroviruses (NV) and hepatitis A viruses (HAV) have been observed. Several frozen fruits were found as contamination source. This development represents a major challenge for public health and food production, as very small infectious doses lead to infection. Very sensitive analytical methods are necessary for the reliable detection of viruses, even in lowest concentrations, in the complex food matrix. This master thesis was performed at the AGRANA Research and Innovation Center (ARIC) within the project framework "Decontamination in the First Transformation". The aim of the work was to develop a methodology for the successful detection of viruses in berry fruits. The main focus was on the optimization of an efficient extraction protocol and the comparison of molecular detection methods based on real-time RT-PCR and digital RT-PCR. The combination of a variety of purification steps ultimately resulted in improved virus extraction and lower PCR inhibition. Comparing the two methods, digital RT-PCR proved to be the most sensitive and the more efficient detection methodology. This is due to the 10-fold better detection limit and the low susceptibility to false-positive results, e.g. by the presence of Bacillus pumilus DNA. In a pilot experiment the microbial load of process water samples from the strawberry processing by means of Next Generation Sequencing (NGS) was investigated and presented in connection with plant hygiene and product quality. The results of this research contributed to the introduction of a detection method for viruses into the routine control of berry fruits with the purpose, to reduce the risk for future outbreaks.
- Beurteilende(r): Domig Konrad