Molecular biological detection of viruses in food

This work describes molecular detection methods for viruses in foods. The detection systems comprise the following modules: 1(st) virus extraction from foods, 2(nd) RNA extraction, 3(rd) reverse transcription and PCR and 4(th) detection and molecular verification reactions. The modular methods enable the use of the extracted RNA for several detection systems. Here we describe detection systems for Norovirus and Rotavirus, both causing viral gastroenteritis. For each pathogen a conventional RT-PCR and a real-time RT-PCR method is described. DNA-sequencing is applied as molecular verification procedure for conventional RT-PCR whereas the fluorescence labelled probe serves as detection and verification system for real-time RT-PCR. The apathogenic phage MS2 is used as process control. This allows monitoring the efficiency of virus extraction from food, of RNA extraction and the possible presence of inhibitors of the molecular reactions.