MOL-PCR and xMAP technology - a multiplex system for fast detection of food- and water-borne viruses.

Abstract Viruses are common causes of food- and water-borne diseases worldwide. Conventional identification of these agents is based on cultivation, antigen detection, electron microscopy or real-time polymerase chain reaction. Since recent technological advancements in detection methods are focused on fast and robust analysis, a rapid multiplexing technology, which can detect a broad spectrum of pathogenic viruses connected to food or water contamination, was utilized. A new semi-quantitative magnetic bead - based multiplex system has been designed for simultaneous detection of several targets in one reaction. The system includes adenoviruses 40/41 (AdV), rotavirus A (RVA), norovirus (NoV), hepatitis E virus (HEV), hepatitis A virus (HAV) and target for external control of the system. To evaluate the detection system, interlaboratory ring tests were performed in four independent laboratories. Analytical specificity of the tool was tested on a cohort of pathogenic agents and biological samples with the quantitative polymerase chain reaction as a reference method. Limit of detection (analytical sensitivity) of 5 × 100 (AdV, HEV and RVA) and 5 × 101 (HAV, NoV) of genome equivalents per reaction was reached. The utilized rapid multiplexing technology represents an acceptably robust and sensitive method, which application could be useful for routine monitoring and control of viruses in food and water safety management.