Informace o publikaci
xMAP TECHNOLOGY - MULTIPLEX SYSTEM FOR FAST DETECTION OF VIRUSES ASSOCIATED WITH FOOD-/WATERBORNE INFECTIONS
| Autoři | |
|---|---|
| Rok publikování | 2019 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | The microbiological safety of food and environment in which we live is currently an intensely discussed topic. Increasing production and the global market exert pressure on ensuring sufficient quality of food and drinking water. Compared to the past, increased attention is paid to important viral agents associated with food/water contaminations, both in the case of intensive research and in routine diagnostics. However, currently used molecular methods for detection of viruses have a major disadvantage in limited possibility to create multiplex systems for rapid detection of multiple pathogens. Therefore, our goal is to develop and validate a new robust detection method that will allow to overcome this limitation. We used a combination of commercially available xMAP technology (Luminex, Texas, USA) with a multiplex ligation reaction. The open architecture of this system allows to create detection panels with the ability to detect up to 50 different targets in one reaction. The first step of the analysis is reverse transcription, followed by ligation of specific probes in the presence of the target sequence. The next step is amplification of the ligated probes. These amplicons carry a fluorescent signal which is detected in the final step of the analysis by hybridization to magnetic microspheres and using a MAGPIX instrument (Luminex, Texas, USA). We have managed to create a system to detect 8 different targets in one reaction. It allows the simultaneous detection of hepatitis A and E viruses, human noroviruses, adenoviruses, rotaviruses and includes positive internal control of the system. Currently, we have a detection panel focused on viral pathogens that are most commonly associated with cases of contaminated food or water sources. In the near future, it is planned to add other relevant viral agents to create a truly robust system with the required sensitivity, specificity and reliability. |