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Real-time PCR a digital PCR v diagnostice parazitů v potravinách
Title in English | Real-time PCR a digital PCR in disgnostics of parasitological agents in food |
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Authors | |
Year of publication | 2016 |
Type | Appeared in Conference without Proceedings |
MU Faculty or unit | |
Citation | |
Description | In recent years, the increase of reported outbreaks causing human-diseases associated with food-borne parasitic infections (uni/multi-cellular) arising from meat consumption has been recorded. This trend could be affected e.g. by the changes in farming management towards bio-production, globalization of food market, global climate change and also by the meat processing. This situation is consequently inducing the need for improvement of the diagnostic applications. Our work is focused on development of a reliable comprehensive molecular diagnostic method useful for rapid control of meat products. We adopted high sensitive multiplex oligonucleotide ligation-PCR assay - MOL-PCR, enabling direct and simultaneous detection of multiple nucleic acid signatures of parasites from complex samples and xMAP technology representing a novel detection platform based on magnetic microspheres. Visualization of the products is then realized via qualitative and quantitative MAGPIX instrumentation. Up to this date, the specific molecular probes allowing the detection and capturing of targeted DNA originating from two parasitic worms - Trichinella spiralis (nematode; partial sequence of 18S rRNA gene) and Taenia saginata (cestode; partial sequence of mitochondrial COX1 gene) were designed. Reaction conditions of MOL-PCR were optimized for simultaneous duplex assay. Currently, the calibration of MAGPIX system is in process. |
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