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Detekce patogenů lilku bramboru přežívajících v půdě
Valkovičová, Nikola
The bachelor's thesis dealt with the detection and sequencing of soil-borne pathogens of pota-to eggplant. The focus is mainly on pathogens of the genus Fusarium. These pathogens cause such potato rot, which forms white mycelium coatings and subsequently mummifies, that it is not suitable for consumption and the yield is reduced. One of the most accurate methods was chosen for the detection of pathogens – PCR. For the detection of Fusarium pathogens from other potato tubers, conventional PCR was used with non-specific primers ITS4 and ITS5 annealing to the region between the genes that code for ITS4 and ITS5 RNA. For the detection of pathogens from the soil, real–time PCR was chosen with primers ITS1F and AFP346 sitting in the region that encode nRNA and space bars ITS1 and ITS2. The tested soil was artificially inoculated with an unknown isolate of the genus Fusarium obtained from a potato tuber and a collection isolate of Fusarium solani var. coeruleum. Se-quencing confirmed infection of the potato tuber with Fusarium culmorum, clarifying the dif-ficulty of quantification.
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Alternative beverages of probiotic character
Moravec, Štěpán ; Langová, Denisa (referee) ; Trachtová, Štěpánka (advisor)
Bachelor thesis deals with explanation of therm probiotics, properties of each probiotic bacteria genuses. preparation of coffee and tea, and also deals with probiotic coffee and tea and molekular diagostic methods which were used during bachelor thesis. There were used mikrobiological and molecular diagnostic methods for analysis of probiotic coffee and teas. Namely for confirmation of bacterial genus Bacillus and for confirmation of survavibility of Bacillus during high temperatures. Presence of bacterial DNA in samples of coffee and teas was confirmed by conventional PCR method and by real-time PCR method. Survibility of genus Bacillus during high temperaturewas confirmed by PMA-PCR method.
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qPCR diagnostika střevního prvoka \kur{Blastocystis} sp. v souboru vzorků od zdravých lidí
ŠLOUFOVÁ, Martina
The main aim of this study was to introduce and optimize the qPCR diagnostic protocol for detection of intestinal protist, Blastocystis sp. We compared the sensitivity of conventional PCR (cPCR) and real-time PCR (qPCR) in a set of 288 human samples from gut-healthy individuals and subtype diversity as detected by Next-generation sequencing (NGS) versus Sanger sequencing. The overall prevalence of Blastocystis sp. was 29 %. Based on the results, we found out that qPCR is a more sensitive method than cPCR. In subtype detection, NGS was completely in agreement with Sanger sequencing but showed higher sensitivity for mixed subtype colonization within one host. A combination of these two approaches could be beneficial for future epidemiological studies.
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