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Application of flow cytometry for multiplex analyses in clinical biochemistry
Babušíková, Lucie ; Tremlová,, Bohuslava (referee) ; Beňovská,, Miroslava (advisor)
This thesis discusses the technique of flow cytometry for multiplex analysis and its use in conjunction with imunochemical methods. As part of this work was carried out clinical studies dealing with secondary prevention of myocardial infarction and atherosclerosis in 186 pacientů. In this time represents myocardial infarction worldwide civilizational problem. A number of possible parameters for monitoring atherosclerosis in the world is still an unresolved issue. In the practical part of this work we performed an analysis using Luminex xMAP technology for new parameters (adiponectin, resistin, osteopontin) to predict atherosclerotic disease associated with myocardial infarcion. Also we wanted to see how these parameters are changed in patients after increasing the dose of therapeutic drugs.
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Implementation and optimization of ELISA method
KOPAČKOVÁ, Tereza
Enzyme-Linked ImmunoSorbent Assay or ELISA is a method that has a wide application, especially in immunology. Antigens and antibodies can be detected by the method. The basic principle of the method is the reaction of antigen and antibody and the subsequent formation of an immunocomplex. The immunocomplex is detected by a conjugate that causes a color change of the substrate in the presence of antigen or antibody in the test sample. The ELISA test is a simple method important fot the diagnosis of various diseases or laboratory research. This method is a special type of enzyme immunoassay. The bachelor thesis is focused on the general implementation and optimization of the ELISA method. The aim of the theoretical part of the work is a general introduction to immunological methods. The division of individual types of ELISA tests, including the principles and use in practice, is elaborated in detail. In the practical part, the ELISA method is introduced and optimized using a DYNAREAD photometer for reading 96well microtiter plates and AlaDYN software. In this way, a total of 8 test samples of sera are examined and subsequently the obtained data are processed. The test measurement was performed using the QuatiVac ELISA test kit and human IgG antibodies against SARS-CoV-2 in serum were determined. The peaks of SARS-CoV-2 spike proteins expresses recombinantly in the human cell line HEK 293 were used as antigens. The measurement was performed using a DYNAREAD instrument and data processing was performer using AlaDYN software. 6 calibrators, a positive control, a negative control and then individual serum samples were measured. The measurement results are 5 serum samples in which IgG class anti-SARS-CoV-2 antibodies were detected and 3 samples in which no antibodies were detected.
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Parasitic protease SmCB2 as a target for the treatment of schistosomiasis
Bakardjieva, Marina ; Mareš, Michael (advisor) ; Mikeš, Libor (referee)
Blood flukes of the genus Schistosoma are parasitic trematodes that cause schistosomiasis, a serious disease afflicting more than 240 million people. The proteolytic system of schistosomes is essential for their viability: it participates in important processes during host-parasite interactions such as food digestion, invasion and tissue migration. Thus, schistosomal proteases are promising molecular targets for therapeutic intervention in schistosomiasis treatment. The thesis focuses on the protease cathepsin B2 from S. mansoni (SmCB2) which has not been studied in detail so far in terms of biochemical properties and biological function. Recombinant SmCB2 was prepared using yeast and bacterial expression systems and was chromatographically purified. Using an in vitro activity assay, the first effective inhibitors of SmCB2 were identified which inhibited its proteolytic activity in submicromolar concentrations. Specific polyclonal antibodies against SmCB2 were prepared and used for immunomicroscopic localization of this protease on the surface of the parasite. ELISA analysis demonstrated that SmCB2 is a parasite antigen recognized by the host immune system in the mouse model of schistosomiasis. The thesis provides valuable information about SmCB2 as a potential target molecule for synthetic...
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Determination of cortisol in saliva
Žánová, Magdalena ; Martínková, Markéta (advisor) ; Milichovský, Jan (referee)
This thesis deals with salivary cortisol levels measured by the ADVIA: Centaur CP Immunoassay System. Experimental saliva sampling was performed on 141 pro- bands from age 7 to 76. Volunteers were divided to groups according to their sex: males, females using hormonal contraceptives (HC) and females not using HC. Re- ference intervals of morning and evening salivary cortisol were defined: females not using HC 13,2 - 55,5 nmol/l and females using HC 15,5 - 44,2 nmol/l for mor- ning salivary cortisol. Females not using HC 4,0 - 16,6 nmol/l and females using HC 7,9 - 22,6 nmol/l for afternoon salivary cortisol. Males 15,8 - 47,7 nmol/l for morning salivary cortisol and 5,2 - 25,4 nmol/l for afternoon salivary cortisol. Dif- ferences in stated intervals were imperceptible in all monitored groups. However, it is necessary to maintain different reference intervals for both morning and evening sampling. Reference interval for morning sampling was 14,3 - 46,2 nmol/l and refe- rence interval for afternoon sampling was set at 4,0 - 22,2 nmol/l. Daily profiles of salivary cortisol were determined in 6 females and 4 males in four different parts of a day. The course of salivary cortisol levels corresponded in females not using HC, females using HC and males with circadian rhythm, which is subject to cortisol. Sa-...
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Application of flow cytometry for multiplex analyses in clinical biochemistry
Babušíková, Lucie ; Tremlová,, Bohuslava (referee) ; Beňovská,, Miroslava (advisor)
This thesis discusses the technique of flow cytometry for multiplex analysis and its use in conjunction with imunochemical methods. As part of this work was carried out clinical studies dealing with secondary prevention of myocardial infarction and atherosclerosis in 186 pacientů. In this time represents myocardial infarction worldwide civilizational problem. A number of possible parameters for monitoring atherosclerosis in the world is still an unresolved issue. In the practical part of this work we performed an analysis using Luminex xMAP technology for new parameters (adiponectin, resistin, osteopontin) to predict atherosclerotic disease associated with myocardial infarcion. Also we wanted to see how these parameters are changed in patients after increasing the dose of therapeutic drugs.
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Search for new biomarkers for screening and early diagnosis of cancer
KALČÍKOVÁ, Kateřina
Thesis on the topic "Search of New biomakers suitable for screening and early diagnostics of cancer" is dedicated to the issue of cancer, a cancer of the breast, prostate, ovarian and after the new findings out as melanom determination of tumor markers and their implications for clinical practice. Their aim is skouted up biomaker suitable for screening and early diagnostics of carcinoma( cancer ) of the breast, ovarian, prostate and malignant melanoma with utilization of various kinds of immunoassay. The theoretical part gives a brief and comprehensive overview of the epidemiology of selected tumors and tumor markers used and their qualities. In the methodological part are given principles applied iminoanalytics methods a list of the statistical methods and a detailed description of the groups of patients. Results section contains tables with results of biomarkers diveded by sort of avalueted tumors. In the discussion are then analyzed facts that follow from the results. It discussed the process and actualy status of searching convenients biomekers and their combinations that can distinguish a population of benign tumors of the population with malignant tumors and markers which are able to distinguish healthy population from the population with malignant tumors of the breast, ovary, prostate and melanoma. This work may serve as a source of knowledge and results to continue in search of such marker that could be used for screening and early diagnosis of cancer and so to improve the prevention of these illneses in the population.
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