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Laboratory methods using in diagnostic morbus Bechterew
FAROVÁ, Veronika
The aim of my bachelor thesis was to get familiar with laboratory methods, which are used in the diagnostics of morbus Bechterew. Very important for the diagnostics of morbus Bechterew is the marker HLA-B27, which is identified by screening, with the help of flow cytometer and then is the presence confirmed or refuted with the help of molecular-biological methods. Other laboratory examination, which is essential when it comes to interception and observation of the activities of morbus Bechterew, is the determination of the C-reactive protein level, which is an indicator of inflammatory reaction in the body of the patient. The last examination I got familiar with was the determination of the sedimentation speed of erythrocytes, which is used as an indicator of ongoing inflammation in the body. In the theoretical part of the thesis I focused primarily on the definition of morbus Bechterew. I dealt with the question what are the symptoms of the disease, then also according to which criteria is the disease diagnosed and what treatment is applied. In the second part of the theoretical part I focused primarily on the major histocompatibility complex, which is highly connected with morbus Bechterew. I dealt mostly with the structure and presentation of antigens by the means of MHC molecules of class I. I also mentioned the structure and the function of C-reactive protein. In the practical part I described laboratory methods used in diagnostics of morbus Bechterew. The first method I tried was screening examination of HLA-B27 marker by the means of flow cytometry. I introduced how the following processes should look like: collection of biological material, preparation of the sample and the analysis made on the flow cytometer. At the end of this part I put an example of the analysis results, how do the results look like when they are positive, negative, or in the grey zone. As a second method I introduced genetic typing of HLA-B27 molecule, which is made by the means of real time PCR. I mentioned the preanalytical part and the process of preparation. Moreover I dealt with the measuring of C-reactive protein by the means of nephelometry and then I also mentioned examination with the help of a test strip. The last method I tried was measuring the sedimentation speed of erythrocytes, I mentioned the principle of method, the process of measuring and the measured results as well. In the last part of my bachelor thesis I worked with the data I got from Motol University Hospital, more precisely from the Department of Immunology, 2nd Faculty of Medicine of Medicine, Charles University in Prague and Motol University Hospital. During the year 2017, overall 452 patients were tested on the presence of HLA-B27 marker and 75 of them tested positive. Then I worked only with HLA-B27 positive patients. I researched how much is the disease represented in both sexes, what age do the HLA-B27 positive patients have and what is their diagnosis.
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