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Lecturer(s)
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Korecká Lucie, doc. RNDr. Ph.D.
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Slováková Marcela, doc. Mgr. Ph.D.
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Course content
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Lecture topics 1st week Introduction to immunoanalysis, structure and properties of antigen, hapten, antibody, immunochemical reaction of antigen and antibody, reaction equilibrium. 2.-3. Immune system reactions to the presence of antigens, overview of immunological tissues, cells and their function, immune mechanisms. 4th week Development of antigens and antibodies as diagnostic tools. Monoclonal and polyclonal antibodies. 5th week Overview of serological methods, quantification methods, titer definition. Agglutination, haemagglutination methods. 6th Precipitating and immunodiffusion techniques for detection and quantification of analytes. 7th week Serological haemolytic methods (antistreptolysin O, complement functional tests). 8th week Immunoanalytical methods with indicators, classification by indicator type. Principles, methods EIA, RIA, FIA, DELFIA, LIA, MEIA. 9th week Screening field immunochromatographic tests - arrangement of tests for low-molecular and high-molecular analytes, concrete examples of applications. Biosensors in immunoassays. 10th week Electromigration methods - immunofixation, immunoelectrophoresis, native and denaturing electrophoresis for evaluation of quality of antigens and antibodies. 11th week Blotting techniques - western blotting, dot blot, different detection methods according to sensitivity. Immunofluorescence techniques for detection of (auto) antibodies, detection of antigens in tissues. 12th week Functional tests in immunology - examination of cellular immunity. Techniques of separation of cell populations. Method of flow cytometry, immunophenotyping, immunoregulatory index. 13th week Examination of phagocytosis functions. Examination of the HLA system. Laboratory diagnostics of allergies.
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Learning activities and teaching methods
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unspecified, Dialogic (discussion, interview, brainstorming)
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Learning outcomes
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The aim of the course is to introduce immunoanalytical methods, including explanations of terms as antigen, antibody, immunocomplex and diagnostic reagents, and principles of methods - serological agglutination and precipitation methods, immunoanalytical methods with indicators, immunochromatographic, electromigration and blotting techniques and cellular immunochemical methods.
After completing the course, the student is able to choose a suitable immunoanalytical method for examination of immunological parameters, including explanation of the principle of the method and evaluation of results.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Written examination
Attendance at seminars. written exam
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Recommended literature
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Bartůňková, Jiřina. Vyšetřovací metody v imunologii. Praha: Grada, 2011. ISBN 978-80-247-3533-7.
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Kopecký Otakar. Laboratorní vyšetření v klinické imunologii a alergologii. Hradec Králové. 2004.
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Slováková, Marcela. Podklady k seminářům.
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Slováková, Marcela. Vybraná laboratorní cvičení z imunoanalytických metod. Pardubice: Univerzita Pardubice, 2013. ISBN 978-80-7395-610-3.
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