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Lecturer(s)
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Slováková Marcela, doc. Mgr. Ph.D.
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Královec Karel, RNDr. Ph.D.
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Matuchová Inka, Ing. Ph.D.
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Morávek Ondřej, Mgr.
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Korecká Lucie, doc. RNDr. Ph.D.
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Course content
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Week 1: Overview of immunological and immunochemical investigation methods, quantification of components, functional tests, antigen properties. Serological methods agglutination, precipitation. Tasks: Rapid agglutination test for the determination of antibodies against Salmonella Typhimurium in human serum. Detection of rheumatoid factor in serum by the latex agglutination method. Detection of CRP in serum by the latex agglutination method. Week 2: Techniques based on the precipitation reaction, turbidimetry, nephelometry, immunodiffusion methods (double, radial), overview of determined analytes. Tasks: Microturbidimetric determination of serum immunoglobulins in PEG environment. Detection of CRP in human blood. Week 3: Cell isolation techniques, principles of cellular immunity examination (cell counts, functional tests). Task: Isolation of the lymphocyte fraction from blood. Week 4: Flow cytometry technique. Task: Preparation of whole blood sample for flow cytometry and analysis of blood cells using FSC/SSC histogram. 5th week: Functional tests of granulocytes, lymphocytes. Task: Monitoring the phagocytic abilities of leukocytes in vitro using Saccharomyces cerevisiae. Week 6: Overview and principles of complement testing. Task: Determination of CH50 in a human serum sample. Week 7: Assignment: Determination of antinuclear antibodies by ELISA method, evaluation of results in a clinical context. Excursion: Examination of tissue autoantibodies by indirect immunofluorescence. 8th week: Immunochromatographic tests. Examples of case studies with a proposal for an examination panel. 9th week: Ing. Matuchová, Ph.D.: Immunoblotting techniques for the detection of autoantibodies ANA, ENA, dg. myositis, rheumatoid arthritis, pANCA and cANCA antibodies, antiphospholipid, organ non-specific, specific; dg. liver disease, diabetes, neuropathy. Immunoblotting for detection of ANA antibodies in human serum. Week 10: Detection of monoclonal Ig and light chains in serum and urine. Examples of detection in serum by the immunofixation method. 11th week: Excursion to a medical facility (Interimun, Medila). Quality control processes, control materials and procedures.
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Learning activities and teaching methods
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Skills training, Laboratory work
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Learning outcomes
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The aim of the subject is to obtain an overview of all the most important methods that are commonly and newly used today in the field of medical immunology and immunohematology. The methods will be discussed comprehensively, from their principle, work procedures and necessary equipment to the evaluation of results and quality control.
After completing the course, the student is able to choose a suitable method for clinical immunological investigation, describe the principles of the methods and evaluate the results.
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Prerequisites
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The subject is closely related to theoretical teaching in immunology and immunohematology and to the student's laboratory skills, which he acquired in previous semesters in the laboratory subjects of biochemistry, hematology and molecular biological methods.
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Assessment methods and criteria
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Written examination, Home assignment evaluation
Laboratory assignments will take place at the department. Introductory seminars are required for several laboratory assignments, and for some only a seminar with demonstrations or case reports is planned, including those taught by external health professionals. The seminars will take place together for all groups of students. Written credit (50%) Laboratory protocols (50%)
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Recommended literature
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Slováková M. Pracovní návody a materiály ve Stagu.
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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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