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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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1. Laboratory safety. Pipetting using micropipets. 2. Rapid agglutination test for detection of antibodies against Salmonella typhimurium in human serum. Detection of rheumathoid factor (RF) in serum by reverse passive latex agglutination method. 3. Precipitation methods - Quantification of IgG by microturbidimetry. Detection of C reactive protein by immune-diffusion method (Ouchterlony). 4. Quantification of proteins by single radial immune-diffusion method (Mancini). 5. Examination of complement activation by classical way in gel (CH100) and in solution (CH50). 6. Hemolytic tests - determination of antistreptolysin (ASLO) antibody activity in patient serum. 7. Detection and quantification of serum specific IgG antibodies against cows' milk proteins by ELISA. 8. Heterogeneous non-competitive ELISA for CEA quantification. 9. Immunochromatographic test for qualitative detection of cotinine in human urine. Detection of Streptococcus A from swab tonsil. Detection of hCG from human urine. 10.-11. Detection of autoantibodies (ANA) by dot blot. 12. Laboratory detection of monoclonal immunoglobulins, free light chains, 2-microglobulin in serum and urine by immunofixation method. 13. Analysis of human blood sample on flow cytometer.
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Learning activities and teaching methods
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Laboratory work
- Participation in classes
- 39 hours per semester
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Learning outcomes
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The aim of this subject focused on laboratory work is to introduce immunoanalytical methods to students, its principle and practical applications. Mentioned methods include serological agglutination and precipitation methods, haemolytic methods, enzyme-linked immunoassays, immunochromatographic, electromigration and blotting techniques. Also, cellular immunochemical methods will be presented.
After completing all laboratories is student able to choose a suitable immunoanalytical method for the analysis of biological samples depending on the requirement of detection or quantification, including the results evaluation.
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Prerequisites
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unspecified
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Assessment methods and criteria
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Student performance assessment
Participation in laboratory exercises. Test.
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Recommended literature
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Kopecký Otakar. Laboratorní vyšetření v klinické imunologii a alergologii. Hradec Králové, 2004.
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Slováková M, Bílková Z, Korecká L. Vybraná laboratorní cvičení z imunoanalytických metod. Pardubice, 2013. ISBN 978-80-7395-610-3.
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