CLINICAL AND BIOCHEMICAL ANALYSIS
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- Versione italiana
- Academic year
- 2022/2023
- Teacher
- ILARIA LAMPRONTI
- Credits
- 6
- Didactic period
- Primo Semestre
- SSD
- BIO/10
Training objectives
- The objective of the “Biochemical and clinical analysis” part is analyze the principal biochemical technologies and several modern diagnostic methodologies, with reports about available techniques for the bio-molecules study. The course is based on the examination of different biochemical and bimolecular methodologies (chromatography, electrophoresis, spectroscopy, etc) with particular attention to real and clinical examples.
Prerequisites
- Basic knowledge of Biology, General and Organic Chemistry, Biochemistry and Molecular biology.
Course programme
- The course is divided in two parts of 6 CFU each, corresponding to 40 hours of lectures and 12 hours of exercitations and seminars.
Topics of the lessons are:
Introduction (2 hours): Lab safety: safety for patients, visitors and personnel. Analytical variability. Qualitative, semi-quantitative and quantitative kind of analysis. Test reliability and errors. Detailed description of the course program.
Emocromo cytometric analysis (2 hours): Hematopoiesis and blood cells. The blood sampling. Cell counts and analysis (multiparameter counters), Hct value, Hb evaluation, MCV value (flow cytometry), blood film.
Cytofluorimetric assays (2 hours): Description of the dedicated instrumentation; cell cycle and apoptosis analysis.
Electrophoresis (8 hours): General principles. Agarose gel, cellogel, polyacrylamide gel, 2D gel, capillary electrophoresis, northern blotting, southern blotting, western blotting. Examples and biomedical relevance: Hb analysis, serum proteins dosage, new markers, proteomic maps, DNA analysis.
Chromatography (6 hours): General principles. Classic liquid chromatography (normal and inverse phase). Retention time and chromatographic resolution. Adsorption, dimensional exclusion, ionic exchange and affinity chromatography. HPLC (High Performance Liquid Chromatography): pumps, injectors, columns, detectors. Clinical applications: nucleic acid and protein separation, analysis of plasma-proteins, monitoring of normal and pathological hemoglobins.
Spettroscopy (2 hours): Spectrophotometry: absorbance, Lambert-Beer equation. Applications: nucleic acid and protein concentration, Hb blood concentration, use of spectrophotometer in ELISA and in enzyme-colorimetric reactions.
Mass spectrometry (2 hours): Function and characteristics of mass spectrometers. Applications: differential expression and identification of proteins (proteome study; biomarker's research) in the proteome analysis.
Immunochemistry (6 hours): Antibodies and antigens (structure and function). Radio Immuno Assay (RIA), Enzyme Linked Immuno Sorbent Assay (ELISA and BIOPLEX). Clinical applications: tumor and inflammation biomarkers, cardio-vascular markers, diagnosis of diabetes mellitus type 1 and celiac disease, fast commercial assays (pregnancy test).
Molecular biology techniques (10 hours): Polymerase Chain Reaction (PCR), DNA sequencing. Clinical applications: diagnosis of genetic diseases and cancer; non invasive pre-natal diagnosis.
Exercitations and seminars (12 hours): In lab will be performed 1) Electrophoretic analysis (agarose gel and DNA or RNA separation); 2) culture and treatment of human erythroleukemic K562 cells; 3) study of cell viability and apoptosis in immortalized human cells.
The following seminars will be also organized: "The importance of pre-analytics in the lab analysis", "COVID clinical assays: from molecular biology to new serological tests", "Possible interferences in analytical results", held by experts in the sector, to illustrate their work by reporting experience and practical examples. Didactic methods
- The course is organized in lessons on all the course’s topics. Moreover, lab exercitations and seminars will be organized.
Learning assessment procedures
- Oral examination. The aim of the oral examination is to test knowledge and deepening of the topics of the course program, and the reasoning skills developed by the student on issues related to biochemistry and applied biochemistry. The student will be asked three questions on three different topics; each answer will be evaluated with a score ranging from 1 to 10, the individual votes will then be added to obtain the final score expressed in thirtieths.
Reference texts
- Scheme and Figures provided by the docent.
Books:
“Biochimica applicata” di Stoppini e Bellotti, EdiSES
“Metodologie biochimiche e biomolecolari” di Maccarone, Zanichelli
“Metodologie biochimiche” di Bonaccorsi di Patti, Casa Editrice Ambrosiana
