Learning objectives
This course aims to provide the informations and the scientific background to approach correctly the practical aspects in Clinical Biochemistry by students of Techniques of biomedical laboratory.
This knowledge is especially needed to understand the complexity connected to the biological sample and to recognize the possible sources of error and to undertake the suitable remedy; moreover, the knowledge of general principles and the methodological steps by which an analytical method is fitted and carried on it provides the cultural background to comprehend the Diagnostic techniques of Clinical Biochemistry.
Prerequisites
There are no obligatory prerequisites, but these topics of Clinical Biochemistry are an interdisciplinary matter and the student needs the basic knowledge in Biology,Chemistry and Biochemistry
Course unit content
- Kind of laboratory test: purposes and usefulness.
- Biological specimens consideration: instruction for optimal collection, treatment and conservation (avoiding pre-analytical variation). Principles of exclusion for collected biological samples.
- Blood, plasma or serum. Coagulation; anticoagulants. Centrifugation - Sedimentation theory. Centrifugal force and centrifugation times. Types of centrifugal separations: differential centrifugation, grade centrifugation, rate zonal centrifugation, isopicnic centrifugation. Centrifuge. Centrifuge rotors
- Blood: composition and homeostasis. More common chemical analysis: Quantitative and qualitative measurement of blood proteins. UV-VIS absorption spectroscopy - Properties of electromagnetic radiation. Interaction electromagnetic radiation-matter. Chromophores and transitions. Absorption intensity.Spectrophotometers.
Separation and quantification of blood proteins: Gel-electrophoresis; Capillary electrophoresis.
Immunoasays. Antigen and antibody. Protein labeling.Principle of the immuno assays.
Detection of radioactivity, flurescence and chemiluminescence. Turbidimetric and nephelometric assays.
- Oxygen transport and utilization. Outlines on bioenergetics. Normal and pathological hemoglobins: detection and quantification.
Chromatography - Retention in chromatography. Distribution coefficient. Classification of chromatographic techniques.Column efficiency. Selectivity and resolution. Separation modes: absorption chromatography, partition chromatography, bonded phases: reverse-phase, ion-exchange, ion-pair, size exclusion and affinity chromatography. High performance liquid chromatography (HPLC): pumps, valve injection, columns, detectors.
- Enzymes and vitamins. Catalysis. Enzyme Kinetic. Control of enzyme reaction. Enzyme assays. Isoenzymes. Principles of isoenzyme detection and measure.
- Analytical and post-analytical variability affecting the laboratory data. Analytical and clinical evaluation of methods. Receiver Operating characteristic (ROC) curves. Reliability on data analysis: quality assurance
- Blood glucose homeostasis. Outlines on digestion, absorption and metabolism of carbohydrates. Bioenergetics. Hormone regulation of carbohydrates homestasis.
More common chemical analysis: blood glucose assays. Sample. Spectrophotometry. Lambert-Beer law and quantittive analysis.
- Blood lipids. Outlines on digestion, absorption and metabolism of the main lipids. Chetogenesis. Lipoproteins and atherosclerosis.
Analytical interferences of lipids. Methods of lipid assay. Ultracentrifugation.
- Blood nitrogen. Outlines on digestion, absorption and metabolism of proteins and aminoacids.
- Electrolyte profile. Outlines on water, electrolyte and acid-base metabolism. Principles of electrochemistry and chemical sensors. Emogasanalysis.
- Proteins of extracellular matrix. Outlines on mineral and bone metabolism. Bone remodeling and fibrotic processes.
- Outlines on porphyrin mtabolism.
- Reactive oxygen and nitrogen species. Cycle of exose monophosphate in the red blood cell.
- Catabolism of purines.
- Urina analysis. Reflectance photometry.
- Reference values. the concept of normality in biology. Decision values.
- Point of care testing (POCT): use and application. Criteria for test waiving. Quality control in POCT. Implementation and monitoring of POCT. Specimen and their treatment for POCT.Current waived tests and kits. Auto-monitoring of some biochemical parameters.
Bibliography
J.W.Baynes, M.H.Dominiczak. Biochimica per le discipline biomediche. Casa Editrice Ambrosiana
L. Spandrio.Biochimica Clinica.Ed. Sorbona ¿ Milano.
Teaching methods
The final oral examination is about the topics of the course; the student needs to demonstrate to have understood and to be able to manage the essential meanings of every section of the program.
