Clinical Chemistry

1.Introduction to the Laboratory: definition of the pre-analytical phase, blood draw and not acceptability criterion of the sample; type of samples; anticoagulants; treatment, transportation, centrifugation and storage of samples.

2. Instrumental analytical techniques: photometric characteristics of light, interaction between matter and radiant energy, Lambert – Beer law, absorbance and calibration curve, zeroing against blank; general principles of photometry and spectrophotometry; single beam spectrophotometer, double beam spectrophotometer; turbidimetry; nephelometry.

3. Clinical applications: spectrophotometry with direct methods, spectrophotometry with indirect methods; colorimetric reactions; enzymatic reactions, primary reactions and couple reactions, NAD and NADP dependent enzymatic reactions; enzymatic- colorimetric reactions, Trinder end point method.

4 Indirect potentiometry: principle of this method.

5. Methods of determination of important tests about clinical relevance (SGOT, SGPT, gGT, PCE and dibucaine number, LDH, Total and Direct Bilirubin, CPK, Alkaline Phosphatase, Amylase and Pancreatic Amylase, Lipase, Urea and BUN, Creatinine, Uric Acid, calculation of clearance endogenous, Iron, Ferritin, Glucose, total Cholesterol, HDL-C and LDL-C, Triglyceride, APO A B, Sodium, Potassium, Chloride, Calcium, phosphorous, Magnesium): Methods, interpretation of clinical data and interferences.

6. Quality assessment: precision and accuracy of the analytical date; specificity and sensitivity of the method; types of error; calibration: definition of detection limit and levels detectable; internal quality assessment, external quality assessment; dilutions.

Electrophoresis

• Electrophoresis and specific proteins, biological samples  and their collections. General characteristics of proteins, overview of their structure and function, methods for their assay• automatic equipments and their functioning. Electrophoretic separation of  serum proteins•  Electrophoretic standard profile: protein fractions (albumin-α1-α2-β1-β2-γ globulins) •  specific proteins of the electrophoretic pattern.  Gammapathies, Electrophoretic Immunofixation  (EIF): principles and methods• Interpretation of the results, evaluation of normal and pathological patterns. Urinary electrophoresis. Search for Bence Jones protein • Methodology • Interpretatione of the results • Immunosubtraction and interpretation of the results. Definition of electrophoresis, electrophoretic techniques: principles, equipments and accessories (power supply,  electrophoretic chamber, buffer, dyes, densitometer) • Protein migration and conditioning factors (voltage,  frictional force,  migration rate, electrophoretic mobility; sample;  buffer solution; ionic strength;  Joule effect; electroendosmosis) • implementation methods: free or front phase; on sifting and non sifting support or zonal • zonal electrophoresis of serum proteins on paper,  cellulose acetate, agarose gel and in capillary • Methodology • Interpretation of migration and of graphics in capillary •evaluation of normal and pathological patterns• Comparison between methods.