ANALYTICAL BIOCHEMISTRY

Chromatographic and Electrophoretic Techniques, both quantitative and qualitative applications. Various supporting media, for example, paper, membranes, gels, thin layer, ion exchange resins molecular sieves, disc, column, continuous flow, and others. Immunoelectrophoresis, conventional, medium and high voltage electrophoresis. Isoelectric focusing. Gas and high-pressure liquid chromatography. Sample preparation, dialysis, desalting, concentration, preparation of derivates. Capillary electrophorsis.


Isotope Techniques
Basic physics of stable and radio-isotopes. Counting techniques and their statistical appraisal. Units of radioactivity. Concepts of physical and biological half-life. Laboratory and patient hazards. Safety standards. Principles of radioactive detection and counting.
Legal requirements for storage and disposal and permission to administer to humans. Use in quantitive and qualitative analysis, for example, single and double isotope dilution techniques, autoradiography. Radio-ligand assay. Use in radioimmunoassay. Problems of purity of labelled compounds, storage and specific activity. Immunoradiometric assay. Labels. Functional Sensitivityof an assay. Interference in assays -e.g. heterophile antibodies.

Osmometry
Methods of measurement, for example, depression of freezing point.

Enzymology
Enzyme kinetics (Km) Fixed incubation and kinetic methods. Enzyme units. Enzyme standards; standardisation of methods. Enzyme immunochemistry. Enzyme multiplied immunoassay. Enzyme linked immunosorbent assay.

Mechanised
Techniques and Work Simplification Robotics Principles of continuous flow systems, single and multichannel. Discrete analysis systems, fast centrifugal analysers. Use of recording and digital output instruments.

Laboratory Data Processing and Computing
Use of computers for data collection, processing, and as management tools. Expert Systems

Analysis of Laboratory Error/Statistics
Concepts of reference range, analytical error, biological variation, and various simple parameters for describing data (mean, mode, standard deviation, confidence limits, variance, tests of significance). Applications to: assessment of inaccuracy and imprecision, errors of instruments, pipettes, and other equipment. Quality control methods.

Quantities and Units
SI Units -their advantages and disadvantages.

Specimen Collection, Preservation, and Preparation for Analysis
Constituent stability. Documentation and specimen flow systems. Interferences in the collection process.

CLINICAL BIOCHEMISTRY
The clinical biochemist should have an understanding of the major biochemical abnormalities found in disease and their methods of detection in the laboratory. They should have a general knowledge of the interpretative aspects of clinical biochemistry.
A Clinical Biochemist should understand the principles of testing the more important biochemical and physiological functions of organs or organ systems and be able to advise clinicians on their performance. They should take every opportunity to consult with clinicians to improve their understanding of the clinical manifestations of disease.

Water and Electrolytes
Distribution of water and electrolytes Measurement of plasma volume, total body water, sodium and potassium spaces Causes of hyper and hyponatraemia and hyper and hypokalaemia. Osmolality. Hyperosmolar coma Plasma and urine osmolality Shock. Metabolic effects of trauma Diagnosis and quantitative assessment of water and electrolyte loss Diuresis; pharmacological -osmotic Measurement of intracellular electrolytes Distinction between diabetes insipidus and compulsive water drinking Syndrome of inappropriate ADH.

Renal Function
Clearance tests: measurement of glomerular filtration rate and renal plasma flow. Tubular function tests: concentration tests, ammonium chloride loading tests, amino acid chromatography, and renal glycosuria. Normal and abnormal urine composition including abnormal pigments. Proteinuria, differential protein clearance. Renal failure. The nephrotic syndrome. Renal Calculi.

Diabetes Mellitus and Hypoglycaemia
Substrate, neural, and endocrine regulation of insulin and glucagon secretion -the gut as an endocrine organ. Diagnosis of diabetes mellitus and monitoring by glycated haemoglobin and urine albumin excretion. Somatostatin. C-peptide. Glucose tolerance, glycosuria. Plasma insulin and glucagon measurement. Differential diagnosis of coma: ketoacidosis, lactate acidosis, hyperosmolar coma, and hyperglycaemia. Diagnosis of insulinoma and other causes of hypoglycaemia; use and dangers of provocative tests, for example, tolbutamide and glucagon. Classification and diagnosis of DM Value and limitations of plasma insulin assays in hypoglycaemia and hyperglycaemia. Glycosylated haemoglobins and proteins.

Diagnostic Enzymology
Types of assays, enzyme determination in serum, urine, and cells. Stability of enzymes. Diagnostic use of enzymes. Use of isoenzymes.

Proteins: The Serum Proteins in Health and Disease
Interpretation of electrophoretic protein patterns, recognition of paraproteins and their further investigation by immunological techniques. Causes of hypoalbuminaemia. Assessment of protein-losing enteropathy and renal loss of protein.
Effects that malnutrition and malabsorption can have on protein state. Disorders of immunoglobulins and changes in specific proteins, for example, alpha 1 -antitrypsin, transferrin, and their use in diagnostics. Urine proteins, including Bence Jones protein.
Protein selectivity as an indication of renal damage. Hypothalmic releasing and inhibitory hormones. Steroid synthesis and metabolism. Congenital adrenal hyperplasia. Mechanisms of hormone action, receptors, cyclic AMP, and cyclic GMP. Transport of hormones Renin-angiotensin-aldosterone system Hormones of the gastro-intestinal tract

Nutrition
The digestion of proteins, carbohydrates, and lipids and the biological role of vitamins and the trace elements. The nutritional concept of protein quality and its assessment by measurement of biological value and nitrogen balance; also the importance of these factors in patients on synthetic diets. The laboratory methods of assessing vitamin status and measuring trace elements.

Inborn Errors of Metabolism
Possible defects in protein biosynthesis arising from genetic mutations.
Quantitative and qualitative enzyme abnormalities occurring in genetic disorders. The biochemical consequences of a primary enzyme block in a metabolic pathway and the ways in which clinical and pathological signs may be produced. Methods of detecting metabolic disorders, with particular consideration of screening selected clinical groups, for example, the mentally subnormal and the newborn. Evaluation of detection programmes. Antenatal diagnosis. Methods of treatment, particularly by dietary restrictions and vitamin supplementation, and the biochemical monitoring of the treatment. Consideration of the following conditions: Amino acid disorders, especially those involving phenyl alanine, tyrosine, methionine and homocystine metabolism, and the transport disorders, cystinuria and Hartnup disease. The organic acidaemias, particularly methylmalonic and propionic acidaemia. Glycogen storage disease, galactosaemia, and heritary fructose intolerance. The porphyrias. Enzyme deficiencies resulting in haemolysis, especially glucose-6-phosphate dehydrogenase and pyruvate kinase. Adrenogenital syndrome, fibrocystic disease. Wilsons disease. Mucopolysaccharide disorders, cerebral lipidoses, and metachromatic leukodystrophy. Disorders of purine metabolism, hyperuricaemia, gout and the Lesch-Nyhan syndrome. Cystic fibrosis.

Haem and Porphyrins
Structure and chemistry of porphyrins. Biosynthesis -delta-aminolaevulinic acid, ALA synthetase. Classification of porphyrias -hepatic, erythropoietic. Other causes of porphyria (lead poisoning, anaemia). Catabolism of haem, formation of bile pigments. Measurement of porphobilinogen, uro and coproporphyrins. Spectroscopy for haem pigments. Measurement of haemoglobin and detection of abnormal forms. Investigation of porphyria including appropriate specimen collection and preservation. Haemochromatosis.

Toxicology and Drugs
Detection and quantification of common drugs in therapy, for example, digoxin, lithium, and anticonvulsants. Overdosage, for example, salicylates, paracetamol, and barbiturates, and suspected addiction, for example alcohol, morphine, morphine derivatives, and amphetamines. Differential diagnosis of coma. Metabolic effects of ethanol. Environmental hazards, for example, lead, mercury. Cerebrospinal Fluid Glucose, differential proteins, enzymes. Oligoclonal bands. Tau-transferrin.

Amniotic Fluid
Bilirubin, creatinine content Presence and significance of alpha-fetoprotein. Lecithin/spingomyelin ratio, palmitate, and other tests of foetal lung maturity. Screening for Down syndrome.

The Biochemical Effects of Neoplasia
Effects of tumors, both anatomical and pathological. Tumor markers, their biochemical and pathological significance and their use in management of benign and malignant tumors. Some examples of this are: alpha fetoprotein, hCG, CEA, ectopic production of hormones and the syndromes these cause.

Molecular Biology
Principles of PCR, Northern, Southern and Western Blots. Testing for common diseases (HFE, CF)

LABORATORY MANAGEMENT
It is not expected that the candidate will have had the opportunity to become fully conversant with all details of laboratory management. The Clinical Biochemist should however, have a reasonable knowledge of the important aspects of the following: Organisation of a clinical biochemistry laboratory, including routine and emergency services. Screening and profiling.
Staff training, performance management, and work assignment.
Laboratory safety including chemical, radiation, physical and biological hazards.
Reagents and apparatus, their selection, sources of supply, and techniques for assessing the quality of equipment and reagents.
Budget preparation and monitoring.

SPECIAL PROJECTS
Short-term projects involving a considerable number of skills required of a laboratory supervisor (analytical, instrumental, evaluative, managerial, organisational) should be undertaken to encourage initiative and independence. The candidate should acquire an ability for clear report writing and should be encouraged to write assays on topics for discussion with his supervisor.

EDUCATIONAL ACTIVITIES
The candidate should plan his course of study in preparation for the Membership examination in consultation with their supervisor and the State Branch Education Representative.
The candidate should attend the following activities as an adjunct to higher own studies and practical experience.