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Clinical diagnosis

Clinical diagnosis. Dr. B.S. Vishwanath Professor, DOS in Biochemistry University of Mysore Mysore. Easy diagnosis Right treatment Increased life span Better quality life. Biased opinion Viewed as business rather than service Expensive Room for exploitation Results in legal issues.

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Clinical diagnosis

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  1. Clinical diagnosis Dr. B.S. Vishwanath Professor, DOS in Biochemistry University of Mysore Mysore.

  2. Easy diagnosis Right treatment Increased life span Better quality life Biased opinion Viewed as business rather than service Expensive Room for exploitation Results in legal issues Pros & cons of clinical diagnosis

  3. Visit to Diagnostic Center Infectious disease – Fever Vaccinations Bruises – Fractures (Painful) Inflammatory disorders Acute / Chronic Allergy Metabolic diseases – Lifestyle General Checkups

  4. Defense mechanism – Healing power • Bruises and Fractures – With time they heal • Infectious disease – Immune system • Stress – Hormone regulations Free radicals: Antioxidants – Enzymes (SOD etc) Current problem – We are in the fast world (notime) we need everything quickly

  5. Are investigations necessary!? • Without investigations the treatments are trial and error

  6. Simple investigations Fever – Likely cause is infection Virus or bacteria TC – DC from whole blood sample TC - Total blood cells DC – Lymphocytes (T and B lymphocytes) Neutrophiles Eosinophiles Monocytes Basophiles Platelets

  7. Blood cell counter

  8. Purpose of cell count • To differentiate bacterial and viral infection • Increased lymphocyte – Viral infection • Increased neutrophiles – Bacterial infection • Decreased platelet – Dengue fever

  9. Tips - Fever • Never allow body temperature to reach above 104 f • By applying cold cloth on forehead region • Para acetamol (tablets/injectables)

  10. Dengue fever • Total cell count – Less • Platelets – Significantly less • Hemorrhagic spots • Platelets supplement • Treatment - Glucocorticosteroids

  11. Lung infection • Presence of Sputum • Colour (white-yellow-green) • Tuberculosis • Any other bacterial infections

  12. Tuberculosis - AFB • Presence of bacteria in the sputum – Confirmatory test

  13. Wound infection • Treatment – Antibiotic • Culture and Sensitivity

  14. Culture and Sensitivity • Antibiotic discs (10 – 12) Resistant - R Sensitivity: + ++ +++ ++++

  15. Alimentary Canal • Gastritis • Ulcers • Barium meal X-ray • Endoscopy • Biopsy/Lavage

  16. Robin Warren and Barry Marshall Nobel prize 2005 Helicobacter pylori

  17. Metabolic disorders • Carbohydrate metabolism – Water soluble, Easily execrated. • Lipid metabolism – Not execrated instead accumulates in the body

  18. Glucose metabolism • Glucose is the immediate energy source • Its metabolism is regulated by many factors Insulin Glucagon Glucose transporters Impaired metabolism – Diabetes mellitus

  19. Chronic diabetic complications • Retinopathy • Nephropathy • Neuropathy

  20. Metabolic syndromes - Diabetes mellitus IDDM Type – I, NIDDM Type - II • FBS & PPBS • Urine sugars (FUS & PPUS) • GTT • Glycosylated hemoglobin • Microalbuminuria

  21. Diabetes

  22. Normal values • FBS: 70 – 120 mg% • PPBS: 120 – 180 mg% • Glycoslylated hemoglobin (HbA1c) > 5% • Microalbuminuria < 300 mg (0-25 mg) • Insulin & C-peptide

  23. Glycoslylated hemoglobin (HbA1c) > 5%

  24. Lipid Metabolism • Hypertension – High Blood pressure • Cardiovascular disease – Heart attack.

  25. Lipids Triglycerides Cholesterol Lipid transfer proteins Chylomicrons HDL LDL VLDL Diagnostic tests

  26. Lipid transfer proteins

  27. Good and Bad cholesterol

  28. Lipid profile

  29. Lipid transfer protein • HDL (30 – 75 mg%) • LDL (up to 150 mg%) • VLDL (up to 40 mg%) • Chylomicrons • Ratio of HDL/LDL gives the risk factor associated with heart. (< 4.5)

  30. Methods HDL: Chylomicrons, LDL and VLDL: PPtation with phosphotungstic acid and MgCl2 Derived values: VLDL: TG/5 LDL: Chol-HDL-VLDL

  31. Liver • Site of detoxification – Toxicity, liver cirrhosis (alcohol) • Hepatitis infection & Jaundice.

  32. Diagnostic methods • Total protein Albumin / Globulin & A:G ratio • Enzyme analysis SGOT, SGPT, LDH, GGT & Alk.phostase • Bilirubin: Total and Direct.

  33. Hepatitis B virus

  34. Jaundice • Obstructive Jaundice • Hepatic Jaundice • Pre-hepatic Jaundice Total protein Albumin / Globulin & A:G ratio Enzyme analysis SGOT, SGPT, LDH, GGT & Alk.phostase Bilirubin: Total and Direct.

  35. Structure of a Nephron

  36. Kidney • Blood Urea: (14 – 40 mg%) • Serum Creatinine: (0.8 – 1.4 mg%)

  37. Heart • Blood pressure • ECG • Echocardiogram • TMT Biochemical: CK, CKMB & LDH1

  38. ECG TMT

  39. Echo cardiogram

  40. Enzyme analysis • Creatine Kinase : BB, MM, MB • CKMB – Specific to Heart muscle • LDH: LDH1-5 (LDH1 is specific to Heart muscle.

  41. Biochemical methods Phenol + H2O2 + 4-aminoantipyrine = Red quinone Glucose: GOD-POD TG: Lipase, glycerokinase, G3Poxidase. Cholesterol: Cho.esterase, Cho.oxidase

  42. Immunotechniques • RIA - ELISA • Direct, Sandwich and competitive ELISA • Precipitation reaction - Immunodiffusion • Western blot

  43. Immunodiagnosis • ELISA – HIV, HbsAg • Westernblot for HIV • TORCH - IgM & IgG • Competitive ELISA for Hormone analysis • Agglutination test • Immunodiffusion

  44. Structure of an antibody Primary antibody Secondary antibody

  45. Direct ELISA for HIV Antigen coating Blocking Primary Ab Washing Secondary Ab Washing Substrate addition Colour development

  46. Monoclonal antibody binding with antigen

  47. Sandwich ELISA for HBsAg

  48. Western blot • ELISA is similar to Western blot but is more informative. • HIV detection: ELISA and Western blot • HIV - I and HIV - II

  49. Western blot

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