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Basic Concepts in Clinical Biochemistry-A Practical Guide.7z

(Chris Devlin) #1

  • 1 Common Clinical Laboratory Hazards and Waste Disposal......

    • 1.1 Waste Disposal in Laboratory



  • 2 Blood Collection and Preservation.........................

    • 2.1 Blood Collection



  • 3 Quality Control in Laboratory............................

    • 3.1 Types of Laboratory Errors

    • 3.2 Methods to Minimize the Laboratory Errors..............



  • 4 Automation in Clinical Laboratory.........................

    • 4.1 Types of Autoanalyzers



  • 5 Photometry: Colorimeter and Spectrophotometer..............

    • 5.1 Colorimeter and Spectrophotometer



  • 6 Preparation of General Laboratory Solutions and Buffers.......

    • 6.1 Molar Solutions

      • 6.1.1 Molarity (M)..............................



    • 6.2 Normal Solutions.................................

      • 6.2.1 Normality (N).............................



    • 6.3 Percent (%) Solutions

    • 6.4 Buffer Solutions

    • 6.5 Physiological Buffers in the Human Body................

    • 6.6 Preparation of Common Laboratory Buffers..............

      • 6.6.1 0.2 M Acetate Buffer (pKa 4.86)

      • 6.6.2 0.2 M Sodium Phosphate Buffer (pKa 6.86)

      • 6.6.3 0.2 M Tris-HCl Buffer (pKa 8.1)................





  • 7 Examination of Urine for Normal Constituents................

    • 7.1 Preservatives Used for Urine Collection

    • 7.2 Physical Examination of Urine

      • 7.2.1 Color and Odor

      • 7.2.2 Appearance...............................

      • 7.2.3 Specific Gravity

      • 7.2.4 Volume

      • 7.2.5 pH......................................



    • 7.3 Chemical Examination of Urine

    • 7.4 Tests for Inorganic Constituents of Urine

    • 7.5 Tests for Organic Constituents

    • 7.6 Detection of Urea



  • 8 To Perform Qualitative Tests for Urinary Proteins.............

    • 8.1 Theory

    • 8.2 Tests for Urinary Proteins

      • 8.2.1 Dipstick Test

      • 8.2.2 The Boiling Test for Coagulable Proteins..........

      • 8.2.3 Sulphosalicylic Acid Test

      • 8.2.4 Nitric Acid Ring Test (Heller’sTest).............

      • 8.2.5 Bence-Jones Proteins........................



    • Using Biuret Reaction................................... 9 To Determine the Quantity of Proteins in Urine Sample

    • 9.1 Theory

    • 9.2 Specimen Requirements

    • 9.3 Principle

    • 9.4 Reagents........................................

    • 9.5 Calculations

    • 9.6 Clinical Significance

    • and to Find A/G Ratio................................... 10 To Estimate the Amount of Total Protein and Albumin in Serum

    • 10.1 Theory

    • 10.2 Principle........................................

    • 10.3 Specimen Requirements

    • 10.4 Reagent

    • 10.5 Procedure

    • 10.6 Calculations

    • 10.7 Estimation of Albumin in Serum

    • 10.8 Principle........................................

    • 10.9 Reagents........................................

    • 10.10 Procedure

    • 10.11 Calculations

    • 10.12 Clinical Significance

    • 10.13 Precautions



  • 11 To Perform Qualitative Test for Reducing Substances in Urine...

    • 11.1 Theory

    • 11.2 Qualitative Test for Reducing Sugars...................

      • 11.2.1 Benedict’sTest............................

      • 11.2.2 Benedict’s Qualitative Reagent

      • 11.2.3 Procedure



    • 11.3 CLINISTIX/Uristix................................

      • 11.3.1 Procedure



    • 11.4 Precautions

    • 11.5 Clinical Significance...............................



  • 12 Quantitative Analysis of Reducing Sugars in Urine.............

    • 12.1 Theory

    • 12.2 Specimen Requirement

    • 12.3 Principle........................................

    • 12.4 Reaction

    • 12.5 Reagents........................................

    • 12.6 Procedure

    • 12.7 Precautions

    • 12.8 Calculation

    • 12.9 Clinical Significance...............................



  • 13 Estimation of Blood Glucose Levels by Glucose Oxidase Method..

    • 13.1 Theory

    • 13.2 Principle........................................

    • 13.3 Specimen Requirements

    • 13.4 Reagents........................................

    • 13.5 Procedure

    • 13.6 Calculations

    • 13.7 Clinical Significance...............................

    • and Wu Method....................................... 14 To Determine the Blood Glucose Levels by Folin

    • 14.1 Principle........................................

    • 14.2 Reagents........................................

    • 14.3 Procedure

    • 14.4 Calculations



  • 15 To Perform Glucose Tolerance Test........................

    • 15.1 Method of Carrying GTT............................

    • 15.2 Specimen Requirements

    • 15.3 Principle........................................

    • 15.4 Reagents........................................

    • 15.5 Procedure

    • 15.6 Calculations

    • 15.7 Clinical Significance...............................



  • 16 Estimation of Urea in Serum and Urine.....................

    • 16.1 Theory

    • 16.2 Specimen Requirements

    • 16.3 Principle........................................

    • 16.4 Reagents........................................

    • 16.5 Procedure

    • 16.6 Calculations

    • 16.7 Clinical Significance...............................



  • 17 To Determine Urea Clearance.............................

    • 17.1 Theory

    • 17.2 Maximum Urea Clearance

    • 17.3 Standard Urea Clearance

    • 17.4 Procedure

    • 17.5 Precautions

    • 17.6 Clinical Significance...............................

    • by Jaffe’s Reaction..................................... 18 To Estimate Creatinine Level in Serum and Urine

    • 18.1 Theory

    • 18.2 Specimen Requirements

    • 18.3 Principle........................................

    • 18.4 Reagents........................................

    • 18.5 Preparation of Protein-Free Filtrate

    • 18.6 Procedure

    • 18.7 Calculations

    • 18.8 Clinical Significance...............................



  • 19 To Determine Creatinine Clearance........................

    • 19.1 Theory

    • 19.2 Specimen Requirement and Procedure

    • 19.3 Clinical Significance...............................



  • 20 To Determine the Uric Acid Concentration in Serum and Urine...

    • 20.1 Theory

    • 20.2 Specimen Requirements

    • 20.3 Principle........................................

    • 20.4 Reagents........................................

    • 20.5 Procedure

    • 20.6 Calculations

    • 20.7 Clinical Significance...............................

    • 20.8 Precautions



  • 21 Estimation of Total Calcium in Serum and Urine..............

    • 21.1 Theory

    • 21.2 Specimen Requirements

    • 21.3 Method

    • 21.4 Principle........................................

    • 21.5 Reagents........................................

    • 21.6 Procedure

    • 21.7 Calculations

    • 21.8 Clinical Significance...............................



  • 22 Estimation of Inorganic Phosphorus in Serum and Urine........

    • 22.1 Theory

    • 22.2 Specimen Requirements

    • 22.3 Methodology....................................

    • 22.4 Principle........................................

    • 22.5 Reagents........................................

    • 22.6 Preparation of Protein-Free Filtrates

    • 22.7 Procedure

    • 22.8 Calculations

    • 22.9 Clinical Significance...............................

    • 22.10 Precautions



  • 23 To Estimate the Amount of Total Cholesterol in Serum.........

    • 23.1 Theory

    • 23.2 Specimen Requirements

    • 23.3 Methodology....................................

    • 23.4 Principle........................................

    • 23.5 Reagents........................................

    • 23.6 Procedure

    • 23.7 Calculations

    • 23.8 Clinical Significance...............................



  • 24 To Estimate Total and Direct Bilirubin in Serum..............

    • 24.1 Theory

    • 24.2 Specimen Requirements

    • 24.3 Principle........................................

    • 24.4 Reagents........................................

    • 24.5 Procedure

    • 24.6 Calculations

    • 24.7 Clinical Significance...............................

    • Serum............................................... 25 To Determine Alanine and Aspartate Transaminase Activity in

    • 25.1 Theory

    • 25.2 Specimen Requirements

    • 25.3 Principle........................................

    • 25.4 Reagents........................................

    • 25.5 Procedure

    • 25.6 Calculation

    • 25.7 Clinical Significance...............................



  • 26 To Estimate the Activity of Alkaline Phosphatase in Serum......

    • 26.1 Theory

    • 26.2 Specimen Requirements

    • 26.3 Principle........................................

    • 26.4 Reagents........................................

    • 26.5 Procedure

    • 26.6 Calculations

    • 26.7 Clinical Significance...............................



  • 27 To Estimate the Activity of Acid Phosphatase in Serum.........

    • 27.1 Theory

    • 27.2 Specimen Requirements

    • 27.3 Principle........................................

    • 27.4 Reagents........................................

    • 27.5 Procedure

    • 27.6 Calculations

    • 27.7 Clinical Significance...............................



  • 28 To Determine Serum and Urinary Amylase Activity............

    • 28.1 Theory

    • 28.2 Specimen Requirements

    • 28.3 Principle........................................

    • 28.4 Reagents........................................

    • 28.5 Procedure

    • 28.6 Calculation

    • 28.7 Clinical Significance...............................



  • 29 To Estimate the Activity of Lipase in Serum..................

    • 29.1 Theory

    • 29.2 Specimen Requirements

    • 29.3 Principle........................................

    • 29.4 Reagents........................................

    • 29.5 Procedure

    • 29.6 Calculation

    • 29.7 Clinical Significance...............................



  • 30 Qualitative Analysis of Ketone Bodies in Urine................

    • 30.1 Theory

    • 30.2 Rothera’s Test for Acetoacetic Acid and Acetone

      • 30.2.1 Principle.................................

      • 30.2.2 Reagents

      • 30.2.3 Procedure

      • 30.2.4 Result



    • 30.3 Gerhardt’s FeCl 3 Test for Acetoacetic Acid

      • 30.3.1 Reagent

      • 30.3.2 Procedure



    • 30.4 Ketostix Test for Acetone and Acetoacetate

      • 30.4.1 Procedure



    • 30.5 Detection ofβ-Hydroxybutyrate

      • 30.5.1 Principle.................................

      • 30.5.2 Procedure



    • 30.6 Clinical Significance...............................



  • 31 Qualitative Test for Bile Pigments and Urobilinogen in Urine.....

    • 31.1 Theory

    • 31.2 Tests for Bile Pigments in Urine

      • 31.2.1 Fouchet’sTest.............................

      • 31.2.2 Hunter’s Test..............................

      • 31.2.3 Gmelin’s Test



    • 31.3 Urobilinogen in Urine

      • 31.3.1 Ehrlich’s Test..............................

      • 31.3.2 Saturated Sodium Acetate Solution



    • 31.4 Precautions

    • 31.5 Clinical Significance...............................

    • in Serum Sample....................................... 32 Determination of Total Lactate Dehydrogenase Activity

    • 32.1 Theory

    • 32.2 Specimen Requirements

    • 32.3 Principle........................................

    • 32.4 Reagents........................................

    • 32.5 Procedure

    • 32.6 Calculation

    • 32.7 Clinical Significance...............................



  • 33 To Measure Activity of Creatine Kinase in Serum.............

    • 33.1 Theory

    • 33.2 Sample Requirement...............................

    • 33.3 Principle........................................

    • 33.4 Enzyme Reagents

    • 33.5 Procedure

    • 33.6 Calculation

    • 33.7 Clinical Significance...............................



  • 34 Analysis of Cerebrospinal Fluid for Proteins and Sugars........

    • 34.1 Theory

    • 34.2 Analysis of Proteins in CSF

      • 34.2.1 Pyrogallol Dye-Binding Method

      • 34.2.2 Turbidimetry Method........................



    • 34.3 Analysis of CSF Glucose

    • 34.4 Clinical Significance...............................



  • 35 To Measure Lipid Profile in Serum Sample...................

    • 35.1 Sample Requirement...............................

    • 35.2 Total Cholesterol Estimation

      • 35.2.1 Principle.................................

      • 35.2.2 Reagents

      • 35.2.3 Procedure

      • 35.2.4 Calculations

      • 35.2.5 Precautions



    • 35.3 Triglycerides Estimation

      • 35.3.1 Principle.................................

      • 35.3.2 Reagents

      • 35.3.3 Procedure

      • 35.3.4 Precautions



    • 35.4 HDL Estimation

      • 35.4.1 Principle.................................

      • 35.4.2 Reagents

      • 35.4.3 Procedure

      • 35.4.4 Precautions



    • 35.5 LDL Estimation..................................

      • 35.5.1 Principle.................................

      • 35.5.2 Reagents

      • 35.5.3 Procedure



    • 35.6 Clinical Significance...............................

      • 35.6.1 Cholesterol Levels

      • 35.6.2 LDL Values

      • 35.6.3 HDL Values...............................

      • 35.6.4 Triglycerides..............................



    • Photometer........................................... 36 To Estimate Sodium and Potassium in Serum by Using Flame

    • 36.1 Theory

    • 36.2 Specimen Type, Collection, and Storage

    • 36.3 Principle........................................

    • 36.4 Reagents........................................

    • 36.5 Procedure

    • 36.6 Clinical Significance...............................



  • 37 To Perform Radioimmunoassay...........................

    • 37.1 Principle........................................

    • 37.2 Advantage

    • 37.3 Applications



  • 38 To Perform Enzyme-Linked Immunosorbent Assay............

    • 38.1 Principle........................................

    • 38.2 Sandwich ELISA

    • 38.3 Competitive ELISA................................

    • 38.4 Direct ELISA

    • 38.5 Applications of ELISA



  • 39 Some Important Case Studies.............................

    • 39.1 Case Studies of Sugar Impairment.....................

    • 39.2 Case Studies of Diabetic Ketoacidosis..................

    • 39.3 Case Studies of Calcium and Phosphate Impairments

    • 39.4 Case Studies of Protein Energy Malnutrition..............

    • 39.5 Case Studies of Gout/Uric Acid.......................

    • 39.6 Case Studies of Liver Functions.......................

    • 39.7 Case Studies of Kidney Functions

    • 39.8 Case Studies of Cardiac Functions



  • Suggested Readings........................................

  • Index...................................................

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