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Analytical Methods of Diagnosing Diabetes Mellitus

Analytical Methods usually use in detecting Diabetic Patiet

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Analytical Methods of Diagnosing Diabetes Mellitus

  1. 1. Analytical Methods of Determining Diabetes Mellitus HANISHA ERICA P. VILLAESTER
  2. 2. A. Blood Test  Sample Collection  Venipuncture  Capillary Blood by Skin Puncture  Done when venipuncture cannot be performed or when reducing frequency of needle sticks is desired and less painful
  3. 3. A.1 FASTING BLOOD GLUCOSE TEST  Person to be tested should be on normal diet for at least 3 days prior to testing  Fasting for 8 – 12 hours  Plasma or serum is used instead of whole blood  More readily measured on automated equipment  Glucose concentration is 10 – 15% higher than in whole blood
  4. 4. A.1 FASTING BLOOD GLUCOSE FASTING BLOOD GLUCOSE < 110 mg/dL Normal 110 – 126 mg/dL Pre-Diabetes > 126 mg/dL Diabetes
  5. 5. A.2.a ORAL GLUCOSE TOLERANCE TEST  Fasting for 8 – 12 hours  Patients requested to drink anhydrous glucose dissolved in 250 – 300mL water  Adults: 75g anhydrous glucose  Children: 1.75g/kg of body weight anhydrous glucose  Blood tested regularly in 30 minutes interval  In conditions of insulin defiency, blood glucose levels got elevated due to impaired utilization of glucose  Required for confirmation  Glucose Tolerance Curve is then plotted
  9. 9. A.2.b Intravenous Glucose Tolerance Test  Undertaken for patients with malabsorption  Patient is given 25g of glucose dissolved in 100mL distilled water as intravenous injection within 5 minutes  Blood samples drawn at 10 minutes interval for the next 2 hours  Interpretation:  Normal individuals, blood glucose level returns to normal within 60 minutes  Diabetes mellitus – decline is slow
  10. 10. A.2.b Intravenous Glucose Tolerance Test  Factors Affecting Glucose Tolerance Test  Acute Infection  Liver Disease  Hyperthyroidism  Hypothyroidism  Starvation
  11. 11. A.2.c Mini or Modern GTT  Two samples are collected  Fasting (zero hour)  2 Hour Post Glucose Load Zero Hour After 2 Hours Normal Person < 110 mg/dL < 140 mg/dL Diabetic Person > 126 mg/dL > 200 mg/dL Increase Glucose Tolerance 110 – 126 mg/dL 140 – 199 mg/dL
  12. 12. A.3 Post Prandial Blood Sugar  Patient advised to have normal meal  Blood collected 2 hours after meal Glucose Level Normal < 140 mg/dL Pre-diabetic 140 – 200 mg/dL Diabetic > 200 mg/dL
  13. 13. A.4 Random Blood Glucose  Only required during emergency  Diabetis Mellitus: > 200 mg/dL
  14. 14. A.5 HbA1c  Test that measures amount of glycated haemoglobin in your blood  Glycated haemoglobin is a substance in red blood cell that is formed when blood sugar attaches to haemoglobin  Generally reflect the state of glycemia over the preceding 8 – 12 weeks thereby providing an improved method of assessing diabetic control
  15. 15. Analytical Methods of Determining Diabetes Mellitus KEZIA AIJELETH SHAHAR CABANGAL
  16. 16. Self monitoring blood glucose (SMBG) People who take insulin should regularly self monitor blood glucose. For people with non-insulin treated type 2 diabetes testing is most useful if patients use the results to learn and alter behaviour, or medication. “...SMBG is most useful if patients use the results to learn, as part of an overall diabetes education package….”
  17. 17. Laboratory tests to prevent and delay complications of diabetes People with diabetes usually die from macrovascular complications of their diabetes; namely cardiovascular disease. This is influenced by all of the commonly recognised risk factors for cardiovascular disease as well as glycaemic control. Fasting lipid levels are measured three monthly until stable and then 6 - 12 monthly thereafter. It is important that management should be individualised Parameter Optimal value Total cholesterol < 4 mmol/L LDL cholesterol < 2.5 mmol/L HDL cholesterol > 1 mmol/L TC:HDL ratio < 4.5 Triglycerides < 1.7 mmol/L HbA1C < 7 mmol/L
  18. 18. Diabetic renal disease The best way of testing for diabetic renal disease is by urinary albumin:creatinine ratio (ACR) and serum creatinine with estimated glomerular filtration rate (eGFR). These tests are performed on everyone with diabetes at diagnosis and repeated at least annually – more frequently if there is proteinuria, microalbuminuria or reduced eGFR. Albumin:creatinine ratio  ACR provides an estimate of daily urinary albumin excretion.  Microalbuminuria cannot be detected on a conventional urinary protein dip stick.  Microalbuminuria is urinary albumin excretion between 30 and 300 mg/day; above 300mg/day represents proteinuria.  ACR is best measured in the laboratory using a first morning urine sample where possible when the patient is well.  An abnormal initial test requires confirmation by testing on two further occasions. If at least one of these tests is positive microalbuminuria has been confirmed.
  19. 19. Renal testing in diabetes *Non-diabetic renal disease is suspected when there is absence of diabetic retinopathy in a person with renal disease, there are urinary abnormalities such as haematuria or casts, or when there is renal disease without microalbuminuria or proteinuria. ACR mg/mmol (confirmed) eGFR mL/min/1. 732 Risk Management men < 2.5 women < 3.5 and > 60 2 - 4% per year progress to microalbuminuria. Annual ACR and eGFR. Good diabetes & BP management. men ≥ 2.5 women ≥ 3.5 or < 60 One third progress to overt nephropathy. CVD risk doubled. Review ACR and eGFR at each visit. Intensive management of glycaemia and CVD risk factors. Use ACE inhibitor and low-dose aspirin. Avoid nephrotoxic drugs. Investigate if suspicious of causes other than diabetes* > 30 or < 30 Almost all proceed to end stage renal disease or die prematurely of CVD. Overt nephropathy Refer specialist
  20. 20. Other tests Testing of LFTs is recommended for people with diabetes:  at diagnosis,  at the start of antidiabetic drug therapy, and  at any other time indicated by clinical judgement Other laboratory tests In patients with type 1 diabetes, intermittent checks for other autoimmune conditions may be useful. This could include testing for thyroid dysfunction or coeliac disease.
  21. 21. 22Urinary Glucose  -Glucose can be detected in urine using the specific test strips that contain glucose  oxidase, peroxidase, and a chromagen.  -Other carbohydrates using Benedict's and Febling's reagents.
  22. 22. 23Urinary Ketones  -Acetone and acetoacetic acid can be detected in urine using the AcetesTM or  KetostixTM systems.  -These tablets or strips use nitroprusside (sodium nitroferricyanide) to detect ketones.
  23. 23. 24Urinary Ketones  -Because beta-hydroxybutyric acid lacks a ketone group is not detected by this assay.  -Quantitative assays for acetoacetate and beta-hydroxybutyric acid are available using beta-hydroxybutyrate dehydrogenase and either NADH or NAD.
  24. 24. 25Urinary Ketones  -If NAD is used as the cofactor and the reaction is buffered at around pH 9.0, beta-hydroxyburyric acid is measured.  -On the other hand, a separate reaction using NADH and buffered around pH 7.0 would measure acetoacetic acid.
  25. 25. 26Microalbuminuria  -Diabetes mellitus causes progressive changes to the kidneys and ultimately results in diabetic renal nephropathy.  -This complication progresses over a period of years and may be delayed by aggressive glycaemic control.  -An early sign that nephropathy is occurring is an increase in urinary albumin.
  26. 26. 27Microalbuminuria  -Microalbumin measurements are useful to assist in diagnosis at an early stage and  prior to the development of proteinuria.  -Microalbumin concentrations are between 20 to 300 mg/d.  -Proteinuria is typically greater than 0.5 g/d.
  27. 27. 28Proteinuria in Diabetes  - Many people excrete small quantities of protein in urine, typically around 10  mg/day of mainly low molecular weight proteins such as albumin.  -Some diabetic patients develop albumin excretion rates 30 µg/min this range  classed as microalbuminuria.
  28. 28. 29 METHODS FOR THE DETERMINATION OF GLUCOSE  The most used  methods of glucose analysis employ the enzymes glucose oxidase or hexokinas.  A) Glucose Oxidase  B) Hexokinase
  29. 29. 30 SELECTED METHODS FOR THE MEASUREMENTS OF GLYCATED HAEMOGLOBINS InterferenceMeasurementMethod Carbamyl Hb HbF Temperature- sensitive HbA1cCation exchange HbA1cMonoclonal antibody Glycated HbAffinity chromatography Phenyl boronate matrix Latex agglutination Fluorescence quenching
  30. 30. ReferencesColagiuri, S., et al. (2003). Comparability of venous and capillary glucose measurements in blood. Diabetic Medicine, 20,953-956. Franciosi, M., et al. (2001) The impact of blood glucose self-monitoring on metabolic control and quality of life in type 2 diabetic patients. Diabetic Care, 24,1870-77. Harris, H. (2005). Elevated liver function tests in type 2 diabetes. Clinical Diabetes, 23, 115-119. Kenealy, T et al. (2004). Diabetic care by general practitioners in South Auckland: changes from 1990 to 1999. The New Zealand Medical Journal,115,219. Kjos, S., Buchanan, T. (1999). Gestational Diabetes Mellitus. New England Journal of Medicine, 341, 1749-1756. NZGG: New Zealand Guidelines Group. (2003). Management of Type 2 Diabetes. Available at: (accessed March 2006) NZGG: New Zealand Guidelines Group. (2003). Assessment and Management of Cardiovascular Risk. December 2003. Available at: (accessed March 2006). Schwedes, U., Siebolds, M., and Mertes, G. (2002). Meal-related structured self-monitoring of blood glucose. Diabetes Care, 25,1928-32.

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    Nov. 21, 2018

Analytical Methods usually use in detecting Diabetic Patiet


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