Biochemical Tests in Diabetes: DR Joe Fleming PHD MCB Frcpath Dept of Clinical Biochemistry CMC Vellore
Biochemical Tests in Diabetes: DR Joe Fleming PHD MCB Frcpath Dept of Clinical Biochemistry CMC Vellore
Biochemical Tests in Diabetes: DR Joe Fleming PHD MCB Frcpath Dept of Clinical Biochemistry CMC Vellore
UK Consensus Statement
Glycemic control is best measured by HbA1c
The method should be a DCCT –aligned HBA1c method
The assay should have acceptable within assay precision
<3% and between assay imprecision <5%
CMC METHOD BIORAD VARIANT HbA1c
PROGRAM
Utilizes the principles of ion-exchange HPLC , without
interference from labile A1c, lipaemia or temperature
fluctuations.
Certification/traceability of reference material
Certified by the NGSP as having documented traceability to
the DCCT reference method. The haemoglobin A1c
calibrators provided in the kit are traceable to the Kyoto
2002 Calibrator set prepared by the IFCC working
group on standardization of HbA1c. The specimens
were prepared in the Netherlands at a hospital with ISO
9001:2000 certificate.
NGSP = 0.906(IFCC) + 2.21.
This method reports performance data and
reference ranges as NGSP values. The
calibrator/diluent set includes both NGSP and
IFCC values.
Refs:
Trinder P Ann Clin Biochem 1969, 6: 24-27
Barham D, Trinder P. Analyst 1972; 97: 142-145.
Higher concentrations of bilirubin interfere in the
peroxidase part of the assay causing a decrease in values
So do uric acid, ascorbate, haemoglobin, tetracycline,
glutathione.
Hexokinase assay
Uses hexokinase and G6PDH enzymes, ATP and NADP+
cofactors
Haemolysis 0.5 g/dL,
lipaemia > 500 mg/dL, positive effect
bilirubin > 5 mg/dL negative effect
Reference Values
ADA 2 fasting plasma values ≥ 126 mg/dL (7.0 mmol/L)
Impaired fasting glucose 101- 124 mg/dL (5.6-6.9 mmol/L)
Glucose AC fasting 70-110 mg/dL
Glucose PC (2 hours) 80-140 mg/dL
Glucose random 70-140mg/dL
Semi-quantitative measurement of urine glucose
Benedicts test based on reduction of copper ions by
glucose to give green to brick red colour. Protein free
urine
All other urine reducing substances interfere.
Analytical sensitivity 250 mg/dl
Dip-stix method GOD/POD
Analytical sensitivity 100 mg/dL
Ketones, ascorbic acid, salicylates false negative
Bleach false positive
ESTIMATION OF SERUM CREATININE
Specimen type, collection and storage
Serum or plasma can be analysed and can be stored at 40C, for 24
hrs.
Collect 24 hr urine in a plastic container with thymol as a
preservative. Stable at 40C for 24 hr.
Centrifuge all urines before analysis.
Principle of the method
NaOH
Creatinine + picric acid -------------- Creatinine picramate (red
colour) at 505 nm
Summary:
We require better standardization and improved accuracy
(trueness) of serum creatinine including the use of the
estimating equation for GFR from the Modification of Diet
in Renal Disease Study (MDRD). The current variability in
SCr estimation renders all equations for GFR less accurate in
the normal and slightly increased range < 1.5 mg/dL (<133
mol/L) which is the relevant range for detecting chronic
kidney disease (CKD). Defined as GFR < 60 ml.min-1
(1.73m2)-1.
SCr should be reported in mg/dL to 2 decimal places ie 0.92
mg/dL not 0.90, mol/L will still be reported to whole
numbers.
Reference
Izawa S, Okada M, Matsui H, and Horita Y. J Med and Pharm Sci
1997; 37: 1385-88
Target Values
< 100 mg/dL (2.59 mmol/L) therapy target in
diabetic patients
<130 mg/dL diabetics
<160 mg/dL non diabetics
160 – 189 mg/dL high
> 190 very high
NATIONAL CHOLESTEROL EDUCATION PROGRAM
(NCEP SEPT 2002)
LDL-C the primary determinant in hypercholesterolaemia
Estimated by a direct LDL-C method. Friedewald formula
cannot provide values with the recommended precision and
accuracy limits
ie total error =/< 12%, accuracy ± 4%
CV =/< 4%
LDL-C value for calibration and QC material traceable
to the reference method for LDL-C
Friedewald formula overestimates LDL in the presence
of Type II hyperlioproteinaemia (increased -VLDL)
Estimation of serum triglycerides
Principle of the method
Triglycerides glycerol using the enzyme
lipoprotein lipase
Glycerol glycerol –3-phosphate using
glycerol kinase
Glycerol-3-phosphate dihydroxyacetone
phosphate + H2O2 using glycerolphosphate oxidase
Reporting of results
Reference Range 45—190 mg/dL