Hemoglobin A1c (HbA1C) Turbidimetric Immunoassay kit
HbA1c is the term known as glycated hemoglobin. Throughout the whole life cycle of red blood cell, HbA1c is formed continuously by the adduction of glucose to the N-terminal of the hemoglobin beta chain. Formation of the sugar-Hb linkage indicates the presence of excessive sugar in the bloodstream, often indicative of diabetes. Therefore, measurement of %HbA1c can also be used to determine three-month average blood sugar level since the average lifespan of a red blood cell is four months. HbA1c level in diabetic subjects will be elevated by 2-3 fold over the levels found in normal individuals. The cut-off value for %HbA1c is 6.5%. The higher the %HbA1c, the higher chance the diabetic subjects are going to develop complications.
HbA1c PETIA kit developed by IMD can accurately measure %HbA1c in human whole blood sample.
PRINCIPLE OF THE ASSAY
This assay is a turbidimetric immunoassay for the quantitative measurement of %HbA1c in human whole blood. A standard or sample is added into a cuvette and mixed with the reaction buffer R1. Total hemoglobin and HbA1c have the same unspecific absorption rate to latex particles. After a short incubation, the test reagent R2 is added into the cuvette and mixed. Latex-HbA1c-mouse anti-human HbA1c antibody complex is formed. Agglutination is formed when goat anti-mouse IgG polyclonal antibody interacts with the monoclonal antibody. The extent to which the microparticles aggregate is quantified by the amount of light scattering measured as absorbance by a chemistry analyzer. The %HbA1c in unknown samples can be interpolated from a reference curve using the standards provided.
Assay procedures may vary depending on the automated chemistry analyzer to be used. A general example of assay procedures is stated as follow:
- Dispense 150µl of R1 into a clean cuvette
- Add 4µl of sample and incubate at 37˚C for 5 minutes
- Further add 50µl of R2
- Read change of absorbance at 660 nm for 5 - 8 minutes after the addition of R2
- Calculate the %HbA1c in unknown sample by interpolation from a reference curve using the standards provided
Caution: Different lot of R1, R2, standard and control should not be mixed for use
R1: 15ml, R2: 5ml
R1: 30ml, R2: 10ml
R1: 60ml, R2: 20ml
R1: 150ml, R2: 50ml
R1: 300ml, R2: 100ml
OTHER MATERIALS REQUIRED
Product Catalog Number Format Method Calibrator 51Ac1-S1 4 x 0.5 ml Control 51Ac1-C1 2 x 0.5 ml