HEMOGLOBIN A1C-DIRECT (HbA1C-DIR)

COD 13047 1 x 60 mL

For in vitro use in the clinical laboratory only

HEMOGLOBIN A1C-DIRECT (HbA1C-DIR)
DIRECT

INTENDED USE METROLOGICAL CHARACTERISTICS

Reagents for the measurement of hemoglobin A1C (HbA1C) concentration in human blood. The obtained values The metrological characteristics described below have been obtained using an A25 analyzer. Results are similar
are useful as an aid in the diagnosis and monitoring of diabetes mellitus. with A15.
These reagents are for use in the BioSystems A25 and A15 analyzer or in other analyzer with similar − Detection limit: 6 mmol/mol.
performance characteristics. The reagents may also be used by a manual procedure. − Measurement interval: (approximate value dependent on the highest standard concentration): 6 - 140
mmol/mol.
CLINICAL SIGNIFICANCE
− Precision:
HbA1C is the product of the irreversible condensation of glucose with the N-terminal residue of the -chain of Mean concentration Repeatability (CV) Within-laboratory (CV)
hemoglobin A.
The HbA1C concentration in blood is directly proportional to the mean concentration of glucose prevailing in the 37 mmol/mol 1.8 % 3.1 %
previous 6-8 weeks, equivalent to the lifetime of the erythrocytes 1, and the estimated average glucose (eAG) 78 mmol/mol 1.6 % 3.0 %
during this period can be calculated with the formulas below2.
eAG (mg/dL) = 28.7 x HbA1C-NGSP-DCCT (%) – 46.7 − Trueness: Results obtained with this reagent did not show systematic differences when compared with
eAG (mmol/L) = 1.59 x HbA1C-NGSP-DCCT (%) – 2.59 reference reagents. Details of the comparison experiments are available on request.
eAG (mg/dL) = 2.64 x HbA1C-IFCC (mmol/mol) + 15.0 LIMITATIONS OF THE PROCEDURE
eAG (mmol/L) = 0.146 x HbA1C-IFCC (mmol/mol) + 0.843 − Interferences: Bilirubin (up to 10 mg/dL) and Lipemia (triglycerides up to 400 mg/dL) do not interfere. Other
HbA1C levels are a valuable adjunct to glucose determinations in the assessment and follow up of individuals drugs and substances may interfere6.
with diabetes mellitus, providing much more reliable information for glycaemia monitoring than do determinations In the immunoassay methods, the presence of acetylated-Hb, carbamylated-Hb, labile HbA1C HbE and HbD
of glucose. Numerous studies have shown that diabetes related complications may be reduced by the long term do not affect the results7,8. Other hemoglobin variants like HbS, HbF or HbC can interfere 7.
monitoring and tight control of blood glucose levels. The HbA1C concentration may also be a useful tool in the In hemolytic anemia, iron deficiency anemia and transfusion, the average age of erythrocytes is altered.
diagnosis of diabetes3. Caution should be used when interpreting the HbA1C results from patients with these conditions.
Clinical diagnosis should not be made on the findings of a single test result, but should integrate both clinical and NOTES
laboratory data.
1. To avoid possible interferences by other assays performing HbA1C determinations in different runs of the
PRINCIPLE OF THE METHOD other test is recommended.
After preparing the hemolysate, the Hemoglobin A1C (HbA1C) concentration is quantified by a latex turbidimetric BIBLIOGRAPHY
assay. The different hemoglobins present in the hemolysate are unspecifically adsorbed on the latex particles 1. Tietz Textbook of Clinical Chemistry and Molecular Diagnostics, 6th ed. Rifai N, Horvath AR, Wittwer CT.
surface in a ratio equivalent to their concentration in the sample. The addition of an anti-human HbA1C antibody
WB Saunders Co, 2018.
causes agglutination that is proportional to the concentration of hemoglobin A1C and can be measured by
turbidimetry. 2. Nathan DM, et al. Translating the A1C assay into estimated average glucose values. Diabetes Care 2008;
31: 1473-1478.
CONTENTS AND COMPOSITION 3. Nathan DM, et al. International Expert Committee report on the role of the HbA1C assay in the diagnosis of
A. Reagent. 1 x 50 mL. Suspension of latex particles, sodium azide 0.95 g/L, pH 8.0. diabetes. Diabetes care 2009: 32: 1327-1334.
B. Reagent. 1 x 10 mL. Anti-human HbA1C antibody, stabilizers, pH 6.0. 4. Hoezel W et al. IFCC reference systems for measurement of hemoglobin A1C in human blood and the
national standarization schemes in the United States, Japan and Sweden: a method-comparison study. Clin
STORAGE AND STABILITY Chem 2004;50:166-174.
Store at 2-8ºC. 5. American Diabetes Association Professional Practice Committee. 2. Classification and Diagnosis of
Components are stable once opened until the expiry date marked in the label if they are stored well closed and Diabetes: Standards of Medical Care in Diabetes-2022. Diabetes Care. 2022 Jan 1;45(Suppl 1):S17-S38.
care is taken to prevent contamination during their use. doi: 10.2337/dc22-S002. PMID: 34964875.
On board stability: Reagents open and kept in the refrigerated compartment of the analyzer are stable 30 days. 6. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.
Indications of deterioration: Absorbance of the blank over the limit indicated in “Test Parameters”. 7. Bry L, Chen P, Sacks D. Effects of hemoglobin variants and chemically modified derivates on assays for
glycohemoglobin. Clin Chem 2001;41:153-163.
ADDITIONAL MATERIALS REQUIRED (NOT PROVIDED) 8. Little R.R., Rohlfing C; Hanson S, Connolly S, Higgings T, Weykamp C, D’Costa M, Luzzi V, Owen W,
S. HbA1C Direct Standards (Cod. 31048). 4 levels for 0.5 mL Human blood. HbA1C concentration is given on Roberts WL. Effects of Hemoglobin (Hb) E and HbD traits on measurements of Glycated Hb (HbA1C) by 23
the label. methods. Clin Chem 2008, 54: 1277-1282.
Human blood used in the preparation of the standard has been tested and found to be negative for the TEST PARAMETERS (Notes 1 & 2)
presence of antibodies anti-HIV and anti-HCV, as well as for HBs antigen. However, the standard should be
These reagents may be used in several automatic analyzers. Specific instructions for application in many of
handled cautiously as potentially infectious.
them are available on request.
Reconstitute with 0.5 mL of distilled water. Stable for 30 days at 2-8°C R1: Use Reagent A, R2: Use Reagent B.
REAGENT PREPARATION A25 A15
Reagents are provided ready to use.
GENERAL
SAMPLES Name HbA1C-DIR HbA1C-DIR
Venous blood collected by standard procedures and with EDTA as anticoagulant. Sample type WBL WBL
HbA1C in blood is stable 7 days at 2-8ºC.
Analysis mode fixed-time bir. fixed-time bir.
PROCEDURE Units mmol/mol mmol/mol
Hemolysate preparation Turbidimetry test yes yes
The calibrators should be treated as patient samples.
Decimals 0 0
1. Pipette into a test tube:
Type of reaction increasing increasing
Blood 50 L
Distilled water 5.0 mL PROCEDURE
Reading mode monochromatic monochromatic
2. Mix gently. Avoid the formation of foam. Incubate at room temperature for 5 minutes.
Main filter 670 670
The hemolysate is stable 72 hours at 2-8ºC.
Test according to Test Parameters. Reference filter - -
Sample 3 3
CALCULATION
The HbA1C values obtained are traceable to IFCC Reference Method. Vol. R1 190 190
The traceable values to Reference Method as described by the US National Glycohemoglobin Standardization Vol. R2 40 40
Program (NGSP) are calculated using the following general formula 4. Washing 1.2 1.2
%HbA1C-NGSP-DCCT (%) = 0.0915 x HbA1C-IFCC (mmol/mol) + 2.15 Reading 1 (cycle) 10 7
CALIBRATION Reading 2 (cycle) 30 20
A reagent blank should be done every day and a calibration at least every 30 days, after reagent lot change or Reagent 2 (cycle) 9 6
as required by quality control procedures. Predilution factor - -
QUALITY CONTROL
CALIBRATION AND BLANK
It is recommended to use the Hemoglobin A1C Controls, Normal (cod. 18001) and Elevated (cod. 18002) to
verify the accuracy of the measurement procedure. Calibration type specific specific
Each laboratory should establish its own internal Quality Control scheme and procedures for corrective action if Number of calibrators 4 4
control results are not within the acceptable limits. Calibration curve increasing poligonal increasing poligonal
REFERENCE VALUES
OPTIONS
The following cut-off points have been established and adopted by many countries for a reference population
(Non diabetic) and for the evaluation of the degree blood glucose control in diabetic patients 1,5. Blank absorbance limit 0.700 0.700
Kinetic blank limit - -
IFCC mmol/mol) NGSP-DCCT (%) Reference values/Degree of control Linearity limit - -
Substrate depletion - -
20 - 38 4.0 - 5.6 Non Diabetic
39 - 47 5.7 - 6.4 Pre-diabetic
> 48 > 6.5 Diabetic
< 53 < 7.0 Treatment Goal

This range is given for orientation only; each laboratory should establish its own reference range.

M13047i-06 06/2023
The lateral lines mark the modifications in the current version.