HbA1c liquidirect Pipetting Scheme

Mix [RGT1] and [RGT2] gently before use to suspend the latex particles completely.
Immunoassay for the Direct Photometric Pipette into cuvettes Reagent blank (RB) [CAL1-4]/ sample /
[CBN/A]
Determination of Glycohemoglobin A1c [RGT1] 750 μl 750 μl
Package size Hemolysate(Sample/[CAL1-4]/ ---- 20 μl
[CBN/A])
[REF] 10770 40 ml Test Kit
[IVD] Mix and incubate for 5 min. at 37°C.
[RGT2] 250 μl 250 μl
Clinical Significance
Mix, incubate minimum 5 min. at 37°C. Read absorbance of the sample (Asample),
The determination of HbA1c is performed for the long term control in diabetes [CBN/A] (A[CBN/A])and of [CAL1-4] (A[CAL1-4]) against the reagent blank (ARB) within
mellitus. HbA1c values provide an indication of the average glucose levels over the 30 min; use exactly the same incubation time for [CAL1-4] and sample.
preceding 4-8 weeks. A high HbA1c value indicates poor glycemic control. Long term Keep ratio [RGT1] : sample : [RGT2] constant if total volume should be reduced.
therapy of the disease requires control of blood glucose levels to prevent the acute
Calculation
complications of ketosis and hyperglycemia. In addition, effective control of blood
glucose levels can help minimize long term complications such as retinopathy, Calculate the absorbance difference (A [CAL1-4] = A [CAL1-4] - ARB) of each calibrator and
plot the values (y-axis) against the respective concentrations (x-axis). The HbA1c
neuropathy, and cardiovascular disease.
concentration in the sample and control is calculated from the multipoint-calibration
Intended Use curve.
Immunoassay for the direct photometric determination of HbA1c in human whole Values of the HbA1c liquidirect Calibrator Set are traceable to the IFCC HbA1c system.
blood with EDTA. HbA1c liquidirect must be used in conjunction with HbA1c Calculating the results in NGSP units follows the formula:
Calibrator Set, [REF] 10776. NGSP HbA1c [%] = HbA1c [mmol/mol Hb] *0.0915 + 2.15
Method Performance Characteristics
This method utilises the linkage of antigen and antibody to directly determine the Linearity: The measuring range is defined by the concentration range of the
concentration of HbA1c in whole blood. Both total Hb and HbA1c have the same calibrators and may encompass 2.2 - 15.0 % HbA1c or 1 – 145 mmol/mol Hb.
unspecific absorption rate to latex particles. Monoclonal anti-human HbA1c (mouse) Analytical sensitivity: approximately 2.2 % HbA1c or 1 mmol/mol Hb.
are cross-linked by anti-mouse IgG (goat), and react specifically with HbA1c, resulting Typical performance data can be found in the Verification Report, accessible via:
in an agglutination of the latex particles. The degree of agglutination depends on the www.human.de/data/gb/vr/su-hba1c-2.pdf or
amount of bound HbA1c. The increase of turbidity in the reaction mixture is www.human-de.com/data/gb/vr/su-hba1c-2.pdf
measured photometrically. Recommended Values
Non diabetic: < 6 % HbA1c or < 42 mmol/mol Hb
Contents
Glycemic control of a person with diabetes:
[REF] 10770 < 7 % HbA1c or < 53 mmol/mol Hb
[RGT1] 1 x 30 ml Reagent 1 Each laboratory should establish its own recommended values. In using HbA1c to
Suspended latex material < 0.13 % monitor diabetic patients, results should be interpreted individually. That is, the
Sodium azide < 0.09 % patient should be monitored against him or herself.
[RGT2] 1 x 10 ml Reagent 2 Quality Control
anti-human HbA1c (mouse, monoclonal) 0.05 mg/ml We recommend using our HbA1c Control Blood normal/abnormal, [REF] 10775, or
anti-mouse IgG (goat, polyclonal) 0.08 mg/dl other control bloods with HbA1c values determined by this method. If controls do not
TRIS buffer 0.21 % fall into the assayed range patient values from that run should not be reported.
Gentamicin sulphate 0.015 % Repeat the run, making sure that all instructions are strictly followed.
Sodium azide 0.09 % Automation
[LYS] 2 x 100 ml Hemolysis reagent Proposals to apply the reagents on analyzers are available on request. Each laboratory
Lysing reagent has to validate the application in its own responsibility.
Sodium azide 0.05 %
Notes
Additional material not supplied with the kit 1. There is a potential risk of reduced values of HbA1c in combination with the
[REF] 10776 washing solutions designated for a particular analyzer. This interference is
[CAL1-4] 4 x for 0.5 ml HbA1c Calibrator Set basically caused by the detergent Tween or Triton X, which might be included in
relatively high concentration in washing additives of other manufacturers. Please
Human blood (lyophilised)
check your wash solution additives.
[REF] 10775 2. It has been reported that results may be inconsistent in patients who have the
[CBN] 2 x for 0.5 ml HbA1c Control Blood normal following conditions: opiate addition, lead-poisoning, alcoholism, intake of large
[CBA] 2 x for 0.5 ml HbA1c Control Blood abnormal doses of aspirin.
Human blood (lyophilised) 3. It has been reported that elevated levels of HbF may lead to underestimation of
HbA1c.
Mandatory material but not supplied with the kit
[REF] 18971 Safety Notes
[WAD] 4 x 25 ml WASH ADDITIVE · Precautionary statements
The usage of HUMAN WASH ADDITIVE ([WAD], [REF] 18971) is mandatory as [RGT1] [RGT2] [LYS]
system wash solution additive for HumaStar Automated Analyzers and P234 Keep only in original container.
Systems as it has been shown not to cause the interference described in Note. P260 Do not breathe dust/fume/gas/mist/vapours/spray.
P262 Do not get in eyes, on skin, or on clothing.
Reagent Preparation and Stability P281 Use personal protective equipment as required.
[RGT1], [RGT2] and [LYS] are ready to use. Store reagents at 2...8°C. All reagents are P303+P361+P353 IF ON SKIN (or hair): Take off immediately all contaminated
stable to the expiration date stated on the labels. clothing. Rinse skin with water/shower.
[RGT1], [RGT2], and [LYS] are stable for 4 weeks after opening when stored at 2...8°C P305+P351+P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove
and tightly closed. contact lenses, if present and easy to do. Continue rinsing.
Specimen Collection and Preparation P337+P313 If eye irritation persists: Get medical advice/attention.
Special preparation of the patient or fasting is unnecessary. Collect venous blood with P401 Store in accordance with local/regional/national/international regulations.
EDTA using aseptic technique. P501 Dispose of contents/container in accordance with local / regional / national /
international regulations.
Stability: 1 week at 2...8°C.
To determine HbA1c, a hemolysate must be prepared for each sample. References
1. Thomas L., HbA1c, Labor & Diagnose, 8 ed., 236-242 (2012)
Hemolysis 2. Nathan D.M. et al., Effect of Aspirin, Clin. Chem. 29, 466-469 (1983)
Pipette into tubes 3. Engbaek F. et al., Analytical characteristics for patients with/without uremia, Clin.
[LYS] 1000 μl Chem. 35, 93-97 (1989)
Sample/[CAL1-4]/[CBN/A] 20 μl 4. Little R.R. et al., HbA1c NGSP standardisation, Clin.Chem. 47, 1985-1992 (2001)
5. Mongia S.K. et al., Effects of Hemoglobin C and S Traits, Am.J.Clin.Pathol. 130, 136-
Mix, allow to stand for 5 min. or until complete hemolysis is evident. 140 (2008)
Use lysates for the assay. Stability: 10 days at 2...8°C 6. Little, R.R. et al., A Review of Variant Hemoglobins Interfering with HbA1c, J.
Diabetes Sci Technol 3 (3), 446-451 (2009)
Assay
7. Little R.R. et al., Effect of Hb E and HbD, Clin.Chem. 54, 1277-1282 (2008)
Wavelength: 600 - 660 nm 8. Ceriello A. et al., Depressed ATIII biological activity in opiate addicts, Diabetologia
Optical path: 1 cm 22, 379 (1982)
Temperature: 37°C
Measurement: against reagent blank (RB), only one reagent blank per series is
required.
SU-HBA1C-2 INF 1077001-2 GB 04-2018-11 |

Human Gesellschaft für Biochemica und Diagnostica mbH