Hemoglobin A1c (HbA1c) Test Kit (Homogeneous Chemiluminescence Immunoassay)

【Packing Specification】

25 servings/?box

【Intended Use】

In vitro test for the quantitative determination of mmol/mol hemoglobin (IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood or hemolysate. HbA1c determinations are useful monitoring of long-term blood glucose control in individuals with diabetes mellitus. Moreover, this test is to be used as an aid in diagnosis of diabetes and identifying patients who may be at risk for developing diabetes.

HbA1c is formed in two steps by the non-enzymatic reaction of glucose with the N-terminal amino group of the β-chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.

In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte's life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.

【Principle】

Sandwich principle.

The Hemoglobin A1c (HbA1c)?test kit employs a wash-free and homogeneous strategy based on proximity hybridization-regulated CRET(Chemiluminescence resonance energy transfer). The AE demonstrated strong chemiluminescence (CL) in the presence of H2O2. Graphene oxide (GO) as an excellent quencher was used to produce the “Signal off” mode that little CL emission was observed through CRET between GO and the AE-labelled DNA3. Once the antigen was introduced, the target-induced proximity hybridization occurred to form a proximate complex, which inhibited the CRET by preventing GO from absorbing AE-labelled DNA3. The CL intensity increased with the increasing HbA1c?concentration.

【Main Components】

Lyophilized spheres reagents: DNA1-labeled HbA1c antibody 1 (mouse monoclonal antibody) (about 20ng/human), DNA2-labeled HbA1c antibody 2 (mouse monoclonal antibody) (about 10ng/human), DNA3-labeled acridine ester (AE), endonuclease, graphene oxide (GO) conjugated antioxidant (AOD), preservative (P300) (0.1%).

Buffer: phosphate buffer (10 mmol/L), preservative (0.1%) and other components; packaging specifications correspond to the quantity.

Lysis solution: composed of sodium hydroxide (40mmol/L), sodium chloride (0.45%), preservative (0.1%), etc.; packaging specifications correspond to the quantity.

Calibration card: containing standard curve for product calibration.

Control Level 1: composed of HbA1c, Haemoglobin and Sucrose. Range of target value: 4.1-6.1%

Control Level 2: composed of HbA1c, Haemoglobin and Sucrose. Range of target value: 12.4-14.0%

※The components in the kits of different batches are not interchangeable.

2?Reaction cups

2?Buffer?solution

2?Substrate solution

2?Pipette

2?Disposable tip

【Storage Conditions and Expiration Date】

Stable for 18?months when stored at 2-8℃?away from light. Reagents are used immediately upon opening.

【Applicable Instruments】

Chemiluminescence immunoassay analyzer (HSCL-5000)?produced by Nanjing Poclight?Biotechnology Co.?Ltd.

【Sample Requirements】

1. Whole blood is recommended to be anticoagulated with heparin or EDTA, and the test must be completed within 4 hours after sample collection; if it cannot be used immediately it is recommended to be stored at 2-8℃ and the test completed within 24 hours, and freezing of whole blood samples is prohibited.

2. Used for human whole blood samples, other body fluids and samples may not get accurate results.

3. The sample must be returned to room temperature (15-30℃) before testing, and the sample should be gently mixed before use.

4. Heat inactivated samples, hemolysis and other abnormal samples should be discarded.

【Test Method】

The test method is homogeneous chemiluminescence?immuoassay

1. Preparation before testing.

1.1 Please read this manual and the instrument manual carefully before use.

1.2 Check whether the instrument can work normally, whether the reaction cup or sample tube, substrate solution and buffer are prepared adequately, and prepare other related consumables.

2. Test procedure

2.1 Turn on the chemiluminescence?immunoassay analyzer.

2.2 Check whether the calibration card is consistent with the lot number of the kit and scan the calibration card.

2.3 Take 200μL of buffer into the reaction cup and mix well

2.4 Take 5μL of the sample to be tested into the lysate and mix upside down, then take 20μL of the mix into the reaction cup, put it into the instrument and start incubation, measurement, processing data and deriving the assay results.

2.5 Total assay time is 5 minutes.

3.Result analysis.

The measured chemiluminescence intensity is converted from the standard curve to obtain the concentration of HbA1c in the unknown sample.

4. Calibration

Each kit batch contains its specific calibration information, which is stored on a calibration card.

【Reference Interval】

The following reference range was determined by statistical analysis of the 95% distribution range of HbA1c levels in a sample of 200 healthy individuals.

Normal reference value: 4.27-6.07%

Clinical diagnostic threshold (Cut-off value): HbA1c ≥ 6.5% as a diagnostic indicator of diabetes

Note: Due to differences in geography, ethnicity, gender and age, it is recommended that each laboratory establish its own reference range.

【Interpretation of Test Results】

1. When the test result of the sample is greater than 18.5%, the instrument shows >18.5%, and when the test value of the sample is less than 3.8%, the instrument shows <3.8%.

2. The test results are for clinical reference only and should be considered in conjunction with clinical symptoms, medical history, other laboratory tests and other circumstances, and are not used as a basis for clinical diagnosis.

3. If the test result is abnormal or differs from the clinical diagnosis, verification is recommended.

Product Performance Index】

1. Appearance: complete components, lyophilized balls in complete form.

2. Accuracy: relative deviation not more than 8%.

3. Blanking limit: not more than3.8%

4. Linearity: 3.8%-18.5%, within this linear range, the linear correlation coefficient r should be not less than 0.990.

5. Repeatability: CV≤5%.

6. Inter-batch variation: CV≤8%.

【Caution】

1. This product is a single-use in vitro diagnostic reagent, please do not reuse it, and do not use the expired product.

2. The reagent can only be used on the supporting instrument analysis system.

3. The test procedure must be followed as indicated, improper operation will lead to wrong results.

4. Components of kits with different lot numbers cannot be mixed.

5. The calibration card used for calibration should not be exchanged with kits of different lot numbers.

6. Do not swallow the reagents or contact with skin, eyes or mucous membranes as the reagents contain preservatives; once contacted, rinse the contaminated area with water.

7. When using the instrument for testing, each sample and the corresponding reagents should be replaced with a new disposable tip to avoid cross-contamination, which may affect the accurate determination of HbA1c?concentration in the sample, and all samples should be aspirated directly to the bottom of the sample tube.

8. The use of fresh samples is recommended. All samples from patients should be treated as potential sources of infection and all waste generated during the test must be disposed of in accordance with local regulations.

9. If the reagent has exceeded its expiration date, it should not be used again.

10. This kit contains animal-derived components that are potentially infectious.

【Production Date and Expiration Date】

See labels for details