Atlas sickle kit is a qualitative solubility test for Sickle Haemoglobin. The test can be performed in two ways:
1. A screening test to detect sickle haemoglobin (HbS)
2. A centrifugation test to differentiate the sickle cell trait (AS) from sickle cell anaemia (SS).
Sickle cell disease (also called sickle cell anemia) is an inherited blood disorder that affects red blood cells. The sickle cell gene causes the body to produce abnormal hemoglobin. In sickle cell disease, the hemoglobin clumps together, causing red blood cells to become stiff and develop a C-shaped (“sickle”) form. These sickled red blood cells can block blood vessels, reducing blood flow in many parts of the body. This process results in tissue and organ damage.
Each red blood cell contains about 280 million hemoglobin molecules. Hemoglobin is the most important component of red blood cells. It is composed of protein (globulin) and a molecule (heme), which binds to iron.
In the lungs, the heme component takes up oxygen and releases carbon dioxide. The red blood cells carry the oxygen to the body’s tissues, where the hemoglobin releases the oxygen in exchange for carbon dioxide, and the cycle repeats. The oxygen is essential for all cells in the body to function. Sickle cell disease reduces or denies adequate oxygen to many parts of the body. This contributes to the severe pain experienced as a sickle cell crisis and both short and long-term organ damage.
- Reagent A (Buffered Saponin)
- Reagent B (Reducing agent, Sodium Dithionite).
Preparation of the working solution
Bring 1 bottle of Reagent ‘A’ and 1 vial Reagent ‘B’ up to Room Temperature. Add Reagent ‘B’ to Reagent ‘A’ and mix well for 5minutes. Rec ord date on bottle.
1. Using the solution prepared as above, place 2 ml quantities into the required number of 75 x 12mm tubes.
2. Using whole anti coagulated blood (EDTA), add (20 μl) to each tube. Mix well and stand for 3 – 5 minutes. Hold against viewer or 1cm away for best results.
3. Always use known positive and negative controls.
- Negative: Clear haemolysed solution.
- Positive: Turbid red solution, partially or completely obscuring the lines on the viewer.
- Centrifugation: Positive results should be centrifuged for 5minutes at 1000RCF.
- HETEROZYGOUS: red-pink supernatant with a dark red band at the top.
- HOMOZYGOUS: Yellowish supernatant with a dark red band at the top.
- NEGATIVE: Slight greyish matter on top of deep red haemolysate.
1. The working solution should be kept refrigerated and will remain stable for up to 2 weeks. Allow the reagent to reach to room temperature before use.
2. ANAEMIC SAMPLES Adjust haematocrit to approximately 50% by removal of plasma. Do not add double volume of sample.
3. False Positives may be caused by abnormal plasma protein or when patients are receiving parental nutrition.
4. False Negatives may be found if old or outdated reagents are used, or the blood of small children under the age of 6 months if the proportion of HbS is less than 20%, or following Blood transfusion in severe anaemia.
Storage and stability
- The reagents should be stored refrigerated between2-8°C.
- Never Freeze or expose to elevated temperature.
- The reagent is stable until the expiry date stated on the product label. Do not use the reagents past the expiry date
Precautions and limitations
- The reagents are intended for in vitro diagnostic useonly.
- Do not use reagents if it is turbid or contain particles as this may indicate reagent deterioration or contamination.
- Protective clothing should be worn when handling the reagents.
- Don’t use these reagents if the label is not available or damaged.
- Don’t use the kit if damaged or the glass vials are broken or leaking and discard the contents immediately.
- Test materials and samples should be discarded properly in a biohazard container.
- Wash hands and the test table top with water and soap once the testing is done.
- Heamolysed blood sample should not be used for testing.
- The test should be performed at room temperature in a well let area with very good visibility.
- Failure to follow the procedure in this package insert may give false results or safety hazard.
- Close the vial tightly after each test.
- The reagent is considered toxic, so don’t drink or eat beside it.