Sickle Cell Procedures and Interpretation

Preparation of working sickle cell buffer

Add the entire contents of one vial of Sickle Cell Lysing Reagent to 50ml of Sickle Cell Buffer and mix completely by swirling. The working buffer may be used for 30 days from date of preparation if stored at 2° to 8° C.

Assay Protocol

1. Transfer 2.0ml of the Working Sickle Cell Buffer to a 12 x 75 mm tube.
   
2. Add 20μl of well-mixed whole blood or control specimen and mix thoroughly and gently. If testing a specimen with a hemoglobin below 7 g/dl, add 40 μl of whole blood.
   
3. Allow to stand for 5 minutes but no longer than 30 minutes at room temperature (15° to 30° C).
   
4. Read results by looking through the test tube at the Line Scale (see below). Hold the test tube upright and perpendicular approximately 3 cm from the Line Scale. Adequate illumination is necessary.
   

Interpretation

• Reactive - If any sickling hemoglobin is present, the solution will be sufficiently turbid to prevent reading the Line Scale through the test tube.
  
• Nonreactive - If a sickling hemoglobin is not present, the solution will be clear enough to allow the Line Scale to be seen through the test tube.