Screening method for erythrocyte osmotic fragility and application of the same
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A technology of osmotic fragility and screening methods, applied in the field of medical testing, can solve the problems of expensive operation and complexity of equipment, achieve good repeatability, fast and accurate experiments, and reduce the screening workload
Inactive Publication Date: 2017-10-24
杭州金域医学检验所有限公司
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[0005] The purpose of the present invention is to provide a method for detection and screening of erythrocyte osmotic fragility, which solves the defects that the existing erythrocyte osmotic fragility test needs expensive equipment or complex operation, and needs manual conversion of optical density values into detection results
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Embodiment 1
[0062] Experimental operation method:
[0063] 1. Take a 48-well ELISA plate, add 0.5ml of erythrocyte osmotic fragility test solution, and preheat at 37°C for 1 minute.
[0064] 2. Add 5 μl of anticoagulated whole blood to be tested (add up to 24 samples to be tested in each 48-well plate), mix immediately, and continue to incubate at 37°C for 5 minutes.
[0065] 3. After 5 minutes, add 0.5 ml of test solution for terminating red blood cell hemolysis, mix well, and centrifuge at 3000 rpm for 5 minutes.
[0066] 4. Use a pipette gun to take 100 μl of the supernatant and add them to the other 24 wells of this plate in sequence, and add water to 200 μl / well. Then discard the remaining supernatant (do not disturb the red blood cells at the bottom of the tube).
[0067] 5. Add 200 μl / well of erythrocyte hemolysis test solution to the original wells, mix well, and after the blood is completely hemolyzed (transparent red), the enzyme-linked colorimeter is used for colorimetry on t...
Embodiment 2
[0072] Experimental operation method:
[0073] 1. Take a 48-well ELISA plate, add 0.5ml of erythrocyte osmotic fragility test solution, and preheat at 40°C for 2 minutes.
[0074] 2. Add 5 μl of anticoagulated whole blood to be tested (add up to 24 samples to be tested for each 48-well plate) and mix immediately, and continue to incubate at 40°C for 10 minutes.
[0075] 3. After 5 minutes, add 0.5 ml of test solution for terminating red blood cell hemolysis, mix well, and centrifuge at 4000 rpm for 6 minutes.
[0076] 4. Use a pipette gun to take 100 μl of the supernatant and add them to the other 24 wells of this plate in sequence, and add water to 200 μl / well. Then discard the remaining supernatant (do not disturb the red blood cells at the bottom of the tube).
[0077] 5. Add 200 μl / well of erythrocyte hemolysis test solution to the original wells, mix well, and after the blood is completely hemolyzed (transparent red), the enzyme-linked colorimeter is used for colorimetr...
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Abstract
The invention provides a screening method for erythrocyte osmotic fragility, which comprises the following steps: S1, test solutions of the erythrocyte osmotic fragility are added into an enzyme plate and preheated at a temperature of 37 to 40 DEG C for 1-2 minutes; S2, 5-10 Mu l of anticoagulant whole blood to be measured is added into holding sample holes of partial areas of the enzyme plate, and then the test solution of the erythrocyte osmotic fragility and the anticoagulant whole blood are mixed evenly; S3, test solutions for stopping erythrocyte hemolysis are added into the partial areas filled with the anticoagulant whole blood to be measured, all solutions are mixed evenly and centrifugation is carried out; S4, the supernatant is removed and placed in the holding sample holes of the other partial areas of the enzyme plate; S5, erythrocyte hemolysis solutions are added into the base solution, and all solutions are mixed evenly; S6, after the hemolysis process is finished, colorimetry is carried out between the solution to be measured of the partial areas filled with the erythrocyte hemolysis test solution and the solution of the other partial areas not filled with the erythrocyte hemolysis test solution. With the advantages of simplicity, trace amounts, fast speed and large samples, the screening method for the erythrocyte osmotic fragility further has the advantages that the screening workload can be reduced, the screening can be completed fast and accurately, the reproducible results can be obtained and the experimental results can be automatically calculated.
Description
technical field [0001] The invention relates to a detection and screening method for osmotic fragility of red blood cells and an application thereof, belonging to the field of medical detection. Background technique [0002] The osmotic fragility test of red blood cells can screen a variety of anemias. This method mainly detects the resistance of red blood cells to hypotonic saline solutions of different concentrations. , Diagnosis of diseases such as obstructive jaundice and detection of hemolytic diseases such as globinogenic anemia. Thalassemia, also known as thalassemia, is a group of inherited hemolytic anemia diseases. Anemia or pathological condition caused by the absence or insufficient synthesis of one or more globin chains in hemoglobin due to inherited genetic defects. Due to the complexity and diversity of gene defects, the type and quantity of globin chains lacking and the variability of clinical symptoms are large. They are named and classified according to ...
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