Method for determining red blood cell penetration fragility

A technology of penetration brittleness and determination method, applied in transmittance measurement, color/spectral property measurement, biological test, etc., can solve the problems of poor result accuracy, many operation steps, and many measurement steps, saving time and good repeatability , the effect of high accuracy

Inactive Publication Date: 2005-03-02
潘干华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above method requires manual sample addition, centrifugation, colorimetry, etc., with many determination steps and complicated procedures; the conventional method takes more than two hours, and the improved one-tube method takes more than ten minutes; because the whole process is manual operation, and there are many steps, The reaction conditions are difficult to control, which leads to poor repeatability and accuracy of the results; at the same time, due to the complexity of the reaction system, steps such as centrifugatio

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] This example is a dynamic analysis method.

[0013] 1. Add 3 ml of 0.4% NaCl solution to the cuvette of a spectrophotometer with automatic constant temperature and automatic recording, and wait until the solution in the cuvette is kept at the predetermined reaction temperature of 37°C;

[0014] 2. Add 10 microliters of blood sample, and immediately mix within 3 to 5 seconds. Immediately start recording the absorbance value measured with a wavelength of 700nm per second. The end point of the reaction is that the absorbance does not continue to decrease (usually 3 ~5 minutes), the absorbance values ​​measured from the first second, the second second, the third second...to the end point are expressed as a1, a2, a3...aZ in turn;

[0015] 3. The following parameters can be obtained through calculation and analysis:

[0016] Relative hemolysis rate from i second to x second = (ai-ax) / ai×100%

[0017] Relative hemolysis rate in the first minute = (al-a60) / a1×100%

[0018] 4...

Embodiment 2

[0020] This example is an endpoint analysis

[0021] 1. Take two test tubes A and T

[0022] 2. Add 10 microliters of blood sample and 3 milliliters of 0.9% NaCl solution to tube T, mix well, add 10 microliters of blood sample and 3 milliliters of 0.4% NaCl solution to tube A, and mix well; put the two tubes at the same time 37 ℃ water bath for 5 minutes, on the spectrophotometer, with a wavelength of 700nm, measure the absorbance of T tube as t, and the absorbance of A tube as a;

[0023] 3. Calculation: terminal hemolysis rate = (t-a) / t×100%

[0024] t: absorbance in isotonic solution (equivalent to absorbance without hemolysis)

[0025] a: Absorbance after hypotonic hemolysis

[0026] Description to the present invention's turbidimetric analysis method measuring condition:

[0027] 1. About hypotonic fluid

[0028] For red blood cells, a solution with an osmotic pressure equal to the intra-erythrocyte osmotic pressure of 280-320 mOsm / L is isotonic, and a solution with ...

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Abstract

The invention relates to a determining method for the erythrocyte osmotic brittleness. It adopts turbidimetric analytic method as below: mixing the blood sample with the hypotonic solution with specific osmotic pressure, taking the undissolved erythrocyte as the main particle forming the turbidity of the mixed solution., selecting a undissolved erythrocyte which has higher absorbency to it, taking the specific wave length that the soluble coloured substance such as the free hemoglobin which is dissolved from erythrocyte have a weaker absorbency to it as the determining wave length, making turbidimetric analysis on the spectrophotometer: calculating the value of the haemolytic rate which is in direct ratio with the erythrocyte osmotic brittleness by comparing the different absorbency of erythrocyte in different duration of time and on the same determining condition within the hypotonic solution with the said specific osmotic pressure, or by comparing the different absorbency of erythrocyte between in the isotonic solution and in the above said hypotonic solution with specific osmotic pressure. The invention has the advantages of high accuracy, good repeatability of measurement, simple procedure, high velocity of determination, capability of realizing the dynamic determination and the full automatic analysis easily.

Description

technical field [0001] The invention belongs to a method for clinically measuring osmotic fragility of red blood cells. Background technique [0002] Human red blood cells account for about 40% of the total blood volume, and water and hemoglobin, which contain the most water in red blood cells, account for 65% and 32% of red blood cells respectively. In a hypotonic solution, water penetrates the cell membrane into the red blood cells, causing the red blood cells to gradually swell and rupture. , Intracellular hemoglobin overflow is called hypotonic hemolysis. The resistance of red blood cells to hypotonic fluid hemolysis is called osmotic fragility of red blood cells. In patients with thalassemia and iron deficiency anemia, the production of hemoglobin in red blood cells is reduced, the cell volume is smaller, and the osmotic fragility is significantly smaller than that of normal people. Auxiliary diagnostic indicators. [0003] The existing method for measuring the osmot...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N21/59G01N21/82G01N33/49
Inventor 潘干华
Owner 潘干华
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