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Eosinophil analogue, preparation method thereof and whole blood quality control substance

A technology for eosinophils and neutrophils, applied in the field of whole blood quality control substances, can solve the problems of difficult quality control of eosinophils, low content of eosinophils, damage of white blood cell simulants, etc., to achieve convenient The effect of large-scale production, low production cost and simple preparation method

Active Publication Date: 2010-11-17
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] U.S. Patent 5512485 discloses a quality control substance containing leukocyte subclass mimics, which describes the treatment of red blood cells by low osmotic pressure to expand the volume of red blood cells by about 2 times, and change the hemoglobin content in red blood cells, thereby changing the properties of the cells. However, the disadvantages of this method are: (1) hypotonic treatment can easily lead to the lysis of red blood cells; (2) the stability of red blood cells after fixation is worse than that of white blood cells; (3) the volume of red blood cells is small There is a certain distribution width; it is difficult to ensure the uniformity of the volume of leukocyte simulants through hypotonic treatment; (4) the leukocyte simulants prepared with red blood cells have the risk of being destroyed by hemolytic agents
Although the instability of the method described in U.S. Patent Nos. 5,512,485, 4,704,364, and 5,380,664 and the complexity of raw materials have been avoided by using mammalian leukocytes similar in nature to human leukocytes as a mimic of human leukocytes, eosinophils in normal mammals The proportion of white blood cells is only about 1%, and the content of eosinophils in the leukocyte simulants obtained by the methods described in 5270208, 5858790, and 6187590 is low, and eosinophils can be easily divided into Neutrophils, which makes quality control of the already low number of eosinophils difficult

Method used

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  • Eosinophil analogue, preparation method thereof and whole blood quality control substance
  • Eosinophil analogue, preparation method thereof and whole blood quality control substance
  • Eosinophil analogue, preparation method thereof and whole blood quality control substance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] 1. Take 100ml of fresh anticoagulated bovine blood, containing 2g / L of anticoagulant;

[0056] 2. Mix 100ml of anticoagulated bovine blood with 400ml of hemolytic agent, stir for 10 minutes, centrifuge at 2000g for 5 minutes, and harvest white blood cells;

[0057] 3. Centrifuge and wash white blood cells with PBS buffer once or twice;

[0058] 4. Take the white blood cells in step 3 and adjust the white blood cell count to 50×10 with PBS 9 pieces / L, centrifugal force 200g, centrifuge for 10 minutes, harvest the precipitate;

[0059] 5. Get the precipitation in step 4, resuspend (osmotic pressure 260 ± 20mOsm / kgH 2 O), after fixing at room temperature for 5 hours, wash with PBS buffer to remove the fixative.

[0060] The eosinophil simulant prepared in step 5 was detected by a blood cell analyzer that simultaneously detects low-angle scattered light and high-angle scattered light, and the results were as follows image 3 shown.

Embodiment 2

[0062] 1. Take 100ml of fresh anticoagulated bovine blood, containing 2g / L of anticoagulant;

[0063] 2. Mix 100ml of anticoagulated bovine blood with 400ml of hemolytic agent, stir for 10 minutes, centrifuge at 2000g for 5 minutes, and harvest white blood cells;

[0064] 3. Centrifuge and wash white blood cells with PBS buffer once or twice;

[0065] 4. Take the white blood cells in step 3 and use PBS to adjust the white blood cell count to 50×10 9 pieces / L, centrifugal force 200g, centrifuge for 10 minutes, harvest the precipitate;

[0066] 5. Get the precipitation in step 4, resuspend (osmotic pressure 1400 ± 20mOsm / kgH 2 O), after fixing at room temperature for 5 hours, wash with PBS buffer to remove the fixative.

[0067] The eosinophil simulant prepared in step 5 was detected by a blood cell analyzer that simultaneously detects low-angle scattered light and high-angle scattered light, and the results were as follows Figure 4 shown.

Embodiment 3

[0069] 1. Take 100ml of fresh anticoagulated bovine blood, containing 2g / L of anticoagulant;

[0070] 2. Mix 100ml of anticoagulated bovine blood with 400ml of hemolytic agent, stir for 10 minutes, centrifuge at 2000g for 5 minutes, and harvest white blood cells;

[0071] 3. Centrifuge and wash white blood cells with PBS buffer once or twice;

[0072] 4. Take the white blood cells in step 3 and use PBS to adjust the white blood cell count to 50×10 9 pieces / L, centrifugal force 200g, centrifuge for 10 minutes, harvest the precipitate;

[0073] 5. Get the precipitation in step 4, resuspend (osmotic pressure 400 ± 20mOsm / kgH 2 O), after fixing at room temperature for 5 hours, wash with PBS buffer to remove the fixative.

[0074] The eosinophil simulant prepared in step 5 was detected by a blood cell analyzer that simultaneously detects low-angle scattered light and high-angle scattered light, and the results were as follows Figure 5 shown.

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Abstract

The invention discloses an eosinophil analogue, a preparation method of the analogue and a whole blood quality control substance containing the eosinophil analogue, wherein, the eosinophil analogue is prepared from neutrophils. The analogue can accurately simulate the characteristic of eosinophils, thus the analogue can be applied to quality control for analytic determination of the eosinophils; and the analogue takes the neutrophils, especially the neutrophils of mammals as raw materials, thus having the advantages of available raw material sources, simple production process, low production cost, good stability and good repeatability among production batches.

Description

technical field [0001] The invention relates to a clinical detection reagent, in particular to an eosinophil simulant, a preparation method of the simulant and a whole blood quality control substance containing the simulant. Background technique [0002] As an important detection tool for cell biology, blood cell analyzer plays an important role in clinical medicine, teaching, scientific research and other work. The stability and accuracy of the blood cell analyzer test results are the prerequisites for its application, which makes quality control essential in the application process of blood cell analysis. At present, medium and high-end hematology analyzers have become the mainstream trend in the development of hematology analyzers. The parameters related to eosinophils are an important information provided by middle and high-end blood cell analyzers. The number of eosinophils is related to parasitic diseases, urticaria, hay fever, eczema, psoriasis, eosinophilic leukemia...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/96G01N21/49G01N21/59G01N27/04G01N23/00
Inventor 张华利徐祖越谢键周志德
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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