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Reagent and method for classifying leucocyte

A white blood cell and reagent technology, applied in the field of cell classification, can solve the problems of increasing instrument cost, limiting the scope of application, reducing the accuracy of results, etc., to achieve the effect of saving instrument cost, ensuring accuracy, and broadening the application range

Active Publication Date: 2007-11-28
SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In the classification reagents and methods in the above prior art, most of them have specific requirements on the pH value, and the scope of application of the pH value only includes a narrow range.
And in Chinese patent CN95115317.X, U.S. patent 5,618,733, U.S. patent 5,389,549, U.S. patent 5,538,893, classification reagent all must contain non-ionic surfactant, the selection range of material is narrower, and pH value also has certain restriction simultaneously, for example, China In the patent CN95115317.X, the pH value is required to be between 5-11
These all limit the scope of application of the above-mentioned technology
[0013] In addition, the classification technology mentioned in the Chinese patent with application number 88101677 and approval number CN1018586B has the optimum reaction condition of normal temperature, but as the ambient temperature changes, it is difficult to ensure that it is in a constant temperature state of 25 degrees, so the measurement results vary with Temperature changes create fluctuations that reduce the accuracy of results
On the basis of this method, if in order to eliminate the influence of temperature on the reaction, it is necessary to add a constant temperature device, but in order to control it at 25 degrees, not only a heating device is required, but also a refrigeration device is required, which greatly increases the cost of the instrument

Method used

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  • Reagent and method for classifying leucocyte
  • Reagent and method for classifying leucocyte
  • Reagent and method for classifying leucocyte

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] A reagent for the four classifications of white blood cells, which consists of the following components:

[0080] Phthalic acid 1g

[0081] NaCl 0.2g

[0082] 2-Phenoxyethanol 10g

[0083] Brij 35 3.4g

[0084] Acid Blue 0.15g

[0085] Replenish water to 1L

[0086] pH 2.5

[0087] The pH of the reagent prepared according to the above formula was adjusted to 2.5, and the osmotic pressure was 100 mOsm.

[0088] Under the condition that the temperature was kept at 25 degrees, 0.5 ml of the above reagent was added to 30 μl of blood. Laser detection of leukocytes was started after 12 seconds of mixing. The size information of the cells is measured by using the low-angle scattered light with a measuring angle of 2-6 degrees, and the shape information of the cells is measured by using the high-angle scattered light with a measuring angle of 8-20 degrees. The results are shown in Figure 1. The leukocytes were divided into four types: lymphocytes, monocytes, eosinophils...

Embodiment 2

[0090] A reagent for the four classifications of white blood cells, which consists of the following components:

[0091] NaCl 0.2g

[0092] Methanol 2g

[0093] SDS 1.5g

[0094] Sodium dihydrogen phosphate 3.08g

[0095] Disodium hydrogen phosphate 4.87g

[0096] Acid Blue 0.15g

[0097] Replenish water to 1L

[0098] pH 6.5

[0099] The pH of the reagent prepared according to the above formula was adjusted to 6.5, and the osmotic pressure was 100 mOsm.

[0100] Under the condition that the temperature is maintained at 35 degrees, 1 ml of the above reagent is added to 30 μl of blood. Laser detection of leukocytes was started after 20 seconds of mixing. The size information of the cells is measured by using the low-angle scattered light with a measuring angle of 2-6 degrees, and the shape information of the cells is measured by using the high-angle scattered light with a measuring angle of 8-20 degrees. The results are shown in Figure 2. The leukocytes were divided in...

Embodiment 3

[0102] A reagent for the four classifications of white blood cells, which consists of the following components:

[0103] NaCl 0.3g

[0104] Methanol 3g

[0105] Laurylglycine 1.2g

[0106] Tris 3.05g

[0107] HCl0.82g

[0108] Acid Blue 0.15g

[0109] Replenish water to 1L

[0110] pH 7.2

[0111] The pH of the reagent prepared according to the above formula was adjusted to 7.2, and the osmotic pressure was 140 mOsm.

[0112] Under the condition that the temperature is kept at 35 degrees, 1.5 ml of the above reagents are added to 30 μl of blood. Laser detection of leukocytes was started after 20 seconds of mixing. The size information of the cells is measured by using the low-angle scattered light with a measuring angle of 2-6 degrees, and the shape information of the cells is measured by using the high-angle scattered light with a measuring angle of 8-20 degrees. The results are shown in Figure 3. The leukocytes were divided into four types: lymphocytes, monocytes, e...

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Abstract

The invention discloses a reagent for four-classification of leukocyte. The reagent comprises a. at least a surfactant which can dissolve erythrocyte and destroy leukocyte membrane; b. at least an organic compound with anionic group which can combine with cationic in leukocyte and make leukocytes generate configuration difference; c. buffering agent which can adjust PH value in 2-8. The invention also discloses the method for four-classification by said reagent. When four-classification of leukocyte is carried out by using the reagent and the method in invention, reaction can be carried out under the condition from acidity to weak alkaline PH and temperature and class of main component surfactant all have very wide selecting range so that the application range of invention is developed greatly. Compared with prior technology it has a great improvement.

Description

technical field [0001] The invention relates to a reagent and a classification method used for cell classification, in particular to a reagent and a classification method used for four classifications of white blood cells in blood. Background technique [0002] In the field of clinical examination, it is extremely common and very important to use the patient's whole blood to classify and count white blood cells in the diagnosis of various diseases. [0003] So far, many patents on devices and methods for leukocyte classification have been published. Among them, Chinese patent CN95115317.X discloses a method for completing the five classifications of white blood cells by using two reagents. In addition to the two reagents containing nonionic surfactants and buffers, the first reagent contains at least one ionic surfactant and an anionic organic compound, which divides blood cells into lymphocytes, monocytes, and eosinophils. and four types of basophils plus neutrophils, the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/50G01N33/49G01N15/14
CPCY10T436/25125Y10T436/107497Y10T436/25Y10T436/101666G01N33/5094Y10T436/108331Y10T436/10G01N15/1459G01N2015/1006
Inventor 许文娟张利娜刘兵
Owner SHENZHEN MINDRAY BIO MEDICAL ELECTRONICS CO LTD
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