Method for counting leukocytes and leukocyte counter

a leukocyte counter and leukocyte technology, applied in the field of counting leukocytes and leukocyte counters, can solve the problems of inability to obtain exact counted value, long time to count cells, and error in counting value, so as to achieve fast antigen-antibody reaction, eliminate the effect of protease inhibitors, and low cos

Inactive Publication Date: 2004-09-02
ARKRAY INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

0011] It is preferable that the method of the present invention includes bonding the same protease inhibitor as a protease inhibitor contained in the specimen to the liberated elastase, and then adding an antigranulocyte elastase antibody into the specimen. With such a preferable embodiment, when a protease inhibitor is contained in the specimen such as blood etc., the effect by the protease inhibitor can be eliminated. Therefore, it is preferable that when the specimen is whole blood, the protease inhibitor is .alpha.1-antitrypsin, because .alpha.1-antitrypsin is the largest in amount (about 90%) among the protease inhibitors in blood.
0012] In the counting method of the present invention, it is preferable that a latex agglutination immunoassay be employed as the immunological technique. More specifically, it is preferable that the method of the present invention includes causing an antigen-antibody reaction between an antibody particle in which the antigranulocyte el...

Problems solved by technology

In the above-mentioned visual counting method, an error in counting value occurs because of mistake about the kind of cells caused by erythrocytes being deficient in hemolysis or the hemagglutination of blood cells, in homogeneous broadening of leukocytes on the calculating board, or the skill of laboratory technicians.
Moreover, the visual counting method is a manual method, and therefore it takes a long time to count cells.
In the automatic counting method, the exact counted value may not be obtained because of the appearance of erythroblast cells, depos...

Method used

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  • Method for counting leukocytes and leukocyte counter

Examples

Experimental program
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Effect test

example 2

[0050] The number of whole leukocytes was calculated by using the elastase concentration with respect to one neutrophil cell, i.e., 0.9084 ng / ml and the ratio of the neutrophil count to the number of the whole leukocytes, i.e., 55.3%. The determination of the elastase concentration with respect to one neutrophil cell was calculated from only neutrophils counted by the usual method (the visual method) and the elastase concentration measured by the same technique as Example 1 (the enzyme immunoassay) except that a regression linear expression in Example 1 was not used. Table 2 shows the

example 3

[0055] Five kinds of specimens were prepared the same way as in Example 1. The elastase concentration of each of these 5 specimens was measured by the below mentioned latex agglutination immunoassay. More specifically, first, each of the above-mentioned specimens was added to 0.1 weight % of saponin aqueous solution so as to lyse leukocyte. This was diluted in the same way as in Example 1 to prepare test liquid. 10 .mu.l of this test liquid was added to 250 .mu.l of phosphate buffered saline (pH7.4), then 50 .mu.l of latex suspension (concentration: 2.0 mg / ml), which had been sensitized by antihuman granulocyte elastase antibody, was added to the test liquid, and incubated at 37.degree. C. for 15 minutes. The latex suspension was prepared by physically adsorbing the above-mentioned antibody to the latex particles, which have an average particle size of 0.1 to 0.2 .mu.m and in which polyethylene was graft-copolymerized to polystyrene at the rate of about 200 ng / cm.sup.2. Then, the ab...

example 4

[0057] The number of leukocytes in the specimen was calculated in the same way as in Example 3 by using 50 cases of adult whole blood (n=50) as the specimens. Furthermore, as controls, the number of leukocytes of each of the above-mentioned specimens (n=50) was counted by using the automatic blood cell counter (K-4500 produced by TOA MEDICAL ELECTRONICS CO., LTD.). FIG. 1 shows the results. FIG. 1 is a graph showing the relationship between the number of leukocytes counted by the counting method according to the present invention and the number of leukocytes counted by using the automatic blood cell counter concerned in each specification (n=50).

[0058] As shown in FIG. 1, in each specimen (n=50), the number of leukocytes calculated from the elastase concentration and the number of whole leukocytes counted by using the automated analyzer were highly correlated (correlation coefficient r=0.973).

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Abstract

A method for counting leukocytes which comprises liberating elastase from granulocytes contained in a specimen, adding an anti-granulocyte elastase antibody to the thus liberated elastase, measuring the antibody bonded to the elastase to thereby determine the concentration of the elastase, and then calculating therefrom the number of leukocytes contained in the specimen with the use of the ratio of the leukocyte count to the elastase concentration. This method makes it possible to conveniently and less expensively count leukocytes at a high accuracy comparable to the one established by using a conventional automatic blood cell counter, as the figure shows.

Description

[0001] The present invention relates to a method for counting leukocytes contained in a specimen and to a leukocyte counter used therefor.[0002] In a blood test, blood cell counting is frequently carried out and its clinical significance is great. For example, an erythrocyte count permits examining the presence and the degree of anemia and further permits clinical diagnosis of various cases caused by oxygen deficiency in the tissue. On the other hand, a leukocyte plays an important role in a defense function of fighting bacteria or virus entering the body, namely, an immunological function. In particular, granulocytes or leukocytes have a function of liberating protease such as elastase, etc. by the stimulation of bacteria or foreign proteins entering the body, and a function of allowing elastase etc., to respond the bacteria or foreign proteins. Since the leukocyte count is increased or decreased in the case of many diseases, it is important to count leukocytes in a screening for d...

Claims

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Application Information

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IPC IPC(8): G01N33/49G01N33/50G01N33/53G01N33/543G01N33/573
CPCG01N2333/966G01N33/573G01N33/53
Inventor OKUBO, AKIOYAMADA, SHIGEKI
Owner ARKRAY INC
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