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Method of assaying substance with affinity in sample including step of destroying blood-cell ingredient

A determination method and substance technology, which can be used in biological tests, measuring devices, analytical materials, etc., and can solve the problems of inability to detect particles, the effect of small hole clogging, etc.

Inactive Publication Date: 2008-12-03
PULSE IMMUNOTECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This size is susceptible to pinhole clogging
However, particles with a diameter of 0.8 to 1 μm cannot be detected using a small hole with a diameter larger than this

Method used

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  • Method of assaying substance with affinity in sample including step of destroying blood-cell ingredient
  • Method of assaying substance with affinity in sample including step of destroying blood-cell ingredient
  • Method of assaying substance with affinity in sample including step of destroying blood-cell ingredient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0243] [Example 1] Bursting test 1 of whole blood

[0244] (1) Measuring device

[0245] use figure 1 The apparatus of (A) tests the disruptability of blood cell components contained in whole blood. The whole blood sample and the reagent are injected and mixed in 1 minute, and then the mixed solution is moved into the reaction tank 3 (pulse application tank), and pulse voltage is applied through the electrode 4 for several seconds to several minutes to rupture the blood cell components. Reagent 2 (latex reagent) was not used at this time. Thereafter, the mixed liquid is diluted in the dilution tank 5, and the particle size distribution of the blood cell component is measured with a particle size distribution meter 6. Temperature control mechanisms 1 and 2 are set to off.

[0246] figure 1 (B) shows the cross section of the pulse application groove. The distance between the electrodes was 0.8 mm, the thickness of the electrodes was 0.03 mm, and the length of the electro...

Embodiment 2

[0253] [Example 2] Bursting test 2 of whole blood

[0254] Using the same whole blood sample and device as in Example 1, blood cell components were destroyed under the following conditions.

[0255] Pulse voltage: AC voltage (rectangular wave) with a frequency of 400KHz

[0256] Electric field strength: 48V / 0.8mm

[0257] Application time: 0 to 60 seconds

[0258] The results are shown in Table 2. The average volume of the counted particles becomes significantly smaller when the pulse voltage is applied for about 10 seconds. Furthermore, it was also confirmed that the number of particles larger than 3 μm corresponding to blood cell components could hardly be counted by application for 10 to 30 seconds. Therefore, it was confirmed that an application time of about 10 seconds is sufficient for an AC voltage (rectangular wave) with a frequency of 400 KHz and an electrolytic strength of 48 V / 0.8 mm. That is, according to the present invention, blood cell components can be des...

Embodiment 3

[0267] (1) Preparation of anti-CRP antibody sensitized latex reagent

[0268] A latex reagent was prepared as an antibody solution in glycine buffer (containing 50 mM glycine, 50 mM sodium chloride, 0.09% sodium azide, hereinafter abbreviated as GBS) containing 0.15 mg / mL anti-CRP antibody (manufactured by Shibayagi). As for the latex particles, 0.9 mL of GBS was added to 0.1 mL of 1.0 μm latex (manufactured by Sekisui Chemical Co., Ltd., 10% solid content suspension) to prepare a latex suspension.

[0269]1 mL of the antibody solution and the latex suspension were mixed, stirred at 37°C for 2 hours, the sensitized latex was centrifuged, and the supernatant was removed. The precipitate was suspended in 2 mL of glycine buffer (0.5% BSA-GBS) containing 0.5% bovine serum albumin to prepare an anti-CRP antibody sensitized latex reagent.

[0270] (2) Measuring device

[0271] use figure 1 The device in (A) measures biologically specific agglutination reactions (antigen-antibody...

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Abstract

A method of assaying a substance having an affinity based on the pearl chain formation of carrier particles, which includes the step of electrically destroying a blood-cell ingredient. In the method, the blood-cell ingredient which interferes with the count of carrier particles is electrically destroyed. The blood cell ingredient can be destroyed without the need of adding a hemolytic agent, which interferes with immunological reactions. Thus, a whole blood sample can be used as it is as an assay sample without the need of separating the blood serum and blood plasma therefrom. Therefore, there is no need of separating out blood serum and blood plasma in preparation for the assay of a blood ingredient.

Description

technical field [0001] The present invention relates to a method for measuring an affinity substance using an agglutination reaction of carrier particles. Background technique [0002] As methods for detecting or measuring the presence of biologically idiosyncratic substances, for example, enzyme immunoassays or radioimmunoassays have been used. These methods have high sensitivity and high precision. However, the reagents are unstable due to the use of enzymes or radioactive isotopes as labels. In addition, since radioactive isotopes are used, since storage and storage are limited, careful consideration and skilled techniques are required for measurement. Therefore, a simpler measurement method is required. Furthermore, since these methods require a long time for measurement, it is difficult to cope with urgent inspections. Under such a background, researches on highly sensitive and rapid measurement methods have begun to flourish. [0003] After 1970, the analytical me...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/49
CPCG01N33/48G01N33/54313G01N33/49
Inventor 岩田惠助
Owner PULSE IMMUNOTECH CORP
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