JPS5931454A - Aqueous solvent for testing clotting of red blood corpuscles - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

DETAILED DESCRIPTION OF THE INVENTION Technical Field The present invention relates to aqueous solvents for red blood cell agglutination tests.

Antigen-antibody reaction, a certain animal, plant, bacterial cell, or virus-derived aggregate 1. The method of measuring the -10,000 or +t+j of Sowara for a fixed purpose is to measure the aggregation f☆reaction between Bit and living body (white γ1 and red blood cells by 3% i#I・). PHA method), reverse passive hemagglutination reaction (R
It is widely used clinically (in the field of gastronomy) as the PHA method), bacterial agglutination reaction, or latex agglutination reaction.

For example, regarding the P II A method, the first step is to sensitize purified llBs antigen to red blood cells, which are used as a carrier for the Ij11 test, to develop HBs antigen sensitivity. The presence of J('Bs antibodies in the sample was determined by whether or not an agglutination reaction was observed in an aqueous solvent when this was fused with a synthetic sheep red blood cell.) Can be judged.

In addition, if an aggregation reaction is observed, the sample is diluted stepwise and the dilution ratio until the agglutination reaction can no longer be observed or detected is determined.
s antibodies can be measured regularly.

In addition, 411 erythrocytes were sensitized to 411 erythrocytes with RP) (lJ, purified JJ, 'Bs antibody using method A, and 11Bs antibody (11
When 9 red blood cells are prepared and the sample is combined with the sample, whether or not an agglutination reaction is observed in the aqueous solution is used to detect whether or not the sample contains J(I3s antigen). can do.

The divine virus in history and the red blood cells of birds agglutinate red blood cells in an aqueous solvent due to the action of the virus's agglutinin. Depending on the presence or absence and strength of the uF aggregation reaction in this aqueous solvent, it is possible to quantitatively measure the amount of virus. It is well known that I'M is also delicious and is widely used for testing various antigens and antibodies.

However, in the aggregation test in these aqueous solvents, Jt! '1
However, the disadvantage of this method is that it contains antibodies against the carrier (in a single-force system, it is difficult to determine whether a non-specific agglutination reaction has occurred).Therefore, when measuring a sample,
It is necessary to perform the measurement after diluting the IW to such an extent that the engravings of these wall-like reactants do not come out, or to confirm the authenticity of the aggregation reaction in an aqueous bath medium using an appropriate confirmation test method. .

In addition, as a method for reducing these non-specific aggregation reactions in aqueous Mts, a method is already known in which a small amount of animal pigment or acrinol is added to the aqueous solvent used for 1 tll+. However, even in these lIθ-, there is a lack of culm IW b￥; Although the M collection l reaction is μ/j mountain, there are still many non-percent reactions remaining, so the appearance of the first 11th method in history. I'm waiting for Li no Sane.

Also, for non-specific reactions /l! Another way to leave is i'
There is also a method of preparing /lif-like reactant trade in the i13 body and removing the 0 metal adhesion in advance with a tc carrier, but in order to process the sample in a short time at 1 tl, it is necessary to ) It's really *tJAc.

The purpose of the present invention is to eliminate non-specific reactions and store them for use.

The name of the invention is the improvement of the measurement method to eliminate non-specific reactions in this agglutination reaction for many years. By making Class I/j12. minute exist in the measurement system, ?1Ilj is constant'?I
Q- is changed by the customer by f1 ÷, and furthermore, the assembly factory 1
r/min, especially when present at 28 degrees, tII
He completed the invention by getting -7.

That is, the present invention relates to an aqueous bath for red blood cell agglutination test (Iv) characterized by containing an animal-derived placental tissue component or muscle tissue component at a tb degree of 40.01 to 0.4% (w/v).

By using this aqueous solvent to prepare a reagent for a red blood cell collection reaction test, it is possible to improve the sensitivity of two sets of measurements.

As the aqueous solvent, all known aqueous media for agglutination tests can be used. For example, water, 1 ton of menstrual fluid, A water, various slow S solutions (phosphate buffer, borate buffer, chrysin Examples include solutions consisting of aqueous solution containing a surfactant, an albumin solution, a dextrose solution, a stock solution, a solution of normal human and animal serum, a solution of a synthetic polymeric substance, an aqueous solution containing a surfactant, and a combination thereof. The pH of the aqueous solvent is approximately 6.0-8.
5 is preferable, and adjustment to such pH is preferably carried out with a mild #I solution. The salt concentration of the aqueous solvent is preferably a relatively high concentration, more preferably a physiologically equidistant solution.

Furthermore, two sets of conventionally known non-specific agglutination reaction inhibiting substances may be further mixed into this solution. As carriers, those known per se are used, and specifically, for example, sheep, guinea pigs, type 0 humans, chickens (red blood cells fixed with formalin, glutaraldehyde, etc. Of these, particularly preferred are sheep red blood cells and type 0 human red blood cells.It is advantageous to use such carriers with a diameter of about 20μ or less, particularly about 5 to 15μ. The process of loading the antibody or antigen onto such a carrier can be carried out according to a method known per se.

In addition, the composition component is, for example, a finely crushed or decomposed tissue (for example, a finely decomposed product of cell tissue of 1°51i or less), etc. tl obtained by treating with ultrasonic waves after treatment with a blender, or by treating with a holiday-degrading enzyme such as papain after treatment with a blender, the total nitrogen content is 111, while non-protein nitrogen≠21@20-70
% is preferably used. The conditions for ultrasonication are 1. Normally 30"C or less, lO~20KH110~6
It is preferable to use particles that have passed through a membrane with a diameter of 2 μm after being subjected to ultrasonic treatment for 0 minutes.

Example Guinea pigs were immunized with purified 1] 13s antigen to obtain HBs antiserum. This anti-11flll was purified by ammonium sulfate fractionation and ion exchange chromatography.
Anti-HBs antibody 1 was obtained, and this purified IBs antibody was plated onto sheep red blood cells fixed with glutaraldehyde to obtain an HBs antibody (Shimasaku sheep red blood cells).

On the other hand, as an aqueous medium of the present invention, 01μ phosphoric acid (pH 7.5) is used to remove decomposed components of normal animal blood and human or sheep placenta.
4012 (w/v), and a control without additives was prepared. 2 of plasma containing citric acid a;f, which was found to cause non-specific agglutination as a result of confirmation tests.
(The ψ-bodies (1 and 2 in the table) were separated into 2 volumes on a microplate using these 5 extracts of solvent according to the Jj PHA method.
A multiple dilution was performed, and the HEs antibody-inserted sheep erythrocytes obtained above were incubated with this diluted array.

A specimen (3) containing another lζHEs antigen was treated and tested in the same manner. The mixed solution t74 is 25μLH due to body dilution.
'Bs antibody bribed sheep red blood cells 25 μe and agglutination after 2 hours at room temperature agglutination 1! F) is evaluated as 4.8.2.1, and when the non-collection is expressed as 0, the result of Table 1 is 11? I got it.

Placenta 'Ml & as ingredients: 1 minced fetus
After washing f4t with 07Q water, p -H7,0
Okishin was used, which had been suspended in Linhal sodium f'8D, decomposed with a breather, treated with H1 sonication, and passed through a 1.2μ membrane.

(Remaining days below) Table 1 From the results shown in Table 1, non-specific The width has been reduced from 1:64 to 1:128 to l:4 to 1:16. In addition, in the case of positive results, it can be seen that there is no effect even if these substances are added.

As described above, the aqueous l6 medium for flocculation tests according to the present invention is one that elegantly improves the flocculation reaction LW, making it possible to perform the first and sixth tests immediately. It is something.

The present invention will be illustrated below with reference to 11 actual 7Ji examples.

Example 1 Sheep red blood cells were fixed with glutaraldehyde17 and sensitized with anti-H1ls antibody by a known method to obtain 411 cells.

As a water conditioner for Kasuri test soap using this carrier, OIM I
J-acid powder Ih was added to pH 7,8 and human candy was added to the mixture hν5.
01% (W/V) was added to obtain an aqueous solution for hemagglutination test. Composition green component, minute is experimental (in 9+l.;
1. ! 1! It was prepared using the same method as the original one.

Fruit R11, Iv112 Niwa I red blood cells were fixed with geltaraldehyde and HBs antigen was produced using a known method.

Using this living body, we obtained aqueous fabric for testing in the F series by adding 101% idQ to 09% sodium chloride.

Fruit/1ili Ige 113 Fruit 111-410 and 2<4) An aqueous solvent was obtained in which NaNa was added as a preservation bar 11.

Patent issued i+: i person Midori Juji Co., Ltd. agent person
Patent attorney *-as,・: -if before procedure(';':
' (1 茫) 1chi1iifl15hui140月1go1'''I! 1. f'
Director-General of the l Office 1 1: f'l) (, J Miaki (, 57) 1 Patent 11, No. 5141893 2 Title of the invention Aqueous solvent for red blood cell agglutination test The number of inventions made by JJII is increased by 111i1.
, Specification 1, page 6, line 20 σ] "Croup protein 1'h J
Correct to 'k r soluble'0

Claims (1)

[Scope of Claims] +l) 4;f Placental tissue (lid, minute or pano) of animal polished rice as a carrier for Token test JIIO, 01~
A water medium for blush bulb condensation test, characterized in that it is contained at a concentration of 0.4% (w/v). (2) The carrier is antibody + 5 (i, cultivated sheep red blood cells, 1 min is human and non-human animal fetal Nr tissue, non-human animal fetus Nr tissue, , i'f
Qil'1trr of #r'5F-+aqueous f/i'yL described in item (1)