JPS60133368A - Immunological measurement - Google Patents

Abstract

PURPOSE:To perform an immunological measurement handily and quickly by filling a capillary tube with a non-soluble carrier having material (such as antibody) trapping labeled antigen or the like immobilized and an insoluble carrier or the like holding a labeled reaction reagent (to react with a label to cause a color formation or the like) to cause an immunological reaction with the tube. CONSTITUTION:A transparent capillary tube 3-20cm long and 0.5-2mm. across is filled with a combination of an insoluble carrier such as glass bead (GB) holding a labeled antibody and GB having an antigen immobilized, GB holding a labeled antibody and GB having antibody immobilized, GB holding a labeled antibody, GB having an antibody immobilized and GB holding an auxiliary labeled matter or the like. For example, a polyester cotton plug a-1, an o-diamisidine held GB a-4, GB a-4 having an anti-hare IgG bonded peroxidase label bonded and a hare IgG bonded GB a-6 are placed at the lower end of the capillary tube with GB-BSA bonds a-3 and a-5 arranged between a-2 and a-4, and a-4 and a-5, the remaining a-7 filled with GB and a cotton plug put into the uppermost section. Serum or the like to be inspected is sucked up to the a-6 from the lower end in about 1min and then, an H2O2 solution is sucked in close to the upper end in about 15min. With hare IgG present in the solution being inspected, the part a-6 is colored but remains colorless without it.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for detecting or quantifying components in a specimen by immunoreaction, and more particularly to an immunoassay method in which the immunoreaction and capture of a labeling substance are performed in a capillary tube.

Conventionally, in the immunoassay method, an antigen or antibody in a subject is immunoreacted with an antigen or antibody labeled with 9A using a radioactive isotope, a W# prime, a fluorescent substance, etc. Radioimmunoassay, W# immunoassay, or fluorescence immunoassay is used to measure substances.

Therefore, in these immunoassay methods L1, the immunoreaction is conventionally carried out in a test, etc., and it is necessary to separate the reacted labeled substance and unreacted labeled substance obtained as a result. It is. However, this minute I! TVC has disadvantages in that it requires complicated operations, specialized skills, and long hours.

Therefore, the present inventors conducted intensive research to solve these conventional drawbacks, and as a result, the present inventors performed immunoexhaustion in capillary tubes and developed a labeled substance that reacted using capillary action and an immobilized label-capturing substance. We discovered that by moving the unreacted labeled substance from the reaction position and capturing and immobilizing each at a different position, it is possible to detect or quantify components in a sample easily and in a short time. Completed the invention.

That is, the present invention provides a method for detecting or quantifying components in a peripheral body by immune reaction, which includes an insoluble carrier having a labeled substance-capturing substance immobilized thereon, and an insoluble carrier retaining at least the present labeled immunoreaction reagent. is filled into a capillary tube, the analyte is inhaled into the capillary tube, and an immune reaction is performed in the capillary tube,
This is an immunoassay method in which a reaction product or a labeled immunoreagent is immobilized by binding with a labeling substance capturing substance, and the immobilized labeling substance is measured.

Components in the specimen to be measured in the present invention include, for example, immunoglobulin, Ben's Jones protein, α-nee antitridicin, α1-microglobulin, α2-microglobulin, β,-microglobulin, hazotoglobin, ferritin, Proteins in biological acids such as transferrin, cellulozolasmin, antithrombin I, myoglobin, myosin light chain, cryoglobulin, and calmodyulin.

GOT, GPT, MuLP, MuCP, LDH, γ-G
TP, creatine kinase, LAP, avalase, macroamylase, cholinesterase, aldolase,
MAO, 5'-nucleotidase, acidophosphatase, OCT, pancreatic enzyme, plasminogen activator, catalase, L-CAT, re? Protein Suno Q-
enzymes such as enzymes, phospholipase A, and phospholipase A. Carbohydrates such as acidic mucopolysaccharides, inulin, neuraminic acid, gangliosides, and mucopolysaccharides. Cholesterol, lil'fo fin, a? Grotein, triglycerides, free fatty acids, phospholipids, bile acids, lipid peroxides and other lipids. Vitamin A, Dsi', ubiquinone, thiamin, riboflavin, vitamin B6, cotinic acid, fibers, vitamin B125 ascorbic powder, inositol type vitamins. Fiplinogun, F.
DP, Zorasminogun, ■Factor, ■Factor, X Factor,
Coagulation factor, zolotone, bozolasten factor, factor ■, factor ■, factor X, zolotrombin, -tron is the coagulation factor of cape globulin.

Paper long hormone, matomezone, luteinizing hormone, follicle stimulating hormone, adrenocorticotropic hormone, LPHlkA
fiH, β-endorphin, enkephalin, thyroid stimulating hormone, prolactin, pasozorecin, neurophyson, oxytocin-active pituitary gland segment substance. 'rs,
Total thyroxine, free thyroxine index, free thyroxine, triodozyronine, linosase Ts
, long-acting thyroid stimulants, calcitonin, thyroglobulin, and other thyroid shunting substances. Catecholamine, metane 7
Phosphorus, normetanephrine, vanillyl mandel modified, homohanillin & 3,4-zohydroxine enylalanine,
3.4-Zohytroxy7-acetic acid, 3-methoxy-
4-Hydroxyphenylethylene glycol, TONOQ
Adrenal medulla/sympathetic nerve secreted substances such as min-β-hydroxylace. Aldosterone, 11-deoxycortecosterone, cortecosterone, 18-hydroxycortecosterone, cortisol, 11-deoxycortisol, 11-hydroxycorticosteroid, 17-hytrooxycorona carp F, 17-kedozoex steroid , dehydroepiandrostene, dehydroepiandrosterone sulfate, androstenedione,
17- Substances secreted by the adrenal cortex such as kedosteroids.

Testosterone, 5α-zohydrotenutonuterone, anthrostandiol, estrone, estradiol, estriol, entetrol, catechol nistrodiene, zolociesterone, pregnanediol, 1
7α-hydrochloride is a substance secreted by the gonads and placenta, such as zoenuterone, pregnanetriol, tricholic gonadotropin, and placental lactogen.

Insulin, proinsulin, C-pedetide, pancreatic glucagon, ganutrin, secretin, CCK-PZ,
Motilin, enteroglucagon, non-creative drugs? Pancreatic and digestive secretions such as lipezotide, somatostatin, substance P, and neurotensin. Herpes simplex virus, varicella-zoster virus, cytomegalovirus, B
B virus antibody, adenovirus, A-Bl influenza virus, Cff1 influenza virus, NoQ
La influenza virus, respiratory syncytial virus, Munzos virus, measles virus, rubella virus, Japanese encephalitis virus, ? Viruses such as Rio virus, hepatitis B virus, hepatitis B virus, line virus, coronavirus, exotic infectious diseases, and rotavirus. antinuclear antibody, anti-DNum antibody, anti-1NA antibody, rheumatoid factor, antiglobulin,
1. IC cells, anti-mitochondrial antibodies, anti-smooth muscle antibodies,
Anti-gastric wall antibodies, intrapillar factor antibodies, anti-striated muscle antibodies, anti-myocardial antibodies, anti-open renal cortex antibodies, anti-thyroglobulin antibodies, anti-thyroid microseam antibodies, anti-inulin antibodies, anti-inulin receptor antibodies, anti-acetylcholine receptor antibodies, etc. autoantibodies.

βIK globulin, clq complement, Tm cells, B cells,
Cellular material of macrophage fathom. Carcinoembryonic antigen, α-fetoprotein, basic 7 ethoprotein, isoferritin, 15-reamin, CRP tumor marker.

Phenobalmital, zolimidone, phenytoin, carbamazepine, parzoloic acid, lidocaine hydrochloride, zogocin, zogitoxin, theophylline, dinpyramite, mexiletene, zololtunorol hydrochloride, diuretics, synthetic nutecide agents, chloramphenicol drugs , aminoglycoside drugs, anti-tuberculosis drugs, methotrexate, opiates, mesatone, valpital, amphetamines, cocaine metabolites, benzozoazevine metabolites, protoxiphene, phencyfridine, cannapinoids, etc. Examples include antigens such as HLA and blood markers and antibodies against these antigens.

In addition to immunoglobulin, the types of antibodies include fc F(ab')z, Fabζ Fab, etc., which are processed using known methods. #1 (Immunology-1-1 Nakayama Shoten (t981)).The same can be said for monoclonal antibodies.

Examples of insoluble carriers include macromolecular proteins, glass, polyethylene, polyethylene, silica, phenol resin, acrylic resin, cellulose, urethane resin, vinyl alcohol, vinyl chloride resin, vinylidene chloride, etc. Spherical or polygonal particles of lysolobilene, iron, fluorine fibers, polyester fat; or fibrous materials such as cellulose, cotton, wool, glass fiber, nylon fiber, vinylon fiber, acrylic fiber, polyethylene fiber, polyester fiber, etc. Can be mentioned. The spherical particles have a diameter of 0.05 to 0.3 mm, particularly 0.1 to 0.
, 17 mm is preferable, and the polygonal particles are preferably 40 to 17 mm.
300 mesh, preferably 80 to 150 mesh.

Examples of labeled immunoreaction reagents include labeled antibodies or antigens against antigens or antibodies in the specimen to be measured, and labeled antigens or antibodies used in the immunocompetitive reaction. As a labeling agent, for example, iodine 7
(7) -f (Example L #i'"'I), I'c, 1Jy
- radioactive isotopes such as ram; enzymes such as peroxidase, alkaline phosphatase, diaphorase, β-D-galactosidase, glucose oxidase, penicillitase; or fluorescein incyanate,
Fluorescent substances such as fluorescein isothiocyanate and rhodamine are included. A method known per se is employed to label the antigen or antibody with these labeling agents (J
, Hlgto, Ch@w, Cytochsm, 22
m 1084 (1974); Imn+unoch@
m1stry a 6 e 43 (1969); same 8s
871 (1971); J, Bio@h@m.

78.235 (1975);F1aor@scentA
i1t1bodymethod@-mucad@mie
pr+ess).

The labeling substance capturing substance may be one that binds to the labeling substance to be measured (labeling agent binding-reaction product or unreacted @R immunoreaction reagent) generated as a result of the immune reaction described below.
Examples include antibodies, antigens, or substances that have affinity for antigens, antibodies, or labeling agents.

To immobilize the labeling substance-capturing substance on an insoluble carrier,
By utilizing the adsorptive properties of the carrier, the labeling substance-capturing substance is bound to the surface of the carrier by a method known per se. For example, when the carrier is glass beads, identification is performed by adsorbing basic amino acids on the surface, bonding a crosslinking agent thereto, and then bonding a labeling substance capturing substance to the crosslinking agent. As the crosslinking agent, glutaraldehyde and maleimides are used. In addition, if the carrier is cellulose-based, a generally known method is to treat the surface with an oxidizing agent such as iron rolls or potassium chloride, and then bind and fix the S visceral substance-trapping substance to this. It is being

In addition, in zero-relaxation, an insoluble carrier holding an S* immunoreaction reagent, a substrate, a coloring agent, a blocking agent, etc. is prepared. As the substrate and the color former, the following ones (substrate dichromator) are used depending on the enzyme of the mm agent. Peroxidase (
Hydrogen peroxide: O-anisidine, 6-)lysine, 4-chloro-1-naphthol, 2,2'-azono-3-ethylbenzothiazoline sulfonate [5nTg), 3-amino-
9-ethyl carno; sol, 2,7-fluorencyamine, 3-methyl-2-penzothiazolinone hydrozone (MBTH), tyramine), alkaline phosphatase (p-nitrophenolline ml!: 4-aminoantipyrine ) or (4-methylumbelliferyl phosphorus wt
:), β-D-galactosidase (0-nitrophenol-β-D-galactoside:) or (4-methyllamperiphery-β-D-galactoside).

In order to hold these labeled immunoreaction reagents, substrates, color formers, blocking agents, etc. on an insoluble carrier, a water or alcohol solution of the above labeled immunoreaction reagents, etc. is mixed with the carrier, and then dried to coat the surface of the carrier. This can be done by attaching a reagent or the like.

In the present invention, as described above, an insoluble carrier on which a WS'IR labeling substance capturing substance is immobilized, an insoluble carrier on which a labeled immunoreaction reagent is retained, and further a substrate, a coloring agent, and a blocking agent are retained as necessary. Fill the capillary tube with the insoluble carrier.

The capillary tube is made of glass or transparent material, or made of cow-transparent paulownia resin, such as vinyl chloride, broken styrene, acryl phthalate R1K.
A material such as IE1 resin can be used, and the capillary diameter is 0.5 to 2 mm, especially 1.0 to 1.1 mm.
, the length is preferably 3 to 20 α, particularly preferably 5 to 15 cfn.

It is necessary that proteins are not adsorbed to the inner wall of this capillary, and it is preferable to treat the inner wall with a neutral buffer such as protein JiL (eg, albumin).

To fill the insoluble carrier with the identified capillary tK iron substance-trapping substance and the reagent-holding insoluble carrier, cotton,
A stopper containing a drinkable ester or the like is filled, and the pressure inside the capillary tube is reduced from the proximal end using an inhalation device such as an aspirator, and the above-mentioned insoluble carrier is inhaled and filled. It is necessary to fill this capillary with at least 1 l of an insoluble carrier immobilized with a visceral substance-trapping substance and an insoluble carrier holding a labeled immunoreaction reagent. It can be filled with an attached insoluble carrier. It is necessary to fill the insoluble carrier containing the labeled substance-capturing substance in such a way that it arrives after the analyte has been subjected to the immunoreaction. Even if one substance that binds to one or both of 41'J is filled with sand to immobilize one of the labeled substances, it is also possible to fill both of them to immobilize both 1a identification substances. ,
It can also be made immobile. In addition, it is essential to separate and pack the insoluble carrier on which the labeled substance-capturing substance is immobilized and the insoluble carrier on which the labeled immunoreaction reagent, substrate, coloring agent, blocking agent, etc. are immobilized. A carrier is used.

In the present invention, when the antigen is the component to be measured in the specimen, the basic loading order of the insoluble carrier on which the labeled substance-capturing substance is immobilized and the carrier on which the labeled immunoreaction reagent is held is as follows. be.

ω Labeled antibody - immobilized antigen @ Marked line antigen - immobilized antibody 1 Labeled antibody - identified antibody - immobilized antibody (back) ■ Labeled antibody - identified antibody - immobilized antigen 6 Labeled antibody -
Identified antibody - immobilized anti-labeled antibody ■ Labeled antibody - (immobilized antibody + auxiliary labeled substance) - (immobilized anti-labeled antibody - auxiliary labeled substance) In addition, it is also possible to use an appropriate combination of these ■ - [F] Demeru.

In order to detect or quantify the amount in the sample using the measuring capillary prepared as described above, the proximal end of the measuring capillary is immersed in the test liquid, and the sample is inhaled into the capillary. . At this time, if the fraction in the specimen is an antigen, the following reaction occurs in the measurement capillary, and the labeling substance is immobilized.

The antigen in the capillary sample in (2) first reacts with the labeled antibody to become an antigen-labeled antibody, and the antigen-labeled antibody and unreacted labeled antibody move.
When it comes into contact with the immobilized antigen, the unreacted labeled antibody reacts to become labeled antibody-immobilized antigen and is immobilized.

The antigen in the capillary sample of @ comes into contact with the labeled antigen and the identified antibody, and the two react while competing, becoming the labeled antigen-fixed antibody and immobilized.

The antigen in the capillary sample at θ reacts with the labeled antibody to become an antigen-labeled antibody1, and the antigen-labeled antibody and unreacted O-labeled antibody move and come into contact with the immobilized antibody, and the antigen-labeled antibody reacts. The identified antibody-antigen-labeled antibody is immobilized.

The antigen in the 00 capillary specimen reacts with the labeled antibody to form an antigen-labeled antibody, which then comes into contact with the immobilized antibody (front) to become immobilized antibody-antigen-labeled antibody. Additionally, if the antigen in the sample is present in excess of the labeled antibody, the excess antigen competes with the labeled antigen and reacts with the immobilized antibody (later), becoming the labeled antigen-identifying antibody and immobilized. .

The antigen in the capillary specimen reacts with the labeled antibody to form an antigen-labeled antibody, which then reacts with the fixed antibody to form the immobilized antibody.
・Antigen becomes immobilized as a labeled antibody. Further, the unreacted labeled antibody moves along the length and reacts with the immobilized antigen to become immobilized as immobilized antigen-labeled antibody.

The antigen in the capillary specimen of G reacts with the labeled antibody to become antigen-labeled antibody 9, and then comes into contact with the immobilized antibody to form the immobilized antibody-antigen-
It becomes a labeled antibody and becomes immobilized. In addition, unreacted labeled antibodies and some antigens that were not immobilized l! The visceral antibody further moves and reacts with the immobilized anti-labeled antibody to form the immobilized Kitamakura-labeled antibody.
It becomes a labeled antibody and becomes immobilized.

The antigen in the hair 11al tube specimen reacts with the labeled antibody to form an antigen-labeled antibody, and then comes into contact with (immobilized antibody and auxiliary label) and (
Immobilized antibody (auxiliary specimen) - antigen - becomes immobilized as a labeled antibody. In addition, the unreacted labeled antibody further moves and reacts with (immobilized anti-labeled antibody auxiliary labeled substance) to become a labeled antibody (immobilized anti-labeled antibody auxiliary labeled substance) and is immobilized. In this method, label binding alone does not produce a label, but a label can be released by combining it with an auxiliary substance. The auxiliary standard fabric is, for example, a material that is attached to the chest of a NADH1 luminescent substance and its excited substance for glucone oxidase, which produces the substrate of /e-oxidase from glucose by a fermentation kite reaction, and the holoenzyme diafoduse.

If the labeling agent immobilized in this way is a radioactive isotope, the amount of the labeling substance is determined by cutting out the part and measuring it with a gamma ray measuring device or the like. Further, when the labeling agent is an enzyme or a fluorescent substance, the labeling substance is colored by the substrate filled in the capillary, the coloring agent, or the excitation wavelength, so that the labeling substance can be measured based on the coloring. Further, the substrate and the coloring agent can also be inhaled after the test liquid is inhaled.

Incidentally, as the test liquid, blood, serum, plasma, saliva, body fluids such as urine, waste water, etc. can be used. If a component that inhibits this reaction is present in the sample, the inhibitor can be activated or removed in advance, or the sample can be fixed or rewritten by diluting it with frozen IT that does not affect the measurement.

As mentioned above, the method of the present invention allows the immune reaction and the capture of the labeling substance to be carried out in a capillary tube, so it has various advantages as follows.

- Since all reagents are filled in the capillary tube, there is no disadvantage of having to prepare each reagent or discarding excess reagents during measurement as in conventional methods.

■ Components in a sample can be detected or quantified simply by immersing the proximal end of the capillary tube in the test liquid, so the operation is extremely simple and can be performed by individuals without the need for specialized skills. No equipment or instruments are required, and measurements can be made in a very short time.

■ Therefore, measurement at the hospital bedside is possible.

■ Only a very small amount is required, for example, by simply cutting the earlobe, placing the proximal end of a capillary tube on it, and inhaling a small amount.
Components in blood can be measured.

■ Capillary tubes can be stored as they are after measurement, and can be easily disposed of.

Next, an example will be given and explained.

Example 1 (1) Reagents and equipment■ Albumin-sucking galanu capillary glass capillary! (diameter 1mm, length 100mm),
Through 0.01M + 7-phosphate buffered saline (PB8) (PH7,2) of Bovinalpmin 20q/-,
Remove excess liquid by suction and dry.

■ Glass beads (GB,) diameter 0.17 mm ■ Alpine [GB (CB, BS) glass beads (diameter 0.17 mm) (GB), Bovin Alpine 20
Soak the glass peas in 11I/- pnsK with purified water for 2 hours.
After washing ~3 times, dry.

■ Using commercially available polyester fiber ■ Cyanidisin mixture QB, BSA 0-cyanidisine salt 6.25- ethanol solution 2
- GB and BS 1f are mixed and dried.

■ Pile tip IgG-binding peroxidase (αRb-IgG
-HRP) Dissolve 4Mg of peroxidase in water, add 0.1M sodium periodate 0,2-, and react for 20 minutes at the lowest temperature. The reaction is permeated overnight against 1 mM acetic acid lIk buffer pH 4,0. After dialysis, 0.2M carbonate buffer @
pH 9.5 was added to bring the dialysate to pH 9.5. Immediately, goat anti-rabbit IgG, 10'I/wl, 0.01 carbonate buffer pH 9.5 was added and reacted at room temperature for 2 hours.

After the reaction was completed, 0,1- of hydrogen chloride was added and heated to 4°C.
, and reacted for 2 hours. After finishing, Sephacryl 5-200
(7 manufactured by Almacia) (column diameter: 2.5 cm, length: 30 t:m), and the eluate was added to 0.01 MP
Filter through BB. Remove the polymer part that flows out first9.
Lyophilized and stored.

(2) Add 8 g of GB and B8 mu to a freeze-dried product equivalent to αRb-IgG-HRP l- prepared using αRb-IgG-HRP mixture QB, B8 mu and mix uniformly in a mortar.

■ Add rabbit IgG-bound GB (Rh-xga-〇) Glass Peas (GB) 5fK concentrated sulfuric acid 5- and let it cool to room temperature for 5 minutes.
Leave for a minute. This was washed with purified water until the washing solution became neutral, and 30 wI/- of Reem-lysine was added to it.
Mix for 1 hour at room temperature. After washing with H water, 2.5% geltaraldehyde was added and reacted at room temperature for 1 hour. After washing with purified water, rabbit IgG (5 mi/la) 2- is added, reacted at room temperature for 3.5 hours, and dried.

(2) Preparation of capillary for measurement Make five marks along the proximal end of the aldimine sorption glass capillary, first at intervals of 5 mm and then at intervals of 3.5 mm. first 5
Pack the polyester fiber (a-1) into the mm. Vacuum this base end? Connect to the capillary tube to depressurize the inside of the capillary, and add cyanidisin mixed QB to the next 3.5 mm.
Fill B8 mu (a-2) by suction. Similarly, GB.

BAA(a-3), αRb-IgG-HRP mixed G
B.

BAA (a-4), GB, B8mu (a-5), Rb
- IgG-◯ (a-6) is sequentially filled. remaining top 5
Fill with GB (a-7) to the bottom of mm and finally again? Fill the remaining 95 mm+ with Lientel Hirofi.

(Figure 1) (3-1) Determination of human serum and rabbit serum Prepare two measuring capillaries, dip the proximal end of one in human serum and the other in rabbit serum, and perform the test shown in Figure 1. The same serum is inhaled until (a-j). The amount of serum at that time was about sp pt, and the time required for inhalation of VC was 1 minute. Next, the proximal end of this capillary tube is immersed in 0.003% hydrogen peroxide PB8, and this is inhaled to the vicinity of the upper end of the capillary tube (8 to 9 cm from the proximal end). The simulated time was 15 minutes. As a result, in the case of human serum, Rb-l1G-○ [(
--6)] was colored brown, but no coloration was observed with rabbit serum.

(3-2) Determination of human blood and rabbit blood containing anticoagulant (EDTM) The same procedure as in (3-1) was carried out except that 0.03% hydrogen peroxide PB8 was used as the substrate solution.

As a result, the rabbit IgG-bound GB portion was colored brown in human blood, while no coloring was observed in rabbit blood.

Example 2 (1) Reagents and instruments (1) Albumin adsorption glass capillary A reagent was prepared in the same manner as in (1) (2) of Example 1, except that a glass capillary with a diameter of 1.5 mm and a length of 125111 KII was used.

■ Glass peas (GB) (diameter 0.17 mm)
■ Algumin a-wearing G, B, (GB, B8) Example 1
Use the same one as in (1) ■.

■? lyester fiber ■ Cyanidisin mixed GB, B8mu The same material as in (1) (■) of Example 1 was used.

■ Rabbit 1. G-linked peroxidase (lb-IgG-H
RP) Prepared in the same manner as in (1) (■) of Example 1, except that 4C of rabbit IgG was used for the goat stake tip 1gG.

■ Rb-IgG-HRP mixed GB, B Add GB, B to the freeze-dried product equivalent to Rb-IgG-HRP 1- prepared by 8 people ■
Add Bum 16f and mix uniformly in a mortar.

■ Pile tip IgG binding GB (αRb-IgG-○) Rabbit I
It was prepared in the same manner as in Example 1 (1) (■) except that goat stake IgG was used instead of GG.

■ Hole holes? A rubber band whose thickness matches the id capillary? A hole is drilled in the top of the id.

[Phase] IQ Rin, EDT M mixed QB, BS person Heno 9
Lin 10q, EDTM 2 1.15f, GB.

Mix Bgum 8.84F uniformly in a mortar, and
Take 9 and then G11. Add B8m 9 and 8f and mix uniformly in the same way.

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube.

First 5 mm of polyester fiber (b-1) Next 3
．． 5 mo+nihe, e-rin/EDTM mixed GB.

18 people (b-2) Cyanidisin mixture Q in the next 3.5 mm
B, B8mm (b-3), GB, B with 3.5mm each
Person B (b-4), Rb-IgG-HRP mixed GB.

BSA (b-5), QB, BSA (b-6), αRb-
Add IgG-○ (b-7) and add Gn to the remaining 5 points below the top end.
(b-8), and the remaining 5 mm was filled again with polyester fibers (Figure 2).

(3) Measurement of human and Marutake fresh blood Prepare two capillary tubes for measurement, and attach nine rubber droppers with holes at their upper ends.

Attach the proximal end of one to the cut part of the rabbit's ear, and attach the proximal end of the other to the incised part of the rabbit's ear.
Inhale slowly using the id. The amount of blood at that time was approximately 15 μt, and the duration of the test was 20 seconds.

The proximal end of each capillary was then treated with 0.003% hydrogen peroxide PB8.
immersed in K, and soak it near the top of the capillary (9 to 10 minutes from the proximal end).
on) up to Su? Use the id to inhale. It took me 7 minutes to complain about this.

As a result, anti-Rb-IgG ((b) in Figure 2) was detected in human blood.
-7)) was colored brown, but the rabbit blood was colored lighter than the former.

Example 3 (1) Reagents and instruments ■ Albumin-adsorbing glass capillary The albumin-adsorbing glass capillary described in (1) (■) of Example 1 was used.

■ Glass beads (GB) (diameter 0.1 mm) ■
Albumin adsorption GB (QB, 18 people) diameter 0.17
It was prepared in the same manner as in (1) (■) of Example 1, except that 0.1 mm glass beads were used for the replacement of n+m.

■ Absorbent cotton ■ Anti-C-reactive protein-binding peroxidase (αCRP-■
RP) Example 1 except that the goat stake tip IgG was replaced with goat anti-CRP.
Create it in the same way as (1) ■.

■αCRP-Dish P mixed GB, B8 μM αCRP, -HRP 1-Add GB and BS 8f to the freeze-dried product of the phase and mix uniformly in a mortar.

■ αCRP-binding GB (αCRP-〇) This example (1
) Goat αC as a substitute for glass beads and rabbit IgG
It was created in the same manner as in (1) (■) of Example 1, except that RP was used.

■ Differential coloring liquid 0.03% hydrogen peroxide, 0.25% gelatin and 003
%o-) PBXo containing lysine (2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube. At first, 5 mm was degreased m (c-1), and then 3
．． CB of 5 mm each, 18 people (e-2), αCRP-H
RP mixed GB, BS (e-3), GB, BSA (e-
5) Set and optically fill αcup-0(c-5). Fill the remaining soil with GB (e-6) to 5 mm below the edge, leaving 5 mm remaining.
Pack with K-F+) degree absorbent cotton (Figure 3).

(3) Determination of the presence or absence of CRP Prepare four measuring capillaries, and connect the proximal end of each to (1)
Rabbit serum, (II) healthy human serum, (1D son serum, (IV
) Dip each sample in serum containing cup 5 q/at, and inhale the test solution up to (c-'s) in Figure 3.

The amount of liquid to be tested at that time was approximately 5 μt, and the time required for inhalation was 30 to 60 seconds.

Next, the proximal end of this capillary tube is immersed in the substrate coloring solution, and this is inhaled to the vicinity of the upper end of the capillary tube (8 to 9 cm from the proximal end). This took only 20 minutes. the result(
In the case of 1) and 0), αCRP-〇 [(c-5 in Figure 3)
)) No coloring was observed in the area, but green coloring was observed in the same area in I[) and (IV).

Example 4 (1) Reagents and instruments ■ Albumin-adsorbing glass capillary The albumin-adsorbing glass capillary described in (1) (■) of Example 1 was used.

■ Glass beads (GB) (diameter 0. l aim
) ■ Albumin adsorption QB (GB, BS) Example 3
(1) ■Use albumin adsorption GB.

■ Cyanisidine mixed GB, B8 diameter 0.17
Created by the same procedure as in Example 1 (11.■) except that 0.1 mm glass beads were used for the mm diameter.

■ Cotton-bound glucose oxidase (C0D-O) Cotton wool 780111i and sodium periodate 214M
Take 1, add 40 liters of water made in N, and mix for 1 hour at room temperature. After the reaction, wash well with purified water, add 10q of glucose oxidase, mix at room temperature for 3 hours, wash well with ice cubes, and dry.

(2) Glucose mixed with absorbent cotton (Gla-mixed absorbent cotton) Take 100019 absorbent cotton and mix with 1 ooq of glucose. Next, add 2-'t- of water made by N and mix and dissolve well, and then dry.

■ Anti-fibrin degradation product binding peroxidase (αFD
P-HRP) Prepared in the same manner as in Example 1 (1) (■) except that 1gG of goat stake tip was replaced with mouse anti-FDP C monoclonal antibody-1).

■ αFDP-HRP mixed GB, BS A dried product equivalent to αFDP-HRP l- prepared by ■ GB, BS
Add 8g of milk and mix evenly in a mortar.

■ αFDP-bound OB Created by the same procedure as in Example 1 (1) (■) except that rabbit IgG was replaced with mouse αFDP C monoclonal antibody-2).

[Stock] Glutathione Mixed QB, BAA Glutathione 1
Mix 0wg, GB, and 88 people 5f uniformly in a mortar.

■ Absorbent cotton (2) #J of the measuring capillary tube! From the proximal end of the albumin-adsorbing Galanu capillary made by No. 4, first two with a width of 5 mm, then 3.5 am l! Make six marks at l intervals. Fill each tube with two fibrous reagents from the proximal end. First, move GOD-〇(d) from the base end to 10 mm from the back.
-2), then G1m from the proximal end to 95ml1lIi
Fill each piece with mixed absorbent cotton (d-1) using a needle.

3.5 without suctioning from the proximal end! Ill! l length, cyanisidine mixed Gi1. B8mu (d -3), a
n, BsA (a-4) glutathione mixed GBJsA (
d -5), αFDP4RP (d -6), QB, 8
8 people (a-7) and filled with αFDP-〇 (d-8). GB (d-9) was filled up to 5 mm below the top of the remaining part, and 5n+mVc absorbent cotton was tied to the remaining part (Figure 4).

(3) Measurement of urinary FDP FDP (D!) At 50 pf/urine volume, dip one of the nine tubes into MIG human urine and inhale at t near the upper end of the capillary (8 to 9 cm from the proximal end). .

The amount of urine at that time was approximately &5 μt, and the time required to achieve this was 9 minutes. As a result, αFDP was found in FDP-added urine.
-0 ((d-8) in Figure 4) was colored brown, but no coloration was observed in the urine of a healthy person.

Example 5 (1) Reagents and equipment ■ Albumin-adsorbing vitreous capillary #ll tube The Aldazun-adsorbing vitreous capillary #ll of (1) of Example 1 was used.

■ Glass peas (GB) (diameter 0.17!IIm)
■ Albumin adsorption Gn (QB, BSA) Example 3
(1) ■ GB and B8 are used.

■ Broken efthyl fiber ■ Cyanidisin mixture QB, BS Person Example 1 (1) ■ Cyanisine 24 combination Gn/BSA
use.

■ Rabbit IgG-binding peroxidase (Rh-IgG-H
IIP) Rb-4gG-HRP of (1) of Example 2 was used.

(2) Rb-IgG-HRP mixed QB, BSM Rb-IgG-HRP mixed QB of (1) of Example 2.

Use B11.

■ QB-conjugated rabbit IgG (Rb-IgG-○) Example 1
Use of Rb-IgG-○ in (1) (a>).

(2) l# of measurement capillary tube] The following capillary tube was prepared by performing the same operation as in (2) of Example 1.

First 5mmK=! t'liennetel fiber (*-1) followed by 3.5+nm each of cyanisidine mixed QB.

BsA (@ -2), GB, BSA (@ -3),
Rh-IgG-HRP mixed GB, BSA (・-4)
GB, 18 (・-5), and Rh-IgG-○ (1
1-6). GB (
a-7), and fill the remaining 5 mm with ethyl fiber (Figure 5).

(3) Determining the presence or absence of anti-Rh-IgG Prepare two measuring capillaries, and connect the proximal end of each to (1)
nb-IgG immunization, goat serum, (IllαRb-IgG
Dip the sample into goat serum (Ophthalonychia fg) that does not contain any phthalate, and inhale the test solution up to (@-3) in Figure 5. The amount of test solution at that time was about 3 μt, and the waiting time was 20 seconds.

Then, the proximal end of this capillary was filled with 0.03% hydrogen peroxide PB8.
near the upper end of the capillary (8 to 9 from the proximal end).
cm). The time taken for this was 20 minutes. As a result, goat serum (1
), the Rb-IgG-○ [Fig. 5 (・-6)] part was colored brown, but no coloration was observed in goat serum (1), which is ophthalmic and does not contain αlb-IgG. .

Example 6 (1) Reagents and equipment ■ Albumin suction galanu capillary The albumin suction $f27 capillary of Example 1 (1) ■ was used.

■Albumin absorption: 2 Nupies (QB, BS people) Example 3 (1) GB, 18 people were used.

■ Absorbent cotton ■ Anti-human albumin-binding peroxidase (αH8mu-HRP) Prepared in the same manner as αRb-IgG-HRP in (1) of Example 1, except that rabbit anti-human albumin was used instead of goat stake tip IgG.

■ αH8mu-1 (RP mixed GB, B8mu αCRP-H
It was created by operating in the same manner as (1) (2) of Example 3, except that (2) of this example was used instead of RP.

■ αH8mu-binding QB (αHamu-Q) Goat αCRP
The sample was prepared in the same manner as in Example 3 (1) (■) except that the sample was replaced with rabbit α-H8.

■ Substrate coloring solution 0.03% hydrogen peroxide, 0.25% gelatin, and 0.03% hydrogen peroxide.
0.03% containing 2,2'-azonozo(3-ethylbenzthiazoline)-6-nurphonic acid. Q1M phosphate buffered saline.

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube.

Initially, the flirt (t − i )% was set to 5 mm and 7
GB, BS Mu (f-2) in mm length, αHB mu HRP mixed GB in 3.5 mm length, B8 mu (f-3), 7 m
QB + BS person (r-4) with m length, a, α with smm length
Fill J (8 people - 〇 (f-5). Remaining top edge 5 mm
GB, BS (f-6) f, fill to the bottom, 5 mm remaining
Fill with absorbent cotton again. After finishing, mark 5 points along the proximal end (Figure 6).

(3) Determining the presence or absence of human albumin in drainage fluid Prepare three test tubes for measurement, and insert (1) human serum 1 into the proximal end of each tube.
sg/PB8 z, (1) Drainage 1.1) Apply the test solution to the drain wL2 respectively and inhale the test solution up to f-5 in Figure 6. The amount of test liquid at that time was about 9 μt, and the time required for this was 1 minute. Next, the proximal end of this capillary tube is immersed in the substrate coloring solution, and this is inhaled to a distance of about 5 m from the capillary proximal end. This took 15 minutes. The result is (
1) and (...), αH ROM-○ [(f-S in Figure 6)
) had blue coloring fc, and 1) had no coloring. From this result, it was determined that the effluent (1) contained human AYVF.

Example 7 (1) Reagents and equipment ■ Albumin e, vitreous capillary Albumin adsorption vitreous system of Example 1 (1) ■ l1ll
Use t#t.

■ Garnu beads (GB) (diameter 0.1 mm
) ■ Albumin adsorption GB (GB, 18 people) Example 3
Use GB and BSA in (1) ■.

■ Ribbed Ntel fiber ■ Cyanidisin mixed GB, BS people Cyanidisin mixed GB of (1) of Example 4.

Uses B111. , ■ Anti-α7 ethoprotein (homemade monoclonal antibody)
Conjugated peroxidase (alpha murp-unp) goat stake IgG was prepared in the same manner as described in (1) (2) of Example 1.

■ αAFP-HRP mixed GB, B8mu αCRP-H
It was prepared in the same manner as in Example 3 (1) (2) except that RP was replaced with α-am FP-HRp of this actual cell example (2).

■ Anti-α-fetoprotein la-conjugated GB (αAFP −
Q) Goat αcnp'4 Maunu anti-αfetoprotein (
The antibody was prepared in the same manner as in Example 3 (1) (2) except that monoclonal antibody-2 (self-produced) was used.

■α7-tonorotein-bound GB (MuFP-〇) Prepared in the same manner as in Example 3 (1) (■) except that goat αCRP was replaced with α-fetoprotein (manufactured by Batori Jujutsu Co., Ltd.).

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube. First 5 mm K? 'IJ Esther Coffin
(g -1>, hjc and 3.5 mm each) Cyanisidine mixed cn, nsum (g -2), GB, BSA (g -
3), αmu FP-HRP mixed QB, B8mu (g-4),
GB, BSA (g-5), alpha FP-0 (g-6)
, GB , 118mu (r-7), and mu FP-○ (g
-8). GB (g
-9) packed, remaining J) 5mmK again? Stuff with liennetel hilofi (Figure 7).

(3) Titll measurement of α-fetoprotein in serum Prepare 6 capillary tubes for measurement, and connect the proximal end of each to (1) α-fetoprotein Of/− serum, (Illαlllton ethoin 10Qng/ml serum, α-fetoprotein 10μ
2/- serum, M healthy human serum, (Vl)J! Dip it in the serum of person i, and inhale the test solution at (g-8)t in Figure 7. At that time, the test sample was about 10 μt, and the time it took to do so was 2 minutes. Next, the proximal end of this capillary was 0.003
% hydrogen peroxide PB1i1 Vc and inhale this to near the upper end of the capillary (8 to 9 on from the proximal end).

The time I spent on this KW was 25 minutes. As a result CIl#
iα Mupp-○ [Figure 7 (g-6)) is not colored, MuF
P-○ (Figure 7 (g-8)). (Ill
is lightly colored on αmu PP-○ [Fig. 7 (g-6)), and A
Color according to FP-0C Figure 7 (g-8)). (M
Coloring was also observed in α-mu FP-〇 (Figure 7 (g-6)). (IV) is αmu FP-〇 [Figure 7 (g-6)
) was darkly colored, and mu FP-〇 (Figure 7 (g-8)) was lightly colored.

(9) had the same results as (1), and (Vl) had the same results as ■).

According to the above results, α-fetoprotein was not detected in healthy subjects, and the patient's serum contained approximately 100 nf/− of α-fetozolotin.

Example 8 (1) Reagents and instruments ■ Albumin-adsorbing glass capillary The albumin-adsorbing glass capillary described in (1) (■) of Example 1 was used.

■ QB, (j [If diameter Q, l 7 mm) ■ G
B. 88 people used GB and BSA in (1) of Example 1.

■? Lientel fiber ■ Cyanidisin mixed GB, Bllum Cyanidisin mixed GB of Example 1 (1) (■).

Uses BSA.

■ Sodium perborate mixture QB, B8, 12q of sodium perboronate and GB, BS, 10f are uniformly mixed in a mortar.

■ Glutathione mixture QB, BS person Glutathione 40■ and QB, BS Mu 10g are uniformly cashed in a mortar.

■ Anti-fibrin degradation product (monoclonal antibody-1) conjugated peroxidase (αFDP-HRP) goat stake tip I
The procedure was repeated in the same manner as in (1) (2) of Example 1, except that gG was replaced with Maunu anti-fibrin degradation product (self-produced monoclonal antibody-1).

■ αPDP-HRP mixed GB, BB IgG-HRP
(1) of Example 1 except that αJ'DP4RP was replaced with αJ'DP4RP.
Create by operating in the same way as ■.

α-FDP binding CB (αFDP-〇)Rb I g
Produced in the same manner as (1) of Example 1, except that c was replaced with a mouse anti-fibrin degradation product (homemade monoclonal antibody-2).

(lp FDP-conjugated GB (FDP-〇)
Create by operating in the same way as ■.

[Phase] Mix albumin mixture GB, Ba 20cm and QB, B8 1f uniformly in a mortar.

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube. First 5mmK polyester fiber (h-1),
Next, 3.5 mm each of albumin mixed GB and BS people (h
-2), cyanidisin mixed GB, BSA (h -3)
, perborated bulk sodium mixed CB, B8A (h −4
), glutathione mixed GB, BSum (h-s), GB,
BSA (h -6), αFDP-HRP mixed QB, B
8 people (h-7), GB, 88 people (h-8), αFDP
-○ (h-9), GB, 88 people (h-10), and F
DP-〇 (h-11) is filled, and finally Gn (h-1
2) to the top end of the capillary tube + 5 m below, and fill the remaining 5 m again.
Fill with m K polyester fibers (Figure 8).

(3) Measurement of urinary FDP Prepare four measurement capillaries, and connect the proximal end of each to (1).
Normal urine, (il) FDP s Oμ9/− spiked urine, (1
) rDpl cape/-added urine, (■) Immerse in patient urine, inhale the test solution, and place near the upper end of the capillary (8 to 9 cm from the proximal end).
). The amount of urine at that time was approximately 25 pt, and the time required was 20 minutes. The result (D is
αPDP-〇 [8th ward (h-9)) without coloring, FDP
-〇 [Figure 8 (h-11)] A thick brown coloration appeared.

In (...), a brown color appeared in αFDP-○ [Figure 8 (h-9)), and a brown color appeared in FDP-0 (Figure 8 (h-11)). In (I), αFDP-〇[弗8(h-9)
) has a rough coloring, and FDP-〇 [Figure 8 (h-11
)) was slightly colored. It was determined that (P/) had an FDP of about 50 μf/gd from the result of (It).

Example 9 ■ Albumin adsorption capillary tube The albumin adsorption capillary tube (1) of Example 1 was used.

■Glass beads (GB (diameter 0.17 mm)■
Albumin-adsorbed galanu beads (Gll, BOA) Albumin-adsorbed CB of (1) in Example 1 was used.

(4) Z-liester fiber ■ Cyanidisin mixed GB, BSA Cyanidisin mixed GB of Example 1 (1) (■).

Uses BSA.

■ Anti-α-fetoprotein-binding peroxidase (αA
FP-HRP) α7 Etozo p-9 of (1) of Example 7 was used.

■ αAFP-HRF mixed GB, BS person αRb-I g
It was prepared in the same manner as in (1) (2) of Example 1, except that G-HRP was replaced with αmuFP-HRP.

(2) Anti-α-fetoprotein binding GB (α5FP-〇) was prepared in the same manner as (1) in Example 1, except that rabbit IgG was replaced with goat FDP.

■ Antiperoxidase-conjugated GB (αHRP-○) rabbit I
Example 1 except that gG was replaced with goat antiperoxidase
Create it in the same way as (1) ■.

(2) Measuring capillary i! 14iIR Example 1 (2
) to create the following capillary tube. First 5mmKf! Lientel fiber (1-1), thread 3.5
mm each, cyanidisin mixed GB, B8 mm (s-2),
GB, BOA (1-3), alpha FD4Rf' mixed QB
, B8mu (1-4), GB, BOA (i-5),
α FP-0 (1-a), QB, BOA (t-7)
, and filled with αHRP-〇(1-8). Pack QB (1-9) to 5 mm below the top edge, leaving 5 mm remaining.
again? Fill with liennetel rut fibers (Figure 9).

(3) Measurement of α-fetoprotein in serum Prepare three measuring capillaries, and connect the proximal ends of each to (1) α-fetoprotein Qf/d, [[jl, (It) α-fetozolotin 500nf/-serum]. , (ill) alpha-fetoprotein 10 μt/- serum, and the test solution is inhaled up to the 9th N (i-8). The test solution at that time was about 10 It,
The time lost was one minute. Next, the proximal end of this capillary tube is immersed in 0.003% hydrogen peroxide PB8, and this is inhaled to the vicinity of the upper end of the capillary tube (8 to 9 mm from the proximal end). The time we met was 20 minutes. As a result (1), there was no coloration in α-muFP-0 [Fig. 9 (1-6)], and α-HRP-0 was not colored.
0 [Fig. 9 (*-8))] Dark brown coloring was observed. (I
l) Color α person FP-〇 [Figure 9 (1-6)) and α
HRP-0 (Figure 9 (1-8)) was also colored. 1) is lightly colored on α-mu FP-0 (Fig. 9 (1-6):),
HRP-◯ [Figure 9 (t-8)) was also colored. From the above, it was possible to distinguish the α-fetoprotein moldiness by color between [Figure 9 (1-6)) and [Figure 9 (t-8)].

Example 10 (1) Reagents and equipment ■ Aldimine adsorption glass capillary Example 11/7 Aldimine adsorption glass 2: x capillary was used.

■ Galanupees (QB) (0.1mm diameter) ■
Albumin adsorption QB (GB, BOA) Example 3 (
1) Use GB and BBmu of ■.

■ Forgiving Ntel Fiber ■ Cyanidisin mixed GB, B8mu Cyanidisin mixed Gb of (1) of Example 4.

Use BS people.

■ Anti-α-fetoprotein (monoclonal antibody-1,
Homemade Fab')-conjugated peroxidase (alpha Fp-n
np) peroxidase 10 mg/l, 5 mg, 0.1 M phosphate buffer (p7.0) and N-succinii 4-(
N-Maleitomethyl) cyclohexane-1-carbone-) (SMCC) l 0 ■ The 10,2-zomethyl nulphoxide solution is added dropwise with stirring. Mix and react at 30°C for 1 hour. Centrifugation (3000 rpm 1
0 minutes) Remove excess reagent that precipitates. Maleimidoperoxidase is separated by Glu filtration (Sephadex G-25
0.1M phosphate buffer pi6.0). Separately reduced N1F
(ab') is prepared and prepared according to a conventional method. n made F(a
b')! (Derived from homemade monoclonal antibody-1) 10w
Add mercazotoethylamine chloride to a final concentration of 0.01 M to I/d 0.1 M phosphate buffer (CPH 6.0), and react at 37° C. for 90 minutes. Reduced Fab'
Sephadex G-25, 0 containing 5mM 1DTM
．． Separate by filtration with 1M phosphate buffer pua,o. Enzyme-labeled Fab' was converted into Ultroguru C-44 (LK
(manufactured by B)) Glue filtration with 0.1M phosphate buffer pH 6.5. Separationffl! ! ! Freeze-dry things.

■ αmu FP-HRP mixed QB, B8mu αCRP-HR
Example 3 ((
Create by operating in the same way as in 1) ■.

■ Anti-α-fetoprotein binding cn (α-muFP-0)
Example 7 (1) (2) is used.

■ Albumin peas-bound gluco-7 oxidase mixed GB, 18 people (COD-0) Take 100 sv of glucone oxidase and 400 sv of Davin Alpmin, 0. Q2M acetate buffer pH5, o
Dissolve in 5tnt. Add two parts of 2.5% geltaraldehyde to this, mix slowly, and leave to stand for 1 hour. After standing, homogenize well, centrifuge, and collect the precipitate.

Add 0.1M lysine 20- to the collected precipitate and leave it overnight, then wash with water and remove the precipitate. Use after drying. Mix Crystal 7 Misaki, GB, and BS Mu 1f uniformly.

[Phase] Glutathione mixed GB, B8 mu Example 4 (1
)'s [phase] glutathione mixed GB.

Uses B8 mm.

(Glucone 1 Ml/gdPB8Q) Preparation of capillary tube for measurement The following capillary tube was prepared in the same manner as in Example 1 (2). First 5m11BVC & Lientel fiber (ji)
, followed by 3.5 mm each of cyanidisin mixed GB, 18
Mu (j-2), GB, B5 Mu (J-3), α Mu yp
-nRp mixed GB, B8mu (j-4), glutathione mixed GB, B8A (J-5), C0D-〇(j-a
), and fill αmFP-o(j-'y). Fill the remaining GB (J -8) to 5 mm below the top edge, leaving J5
mm again? Fill with Lientel fiber (Figure 10).

(3) Measurement of α-fetozolotin in serum Prepare three measuring capillaries, and connect the proximal end of each to (1) α-fetoprotein Of/v blood i11, ([) α7 ethoprotein 500nf/-serum, (4) α7 Etozo U Tin 10μ
t/- serum, inhale the test solution up to (j-7) in Figure WJ9. The sample solution at that time was about 9 pL, and the time it took to drain was 2 minutes. Next, the proximal end of this capillary tube was immersed in Glucone 1q/5dPB8, and this was placed near the top end of the capillary tube (
8 to 9 points from the proximal end. ). The time lost was 25 minutes. As a result, (1) is αmu FP-0 (
Figure 10 (j-7): ) is not colored, (1) is αmu FP
-○ [Fig. 10 (j-7))], and the button is (tAFP-○ [Fig. 10 (j-7>)) Kllll<
Colored. From the above [Figure 10 (j-7)], α
It was possible to differentiate the concentration of fetzolotine.

Example 11 (1) Reagents and equipment ■ Aldimine adsorption capillary The albumin adsorption capillary described in (1) (■) of Example 2 was used.

■ Galanupease (CB) (0.17 mm in diameter)
(2) Albumin-adsorbing gums (GB, B8) GB and B8 (GB, B8) in (1) of Example 1 were used.

■ Absorbent cotton ■ Pile tip
(αRb-IgG-FITC) Goat stake IgG
5■/- Add 0.1-00.5M carbonate buffered saline (pH 9.5) to the saline and make #M%A. Separately prepared FITC2119/do, 05M carbonate buffered saline (pH 9,5) was placed in a 20-beaker, and the IgG solution was placed in a dialysis tube, immersed, and reacted overnight at 4°C. let Glu filtration after reaction (Sephap+)
Fluorescently labeled compounds are separated in a solution of 0.01 M phosphorus-rR buffered saline).

Freeze-dry the separated ah.

■ αRbIgG-FITC mixed GB, GB in the lyophilized product equivalent to αRb-IgG-FITC1- prepared by BS person ■
, BS Mu 2f and mix uniformly in a mortar.

(2) RbIgG-binding QB (αRbIgG-○) Rb-IgG-○ in (1) of Example 1 was used.

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube. Cotton wool (k-1) for the first 5 mm, QB and BSA (k-2) for the next 3.5 mm, and 3.5 mm for the next 3.5 mm.
αRbIgG-FITC (k-3), GB,
BSA (k-4), and αRbIgG-0 (k-5
), and fill the remainder with c, n, n5A (
k-6), and the remaining 5 mm was filled with absorbent cotton again (
Figure 11).

Two measuring capillaries are prepared, the proximal end of one of them is soaked in human serum, and the other capillary is soaked in rabbit serum, and the capillaries are inhaled to near the upper end of the WJ11 diagram (8 to 9 cM from the proximal end). The amount of serum at that time was approximately 6 μt, and the time required was 30 minutes.

When the inhaled hair Mlt was applied to an excitation pump, fluorescence was observed in the case of human serum (Fig. 11 (k-5)), but fluorescence was observed in the case of rabbit serum.

Example 12 (1) Reagents and equipment The same materials as in Example 1 (1) were used for (1) to (2).

■ Hole hole? IF The same material as in Example 2 (11.■) was used.

■ Use the same [phase] as in (1) of Heno9-rin eEDT human mixed QB and B8-mu Example 2.

(2) Preparation of capillary tube for measurement Make five marks from the proximal end of the Aldavan adsorption capillary at intervals of 5 mm at first and 3.5 mm from then on. first 5
At +nm? Fill with Lientel fiber (a-1). Vacuum this base end? The pressure inside the capillary is removed by connecting it to the capillary tube, and the next 3.5 mm is filled with heno-9-phosphorus-EDTM mixture GB, B.
Cyanidisin mixture GB, B 8 mm (a-2) is filled by suction at a distance of 3.5 mm next to SA (a-1').

Similarly, GB, BSA (a-3) αRb-IgG-H
RP mixed QB, BSA (a-4) GB, BSA,
(11-5) Sequentially fill with Rb-IgG-〇(&-6). Fill (a-7) with the remaining top 5 mm down, and then again? Fill the remaining 95 mm with polyester fiber.

(3) Judgment of human fresh blood and rabbit fresh blood Prepare two measurement capillaries, attach the proximal end of one to a cut in a part of a rabbit's ear, and attach the proximal end of one to a cut in a part of a rabbit's ear. Spread the same blood up to a-6)? Use the pressure to inhale slowly.

The blood volume at that time was approximately 9 μt, and the time required for inhalation was 20 seconds. Next, the proximal end of this capillary was set to 0.0
The capillary is immersed in 3% hydrogen peroxide PB8 and inhaled to the vicinity of the upper end of the capillary (8 to 9 cm from the proximal end). The time required for this was 15 minutes. As a result, in the case of human blood, the Rh-IgG-00(a-6) portion was colored brown, but no coloring was observed in rabbit blood.

Example 13 (1) Reagents and equipment ■Albumin adsorption capillary The albumin adsorption capillary described in Example 1 (1) (■) was used.

■ GB with a diameter of 0.1 mm ■　QB+B8mu GB and BSA in (1) of Example 1 were used.

■? lyester fiber ■ Cyanidisin mixed QB, B8mu Cyanidisin mixed GB of (1) of Example 4.

Uses BSA.

■ Bulk sodium perborate mixture GB, BS 12 q of human sodium perborate and 10 g of GB, BB are uniformly mixed in a mortar.

■ Glutathione mixture QB, B8 Muglutathione 401q and QB, BSA I Of Mix uniformly in a mortar.

■ Anti-α-fetozolotine-binding peroxidase (αA
FP-HRP) αAFP-HRP of Example 7 (11) was used.

■ (IAFP-1 (RP mixed GB, B8A Example 7)
(1) ■ αAFP-11RP mixed QB.

Uses BSA.

Anti-α-fetozolotine-conjugated GB (αAFP-Q
)implementation? ! l 7 (1) ■αm FP -0 for &.

(2) Preparation of capillary tube for measurement The same operation as in (2) of Example 1 was performed to prepare the following capillary tube. First 5'mm Kl? Lyester fiber (
1-1), followed by cyanidisin mixed QB in 3.5 mm increments
, BSmu (t-2), perborated bulk sodium mixed QB,
BBmu (t-3), glutathione mixed GB, B8mu (l
-4), GB, BSA (1-5), αmu FP-HRP mixed GB, B8mu (1-6), QB, BSA (t-7)
, and filled with αAFP-○(r-8). Pack QB (1-9) to 5 mm below the top of the remaining part, and leave 5 mm K
again? Fill with linear chill fiber.

(3) Measurement of α-fetoprotein in serum α-fetozolotin 200Pf/− human serum was diluted 10 times with PH1,
Final degree α-fetozolotine 20μf/- (human serum PB
Supernatant liquid obtained by adding 50 μf of neutral hydroxide to human serum (Sanzol B) and α-fetoprotein 20 μf/- serum (0.8 mixed solution) (Sansol B) and α-fetoprotein 20 μf/- serum. Prepare three test solutions (Sample C).

Prepare three measurement capillary tubes, immerse the proximal ends of each in Samplem, Sample B, and Sump+C, and inhale the test liquid to near the top of the capillary (8 to 9 cm from the proximal end). As a result, Sansolm αmuFP-QC Fig. 12 (t
-S)) brown coloring, Sunsol B is αmu FP-○ [1st
Figure 2 (1-8)) is not colored, and the sump kc is α
Brown coloration was observed in Mu'i'P-0 [Figure 12 (t-8)). According to the above results, if a substance that inhibits this reaction is present in the test liquid, it can be measured by dilution or treatment with an added substance.

[Brief explanation of drawings]

Figures 1 to 12 show measurement capillaries used in the present invention. Applicant Kame - Chemical Products Co., Ltd. Figure 2 Figure 11 Procedural amendment (voluntary) May 15, 1980 J Indication of the case 1988 Patent Application No. 239549 2 Name of the invention Immunoassay method 3 Relationship with the case of the person making the amendment Applicant address 5-113-5 Nihonbashi, Chuo-ku, Tokyo Name Daiichi Chemical Co., Ltd. Representative 6 Takeshi Nai Fuji 4 Name of agent (8632) Attorney Shinfu Ono) j (,:',:j'1ljkj):J 5 Date of amendment order "Detailed explanation of the invention" and "Brief explanation of drawings" columns of the specification of invention, drawing 7, contents of amendment (1) Details In the bottom line of page 78 of the book, after the words ``Noh becomes.'', change the line and insert the written text. Example 14 (1) Reagents and equipment ■ Albumin adsorption glass capillary Pass 20q/- of albumin through a glass capillary (diameter 1 mm, length 100 mm), remove excess liquid by suction, and dry. ■ Glass beads (GB) Diameter 0.17n+m All commercially available products are used. ■GB, BBmu Soak GB in PBB of albumin 2 (1 g/-), wash GB 2 to 3 times with purified water, and then dry. ■ Ribbed ester fiber Use commercially available products. ■tzslαfetozolotine reagent mixed GB. 88A (hereinafter abbreviated as Rnee afp mixed GB, BBm) Commercially available aFP #l constant reagent (Daiichi Radioiso)-manufactured by Zo Co., Ltd.)
Contents of the measurement kit Reagent: Radioactive iodinated AFP (tzie) solution (0.9μC1//signature), G B, B1
1 'i 1. Add it, mix evenly and dry with phosphorus pentoxide. ■Anti-AFP-GB /1QC5IrJ) II-T, -II Guktu (1glq
-a is added and left to stand at room temperature, and then washed with purified water. Geltaraldehyde is added to the surface-treated GB, left to stand, and then washed with purified water in the same manner. Then, A to the crosslinked GB
After adding FP antibody 5119 and reacting at room temperature for 2 hours, the mixture was dried to prepare anti-mutant TP-GB. ■PBB Used for PBB7 in Example 1 (■). (Creating a capillary for measurement) Make five marks at 3.5 m intervals from the proximal end of the albumin-adsorbing glass capillary.
l-1) t-pack. This proximal end was connected to a vacuum tube to remove pressure inside the capillary tube, and the next 3.5 mm KGB, B
11 (m −2) k Suction and fill. Similarly, the next 3
．． 5mm [R knee afp mixed GB, sea (m-3),
Subsequently, GB, BBA (m-4), anti-alP-GB (m-
5) Fill f sequentially. GB to 5 mm below the top edge of the remaining part,
Fill with BBA (m-6'yk) and finally fill with Lyester fiber (m-7) to a remaining 5 mm. (Figure 13) (3) Measurement of α-fetozolotine in serum 5 measurement capillaries , and the proximal ends of four of them are coated with a known concentration.
: Q, 3.125125. 50 μf/-1 and the test serum was added to the remaining bottle of factor 13 (
m-3) At t, soak each substance uniformly and inhale. The amount of sample collected at this time was 5 μm. Next, the proximal end of this capillary '1 is immersed in PBS and sucked up to the upper end of the measuring capillary. The time required for this measurement was 10 minutes. After measuring the above capillary with its proximal end down in a gamma counter, the immobilized affinity (m-5
) and measure it using a gamma counter. (4) Result Capillary gamma ray measurement value (OPM) Concentration μt/-Measure capillary as it is Cut and measure F
P 0 1226 1211 3.115 1219 1181 25 1076 1071 50 714 723 Subject 768 725 As a result of the above, the amount of immobilized radioactivity decreases as the AFPII degree increases, and the mu-yp concentration in the subject is 55-60 μt.
/d, and the measurement is t. Example 15 (1) Reagents and equipment ■ Albumin adsorption glass capillary Albumin 20n7m PBB is passed through a glass capillary (diameter 1 mm, length 100 mm), and excess liquid is removed by suction and dried. ■Use commercially available glass beads (GB) with a diameter of 0.17 mm. ■GB, PBB Immerse GB in PBB containing 20 pounds/- of albumin, wash GB with purified water 2 to 3 times, and then dry. ■Uses commercially available fiber products. ■1 5 engineering anti-α-fetozolotine reagent stand GB. BBA (hereinafter abbreviated as R knee anti-a, fp mixed GB, BBA) Measurement kit contents of commercially available AFF measurement reagent (manufactured by Daiichi 2 Zoisotozo Co., Ltd.) Reagent Radioactive iodinated α-fetoprotein antibody (r26e) ( Add 1.02 GB and BBA to the freeze-dried product (0.9 μO1/) and mix uniformly. ■Antimu FP-GB GF35tK&'L-lysine (1%) 3- was added and left to stand at room temperature, and then washed with purified water.Geltaraldehyde was added to the surface-treated GO, left to stand, and purified water was added in the same manner. Wash with Next, 5 tons of AFP antibody was added to the cross-linked GB and reacted at room temperature for 2 hours, dried, and the anti-F P-GB
It was created. ■PB8 Use of FBB'i- in Example 1. (2) Creation of capillary for measurement Make five marks at 3.5 mm intervals along the proximal end of the albumin-adsorbing glass capillary. At first? Lyester fiber (n
- Fill in 1). Vacuum this base end? Gs, ssm (n-
2) Fill with k suction. Similarly, the next 3.5mm is R knee anti-afp mixed GB, B8mm (Kl-3), followed by GB.
, BBmu (n-4), and anti-1FP-GB (n-5) are all sequentially filled. GB until remaining 5mm below the top edge. B11 (11-6) T-filling and again after forging? Reeste Fiber (n-7)'t-Fill to remaining 95mm. (Figure 14) (3) Measurement of α7-nitoprotein in serum Prepare seven measurement capillaries, and connect the proximal ends of six of them to a known concentration ayp.
:0,3e10.100+1000.10000. of
/-, another one is soaked in the test serum and inhaled until (n-4). The amount of inhalation at that time was about 5μ! Met. Next, it is immersed in PB8 above the proximal end of this capillary tube and sucked up to the upper end of the measuring capillary tube. This took less than 10 minutes. After measuring the above capillary with its proximal end facing down using a gamma counter, the immobilized affinity (n-5
) and measure it with a gamma counter in the same way. (4) Results Capillary gamma ray measurement value (CIPM) Todoroki FP O564110 3585112 10628138 100776154 500841191 5000856585 Subject 727 149 As the above results Muyp concentration increases, the amount of immobilized radioactivity also increases, and Atrpa [ Ha 50-100n
f/- and measurement too J (2) same, page 79, line 2, "12" is corrected to "14". (3) Submit attached drawings Figures 13 and 14, Figures 13 and 14 Procedural amendments (voluntary) November 20, 1980 Manabu Shiga, Commissioner of the Patent Office, 1988 Patent Application No. No. 239549 2, Name of the invention Immunoassay method 3 Relationship to the case of the person making the amendment Applicant address 5 Homare, 3-13 Nihonto, Chuo-ku, Tokyo Name
Daiichi Chemical Co., Ltd. Representative Takeshi Naito 4 Agent 6 Column 7 of "Detailed Description of the Invention" of the specification to be amended, Contents of the amendment α) Page 3, line 4 [ The phrase "label-capturing substance" is corrected to "label-capturing substance." ρ) Same, page 4, line 7, "...Protein, α!-Antitrizocine" is corrected to "...Protein, α1-Antichymotrigisin, α1-Antitrizocin." 0) Same, page 4, line 13, ``Calmodulin etc.'' has been replaced with ``calmodulin, brealdimine, alzo-S-transcotin, thyroxine protein, retinol-binding protein, hemovexin, fibronectin, pregnancy/J-specific protein ( SPI ) etc.” (4) Same, page 5, lines 5 to 6, "Phosphodinase A, etc." has been replaced with "Phospholy, e-zeA s DNase, RNas."
e, terminal transferase, bezosin, trizisinodan, chymotorinosi/, enterokinase, aminopeptidase, peroxidase, catalase, enolase, tyrosine hydroxylase, do-Q-decarboxylase, do-no-Q-mine beta hydration #suhiro” is corrected. do. 6) Same, page 5, lines M8 to 9, ``azaprotein'' is corrected to ``Ari?zolotin''. (6) Same, page 6, line 3, “β-thromone is globulin, etc.” is replaced with “β-thrombodaloprine sC1 inhibitor β2 macrolog μzolin,”
α, fssunnin inhibitor, platelet factor 4, platelet membrane protein, protein C, etc.” ri) Same, page 6, line 9, correct "r'rs J" to "T4". (8) Same, page 6, line 13, ``thyroglobulin'' is corrected to ``sideglobulin''. (9) Ibid., No. 8, line 14 [Digestive secretions. '' should be corrected to ``Digestive secretions. Syphilis assay, immunoserological test for pathogenic microorganisms. Mygoderasma antibody, Rickettsia, anti-streptolysin o1, anti-stresotokinase, anti-deoxynucleotide nucleokinase BS J.'' tll Same as above. Page 9, line 7, ``B reduced hepatitis virus,'' is corrected to ``hepatitis B virus, hepatitis virus S, E, C., non-A, non-Bm, J. αυ Same, page 9. In line 8, ``Extra-infectious diseases,'' should be corrected to ``Extra-infectious diseases, rabies, mumps, Koxarachy virus, chlamydia.'' 03 Same, on the bottom line of page 9, the phrase [anti-insulin receptor antibody] is corrected to read "anti-insulin receptor antibody." 0 Same, page 10, line 1, "anti-acetylcholine receptor antibody" is corrected to "anti-acetylcholine receptor antibody." ae Same, page 10, line 2, “Complement of C1q etc.” r C1qSC1ys C11ils ChCps C
4s C51C6 ~ Complement system components such as Cr, Cg, ChβIE globulin, etc.” is corrected. n5 Same, 10th layer, line 5 "...protein, inferitin" is replaced with "...f, atin, ferritin, inferritin"
I am corrected. Tie same, page 10, line 6, rcnp, etc. rCRP, immunoacetic xolotin (IAP),
``Pancreatic embryonic antigen (POA), dysfactor, etc.'' (+7+ Same, page 11, line 2, rHr, A, blood type, etc.'') should be corrected as ``renin, a/zootensin nogun, anzoo f7 si. 7
1% [1% [, Anzoote/Synconparting Enzyme, Kinin, Kininogu/~f2 Zumakarikrein, Glandnote Kallikrein, etc. Renin Anzootense y-based HLA. "Blood type, blood cross-matching test, etc." 081 Same, page 11, line 8 "(Immunology-1-1 Proposal 41: Store (1981)
I am corrected. (l Same as above, page 15, line 11, “0-anisidine” is corrected to “0-cyanidisine.”) Kan, page 17, line 6, “vinyl chloride,” is changed to “vinyl chloride, ?liethylene.” $ IJ Carbone),
=lelJf Robiren,” he corrected. 31) Same, page 28, line 7, correct the statement r6.25tdJ to [6,25 哩]. 0 Same, page 28, line 10, ``IrGJ'' is corrected to ``9G antibody'' on page 28. (c) Same, page 29, line 3. Correct "r Ifo, 10 m/g/g" to r IrG antibody (10 g/*) J. Q4 Same, page 29, line 9, "eluate" is corrected to t [reaction solution]. (c) Same, page 33, line 13, page 34, line 4, and line 5 of the same, ``Correct the word ``fPGJ'' to ``ifG antibody'' on page 34. @ Same, page 38, line 3 □ and line 5 [correct αCRP J to read [αCRP antibody]. (c) Same, page 38, lines 14-15 rGB, BS
A(c-5)J and rGB, BSA(c-4)J
I am corrected. (To) Same, p. 43, line 10, correct "r(d-4) glutathione" to "r(d-4), glutathione." (c) Same, page 45, line 13, ``rGB-combined rabbit IfG'' is corrected to ``rabbit IfG-combined GBJ.'' (To) Ibid., page 46, line 11, ``Anti-Rb-IfGJ'' is corrected to ``anti-Rb-IfG antibody.'' c3]) Ibid., page 48, line 4, ``anti-human alpkin'' is corrected to ``anti-human albumin antibody.'' (To) Same, page 48, line 6 is corrected to read ``Goat-pile-tip IfG antibody is rabbit anti-human alzomin antibody''. (To) Same, page 48, line 13, "αH8AJ" is corrected to "r ffH8A antibody." (b) Same, page 48, line 14 [Goat αCRPt-Rabbit α-H8A J is replaced with “Goat α
Correct CRP antibody to rabbit αH8A antibody. (To) Same, page 51, line 13, "anti-α-fetozolotine" is corrected to "anti-α-fetoprotein antibody." (To) Same, bottom line of page 51 to line 1 of page 52 [Goat stake tip 1tG is Max anti-α-fetozotein” is corrected to “Goat stake tip IfG antibody is Max anti-α-fetozolotine antibody” Suru (b) Same, page 52, lines 7 and 8, ``anti-α-fetoprotein'' is corrected to ``anti-α-fetoprotein antibody.'' (To) Same, page 52, line 12, "Goat αCRPJ" is corrected to "Goat αCRP antibody." ), page 56, line 11, ``anti-fibrin degradation product'' is corrected to ``anti-fibrin degradation product antibody''. (b) Same, page 56, line 13, "Goat stake IfG is a mouse anti-ficillin decomposition product" is corrected to read "Goat stake IfG antibody is a Max anti-fibrin decomposition product antibody." (b) Same, page 57, line 5, "anti-fibrin degradation product" is corrected to "anti-fibrin degradation product antibody."轡 Same, page 60, line 13, ``anti-α-fetoprotein'' is corrected to ``anti-α-fetozolotin antibody.'' 4. Same, on the bottom line of page 60, ``α-fetoprotein'' is corrected to ``peroxidase bound to anti-α-fetozolotin antibody.'' - In the same publication, page 61, line 5, "anti-α7 etozolotein" is corrected to "anti-αfetozolotine antibody." ), page 61, line 6, "Goat FDP J" is corrected to read "anti-α-fetozolotine antibody." (44 Id., p. 61, lines 8 and 9, "anti-peroxidase" is corrected to "anti-peroxidase antibody.") 0 Id., p. 62, line 1, "αAFDJ" is corrected to "αAFPJ." → Ibid., page 64, line 10 and page 66, line 8, “anti-α-fetozolotine” is corrected to “anti-α-fetozolotine antibody.” (To) Ibid., page 70, line 4- Line 5 “Pile tip IfG [combined fluorescein inch] LI
L 咄L'14 Corrected to ``Stake tip IfG antibody-conjugated fluorescein isothiocyanate.'' Tseng, page 70, line 6, ``Goat pile tip IfGJ'' is corrected to ``Goat pile tip fG antibody.'' *1 Same, page 76, lines 5 and 11, "anti-α-fetozolotine" is corrected to "anti-α-fetozolotine antibody." ) No. 3 in the written amendment to the procedure submitted on May 15, 1988.
The words "afp" in the 7th line of the page and the 1st line of the 8th page are corrected to rAFPJ. 0 In the written amendment, the words "rAFP antibody" on page 4, line 4 and page 8, line 13 are corrected to "anti-AFP antibody."轡 In the same procedural amendment, page 4, line 8 and page 9, line 2 [PBS of ■ in Example 1 is used. ” will be deleted. ) In the written amendment to the same procedure, on the bottom line of page 7, the phrase "anti-α fetoprotein" is corrected to "anti-σ fetozopty antibody." Procedural amendment (voluntary) May 29, 1985 Manabu Shiga, Commissioner of the Patent Office, 1988 Patent Application No. 239549 2 Name of the invention Immunoassay method 3 Relationship with the person making the amendment Applicant's address Tokyo 3-13-5 Nihonbashi, Chuo-ku Name
No. 4 - Takeo Naito, representative of Kagaku Yakuhin Co., Ltd. 6, agent 6, next to [Detailed description of the invention] in the specification subject to amendment 7, content of amendment (1) In the specification, page 2, lines 1 to 6 In the second line, ``More specifically, the immune reaction and labeling substance'' should be corrected to ``More specifically, the labeling substance.'' (ii) On page 7, lines 12 and 13, ``tehydroepianodrosten'' is corrected to ``dehydroepia, androsterone''. 0) Same, page 13, line 11, "C J, Histo,..." is replaced with "C J, Histo,..." is adopted.
・” I corrected it. (4) Same, in the second line from the bottom of page 18, the phrase [combined with either one or both] is corrected to read "combined with either one." (5) Same, page 19, line 6, ``It is necessary to do so,'' is corrected to ``It may be necessary to do so.'' (6) Same, page 20, lines 4 to 5, "immobilized Kitamakura-labeled antibody" is corrected to "immobilized anti-labeled antibody." σ) Ibid., line 7, ``(Immobilized Kitamakura-labeled antibody)'' is corrected to ``(immobilized anti-shield antibody.'' (8) Ibid., page 23, line 9, ``Antigen-labeled antibody.'' is corrected to be "antigen-labeled antibody." The line "Becomes a labeled antibody - a labeled antibody" is corrected to "become a labeled antibody - a labeled antibody and an immobilized anti-labeled antibody - a labeled antibody - an antigen". +111 times, page 24, No. 3 In line 1, "immobilized anti-labeled antibody" should be corrected to "immobilized anti-labeled antibody". ” is corrected. α row Same, line 7, “The labeled substance is released” is corrected to “becomes the labeled substance.” I Same, line 10, “Diaphorase of the holoenzyme” is corrected. Correct ``diaphorase.''
H, which is necessary for a luminescent substance to emit light, etc. ” he corrected. 4. Ibid., page 25, line 11, ``activate the inhibitor'' is corrected to ``inactivate the inhibitor.'' (17) Same, page 37, line 9, "anti-C-reactive protein-bound peroxidase" is corrected to "anti-C-reactive protein antibody-bound peroxidase." Same as above, in line 11, ``The stake tip IgG was replaced with goat anti-CRP'' is corrected to ``the antigen-labeled antibody was replaced with a goat anti-CRP antibody.'' (11, page 38, line 7 [■ Base coloring solution] is corrected to [■ substrate coloring solution."). ■ Page 41, line 13, "Anti-fibrin degradation product binding." ``Ficillin degradation product antibody binding'' @ Same, bottom line ``Goat tip IgG mouse anti-FDP J'' is corrected to [Goat tip IgG antibody called Max anti-FDP antibody.'' @ Same, page 42 In line 7, "αFDP-Chouai GBJ" is corrected to "αFDP antibody supercombination GB (αli'DP-○)." Correct. e◆ Same, page 46, lines 13-14 “αRb -I
``There is no αRb-IgG antibody'' should be corrected to ``There is no αRb-IgG antibody.'' (To) Ibid., p. 47, line 9, “αRb-IgG is not recognized.”
Corrected to ``No IgG antibody detected.'' (To) Same, page 57, line 2 r ``IgG-HRP'' is corrected to ``αRb-IgG-HRP''. 0 Same, same line 4 [α-F IJ P M combination GBJ]
Corrected as antibody-bound GBJ. (c) Same, page 71, line 5, "(αRbIgG-○)" is corrected to "r(RbIgG-○)". (Sec.) Same, line 13, "α1%b IgG-○" is corrected to "rRbIgG-○". (To) In the procedural amendment submitted on November 20, 1988, page 7, lines 11 to 12, "Renin anzootency y-based HLA, blood type, blood cross-matching test, etc." was replaced with "Renin "Anzootency system. HLA% blood type, blood cross-matching test, etc." is corrected.

Claims (1)

[Claims] 1. In a method for detecting or quantifying a component in a specimen by immunoreaction, an insoluble carrier on which a visceral substance-trapping substance is immobilized and an indecent carrier holding at least a labeled immunoreaction reagent are mixed into a hair #1tlt. The analyte is inhaled into the capillary tube, an immunoreaction is carried out in the capillary tube, and the reaction product or I! One Shimento reagent! An immunoassay method characterized by combining with a textile capture substance to immobilize it and measuring the immobilized label substance〇