DE2035784A1 - Tubes part-filled with antigens - in gel for antigen-antibody reaction in body fluids visible as a narrow precipitation zone - Google Patents
Tubes part-filled with antigens - in gel for antigen-antibody reaction in body fluids visible as a narrow precipitation zoneInfo
- Publication number
- DE2035784A1 DE2035784A1 DE19702035784 DE2035784A DE2035784A1 DE 2035784 A1 DE2035784 A1 DE 2035784A1 DE 19702035784 DE19702035784 DE 19702035784 DE 2035784 A DE2035784 A DE 2035784A DE 2035784 A1 DE2035784 A1 DE 2035784A1
- Authority
- DE
- Germany
- Prior art keywords
- gel
- antigens
- filled
- antigen
- agar
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000427 antigen Substances 0.000 title claims abstract description 19
- 102000036639 antigens Human genes 0.000 title claims abstract description 19
- 108091007433 antigens Proteins 0.000 title claims abstract description 19
- 210000001124 body fluid Anatomy 0.000 title claims abstract description 11
- 239000010839 body fluid Substances 0.000 title claims abstract description 11
- 238000001556 precipitation Methods 0.000 title claims abstract description 8
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 6
- 239000000499 gel Substances 0.000 claims abstract description 19
- 238000000034 method Methods 0.000 claims abstract description 7
- 238000012360 testing method Methods 0.000 claims abstract description 7
- 229920001817 Agar Polymers 0.000 claims abstract description 6
- 235000010419 agar Nutrition 0.000 claims abstract description 5
- 241000206672 Gelidium Species 0.000 claims abstract description 4
- 241000700605 Viruses Species 0.000 claims abstract description 4
- 229940072221 immunoglobulins Drugs 0.000 claims abstract description 4
- 239000004033 plastic Substances 0.000 claims abstract description 4
- 229920003023 plastic Polymers 0.000 claims abstract description 4
- 239000011521 glass Substances 0.000 claims abstract description 3
- 244000005700 microbiome Species 0.000 claims abstract description 3
- 108060003951 Immunoglobulin Proteins 0.000 claims description 3
- 230000000951 immunodiffusion Effects 0.000 claims description 3
- 102000018358 immunoglobulin Human genes 0.000 claims description 3
- 239000000654 additive Substances 0.000 claims 1
- 239000000872 buffer Substances 0.000 claims 1
- 230000003054 hormonal effect Effects 0.000 claims 1
- GPRLSGONYQIRFK-UHFFFAOYSA-N hydron Chemical compound [H+] GPRLSGONYQIRFK-UHFFFAOYSA-N 0.000 claims 1
- 238000001514 detection method Methods 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 2
- 108010017384 Blood Proteins Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 206010070834 Sensitisation Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 210000000944 nerve tissue Anatomy 0.000 description 2
- 230000008313 sensitization Effects 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 208000025747 Rheumatic disease Diseases 0.000 description 1
- 102000005686 Serum Globulins Human genes 0.000 description 1
- 108010045362 Serum Globulins Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000000845 cartilage Anatomy 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 229920002301 cellulose acetate Polymers 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000000416 exudates and transudate Anatomy 0.000 description 1
- 210000004565 granule cell Anatomy 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 210000000281 joint capsule Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- -1 polyethylene Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 210000002796 renal vein Anatomy 0.000 description 1
- 230000000552 rheumatic effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
- G01N33/559—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody through a gel, e.g. Ouchterlony technique
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Dispersion Chemistry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Description
Verfahren zur Bestimmung von humoralen Antikörner-Immunglobulinen gegen Antigene aus Körperflüssigkeiten, Körnerzellen. Mikroorganismen und Viren durch Präzinitation bei eindimensionaler Immunodiffusion. Method for the determination of humoral antibody immunoglobulins against antigens from body fluids, granule cells. Microorganisms and viruses by precinitation with one-dimensional immunodiffusion.
Das Verfahren besteht in der Verwendung durchsichtiger Röhrchen mit 1 bis 5 mm Durchmesser aus inertem Glas oder Kunststoff zur Durchführung der Präzipitationsreaktion. Dabei werden die nach bekannten Methoden -gewonnenen Antigene in ein geeignetes Gel, z.B. aus Agar-Agar, Agarose, Zelluloseazetat, aufgenommen und in entsprechende Röhrchen von etwa 5 bis lo cm Länge eingebracht, wobei die Füllung nur einen Teil der Röhrchen beträgt, z.B. 2 bis 5 cm, so daß eine Uberschichtung mit den zu untersuchenden Körperflüssigkeiten, z.B. Blutserum, Plasma, Liquor, Cerebrospinalis, Harn, Exsudaten und Transsudaten, Gelenkflüssigkeiten, in etwa dem gleichen Ausmaß möglich ist. Die vorbereiteten Teströhrchen werden an beiden Enden verschlossen und vor Benützung am nicht mit dem Gel gefüllten Ende geöffnet. Dann wird die zu untersuchende Körperflüssigkeit eingefüllt. Diese muß in unmittelbarem Kontakt zum Gel stehen. Die Präzipitation wird dann in üblicher Weise in Wärme bei etwa 370 C und evt. anschließend in Kälte bei 40 C durchgeführt. Sofern in der zu untersuchenden Körperflüssigkeit Antikörper, d.h. Immunglobuline, gegen die Antigene enthalten sind, erfolgt im Gel im Bereich der Grenzschicht eine Antigen-Antikörperreaktionl die als Präzipitatsstreifen sichtbar wird. Die Breite dieses Streifens kann in bestimmten zeitlichen Intervallen mehrmals gemessen werden. Dadurch sind quantitative Vergleiche möglich.The procedure consists of using clear tubes with 1 to 5 mm in diameter made of inert glass or plastic for carrying out the precipitation reaction. The antigens obtained by known methods are converted into a suitable one Gel, e.g. from agar-agar, agarose, cellulose acetate, taken up and in the appropriate Tubes of about 5 to lo cm in length were introduced, with the filling only a part of the tube is, for example 2 to 5 cm, so that an overlap with the Body fluids, e.g. blood serum, plasma, liquor, cerebrospinalis, urine, exudates and transudates, synovial fluids, are possible to about the same extent. The prepared test tubes are closed at both ends and before use opened at the end not filled with the gel. Then the body fluid to be examined filled. This must be in direct contact with the gel. The precipitation is then in the usual way in heat at about 370 C and possibly then in cold carried out at 40 C. If there are antibodies in the body fluid to be examined, i.e. immunoglobulins, against which antigens are contained, takes place in the gel in the area the boundary layer shows an antigen-antibody reaction, which is visible as a precipitate strip will. The width of this strip can be several times in certain time intervals be measured. This enables quantitative comparisons.
Das vorliegende Verfahren unterscheidet sich von der quantitativen Plasmaproteinbestimmug durch eindimensionale Immundiffusion dadurch, daß dort das Gel mit einem Antikörperserum beladen wird und das Antigen in Form der Plasmaproteine überschichtet wird. Hier hingegen wird das Gel mit besonderen Antigenen beladen und mit Körperflüssigkeiten überschichtet, von denen man vermutet, daß sie Anti° körper gegen die genannten Gewebe enthalten. Diese Antikörper können durch Testung mit verschiedenen Teströhrchen, in denen unterschiedliche Antigene enthalten sind, ermittelt werden.The present method differs from the quantitative one Plasma protein determination by one-dimensional immunodiffusion in that the Gel is loaded with an antibody serum and the antigen in the form of plasma proteins is overlaid. Here, on the other hand, the gel is loaded with special antigens and overlaid with body fluids believed to be anti ° body against the tissues mentioned. These antibodies can be tested by testing with different test tubes containing different antigens, be determined.
1. Beispiel: Es soll ein Teströhrchen zur quantitativen Bestimmung der humoralen Antikörper gegen Nierengevebe hergestellt werden.1st example: A test tube should be used for quantitative determination the humoral antibodies against renal veins can be produced.
Hierzu wird eine 1,4 %ige Agar-Agar-Lösung aus 1 %iger NaCl-Lösung, die mit Trispuffer auf pH 7,2 unter Zusatz von 1 : lo ooo Cialit hergestellt ist, verwendete Ein Homogenat aus frischer oder aufgeschwemmter, Lyophylisierter Niere wird mit Ultraschall 3 Minuten behandelt und sedimentiert. Der Üb erstand wird zur Bestiiimung des Äquivalenzbereiches geometrisch verdünnt und die Verdünnungen jeweils im Verhältnis 1 : 1 mit der auf 45° C abgekühlten Agarlösung gemischt und mittels einer Spritze mit Kanüle in vorbereitete Kunststoffröhrchen aus Polyäthylen mit einem Durchmesser von 2,1 mm eingefüllt. Die Röhrchen mit den verschiedenen Antigenkonzentrationen werden im Vakuum bei einer Temperatur von 45° C entgast und danach abgekühlt.For this purpose, a 1.4% agar-agar solution from 1% NaCl solution, which is made with Tris buffer to pH 7.2 with the addition of 1: lo ooo Cialit, used a homogenate from fresh or bloated, lyophilized kidney is treated with ultrasound for 3 minutes and sedimented. The surplus becomes Determination of the equivalence range geometrically diluted and the dilutions in each case mixed in a ratio of 1: 1 with the agar solution cooled to 45 ° C and using a syringe with a cannula in a prepared plastic tube made of polyethylene with filled with a diameter of 2.1 mm. The tubes with the different antigen concentrations are degassed in vacuo at a temperature of 45 ° C and then cooled.
Mit einem Standardserum gegen Nierenantigene wird die optisalze Antigenkonzentration für die Präzipitation getestet, danach werden die weiteren Röhrchen mit Gel, das diese Antigenkonzentration enthält, beschickt. Diese werden ebenfalls durch Vakuum entgast und dann an beiden Enden thermisch verschlossen0 Die Herstellung von Teströhrchen zur Bestimmung von andersartigen Organsensibilisierungen erfolgt in analoger Weise unter Verwendung der jeweiligen Organantigene, z.B. aus Schilddrüse, Leber, Herz, Nervengewebe, Sperma u.a.With a standard serum against kidney antigens, the optimal antigen concentration is determined tested for precipitation, after which the other tubes are filled with gel that contains this concentration of antigen. These are also made by vacuum degassed and then thermally sealed at both ends0 The production of test tubes to determine other types of organ sensitization in an analogous way using the respective organ antigens, e.g. from the thyroid gland, Liver, heart, nerve tissue, sperm, etc.
2. Beispiel= Zum Nachweis von Rheumaantikörpern werden in ähnlicher Weise wie im Beispiel 1 Röhrchen mit Agar-Gel beschickt, dem eine Mischung von Serumglobulin und antigenen Organextrakten aus Synovialschleimhaut, Gelenkkapsel, sowie von Knorpelgewebe aus rheumatisch erkrankten Gelenken zugesetzt ist.2nd example = For the detection of rheumatoid antibodies are in similar In the same way as in Example 1, tubes filled with agar gel containing a mixture of serum globulin and antigenic organ extracts from synovial mucosa, joint capsule and cartilage tissue from rheumatic joints is added.
3. Beispiel: Zum Nachweis von Autoantikörpern gegen Gewebe des ZNS wird das Gel mit Organantigenen aus Nervengewebe beladen. Dabei werden die Lipide durch einen Emulgator (Laurylnatriumsulfat) emuligiert und mitverwendet.3rd example: For the detection of autoantibodies against tissue of the CNS the gel is loaded with organ antigens from nerve tissue. This is where the lipids emulsified by an emulsifier (lauryl sodium sulfate) and used.
4. Beispiel: Zum Nachweis von Antikörpern gegen Tumorgewebe wird das Gel mit den jeweiligen Tumorantigenen beladen.4. Example: For the detection of antibodies against tumor tissue, the Load the gel with the respective tumor antigens.
5. Beispiel: Zum Nachweis einer Sensibilisierung gegen Streptokokken wird ein antigener Extrakt aus Streptokokken in das Gel eingearbeitet.5. Example: To demonstrate sensitization to streptococci an antigenic extract from streptococci is incorporated into the gel.
6. Beispiel: Zum Nachweis von Antikörperbildung gegen Viren werden Virusantigene dem Gel zugesetzt. Als Gele können Agarose, Zellulose, Azetat und alle geeigneten Gele verwendet werden, in denen eine Antigenantikörperreaktion möglich ist.6. Example: To be used for the detection of antibody formation against viruses Virus antigens added to the gel. As gels, agarose, cellulose, acetate and all suitable gels are used in which an antigen-antibody reaction is possible is.
Claims (2)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19702035784 DE2035784A1 (en) | 1970-07-18 | 1970-07-18 | Tubes part-filled with antigens - in gel for antigen-antibody reaction in body fluids visible as a narrow precipitation zone |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19702035784 DE2035784A1 (en) | 1970-07-18 | 1970-07-18 | Tubes part-filled with antigens - in gel for antigen-antibody reaction in body fluids visible as a narrow precipitation zone |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| DE2035784A1 true DE2035784A1 (en) | 1972-01-27 |
Family
ID=5777213
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| DE19702035784 Pending DE2035784A1 (en) | 1970-07-18 | 1970-07-18 | Tubes part-filled with antigens - in gel for antigen-antibody reaction in body fluids visible as a narrow precipitation zone |
Country Status (1)
| Country | Link |
|---|---|
| DE (1) | DE2035784A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2577321A1 (en) * | 1985-02-08 | 1986-08-14 | Lapierre Yves | DEVICE AND METHOD FOR DISTINGUISHING ERYTHROCYTE AGGLUTINATES |
| US5665558A (en) * | 1994-05-17 | 1997-09-09 | Gamma Biologicals, Inc. | Method and apparatus useful for detecting bloodgroup antigens and antibodies |
| US5905028A (en) * | 1994-05-17 | 1999-05-18 | Gamma Biologicals, Inc. | Method and apparatus useful for detecting bloodgroup antigens and antibodies |
-
1970
- 1970-07-18 DE DE19702035784 patent/DE2035784A1/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2577321A1 (en) * | 1985-02-08 | 1986-08-14 | Lapierre Yves | DEVICE AND METHOD FOR DISTINGUISHING ERYTHROCYTE AGGLUTINATES |
| EP0194212A1 (en) * | 1985-02-08 | 1986-09-10 | Fondation pour la recherche de diagnostiques de laboratoire | Method for detecting agglutinated erythrocytes |
| US5665558A (en) * | 1994-05-17 | 1997-09-09 | Gamma Biologicals, Inc. | Method and apparatus useful for detecting bloodgroup antigens and antibodies |
| US5905028A (en) * | 1994-05-17 | 1999-05-18 | Gamma Biologicals, Inc. | Method and apparatus useful for detecting bloodgroup antigens and antibodies |
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