EP0163631A1 - Method for the determination of the results of agglutination reactions - Google Patents
Method for the determination of the results of agglutination reactionsInfo
- Publication number
- EP0163631A1 EP0163631A1 EP19830903791 EP83903791A EP0163631A1 EP 0163631 A1 EP0163631 A1 EP 0163631A1 EP 19830903791 EP19830903791 EP 19830903791 EP 83903791 A EP83903791 A EP 83903791A EP 0163631 A1 EP0163631 A1 EP 0163631A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- agglutination
- reaction
- measurement
- light
- results
- 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
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity
-
- 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
Definitions
- the present invention is concerned with a method for the measurement of the results of agglutination reactions in an analyzer of vertical measurement.
- agglutination tests are used, e.g., for the establishment of bacteria, viruses, antigen components of fungi or of their antibodies as well as of abnormal proteins (e.g. tumour-specific proteins).
- hemagglutination HA
- HI hemagglutination inhibition
- indirect hemagglutination as well as latex agglutination.
- the said tests are used in serological diagnostics, serotyping, as well as in various establishments of antigens. For example, in epidemy studies as well as in the specific diagnostics of infection diseases, the said tests are highly usable.
- a positive result of agglutination may mean either a positive result or a negative result.
- Hemagglutination can be ascertained on the bottom of the reaction vessel as a diffuse covering. For example, in a HA-test, viruses and bacteria have an ability to agglutinate red blood cells under certain conditions on the bottom of the reaction vessel over a large area. If no hemagglutination takes place, the red blood cells are assembled in the reaction vessel usually as distinctly limited area, a so-called button.
- agglutination tests have been read visually.
- the criteria for the interpretation of the results vary.
- the agglutination tests may also be connected with a colour indicator, whose visual interpretation, in a negative or positive test, is, however, laborious.
- Tests of this type are used extensively in different determinations. What is also frequently concerned is tests carried out in massive numbers, such as syphilis and rheumatism tests, blood group determinations, and establishment of pregnancy. In such cases, the performance of the tests must be simple, automated, accurate, and reliably reproducible.
- the agglutinated reaction mixture is shaken before measurement so strongly that the agglutination mix is dispensed in the reaction mixture (GB Pat. No. 1,229,971). 2.
- the reaction mixture is measured by means of two separate beams of measurement (GB Pat. No. 1,229,971 and US Pat. No. 3,883,308), the values yielded by them being compared with two separate threshold values.
- the method in accordance with the present invent ⁇ on and with the said earlier invention permits, e.g., the measurement of the results of agglutination reactions, among other things, in a spectrophotometer, adsorption photometer, fluorometer, and nefelometer.
- the method in accordance with the present invention permits, e.g., the formation, measurement and production of the output of agglutination reactions reliably and rapidly either automatically or partly non-automatically.
- agglutinations are promoted by appropriate centrifuging or shaking.
- the reaction mixture is incubated.
- the shaking of the reaction mixture is performed so that even a weak agglutination mix is not scattered in the reaction mixture but remains on the bottom of the reaction vessel, but, however, strongly enough so that the non-agglutinated part can be placed homogeneously in the reaction mixture.
- the reaction mixture is measured.
- the absorbance of the agglutination produced on the bottom of the cuvette of the reaction mixture is measured (preferably, e.g., by means of a vertical beam of measurement) so that the source of light or the reaction vessel is moved in the horizontal plane and, at the same time, the intensity of the light passing through is measured at several phases along the path of movement.
- the absorbance values measured by means of one beam of measurement at several different points are analyzed and compared in accordance with a program. The production of the output takes place automatically.
- Figure 1a shows an agglutination placed on the bottom of a cuvette for reaction mixture, the result being in this case positive,
- Figure 1b shows an agglutination placed on the bottom of a cuvette for reaction mixture, the result being in this case negative
- Figure 2 schematically illustrates the measurement of a block of cuvettes for reaction mixture
- Figure 3 is an exemplifying embodiment of a system for the determination of blood groups
- Figure 4a illustrates the measurement of the bottom of a cuvette at several different points
- Figure 4b shows the corresponding measurement reaults.
- the red blood cells are examined by means of an agglutination test as follows: 1) A cell suspension of about 2.5 % is made of the red blood cells to be examined, in physiological salt solution.
- a cell suspension of about 2.5 % is made of A- and B-test cells in physiological salt solution.
- the apparatus is reset to zero, e.g., by means of 100 ⁇ l of BSA-solution.
- a suspension of about 2.5 % is made of the red blood cells to be examined, in physiological salt solution.
- 50 ⁇ l of the suspension prepared under 1) as well as 50 ⁇ l of a - D - serum are pipetted into the FP-blood-group cuvettes.
- a cell suspension of 2 .5 % or 5 % is made of washed cells .
- 100 ⁇ l of a - D - serum and 100 ⁇ l of 2.5-% or 5-% cell suspension are pipetted into FP-bloodgroup cuvettes.
- the apparatus is reset to zero, e.g., by means of 100 ⁇ l of a - human - serum.
- Figure 1a shows an agglutination 2 on the bottom of the reaction-mixture cuvette 1 , the result being in this case positive.
- Figure 1b shows a negative result, in respect of agglutination, in cuvette 3, wherein the red blood cells are evenly dispensed in the reaction mixture 4.
- Figure 2 schematically shows how the block 5 of reaction-mixture cuvettes moves through the measurement beam 6 so that each reaction-mixture cuvette 7 in the cuvette block 5 is read by means of the vertical measurement beam 6 at different points.
- the vertical measurement beam 6 starts from an opening 8 of appropriate size facing each reaction-mixture cuvette 7 and passes in the direction of the longitudinal axis of the reaction-mixture cuvette 7 to the corresponding detector 9.
- the agglutination 10 on the bottom of each reaction-mixture cuvette 7 can be measured at several points by appropriately displacing the cuvette block 5 in the direction of the x-axis and/or, if required, also in the direction of the y-axis.
- the measurement beam 6 may also pass in the opposite direction, from the top downwards to a detector placed underneath the reaction-mixture cuvette 7 , or the cuvette may also be stationary and the beam of light be displaced across the cuvette bottom.
- the beam of light may also move across the cuvette bottom at several points.
- Fig. 4a Reference numerals 11, 12 and 13 denote different paths of movement.
- Figure 4b illustrates the corresponding measurement results.
- the curve 15 corresponds, e.g., to the measurement of point 12 as measured photometrically.
- the equipment can be constructed so that the agitator is placed in connection with the photometric measurement head.
- outputs in accordance with the above can be produced, e.g., as a function of shaking intensities, thereby finding out differences between the intensities of different agglutinations.
- the equipment is provided with appropriate computer equipment (Fig. 3 p) .
- Figure 3 illustrates a system as an exemplifying embodiment of the method invention for the determination of blood groups : a. Whole-blood sample b. The blood-sample is centrifuged c. Plasma or serum and red blood cells separated in the tube d. The tube is placed in its position, indicated by the arrow e. 25 ⁇ l of red blood cells are taken out of the tube into a dosage-diluter (1) f. The dosage-diluter is emptied into an empty tube g. By means of a dosage-diluter, 1000 ⁇ l of reagent is added h. 50 ⁇ l of cell suspension is dosed by means of a dosage-diluter into four reaction-mixture cuvettes i.
- 200 ⁇ l of plasma are taken into a second dosage-diluter (2) j.
- 50 ⁇ l of plasma are dosed into each of the following four reaction-mixture cuvettes k.
- 50 ⁇ l of reagent are added into the reaction- mixture cuvettes l.
- the reaction-mixture cuvettes are incubated m. Centrifuging or shaking if required n. Shaking so that agglutinated mix is not dispensed in the reaction mixture.
- non-agglutinated red blood cells are dispensed in the reaction mixture o.
- the shaking is followed by automatic measurement by means of a vertical measurement beam at 5 to 10 different points through the bottom of each reaction-mixture cuvette p. Storage, calculation, analysis, and output of the measurement result.
- systems can be constructed, e.g., with different degrees of automation or with different mechanical embodiments.
- Such systems may comprise one apparatus or several separate apparatuses.
- the measurement results may be compared, e.g., with standard values, with each other, and/or with positive or negative results, or with both.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Hematology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Plasma & Fusion (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
Méthode permettant de mesurer le résultat d'une réaction d'agglutination et selon laquelle le mélange de réaction est secoué après incubation, de sorte que la partie non agglutinée est répartie de manière homogène dans le mélange de réaction. Ensuite le pouvoir d'absorption de l'agglutination restant sur le fond du récipient de réaction est mesuré dans un analyseur qui mesure verticalement de sorte que le faisceau lumineux ou le récipient de réaction est déplacé dans le plan horizontal et le pouvoir absorbant est mesuré en plusieurs points différents. Le résultat de la réaction d'agglutination est obtenu par comparaison des valeurs de pouvoir d'absorption.Method for measuring the result of an agglutination reaction and according to which the reaction mixture is shaken after incubation, so that the non-agglutinated part is distributed homogeneously in the reaction mixture. Then the absorption capacity of the agglutination remaining on the bottom of the reaction container is measured in an analyzer which measures vertically so that the light beam or the reaction container is moved in the horizontal plane and the absorbency is measured in several different points. The result of the agglutination reaction is obtained by comparison of the absorbency values.
Description
Claims
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/FI1983/000073 WO1985002259A1 (en) | 1983-11-21 | 1983-11-21 | Method for the determination of the results of agglutination reactions |
Publications (1)
Publication Number | Publication Date |
---|---|
EP0163631A1 true EP0163631A1 (en) | 1985-12-11 |
Family
ID=8556340
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP19830903791 Pending EP0163631A1 (en) | 1983-11-21 | 1983-11-21 | Method for the determination of the results of agglutination reactions |
Country Status (3)
Country | Link |
---|---|
EP (1) | EP0163631A1 (en) |
JP (1) | JPS61501162A (en) |
WO (1) | WO1985002259A1 (en) |
Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE3422616A1 (en) * | 1984-06-19 | 1985-12-19 | Behringwerke Ag, 3550 Marburg | METHOD FOR DETERMINING A PARTNER OF A REACTION AND DEVICE THEREFOR |
US4730921A (en) * | 1985-10-31 | 1988-03-15 | Genetic Systems, Inc. | Photodensitometer for minimizing the refractive effects of a fluid sample |
EP0229355A3 (en) * | 1986-01-06 | 1988-01-07 | Orion Corporation Ltd | Apparatus and method for carrying out photometric assays |
DE3919260A1 (en) * | 1989-06-13 | 1990-12-20 | Hoechst Ag | METHOD FOR QUANTITATIVELY EVALUATING AGGLUTINATION REACTIONS |
AU686604B2 (en) * | 1993-05-17 | 1998-02-12 | Fujirebio Inc. | Method and apparatus for performing an indirect agglutination immunoassay |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE1598944A1 (en) * | 1966-07-21 | 1971-06-24 | Pfizer & Co C | Method and device for the automatic detection of agglutinations in a reaction zone |
FR95147E (en) * | 1967-05-12 | 1970-07-24 | Centre Nat Rech Scient | Apparatus intended more particularly for the automatic determination of blood groups. |
FI56905C (en) * | 1978-02-28 | 1980-04-10 | Osmo A Suovaniemi | FOERFARANDE OCH ANORDNING FOER AUTOMATISK MAETNING AV AGGLUTINATIONSPROV T EX I SPEKTROPOTOMETER ADSOPTIONSFOTOMETER FLUOROMETER ELLER NEFELOMETER |
JPS6145479Y2 (en) * | 1979-09-10 | 1986-12-20 | ||
EP0056058B1 (en) * | 1980-07-24 | 1985-06-05 | Labsystems Oy | Method and equipment for the measurement of properties of a liquid |
EP0056414A1 (en) * | 1980-07-24 | 1982-07-28 | Labsystems Oy | Method and apparatus for the measurement of the properties of an agglutination |
WO1982000357A1 (en) * | 1980-07-24 | 1982-02-04 | Oy Labsystems | Method and apparatus for the measurement of the properties of an agglutination |
WO1982000354A1 (en) * | 1980-07-24 | 1982-02-04 | Oy Labsystems | Method and apparatus for the measurement of the properties of an agglutination |
FR2488691A1 (en) * | 1980-08-14 | 1982-02-19 | Commissariat Energie Atomique | METHOD AND DEVICE FOR DETECTION AND QUANTIFICATION OF REAL-TIME AGGLUTINATES |
FR2509860A1 (en) * | 1981-07-17 | 1983-01-21 | Louis Serge | Measuring agglutination of red blood cells for blood grouping etc. - to give numerical factor calculated from multipoint opacity determination |
-
1983
- 1983-11-21 JP JP50008783A patent/JPS61501162A/en active Pending
- 1983-11-21 EP EP19830903791 patent/EP0163631A1/en active Pending
- 1983-11-21 WO PCT/FI1983/000073 patent/WO1985002259A1/en not_active Application Discontinuation
Non-Patent Citations (1)
Title |
---|
See references of WO8502259A1 * |
Also Published As
Publication number | Publication date |
---|---|
JPS61501162A (en) | 1986-06-12 |
WO1985002259A1 (en) | 1985-05-23 |
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Legal Events
Date | Code | Title | Description |
---|---|---|---|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
17P | Request for examination filed |
Effective date: 19850627 |
|
AK | Designated contracting states |
Designated state(s): AT BE CH DE FR GB LI LU NL SE Kind code of ref document: A1 Designated state(s): AT BE CH DE FR GB LI LU NL SE |
|
17Q | First examination report despatched |
Effective date: 19870529 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN |
|
RIN1 | Information on inventor provided before grant (corrected) |
Inventor name: SUOVANIEMI, OSMO Inventor name: EKHOLM, PERTTI Inventor name: PARTANEN, PAUL Inventor name: KAUKANEN, ESKO |