FI91808C - Immunological assay system - Google Patents

Description

1 91808

Inununological determination system

The invention relates to an automatic immunoassay system using the inner surface of a reaction vessel as a solid phase. The system of the invention can be used in general for photometric, fluorometric, phosphorimetric or radiological immunoassays.

To determine the antibody (or antigen) in a sample, the so-called heterogeneous immunoassay methods in which an antibody (or antigen, respectively) and a labeled antibody (or antigen, respectively) are allowed to bind to an antigen (or antibody, respectively) placed on a solid surface. These methods have a solid surface on which the inununological reaction has taken place, and the reaction solution is separated from the solid surface before measuring the tracer signal so that the excess tracer in the reaction solution does not mask the signal in the solid phase bound antibody (or antigen, respectively). This signal may be, for example, radioactivity (RIA), fluoro-. 20 reactive activity (FIA) or enzyme activity (EIA).

In all these methods, the solid phase is washed thoroughly before measurement.

The methods currently in use always have manual steps; at least a washing step for which no reliable means of operation have been found in operation.

Automated methods should aim to use reaction and measurement cuvettes formed from several sample vessels. The reactions must generally be carried out accurately at a temperature of 30 ° C. To control the temperature, incubators have been used which have a heated body and a separate cuvette-shaped cavity for each cuvette in the cuvette system.

However, this solution is not satisfactory, since it is thus difficult to obtain the temperature of the various cuvette paths to remain sufficiently exactly as desired. In this way, it is not possible to use cuvettes as large as would be advantageous from the point of view of automation.

The object of the present invention is, above all, to provide a fully automatic heterogeneous immunoassay system. In particular, it is an object to provide a system in which the heating and washing steps of MySs take place automatically and reliably, even when using large cuvette systems.

The present invention includes: a horizontally movable cuvette carriage; above it a horizontally movable dispensing head; a single-candle liquid dispenser attached to the dispensing head and having a tip that can be lowered to the bottom of the cuvette at the bottom; a dispensing apparatus for introducing the desired liquid into the cuvette and removing the solution 10 in the cuvette from the cuvette through the tip; as well as a measuring device attached to the dosing head. The invention and some of its applications are precisely defined in the claims.

A preferred embodiment of the invention is illustrated more closely by means of the accompanying drawings. In them, Fig. 1 15 shows an overall view of the system, Fig. 2 a partial view of the contents of the analyzer, Fig. 3 the tip of the liquid dispenser lowered to the bottom of the cuvette and Fig. 4 the thermostated incubator of the apparatus.

The main parts of the equipment are the analyzer 1, the computer 2 with 20 display units 3 and the printer 4. Other control or communication and storage devices 5 can also be connected to the system. The analyzer also has its own access and output panel 6. The analyzer cover 7 opens. The ignition system is placed in the moving carriage 8. The trajectory of the carriage is 25 straight lines and marked with an arrow at the head of the carriage. Above the carriage 8, a dosing head 9 is moved along a path perpendicular to the trajectory of the carriage, to which a photometer 10 and a liquid dispenser 11 are attached. The direction of movement of the dosing head 9 is indicated by an arrow 30a next to it.

The measurement is performed by passing a light beam from below the hole 12 through the bottom of the sample cuvette through the measuring tip 13 of the photometer 10 for detection.

The liquid dispenser 11 has a main channel 14 connected to the pump 1 and a side channel 15 connected to the piston pump opening inside it (Fig. 3). The tip 15 of the liquid dispenser can be lowered to the bottom of the cuvette below.

The tip 15 of the liquid dispenser 11 is made of a flexible material and is preferably curved (Figure 3). When the curved tip 15 is lowered to the bottom of the cuvette 18 of the cassette system 17, the mouth of the cattle settles in the corner of the cuvette 18. This allows the liquid in the cuvette 18 to be absorbed as completely as possible.

The analyzer uses a vertical measuring cuvette 17, 5 with a plurality of cylindrical measuring cuvettes 18 with a transparent bottom, which are connected from the top to each other into a rectangular matrix on a support plate. An antigen of the antibody to be determined is attached to the inner walls, in particular the bottoms, of the Ky waters 18. The carriage 8 10 has light-blowing holes at the cuvettes 18.

Carriage 8 has a thermostated incubator in which the cuvette system is placed. The incubator has heating pins located between the cuvettes 18. The pins 19 are rectangular in cross-section, the sides of which curve inwardly, mimicking the walls of the cuvettes 18 so that a small gap divides between the pins 19 and the cuvettes (Fig. 4). In this way, air can circulate freely between the cuvettes, and the temperature of all the cuvettes remains as close as desired.

The carriage 8 has a place 20 for the cuvette system 17 as well as a place 21 for the sample vessels 20 and a place 22 for the necessary reagent and liquid removal vessels.

The hardware works as follows:

The lid 7 of the analyzer is opened and the samples, for example serum samples, are placed in the space 21 reserved for them and the cuvette set 17 in the space 20 reserved for it. The lid 7 is closed and the desired program is started. The liquid dispenser 11 picks up and dilutes each sample in its own cuvette 18 and collects and dispenses the labeled antibody and any other reagents that may be needed into each cuvette. Samples 30 are incubated for the desired time at which the immunological reaction occurs.

Now, the liquid dispenser 11 alternately draws the reaction solutions of all the cuvettes 18 into the tank channel 14 and removes them into the container provided for them, while rinsing the channel. Now, 14 washing liquids are dispensed alternately into all cuvettes from the tank duct, after which the cuvettes are again emptied as described above. Finally, the amount of labeled antibody bound to each cuvette is measured and the results of the assay are printed as desired.

The solution is removed from the cuvette by inserting the candy 15 into the bottom of the cuvette 18 and, when the solution is aspirated, the curved candy 15 is moved against the corner of the cuvette. Thus, the tip 15 is always accurately placed in the corner of the cuvette 18 and the solution removed as accurately as possible.

The wash liquid is dispensed into the cuvette 18 so that the liquid jet does not remove unreacted antibody, but nevertheless removes excess labeled antibody and other epa-10 purities in solution.

A preferred photometric application has been described above. The arrangement can also be applied to fluorometric or radiological measurement. Otherwise, the hardware may differ from the above in its individual details. It may have, for example, a separate incubation station to which the trolley is driven. The dosing head can, of course, also be used during incubation, such as photometry.

In one preferred embodiment, several different assay sequences are performed simultaneously.

Claims (8)

An immunological determination device comprising a wave (8) movable in the horizontal plane having a space (20) for a cuvette system (17), a dosing spirit (9) in the horizontal plane above the carriage (8), a liquid dispenser (11) having a tip fixed in the dispensing spirit (9) and the spring tip (15) can be sunk into the bottom of a cuvette (18) on its lower side, 10. a dispensing device for feeding liquid to the cuvette and before removing a solution from the cuvette through the tip (15), a measuring device (10) which is fixed in the dosing spirit (9) and which is intended to measure a signal, characterized in that the cuvette space (20) in the carriage (8) ) is a heated incubator, the cuvettes (18) of the cuvette system on its inside have an antigen fixed for the countermeasure to be determined, and that the measuring device is intended to measure a signal emitted by a marking member stuck to the inner wall of the cuvette (18) . 2. Device according to claim 1, characterized in that the carriage is movable only along a rectilinear path. 3. Device according to claim 1, characterized in that the dosing spirit is mobile only along a linear path. Device according to any one of claims 1-4, characterized in that the carriage has a space (20) for a cuvette system (17), spring cuvettes (18) form a rectangular matrix and year fixed to each other from their upper spirits. with a base plate and where in the incubator there are heating pins (19) which are placed in the space between the cuvettes. 5. Device according to claim 4, characterized in that the cross-section of the heating pins (19) is substantially square. 6. Device according to claim 5, characterized in that,. that the carriage has a space (20) for a cuvette system (17), the spring cuvettes are cylindrical and the side walls of the heating pins (19) are concave to correspond to the curvature of the cuvettes' walls. Apparatus according to any one of claims 1-6, characterized in that the tip (15) of the water dispenser (11) is hooked. 8. A method of determining a counterpart in a liquid sample, in which a method of placing a cuvette system (17) in a horizontal wave (8) places the immunological reaction, 10. with the aid of a measuring device ( 10) which is fixed in the dosing spirit (9) feeds a signal that a marking signal emits, characterized in that the cuvettes (18) of the cuvette system (17) have fixed on their inside an antigen for the object to be determined, - in a thermoregulable wave (8) places the sample vessel containing the samples to be determined, as well as the reagent kMrl containing the necessary reagents, with the aid of a heat meter (11) having a tip (15) extending above the cart (8), the tip (15) can be soaked in the bottom of a cuvette (18) on its lower side, obtaining for each cuvette a sample from the sample and reagents from the reagent vessel. with the aid of the tip (15), the solutions from the cuvettes (18) are removed, with the aid of the tip (15), dispensed with water to the cuvettes, and removed it from them, and feeds as a marking substance which adheres to the internal wall of the cuvette (18). li