JPS6363862B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention provides a method for applying a predetermined amount of a first reactant, such as a biological fluid (eg, serum or plasma) containing the substance to be analyzed, to a biological indicator (eg, an enzyme or The present invention relates to an analytical device of the type using a solid reaction support for each successive addition of a predetermined amount of a second reactant comprising a protein immobilized with a radioactive isotope (in radioimmunology).

Conventionally, a spherical beaded solid reaction support, usually made of polystyrene, coated with a protein having the properties of an antibody (eg, a polypeptide produced from vaccinated animals) has been used.

The support is placed in a container in the form of a test tube, from which a predetermined amount of the first reactant is introduced into the container, and then, after incubation for a predetermined period of time, the excess reactant is removed. Rinse the balls to remove them.

This cleaning requires manual work, such as inverting the tube with the open end and discharging the liquid to prevent the beads from popping out. This operation has to be carried out anew for the second reactant, the remainder of the second reactant being drained off again by a water wash, and after this the tube is, for example, photographed while injecting the colored reactant. Sent to analytical process for colorimetric analysis.

The more simply and completely the principle of trapping beads that successively trap the expected amounts of reactants is formed, the more unsatisfactory the technology used becomes. In fact, manual rinsing is unreliable due to non-uniform rinsing and temperature conditions, and because of the large number of operations required and the inevitable errors associated with it, various liquids can be It is not suitable for analysis based on the combination of

An object of the present invention is to provide a biological fluid analysis device that can analyze biological fluids with simple operation and with high reliability.

According to the invention, said object comprises a container member having a bottom and accommodating a solid reaction support on said bottom for capturing a predetermined substance contained in a biological liquid; a tube member extending into the tube member and having one end connected to the upper portion of the container; and an inlet member connected to the other end of the tube member for introducing a washing liquid for the solid reaction support into the tube member. an inlet provided in the container member for injecting the biological liquid into the container member;
rotating means connected to the assembly for rotating the assembly of the tube member and the inlet member around the inlet member; an outlet member provided at a portion of the container member furthest from the axis of rotation of the assembly for discharging the biological liquid and the introduced washing liquid from the container member. This is accomplished by a liquid analyzer.

According to the apparatus of the present invention, a predetermined substance contained in a biological liquid can be captured on a solid reaction support with simple and efficient operation, and excess organisms attached to the solid reaction support can be removed by a washing liquid. Biological fluids can be washed and analyzes of biological fluids can be performed quickly.

Hereinafter, the present invention will be explained in detail using preferred non-limiting specific examples shown in the drawings.

1 to 3 show an analysis apparatus according to the present invention, which is composed of a rotor 1. FIG.

The rotor 1 is equipped with a plurality of containers 2 each housing one ball 3 as a conventional solid reaction support for capturing solids at the periphery, and a plurality of containers 2 each housing one ball 3 as a conventional solid reaction support for capturing solids, and a center section as an inlet member for introducing a cleaning liquid. Equipped with 4 orifices. The orifice 4 and each container 2 are connected by straight pipes 5 as pipe members arranged radially, and the pipes 5 exist in a plane substantially perpendicular to the rotation axis 6 of the rotor 1. . An annular groove may be provided instead of the orifice 4, and in this case, the annular groove and each container 2
and are suitably connected by radially arranged tubes. At the top of the container 2, an orifice 8 is provided as an inlet for injecting a reactant as a biological liquid into the container 2.

As shown in FIG. 2, the rotor 1 consists of two half bodies 1A and 1B divided in horizontal cross section, and a groove defining a tube 5 is provided in each of the half bodies 1A and 1B. . Here, half-split body 1A, 1
A groove may be provided only on one side of B, and the other side may be flat.

The container 2 is directly defined by the overlapping peripheral edges of the two halves 1A and 1B, and is equipped with an orifice 7 on the side as a discharge port for discharging the liquid in the rotor 1. An orifice 8 for injecting the reactants into the vessel 2 is provided at the top. The bottom part 9 of the container 2 is shaped so that when the rotor 1 is in a rotating state, the balls 3 rise to approximately the height of the orifice 7 due to centrifugal force (see Figs. 4A, 4B and 4B). (described later with reference to FIG. 4C).

The connection means and relative positioning means for the half bodies 1A, 1B constituting the rotor 1 can be appropriately selected by those skilled in the art. In this specific example, the half-split body 1B
A fitting part 10 consisting of an annular protrusion provided on the half body 1A and an annular groove that fits into the protrusion part provided on the half body 1A, and a cylindrical groove provided on the half body 1B. A half body 1 consisting of a protrusion and a recess provided in the half body 1A and fitted into the protrusion
The fitting portions 11 for centering A and 1B are shown. The rotor 1 is rotated by a shaft portion 13 provided at the center and serving as a driving rotation means.

Hereinafter, the features of this specific example related to the shape of the container 2 will be explained along with the uses of this specific example.

(1) Fig. 4A Each container 2 has a ball 3 suitable for a specially formulated liquid.
is stored. The rotor 1 is arranged such that the container 2 preferably receives a measured volume of a first reactant such as any other biological fluid (serum or plasma) containing the substance to be analyzed via an orifice 8. , the container 2 is positioned below the pipette 12 containing the first container. In order to position the orifice 8 of each container 2 below the pipette 12, for example, the rotor 1 is rotated one after another by an angle corresponding to the container 2. After the first reactant is injected into all the containers 2 through the orifice 8, they are left in a warm state for a predetermined period of time.

The container 2 has a volume sufficient to accommodate the ball 3 and a predetermined volume of the first reactant without the first reactant injected into the container 2 reaching the fifth height.

(2) FIG. 4B The rotor 1 is kept in rotation by the shaft 13, and the excess first reactant is discharged through the container 2 or the orifice 7. The ball 3 moves up to the level of the orifice 7 due to the bottom part 9 with sloping walls. Next, a cleaning liquid is introduced into the rotor 1 through the orifice 4. The introduced cleaning liquid reaches the container 2 through the tube 5 due to centrifugal force, and powerfully cleans the balls 3. The presence of the ridge 14 in the area of the orifice 4 facilitates the distribution of the cleaning liquid introduced from the orifice 4 into the tube 5. Moreover, since the shape of the container 2 is tapered particularly toward the periphery, the position of the ball 3 is maintained in a preferred position in the presence of centrifugal force. After the cleaning liquid surrounds the ball 3, it is discharged from the container 2 through the orifice 4 while rotating the ball.

(3) Fig. 4C While rotating the rotor 1, stop introducing the cleaning liquid and dry the balls 3. This step of drying the balls 3 is carried out by the rotation of the rotor 1 in this embodiment, whereas the conventional apparatus requires complicated elevation, and is therefore very easily carried out.

The above operations according to FIGS. 4A to 4C are repeated using the second reactant. The second reactant includes a protein having an immobilized biological indicator, such as an enzyme or a radioisotope.

Next, once the rotor 1 stops, the balls 3 in the container 2 descend again. This provides for direct analysis using a counter if a radioisotope is used, or indirect analysis if a colored reactant or a terminating reactant is used in addition to the radioisotope. Ru. The wall defining the lower part of the container 2 is configured to allow such an analysis to be carried out.

5A and 5C show each container 2 of the rotor 1.
A case is shown in which there is not only one ball 3 but a plurality of small balls 3'. The diameter of the small ball 3' is practically about 10 to 20 microns. Further reference number 2
0 indicates a barrier that prevents the balls 3' from flying out of the container during rotation of the rotor 1, and the barrier 20 is fitted into a cutout provided in the half bodies 1A and 1B forming the rotor 1. or glued. The barrier 20 is preferably of the fine filter type.

Here, each vessel 2 with a collection of beads 3' is placed below a reactant supply device, such as a pipette 12. The ball 3' of each container 2 can correspond to a particular reactant.

FIG. 5B corresponds to the moment when the rotor 1 is rotating and the washing liquid is injected; in this figure, as a result of centrifugation, excess reactant is discharged from the orifice 7 and the ball 3' is tilted. Bottom part 9 with a wall part
It moves until it is stopped by the barrier 20.

The presence of a large number of beads 3' of small diameter can in particular increase the trapping area and trapping power of the reactants.

The container 2 may have a shape different from that depicted in FIGS. 1 to 5B. In particular, container 2 is the sixth
and FIG. 7, in which the inner inner wall 21 and the outer inner wall surface 22 of the container 2 are substantially parallel, and the outer inner wall 22 has an orifice 7 for the discharge of liquid. It has an outlet member 23 defined by the edge.

In FIGS. 6 and 7, the inner wall 2 of the container 2
1 and 22 form a capture support. However, the general configuration of the container 2 is similar to that which can be used when the support is constituted by one or many beads.

This analyzer exhibits the same general form as the one described above.

The capture solid support consists of the lower 3" wall of the container 2 coated with a protein having antibody-type properties. The inner inner wall 21 and the outer inner wall 22 of the container 2 at the height of the lower 3" are parallel. It is. moreover,
The outer inner wall 22 has an outlet member 23 defining the orifice 7 at its edge, the outlet member 23 being positioned at the rear of the container 2 with respect to the direction of rotation as shown.

The rotor 1 has an orifice 8 in each container 2;
a first reactant supply device for injecting a volume, preferably a measured, predetermined volume of the first reactant, such as serum, plasma or any other biological fluid containing the substance to be analysed; placed below. After injection of the first reactant, there is a pause for the required incubation period.

Next, the rotor is rotated once, and the orifice 4
Inject the cleaning solution from the At the outlet of the tube 5 there are provided means 24 forming a baffle plate so as to better direct the cleaning liquid towards the bottom of the container 2. Excess reactant is discharged from the container 2 via the orifice 7 by centrifugal force. Furthermore, the cleaning liquid, whose good distribution is helped by the presence of the ridges 14, passes through the tube 5 and intensively cleans the lower wall of each container 2. The cleaning liquid is discharged from the orifice 7. The drainage of the cleaning liquid is complete due to the presence of the outlet side 23.

After washing, while continuing centrifugation, feeding of the washing liquid is optionally stopped and drying of the container 2 begins.

These operations are then repeated using a second reactant, which includes a protein having an immobilized biological indicator, such as an enzyme or a radioisotope. Once rotor 1 is stopped, direct analysis by counter when using radioisotopes or indirect analysis when using additionally colored reactants or terminating reactants can proceed. .

The parallelism of the inner and outer inner walls of the lower part of the container section facilitates photometric analysis in each container 2.

However, the shape of the container 2 may also be other shapes, in particular shapes corresponding to FIGS. 1 to 5B.

According to the above embodiment, the operation of the device can be simplified and the reliability can be increased because the processing is very homogeneous. By way of example, the embodiment described above makes it possible to easily and quickly obtain the five simultaneous formulations required for the analysis of the thyroid gland.

In particular, the rotor 1 can be divided into two zones and several containers 2 can be used as test references, or the same acquisition support can be used to make it possible to carry out the same test on several patients, or in particular the same It also has wide application due to the possibility of preparing various supports etc. that make it possible to carry out several tests on the patient.

Note the important advantage of small space requirements, since such devices can be manufactured up to approximately 5 cm in diameter.

[Brief explanation of drawings]

FIG. 1 is a partial perspective view of an embodiment of the present invention, FIG. 2 is a partial axial cross-sectional view of the device of FIG. 1, and FIG.
4A, 4B, and 4C are explanatory diagrams of the operation of the device shown in FIG. FIGS. 6 and 7 are explanatory diagrams of the effects of still another specific example of the present invention. 1...rotor, 2...container, 3...ball, 4...
Orifice, 5...tube, 6...rotating shaft, 7...orifice, 8...orifice, 9...bottom.

Claims (1)

[Scope of Claims] 1. A container member having a bottom and housing a solid reaction support on the bottom for capturing a predetermined substance contained in a biological fluid; a tube member connected at one end to the upper part of the container; an inlet member connected to the other end of the tube member for introducing a washing liquid for the solid reaction support into the tube member; an inlet provided in the container member for injecting the biological liquid into the container member; and an inlet provided in the container member for injecting the biological liquid into the container member; rotating means connected to the assembly and said biological liquid in said container member and said introduced washing liquid being discharged from said container member when centrifugal force due to rotation of said assembly is applied; and an outlet member provided at a portion of the container member farthest from the rotational axis of the assembly. 2. The container member includes a plurality of containers, the tube member includes a plurality of straight pipes having the same length, and the assembly and the inlet member are arranged at equiangular pitches around the central axis. 2. The device of claim 1, further comprising: a plurality of containers; and a plurality of tubes connecting the inlet member and each of the containers. 3. Device according to claim 1 or 2, characterized in that the assembly consists of two halves divided in horizontal section. 4. The solid reaction support is a spherical member consisting of one sphere, and the outlet member is configured to retain the sphere within the container member when centrifugal force due to rotation of the assembly is applied. Device according to any one of claims 1 to 3, characterized in that it is configured. 5. The solid reaction support is a spherical member composed of a plurality of microspheres, and the outlet member is configured to retain the microspheres in the container member when centrifugal force due to rotation of the assembly is applied. An apparatus according to any one of claims 1 to 3, characterized in that the apparatus is configured as follows. 6. The device according to any one of claims 1 to 4, wherein the outlet member has one small orifice. 7. The device according to any one of claims 1 to 5, wherein the discharge port member comprises a fine filter. 8. The device of claim 7, wherein the microspheres have a diameter of 10 to 20 microns. 9 A wall of the container member connecting the outlet member and the bottom is such that when the centrifugal force acts, the ball member freely moves from the position of the bottom to the position of the outlet member, and the centrifugal Claims 4 to 8, characterized in that the ball member has a shape such that it freely moves from the position of the outlet member to the position of the bottom part when no force is applied.
Apparatus according to any one of paragraphs. 10. Apparatus according to any one of claims 1 to 3, characterized in that the solid reaction support is constituted by a lower wall of the container member. 11 The inner inner wall and the outer inner wall of the container member are substantially parallel, and the outer inner wall constitutes the outlet member and allows complete drainage of the cleaning liquid when subjected to centrifugal force due to rotation of the assembly. An apparatus according to any one of claims 1 to 3, characterized in that it is configured as follows. 12. The apparatus of claim 11, wherein the outlet member is located after the container member with respect to the direction of rotation of the assembly.