JP3249609B2 - Immunochemical measurement device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an immunochemical measuring apparatus which fully automates the analysis of a sample.

[0002]

2. Description of the Related Art First, FIG.
This will be described with reference to FIG. Note that (S1) to (S1) in FIG.
17) shows each step.

(S1): A liquid medium (for example, a phosphate buffer solution) and a sample (an antibody or an antigen against an antigen or an antibody to be measured) are injected into a reaction vessel made of a nonmagnetic substance.

(S2): A magnetic latex is added as a first reagent.

(S3): After mixing, wait for about 6 minutes.

(S4): The magnets are set on both sides of the reaction vessel, and the magnetic flux passes through the reaction vessel with the opposite surfaces of the two magnets having opposite polarities.

(S5): The liquid medium and the magnetic latex reacted with the sample are separated by the magnetic field.

(S6): Only the liquid medium is drained.

(S7): Reset the magnet.

(S8): Eu (Europume) latex is injected together with a liquid medium as a second reagent.

(S9): Mix.

(S10): After mixing, wait for about 6 to 26 minutes.

(S11): The magnet is set again.

(S12): The reacted magnetic latex, Eu latex and antibody (or antigen) are separated from excess Eu latex and liquid medium.

(S13): The amount of excess Eu latex is measured by the amount of transmitted light (OD) and / or the fluorescence intensity (FI).

(S14): Excess Eu latex and liquid medium are discharged.

(S15): The magnet is reset again.

(S16): A dispersion (for example, water containing a surfactant or physiological saline) is injected.

(S17): After mixing, it is turned to measurement.

As described above, the measurement of one sample is completed. This is described in terms of the reaction method. (A) The first reagent consisting of an antibody or an insoluble magnetic particle carrying an antigen to the antigen or antibody to be measured and the antigen or antibody to be measured are placed in a reaction vessel. React in liquid medium. (B) The insoluble magnetic particles in step (a) are adhered to the reaction vessel wall by the action of a magnetic field, and after removing the liquid medium, the insoluble magnetic particles are further washed. (C) a second reagent comprising an insoluble fluorescent dye-labeled particle carrying an antibody or an antigen against the antigen or antibody to be measured, which is the same as in step (a);
The antibody-reacted insoluble magnetic particles are reacted in a liquid medium. (D) attaching the insoluble magnetic particles of step (c) to the reaction vessel wall by the action of a magnetic field, first measuring the unreacted excess insoluble fluorescent dye-labeled particles, and then measuring the liquid medium and the unreacted insoluble fluorescence; The dye-labeled particles are removed, and then the fluorescence intensity of the insoluble fluorescent dye particles that have reacted with the insoluble magnetic particles is measured. It turns out that.

[0021]

In the conventional immunochemical measurement method, for example, the above-mentioned procedures (S1) to (S17) are required, and it is difficult to fully automate the method. In particular, the setting and resetting of the magnet in the reaction vessel had to be done manually, which was a bottleneck in full automation.

An object of the present invention is to provide an immunochemical measuring apparatus which automates all procedures including setting and resetting of a magnet and has high processing efficiency.

[0023]

In the immunochemical measuring apparatus according to the present invention, a container set for mounting or removing a reaction container is formed over the entire periphery of the turntable, and a space is provided between the reaction containers. A sample portion in which movable magnets movable to a position facing the side surface of each reaction vessel and a position distant from the side surface are arranged in such a manner that the polarities of the movable magnets are sequentially reversed, and
A reaction vessel moving mechanism for attaching and detaching the reaction vessel to and from the vessel setting section, a sample dispensing mechanism, a reagent dispensing mechanism, a cleaning mechanism for the reaction vessel, and a cleaning mechanism for the reaction vessel outside the rotary table. A stirring mechanism for the contents, a fluorescence detection mechanism for measuring the fluorescence intensity of the sample, a transmitted light detection mechanism and a mechanism for inserting and pulling out the movable magnet are arranged and fixed, and the rotary table is rotated according to a predetermined sequence. And an operation unit having a control mechanism that can operate the mechanisms according to timing.

Further, the present invention uses a fixed magnet in place of the movable magnet and moves the reaction vessel from the reset position to the set position or vice versa, so that the fixed magnet can be positioned on the side surface of the reaction vessel. It is configured not to do it.

[0025]

In the present invention, when the rotary table of the sample section is rotated clockwise or counterclockwise by a required step by the control mechanism, the first reagent is dispensed into a predetermined reaction vessel, mixed and stirred, and the movable magnet is moved. Is set, and in that state, the drainage and movable magnet are reset, the second reagent is dispensed, mixed and stirred, the movable magnet is set, drained in that state, the movable magnet is reset, and the dispersion liquid is set. Is injected, and after mixing and stirring, the operation for measuring is continuously performed.

Further, a fixed magnet is used in place of the movable magnet,
The same measurement as described above is performed by moving the reaction vessel.

[0027]

FIG. 1 is a plan view showing an entire configuration of an embodiment of the present invention. Reference numeral 100 denotes an immunochemical measurement device, which is roughly divided into a sample unit 100A and an operation unit 100B. The sample section 100A is mainly composed of a rotary table 10 which is rotated clockwise or counterclockwise by a drive mechanism (not shown), and the reaction vessel 1
2 (not shown) are formed at equal intervals. The illustrated example of the container setting portion 11 is a rectangular hole, into which a reaction container 12 described later is fitted, and which can be detached by lifting. Numeral 13 denotes a movable magnet, which is arranged between the container setting portions 11 so as to be inserted and pulled out.

1 to 40 shown inside the rotary table 10
Indicates the fixed position, and at the same time indicates the number of the reaction vessel 12 corresponding to the position shown. Therefore, assuming that the rotary table 10 is rotated clockwise by two pitches from the state shown in FIG. 1, the reaction container (39) faces the position [1] (the position is [], and the reaction container The number 12 (also referred to as a sample) is shown in parentheses).

Various mechanisms are arranged and fixed outside the sample section 100A to form an operation section 100B. Hereinafter, these mechanisms will be sequentially described.

Reference numeral 20 denotes a reaction vessel moving mechanism, and reference numeral 30 denotes a sample dispensing mechanism, which is provided corresponding to the position [1]. The reaction vessel moving mechanism 20 includes a reaction vessel 12
The sample dispensing mechanism 30 has a function of dispensing a sample into the reaction container 12. A reagent dispensing mechanism 40 is provided corresponding to the position [21], and includes a first reagent (for example, magnetic latex) and a second reagent.
It has the function of injecting reagents (eg, Eu latex), dispersions, and the like.

Reference numeral 50 denotes a cleaning device, which is provided corresponding to [position 18] and has a function of cleaning the inside of the reaction vessel 12. Reference numeral 60 denotes a stirring mechanism, which is provided corresponding to the position [16] and has a function of mixing and stirring the contents of the reaction vessel 12. 70 is a photometric light source such as a xenon flash lamp, a halogen tank stainless lamp with a chopper mechanism, etc., which is provided corresponding to the position [11], and 80A is a transmitted light detecting mechanism, corresponding to the position [11]. Is provided,
The intensity of the transmitted light from the light source 70 is measured. 80B is a fluorescence detection mechanism for measuring the fluorescence intensity of the sample.
90A is an insertion mechanism, which is a magnet insertion portion 91 provided between positions [25] and [26] and a magnet insertion portion 92 provided between positions [24] and [25]. Consists of Reference numeral 90B denotes a drawing mechanism, which includes a magnet drawing portion 93 provided between positions [6] and [5].
It consists of a magnet pull-out portion 94 provided between positions [4] and [3].

FIG. 2 shows the relationship between the details of the rotary table 10 and the magnet insertion portions 91 (92). FIG. 3 is a partial plan view of the rotary table 10, and FIG.
FIG. In these figures, a reaction container 12 is fitted to the container setting section 11. A U-shaped mounting member 16 is provided on the periphery of the lower surface of the turntable 10, and the upper end of the arm 14 is rotatably mounted on the mounting member 16 by a pin 15. The movable magnet 13 is fixed to the lower end of the arm 14. On the other hand, a fixture 17 is provided on the lower surface of the turntable 10 similarly to the fixture 16,
An L-shaped magnetic piece 18 is fixed to this.

Next, referring to FIG. 2, the magnet insertion portion 91 (92)
Will be described. W is a coil, which attracts the plunger P against the spring C by energization, and pulls the upper end of the arm 14 of the movable magnet 13 in the direction of the arrow at the tip end thereof. Therefore, the arm 14 is tilted as indicated by the dotted line, the movable magnet 13 approaches the magnetic piece 18 to be in the attracted state, and the state shown by the dotted line is maintained. This state indicates that the movable magnet 13 has moved to a position facing the side surface of the reaction vessel 12.

In FIG. 1, the magnet insertion portions 91 and 92 are operated at the same time so that the two movable magnets 13 are in contact with both side surfaces of the reaction vessel 12, in other words, the reaction vessel 12 is sandwiched between the movable magnets 13. Shape. Since the movable magnets 13 are arranged so that the polarities of the adjacent magnets are sequentially reversed, that is, the reaction vessel 12 enters between the N-pole and the S-pole, so that the magnetic field works effectively and the separating action is sufficiently performed. Will be The configuration of the magnet extracting portions 93 and 94 is opposite to that of the magnet inserting portions 91 and 92, that is, when the coil W is energized in FIG.
Is driven in the direction opposite to the arrow, and pushes the upper end of the inclined arm 14 with the projection shown by the imaginary line to return the arm 14 from the dotted line state to the solid line vertical state.

FIG. 5 shows an embodiment in which the movable magnet 13 is embedded in the arm 14 by plastic molding.

Next, the overall operation of FIG. 1 will be described. Although the control mechanism is omitted in FIG. 1, the following description is based on the assumption that the control mechanism rotates the turntable 10 clockwise and counterclockwise by a predetermined pitch in accordance with a predetermined sequence. It is operated or stopped at the timing of.

1) First, the sample (1) is located at the position [1].
It is in. Then, the reaction container moving mechanism 20 detaches and takes it out.

2) Next, it rotates counterclockwise by 26 pitches. Sample (4) is at position [18], sample (2)
Is at the position [16], and the sample (11) is
It comes to the position [25] of 1 and 22, stops, and executes each operation.

3) Next, rotate 9 pitches clockwise,
The sample (38) stops at the position [21] for dispensing the first reagent. At this time, the sample (22) is located at the position [5] of the magnet extraction portions 93 and 94, and executes the respective operations.

4) Next, in order to stir the sample (38), the sample (38) is rotated counterclockwise by 5 pitches to a position [16].

5) Next, the sample (9) is rotated counterclockwise by 16 pitches to bring the sample (9) to the position [11]. At this time, the sample (4) is located at the position [6] of the magnet extraction portions 93 and 94, and executes the respective operations.

6) Next, the sample (2) is moved to the position [11] by rotating the sample 7 clockwise by 7 pitches. Then, the sample (9) is in the washing position and the sample (7) is in the stirring position. Perform the action.

7) Next, the sample (41) is rotated counterclockwise by nine pitches and set at the position [1] where the reaction container enters and exits. At this time, the sample (21) is at the reagent dispensing position [21] and receives the second reagent. The sample (26) is located at the position [26] of the magnet insertion portion 91, and performs each operation.

8) Next, the sample (7) is rotated clockwise by 18 pitches to move the sample (7) to the position [25], and the movable magnet 13 is set. At this time, the sample (3) is at the reagent dispensing position [21], and the dispersion liquid R3 is dispensed here.

9) Next, the sample (21) is rotated clockwise by 17 pitches to move the sample (21) to the stirring position [16]. At this time, the sample (23) is placed at the washing position [18].
And the sample (9) is at the position [4] of the magnet extraction, and executes the respective operations. 10) Finally, rotate 4 pitches clockwise to position [1].
Wait for taking sample (2). Repeat the above.

The number of the container setting sections 11 in the above embodiment, or the positions where the respective mechanisms such as the reaction vessel moving mechanism 20, the sample dispensing mechanism 30,... Are provided are not limited to those illustrated. Various positions are possible. Obviously, it can be easily handled by changing the control sequence accordingly. Also, the insertion mechanism 90 of the movable magnet 13
A, the pull-out mechanism 90B and the like are attached and detached by rotation, but it goes without saying that these may be configured to be moved forward and backward.

FIG. 6 shows another embodiment of the present invention.
In the embodiment of FIG. 1, the reaction vessel 12 is fixed, and the movable magnet 13 is moved to set and reset. In FIG. 6, the reaction vessel 12 is moved and the fixed magnet 13A is not moved. It is.

In FIG. 6, reference numeral 11A denotes the reaction vessel 12;
Is a container setting part which can be moved to a set position and a reset position. That is, the reaction vessel setting mechanism 90C and the reaction vessel reset mechanism 90 move from the reset reaction vessel 12a to the position of the set reaction vessel 12b or vice versa.
D can be operated.

In this case, the fixed magnet 13A is provided inside the rotary table 10, but it may be provided outside and the setting and resetting may be reversed. Further, the reaction container setting mechanism 90C and the reaction container reset mechanism 90D for moving the reaction container 12 may be various structures as long as they can move the reaction container 12 forward and backward. In the embodiment of FIG. 6, the reaction vessel setting mechanism 90C and the reaction vessel reset mechanism 9 are provided separately from the reaction vessel moving mechanism 20.
Although 0D is provided, this can be integrated into one.

Further, as a method of using the immunochemical measurement device of the present invention, a sequence for measuring the amount of transmitted light of each reaction vessel 12 passing through the optical path while the turntable 10 is rotating is added to the conventional method. Latex agglutination reaction can also be measured.

In the above embodiment, the shape of the reaction vessel 12 is rectangular, but it can be any shape such as a circle or a square.

[0052]

As described above in detail, the immunochemical measuring apparatus according to the present invention has a container set section for mounting or removing a reaction container around the entire periphery of the turntable, and has the above-mentioned configuration. A movable magnet movable between a position opposed to the side surface of each reaction container and a position distant from the side surface between the reaction containers is sequentially adjacent to each other, and a sample portion arranged by reversing the polarity is provided outside the rotary table. A reaction vessel moving mechanism for attaching and detaching the reaction vessel to and from the vessel setting section, a sample dispensing mechanism, a reagent dispensing mechanism, a washing mechanism for the reaction vessel, and a stirring mechanism for stirring the contents of the reaction vessel. ,
A fluorescence detection mechanism for measuring the fluorescence intensity of the sample, a transmitted light detection mechanism for measuring the transmitted light intensity of the sample, and a mechanism for inserting and extracting the movable magnet are arranged and fixed. An operating unit having a control mechanism for rotating and operating each of the mechanisms in accordance with the timing, so that the insertion and withdrawal of the magnet into and from the reaction vessel can be automatically performed according to the rotation of the rotary table. This has the advantage that full automation, which has been difficult so far, can be realized.

In the present invention, the movable magnet is replaced with a fixed magnet, and the reaction vessel is moved from the reset position to the set position or vice versa so that the fixed magnet may or may not be located on the side of the reaction vessel. Since the same measurement as described above can be performed, and the reaction vessel can be reset and set using the reaction vessel moving mechanism, the configuration can be simplified accordingly.

[Brief description of the drawings]

FIG. 1 is a diagram showing a configuration of an embodiment of the present invention.

FIG. 2 is a side view showing a part of the embodiment of FIG.

FIG. 3 is a partial plan view of FIG. 2;

FIG. 4 is a partial front view of FIG. 2;

FIG. 5 is a perspective view showing another embodiment of the movable magnet of the present invention together with a reaction vessel.

FIG. 6 is a diagram showing a configuration of another embodiment of the present invention.

FIG. 7 is a diagram illustrating an example of a conventional immunoassay procedure using a latex agglutination reaction.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 10 Rotary table 11 Container setting part 11A Container setting part which can move a reaction container 12 Reaction container 13 Movable magnet 13A Fixed magnet 20 Reaction container moving mechanism 30 Sample dispensing mechanism 40 Reagent dispensing mechanism 50 Cleaning device 60 Stirring mechanism 70 Light source 80A Transmitted light detection mechanism 80B Fluorescence detection mechanism 90A Insertion mechanism 90B Extraction mechanism 90C Reaction vessel setting mechanism 90D Reaction vessel reset mechanism 100 Immunochemical measurement device 100A Sample section 100B Operation section

Continuation of the front page (56) References JP-A-60-64255 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) G01N 35/02 G01N 33/553 JICST file (JOIS)

Claims (2)

(57) [Claims] A first reagent comprising an insoluble magnetic particle carrying an antibody or an antigen against the antigen or antibody to be measured, and an antigen or antibody to be measured reacting in a liquid medium of a reaction vessel; After attaching the insoluble magnetic particles to the reaction vessel wall by the action of a magnetic field and removing the liquid medium, further washing the insoluble magnetic particles,
A second reagent consisting of insoluble fluorescent dye-labeled particles carrying an antibody or antigen against the antigen or antibody to be measured, and reacting the insoluble magnetic particles that have undergone the antigen-antibody reaction in a liquid medium, After attaching the insoluble magnetic particles to the reaction vessel wall by the action of a magnetic field, the liquid medium and the unreacted insoluble fluorescent dye-labeled particles are removed, and then the fluorescence intensity of the insoluble fluorescent dye particles reacted with the insoluble magnetic particles and And / or a device for measuring the intensity of transmitted light, wherein a container set for attaching or detaching a reaction vessel is formed over the entire periphery of the turntable, and each reaction vessel is provided between said reaction vessels. A movable magnet that can move between a position facing the side surface and a position distant from the side surface is sequentially adjacent, but the sample portion is arranged with the polarity reversed; A reaction vessel moving mechanism for attaching and detaching the reaction vessel to and from the vessel setting section, a sample dispensing mechanism, a reagent dispensing mechanism, a washing mechanism for the reaction vessel, and a A stirring mechanism for the contents, a fluorescence detection mechanism for measuring the fluorescence intensity of the sample, a transmitted light detection mechanism for measuring the transmitted light intensity of the sample, and a mechanism for inserting and extracting the movable magnet are fixedly arranged. An operation unit having a control mechanism for rotating the rotary table in accordance with a sequence and operating each of the mechanisms according to timing. 2. A reaction between a first reagent comprising an antibody or an insoluble magnetic particle carrying an antigen against the antigen or antibody to be measured and the antigen or antibody to be measured in a liquid medium of a reaction vessel, After attaching the insoluble magnetic particles to the reaction vessel wall by the action of a magnetic field and removing the liquid medium, further washing the insoluble magnetic particles,
A second reagent consisting of insoluble fluorescent dye-labeled particles carrying an antibody or antigen against the antigen or antibody to be measured, and reacting the insoluble magnetic particles that have undergone the antigen-antibody reaction in a liquid medium, After attaching the insoluble magnetic particles to the reaction vessel wall by the action of a magnetic field, the liquid medium and the unreacted insoluble fluorescent dye-labeled particles are removed, and then the fluorescence intensity of the insoluble fluorescent dye particles reacted with the insoluble magnetic particles and And / or a device for measuring the intensity of transmitted light, wherein a container set portion is formed, which is capable of mounting or demounting a reaction container along the entire periphery of the turntable and movable to a set position and a reset position. When the respective reaction vessels are at the set position, the polarities of the adjacent ones are sequentially reversed at positions opposed to the side surfaces of the reaction vessels, and are arranged. A sample section having a fixed magnet placed thereon, a reaction vessel moving mechanism for attaching and detaching the reaction vessel to and from the vessel set section outside the rotary table, a sample dispensing mechanism, and a reagent dispensing mechanism; A washing mechanism for the reaction vessel, a stirring mechanism for stirring the contents of the reaction vessel, a fluorescence detection mechanism for measuring the fluorescence intensity of the sample, a transmitted light detection mechanism for measuring the transmitted light intensity of the sample, and a reset position for the reaction vessel. And a reaction vessel reset mechanism for moving the reaction vessel from the set position to the reset position are arranged and fixed.Furthermore, the rotary table is rotated according to a predetermined sequence, and the mechanisms are timed. And an operation unit having a control mechanism that operates according to the following.