JPS61254858A - Cassette for pretreatment for immunological analysis - Google Patents

Abstract

PURPOSE:To make possible the automation of a pretreatment for an immunological analysis by using a cassette for pretreatment having a reaction chamber to bring a sample and stroma into reaction with each other, filter means, filtrate chamber, etc. CONSTITUTION:The pretreatment is executed in the process for moving intermittently the cassette 11 for pretreatment supplied from a cassette hopper 1 on a platform 2. The sample and stroma are first dispensed from a dispensing hole 13 into the reaction chamber 20, then the hole 13 is closed by pressing a syringe tip 21 by a tip driving means 5. The cassette 11 is turned by a cassette roller 10 to stir the sample and the stroma. The tip 21 is driven by a tip driving means 6 to exert the pressure to the reaction liquid in the chamber 20 after the progression of the pretreatment reaction. The reaction liquid is ejected into the filter chamber 15 by a filter means 14. The reaction liquid from which the stroma is removed is diluted and is distributed to diluent reservoirs 16-18 and thereafter the liquid is subjected to the immunological analysis.

Description

Detailed Description of the Invention [Technical Field of the Invention] The present invention relates to a cassette for immunoassay pretreatment for quantitatively analyzing a specific antigen, antibody, or complement value in a sample. It is.

[Technical Background of the Invention and Problems thereof] Immunoassay methods include radioimmunoassay method and enzyme immunoassay method, and in view of the fact that these analysis methods require a long time for analysis, microcapsules, which were previously proposed by the applicant, have been proposed. An analysis method using rMcIA (hereinafter referred to as rMcIA)
There is a law (abbreviated as "law").

For details of this MCIA method, please refer to JP-A-60-17359@
As disclosed in the specification related to the above, a microcapsule and a complement that bind an antibody or an antigen to the surface and encapsulate a specific substance therein, and when the specific substance is not itself a fluorescent substance, the above-mentioned Antigen-antibody reactions can be achieved by mixing a reagent containing a coexisting substance that causes a reaction product produced by reacting with a specific substance to emit light such as fluorescence or chemiluminescence, and a sample containing an antigen or antibody. cause
The specific substance is flowed out from the microcapsules destroyed by the action of complement activation, and when the specific substance is a fluorescent substance, the emitted fluorescence is spectroscopically analyzed, and when the specific substance is not a fluorescent substance, The principle is to determine the antigen, antibody, or complement value in a sample by spectroscopically analyzing light such as fluorescence or chemiluminescence emitted by a substance generated by the reaction of the specific substance and a coexisting substance. This is an excellent rapid and highly sensitive analytical method. As microcapsules capable of binding antibodies or antigens, blood cells from animals such as sheep are used.

By the way, when animal blood cells are used, the target antigen-
Hemolysis occurs not only due to the antibody reaction but also due to a reaction unrelated to the antibody reaction (hereinafter referred to as a "non-specific reaction"), and the resulting hemolysis causes measurement errors. This is thought to be due to the fact that human serum contains antibodies that identify all animal blood cells as non-self. Therefore MCI
In method A, it is necessary to absorb antibodies that cause this nonspecific reaction.

As a method for absorbing antibodies that cause non-specific reactions, animal blood cells or their ghost membranes (hereinafter referred to as "stroma J") are added to the sample serum, and the reaction is carried out at a low temperature (for example, 4°C) for several minutes to several tens of minutes. After that, the stroma is removed using a centrifuge.The necessity of stroma removal is because the hemolyzed stroma acts as an interfering substance during the measurement.

However, since using a centrifuge to remove stroma requires batch processing, it is extremely difficult to automate the pretreatment process for immunoassays.

Due to the demand for rapid immunoassay processing, automation of the immunoassay preprocessing process is strongly desired.

[Object of the Invention] The present invention has been made in view of the above circumstances, and its purpose is to provide a cassette for immunoassay pretreatment that enables automation of the immunoassay pretreatment process. It is in.

[Summary of the Invention] The outline of the present invention for achieving the above object is to provide a reaction chamber in which a sample and a stroma that absorbs a specific antibody in the sample are reacted, and a dispensing of the sample and stroma into the reaction chamber. a hole, and is driven on the wall surface of the reaction chamber by an external force, and
a syringe tip that closes the dispensing hole and presses the reaction liquid in the reaction chamber according to the amount of sliding; a filtration means that filters the reaction liquid by sliding the syringe tip and removes stroma from the reaction liquid; It is characterized by having a filtrate chamber that receives the filtrate that has passed through the filtration means.

[Embodiments of the invention] Hereinafter, the present invention will be specifically explained with reference to Examples.

FIG. 1 is a schematic perspective view of an immunoassay apparatus including an immunoassay pretreatment cassette according to an embodiment of the present invention. Figure 101
1 is an immunoassay preprocessing section, and 102 is an immunoassay section that determines and analyzes a specific antigen, antibody, or complement value in a sample.

This immunoassay preprocessing section 101 includes a cassette hopper (cassette supply means) 1 that stores a plurality of cassettes for immunoassay preprocessing (hereinafter referred to as "cassettes J") 11 according to the present invention, and A platform 2 for intermittently moving the cassette 11 in the direction of arrow X, and first and second chip drive means 5.6, which will be described in detail later.
and a cassette moving means 7 that moves the cassette 11 intermittently.

The immunization unit 102 also includes a dispensing means 103 for diluting a sample processed by the immunoassay pretreatment device 101, dispensing a diluent, and dispensing a reagent, and a dispensing means 103 for diluting a sample processed by the immunoassay pretreatment device 101 and dispensing a reagent, and a diluting solution dispensed by the dispensing means 103. and reaction photometry means 104 for performing immunoreaction with the reagent and photometry of the reaction solution.

FIG. 2 is a plan view of the immunoassay preprocessing section 101.

The platform 2 has steps a to m, and a groove for receiving and holding the cassette 11 is cut out at each step. Then, the cassette 11 sequentially moves intermittently through steps a to 1, and in each step, the sample is dispensed into the cassette 11, the strain is dispensed,
Stirring 2 Reaction 1 Removal of solid matter (stroma) in the reaction solution,
The sample is diluted.

Said 1st. The second tip drive means 5 and 6 each have a push rod 3. A push rod driving means 8 for driving the push rod 3 and a push rod 4. The device comprises a push rod driving means 9 for driving this push rod 4, and the first tip driving means 5 presses the syringe tip (detailed description in the figure) of the cassette 11 located at step d, Further, the second tip driving means 6 is for pressing the syringe tip of the cassette 11 located at step k.

Further, a cassette roller 10 is attached below the cassette 11 located at step d, and the rotation of this cassette roller 10 causes the cassette 11 located at step d to rotate.

Next, referring to FIGS. 3 to 7, the cassette 1
The configuration of 1 will be explained.

Fig. 3 is a plan view of the cassette 11, and Fig. 4 is Fig. 3A.
The o-A sectional view and FIG. 5 are the Bo-8 sectional views in FIG. 3.

This cassette 11 has a reaction chamber 20 in which a sample pretreatment reaction is performed. Filter support section 22. i11 excess liquid chamber 15, 1st.
Second. Third diluent reservoir 16.17.18 and protrusion 19
a, 19b, and is formed into a substantially cylindrical shape by, for example, resin; a filtration means 14 supported on the reaction chamber 20 side of the filter support portion 22; and an end portion inside the reaction chamber 20. A syringe tip (hereinafter referred to as "chip") 21 is inserted into the cassette.In particular, a dispensing hole 13 communicating with the inside of the reaction chamber 20 is provided on the circumferential surface of the cassette body forming the reaction chamber 20. This dispensing hole 13
The sample and stroma can be dispensed into the reaction chamber 20 via the tube. The chip 21 inserted into the end of the reaction chamber 20 is moved along the inner wall of the reaction chamber 20 by the push rod 3.4 (see FIG. 2) constituting the chip driving means 5.6. It is designed to slide in the Y direction.

The filtering means 14 includes a first filter 14a and a second filter 14b that are in contact with the filter support 22. For example, a glass fiber or cellulose acetate-based ashless bulb can be applied to the first filter 14a. When using glass fiber, it is preferable to compress approximately 0°02° to the inner diameter of the reaction chamber 20, for example, 9IIIΦ, and when using ashless pulp, it is preferable to compress cellulose acetate of 0.05 (I) to a thickness of 3I. Further, as the second filter 14b, a filter made of a mixture of cellulose acetate and cellulose nitrate, or a Teflon filter can be used.

Incidentally, the protrusions 19a and 19b are for holding the cassette 11 when the cassette 11 is moved.

FIG. 6 is an explanatory diagram of the filter support section 22, and the third
This is the view from the direction of arrow Y in the figure. Moreover, FIG. 7 is a sectional view taken along the line Go-C in FIG.

As is clear from FIGS. 6 and 7, this filter support part 22 has an uneven shape on the surface that contacts the first filter 14a, and has a plurality of holes 23 (4 in the drawing) in the recessed part.
) Consisting of holes drilled at equal intervals. The drilled holes 23 are for guiding the reaction liquid (filtrate) that has passed through the filtration means 14 to the first filtrate chamber 15 .

In the above configuration, from the cassette hopper 1 to the cassette 11
is supplied, and the supplied cassette 11 starts to move intermittently by driving the cassette moving means 7. Then, in step b of the platform 2, the sample is dispensed into the reaction chamber 20 of the cassette 11, and in step C, the stroma is dispensed. The sample and stroma are dispensed through the dispensing hole 13 of the cassette 11 using a dispensing nozzle (not shown).

When the cassette 11 containing the sample and stroma is located at step d, the first tip driving means 5 operates;
The chip 21 of the cassette 11 is pressed via the push rod 3. This pressing is performed so that the chip 21 slides almost to the center of the reaction chamber 20 . Then, the tip 21 fills the dispensing hole 13 of the cassette 11. Here, if the sliding movement 1 of the chip 21 is too large, the sample before the pretreatment reaction will flow out into the filtrate chamber 15 through the filtration means 14, so care must be taken.

Then, the cassette 11 with the dispensing hole 13 closed is rotated by the cassette roller 10 in the next step e, and the sample and stroma in the reaction chamber 20 are stirred by this rotation. In order to improve stirring efficiency, it is preferable to rotate the h set 11 in forward and reverse directions at regular intervals.

After the stirring in step e, the pretreatment reaction of the sample progresses during the movement process in steps f to j. During this time, the cassette 11
It is preferable to cool it at a constant temperature of 4°C to 10°C.

The cassette 11 can be easily cooled by constructing the platform 2 of metal and cooling the platform 2 using a known technique such as a Peltier device.

After the pretreatment reactions in steps C to j, cassette 1
1 is located at step Vk, the second chip drive means 6 operates, and the chip 2 of the cassette 11 is moved through the push rod 4.
1 is pressed again. This pressing causes the tip 21 to slide, and pressure is applied to the reaction liquid in the reaction 20. Then, the reaction liquid in the reaction chamber 2O passes through the first filtration means 14.
Second filters 14a, 14b and filter support section 22
into the filtrate chamber 15 through the hole 23 .

At this time, the first. Stroma in the reaction solution is removed by the second filters 14a and 14b.

According to the experiments of the present inventor, the filtering hand 8 as in this embodiment
Stage 14 is the first. When the second filters 14a and 14b have a double structure, there is no clogging caused by stroma, and smooth filtration (stroma removal) of the reaction liquid can be performed.
Moreover, there is no loss of plugged components in the sample, and the recovery rate is 1.
A good result of 00% Jail was obtained. Furthermore, in the above filtration action, if the pressing force applied to the tip 21 by the tip driving means 6 is too large, there is a risk of damaging the filtration means 14, especially the first filter 14a, so care must be taken.

According to experiments, it is necessary to keep it below 10 ka/CI2.

When the cassette 11 is positioned at step l due to the intermittent movement in which the stroma is removed in step k, the first
The sample in the filtrate chamber 15 is absorbed and diluted, for example, 2 times, 30 times, or 300 times with a berronal buffer or the like. Then, this diluted solution is used as the first diluted solution. Second. It is distributed in the third diluent reservoir 16.17.18. This 2nd and 3rd. The diluent in the fourth diluent reservoir 16, 17, 18 is dispensed into the reaction tube of the reaction photometer 104 via the dispensing means 103, and is subjected to immunoassay by the immunoassay section 102. After the diluted solution has been subjected to immunoassay, the cassette 11 is discarded into a cassette disposal box.

In this way, according to the present example, a reaction chamber 2 in which a sample is reacted with a stroma that absorbs a specific antibody in the sample is used.
0, and a dispensing hole 13 for the sample and stroma into the reaction chamber 20, which slides on the inner wall of the reaction chamber 20 by an external force, and depending on the amount of sliding, closes the dispensing hole 13 and closes the dispensing hole 13 of the reaction chamber. a syringe tip 21 that presses the reaction liquid in 20;
The reaction liquid is filtered by sliding the syringe tip 21,
A filtration means 14 for removing stroma from the reaction solution, and a filtrate chamber 15 for receiving the filtrate that has passed through the filtration means 14.
Because it has a
The reaction and the removal of Straw-7 from the reaction solution can be performed quickly and easily, and therefore, by using this cassette 11, it is possible to automate the pretreatment process for immunoassay. especially'
The filtering means 14 is the first filter. second filter 14a,
14b, the reaction solution can be filtered smoothly without clogging due to stroma.

Furthermore, by making the cassette 11 disposable, the effort of cleaning the cassette 11 becomes unnecessary.

Although one embodiment of the present invention has been described above, it goes without saying that the present invention is not limited to the above-mentioned embodiment, and can be modified as appropriate within the scope of the gist of the present invention.

[Effects of the Invention] As detailed above, according to the present invention, it is possible to automate the pretreatment process for immunoassay. A cassette for immunoassay pretreatment can be provided.

[BRIEF DESCRIPTION OF THE DRAWINGS] FIG. 1 is a schematic perspective view of an immunoassay device including an immunoassay pretreatment cassette according to an embodiment of the present invention, and FIG. 2 is a plan view of the immunoassay pretreatment section in FIG. 1. , FIG. 3 is a plan view of the immunoassay pretreatment cassette, FIG. 4 is a sectional view taken along the line Ao-A in FIG. 3, FIG. 5 is a sectional view taken along the line Bo-B in FIG. FIG. 7 is a sectional view taken along line Co-C in FIG. 6. DESCRIPTION OF SYMBOLS 11... Immune analysis pretreatment cassette, 13... Dispensing hole, 14... Filtration means, 15... Filtrate chamber, 20...
Reaction chamber, 21... syringe tip.

Claims (1)

[Claims] a reaction chamber in which a sample reacts with a stroma that absorbs a specific antibody in the sample; a dispensing hole for the sample and stroma into the reaction chamber; a syringe tip that closes the dispensing hole and presses the reaction liquid in the reaction chamber according to the amount of sliding; a filtration means that filters the reaction liquid by sliding the syringe tip and removes stroma from the reaction liquid; An immunoassay characterized by having a filtrate chamber that receives the filtrate that has passed through the filtration means, and in which the filtrate in the filtrate chamber is subjected to an immunoassay for quantitatively analyzing a specific antigen, antibody, or complement. Pretreatment cassette.