JPS61254857A - Pretreatment device for immunological analysis - Google Patents

Abstract

PURPOSE:To execute quickly a pretreatment for an immunological analysis by dispensing respectively a sample and stroma into a cassette for pretreatment while moving the cassette so that both react with each other and filtering the reaction liquid thereby removing the stroma. 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 a reaction chamber 20 and thereafter the hole 13 is closed by pressing a syringe tip 21 by a tip driving means 5. The cassette 11 is then 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 a 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 1] The present invention relates to an immunoassay pretreatment device that performs pretreatment for an immunoassay in which a specific antigen, antibody, or complement value in a sample is quantitatively analyzed.

[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 ゛).

This MCIA method, as disclosed in detail in specification 1 of JP-A No. 60-17359, uses a cyclocapsule that binds an antibody or an antigen to its surface and encapsulates a specific substance therein, and a complement; When the specific substance itself is not a fluorescent substance, a reagent having a coexisting substance that causes a reaction product produced by reacting with the specific substance to emit light such as fluorescence or chemiluminescence, and an antigen or an antibody. by mixing with a sample containing the antigen-antibody reaction,
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 quantify the antigen, antibody, or complement value in a sample by spectroscopically analyzing light such as fluorescence or chemiluminescence emitted by a substance produced by the reaction of the specific substance and coexisting substances. b, which is excellent as a rapid and highly sensitive analysis method.
It is. 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 recognize 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") are added to the sample serum, and the reaction is carried out for several minutes to several tens of minutes at low humidity (for example, 4°C). After that, the stroma can be removed using a centrifuge. The necessity of stroma removal is due to the fact that hemolyzed stroma acts as an interfering substance during measurements.

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 an immunoassay preprocessing device that can quickly perform preprocessing for immunoassay. .

[ll of invention! Summary of the present invention to achieve the above object is an immunoassay preprocessing device that performs preprocessing for an immunoassay for quantitatively analyzing a specific antigen, antibody, or complement in a sample. A pretreatment cassette capable of removing stroma 7 from the reaction solution by reacting it with a stroma that absorbs a specific antibody in a sample and filtering the reaction solution, and a cassette that sequentially supplies this pretreatment cassette. A supply means, a step of dispensing the sample and stroma into a plurality of pretreatment cassettes supplied by the cassette supply means, a step of stirring the dispensed sample and the stroma, and a step of stirring the agitated sample and the stroma. and a step of removing stroma from the reaction solution, and a cassette moving means for intermittently moving a pretreatment cassette onto the platform.

[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 device including an immunoassay pretreatment device according to an embodiment of the present invention. 101 in the figure is an immunoassay pretreatment device according to the present invention, and 102 is an immunoassay section for quantitatively analyzing a specific antigen, antibody, or complement value in a sample.

This immunoassay pretreatment device 101 includes a cassette hopper (cassette supply means) 1 that stores a plurality of pretreatment cassettes (hereinafter referred to as "cassettes") 11 to be described later, and a cassette 11 supplied from the cassette hopper 1 indicated by an arrow It comprises a platform 2 for intermittently moving the cassette 11, first and second chip driving means 5 and 6, which will be described in detail later, and a cassette moving means 7 for intermittently moving the cassette 11.

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 pretreatment apparatus 101.

The platform 2 has steps a to l, and a groove for receiving and holding the cassette 11 is cut out for each step. Then, the cassette 11 sequentially moves intermittently through steps a to -, and in each step, the sample is dispensed into the cassette 11, the stroma is dispensed,
11 Stirring 9 Reaction 1 Removal of solid matter (stroma) in the reaction solution and dilution of the sample are performed.

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. It comprises a push rod driving means 9 for driving this push rod 4, and the first tip driving means 5 presses the syringe tip (described in detail in the section below) 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.
5 is a sectional view taken along line n-A, and FIG. 5 is a sectional view taken along line Bo-B in FIG.

This cassette 11 includes a reaction chamber 20.in which a sample pretreatment reaction is performed. Filter support section 22. Al 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 in the reaction chamber 2011 of the filter support section 22; The inserted syringe tip (hereinafter referred to as “
21 (referred to as "chip"). 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, and a sample and stroma are dispensed into the reaction chamber 20 through this dispensing hole 13. It is designed to be possible. 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 IIH 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, 9 mmΦ, and when using ashless pulp, press 0.05 g of acetic acid cellulose to a thickness of 31 mm. Preferably. Furthermore, the second
As the 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 a view seen from the direction of arrow Y in the figure. Moreover, FIG. 7 is a sectional view taken along line Co-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 (Ill 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 116 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 tip 21 slides almost to the center of the reaction chamber 20. Then, the tip 21 closes the dispensing hole 13 of the cassette 11. Here, care must be taken because if the slide 11Jffi of the chip 21 is not large, the sample before the pretreatment reaction will flow out into the filtrate chamber 15 through the filtration means 14.

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 cassette 11 in forward and reverse directions at regular intervals.

After the stirring in step e, the pretreatment reaction of the sample moves during the movement process of steps f to J, but during this time the cassette 11
is preferably cooled 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 Vertier element.

After the pretreatment reactions in steps C to j, cassette 1
1 is located on the stick k, the second chip driving 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 chip 21 to slide, and pressure is applied to the reaction liquid in the reaction chamber 20. Then, the reaction liquid in the reaction chamber 20 passes through the first filter, which is the filtering means 14. Second
The liquid is pushed out into the filtrate chamber 15 through the filters 14a, 14b and the hole 23 of the filter support part 22. 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 inventor of the present application, as in this embodiment, the filtering means 14 is connected to the first filter. When the second filters 14a and 14b have a double structure, the reaction solution can be smoothly filtered (stroma removal) without clogging by stroma, and there is no loss of plugged components in the sample. Good results were obtained with a recovery rate of around 100%. 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, when the cassette 11 is positioned at a step short position due to intermittent movement after the stroma is removed in step k, which is required to be 10 kMc+2 or less, the first
The sample in the filtrate v15 is absorbed, and the sample in the filtrate v15 is absorbed.
It is diluted, for example, 2 times, 30 times, or 300 times with Ij solution 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 116, 17, 18 is dispensed into the reaction tube of the reaction photometer 104 via the dispensing section ff1103, and is subjected to immunoassay by the immunoassay section 102. Cassette 11 after subjecting the diluted solution to immunoassay
will be discarded in the cassette waste box.

In this embodiment, after dispensing the sample and stroma into the reaction chamber 20 of the cassette 11, the tip 21 of the cassette 11 is pressed by the first tip driving means 5 to perform the dispensing of the cassette 11. Close the hole 13 and remove the cassette roller 1.
0, the sample and the stroma in the chamber 20 are stirred, and after the pretreatment reaction of the sample, the tip 21 of the cassette 11 is pressed again by the second tip driving means 6, and the reaction chamber 20 is stirred. The stroma in the reaction solution is removed by filtering the reaction solution using the filtration means 14, and pretreatment for immunoassay can be quickly performed without batch processing. By combining the device 101 with the immunoassay section 102, it becomes possible to realize an immunoassay processing device that can perform rapid immunoassay processing.

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.

[Advantageous Effects of the Invention 1] As detailed above, the present invention provides an immunoassay preprocessing device that can automate immunoassay preprocessing and quickly perform immunoassay preprocessing. I can do it.

[Brief explanation of the drawing]

FIG. 1 is a schematic perspective view of an immunoassay device including an immunoassay preprocessing device according to an embodiment of the present invention, FIG. 2 is a plan view of the device of this embodiment, and FIG. 3 is a preprocessing device for the device of this embodiment. A plan view of the cullet, FIG. 4 is a sectional view taken along line Ao-A in FIG. 3, FIG. 5 is a sectional view taken along line Ao-A in FIG. The figure is Figure 6G.
It is an o-C sectional view. DESCRIPTION OF SYMBOLS 1... Cassette supply means, 2... Platform, 5... First chip drive means, 6... Second chip drive means, 7... Cassette moving means, 10... Cassette roller , 11... Pretreatment cassette, 13... Dispensing hole, 1
4...filtration means, 15...11 liquid chamber, 20...reaction chamber, 21...syringe tip. No.

Claims (2)

[Claims] (1) An immunoassay pretreatment device that performs pretreatment for immunoassay for quantitatively analyzing specific antigens, antibodies, or complements in a sample, which reacts the sample with stroma that absorbs the specific antibodies in the sample. a pretreatment cassette capable of removing stroma in the reaction solution by filtering the reaction solution; a cassette supply means for sequentially supplying the pretreatment cassette; and a plurality of pretreatment cassettes supplied by the cassette supply means. A step of dispensing the sample and stroma into a pretreatment cassette, a step of stirring the dispensed sample and the stroma, a step of reacting the stirred sample with the stroma, and removing the stroma from the reaction solution. 1. An immunoassay preprocessing device comprising: a platform having a step for moving a preprocessing cassette onto the platform; and cassette moving means for intermittently moving a preprocessing cassette onto the platform. (2) The pretreatment cassette includes a reaction chamber for reacting a sample with a stroma that absorbs a specific antibody in the sample, a dispensing hole for the sample and stroma to the reaction chamber, and a hole for dispensing the sample and stroma into the reaction chamber by 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, and by filtering the reaction liquid by the sliding of this syringe tip, The immunoassay pretreatment device according to claim 1, comprising a filtration means for removing stroma, and a filtrate chamber for receiving the reaction liquid filtered by the filtration means.