System for contamination-free reaction processes

Description: TECHNICAL FIELD [Detailed description of the invention]

【0001】

[Industrial application field]

The present invention relates to suitable reaction vessels and sealing devices, as well as devices and methods for automatically performing reactions in reaction vessels.

【0002】

[Problems to be Solved by Conventional Techniques and Inventions]

The main configuration of the health device is to detect the disease by so-called medical diagnostic methods. This involves detecting a response to a particular sample using a sample taken from a patient who may be affected by the disease. Substrates for enzymatic reactions, enzymes themselves, antigens and, more recently, nucleic acids have also been found to be suitable specimens. In this connection, the analysis of the latter half of the substance has been found to be much more difficult, sometimes due to its very low concentration. Since very small concentrations of interfering substances have a great effect on the measurement results, it is essential that the treatment be free of scattering or carry-over in order to obtain reliable results. There is considerable risk of carryover, especially in the case of tests in which several analyzes are performed consecutively on a single instrument.

【0003】

In nucleic acid diagnostics, the detection limit is significantly reduced by providing amplification methods, such as polymer chain reactions (US Pat. No. 4,683,195). However, instead, the risk of erroneous measurement results due to contamination also increases, especially with the formation of aerosols in the air. Therefore, the preparation and amplification of the sample to be analyzed should be performed in an isolation room. In addition, oil and wax coatings and the use of insensitive internal enzyme decontamination systems have been proposed as further solutions (US Pat. No. 5,035,996). However, these solutions are cumbersome and are not suitable for performing some reactions automatically.

【0004】

In the case of a polymer chain reaction, a special tube is used that can transfer heat well from the heater provided below because of its conical shape. In addition, most of the equipment used to perform thermal cycles utilize cap heaters to suppress evaporation and avoid condensation.

【0005】

Also mentioned is a plate containing several reaction vessels that can be sealed with a regular cover that can be torn by hand. The disadvantage of this configuration is that when the plate is opened, splashes from one reaction vessel can enter the nearby reaction vessel, increasing the risk of carryover.

【0006】

European Patent No. 0 408 208 also describes the reaction vessel in the form of a microtitre plate with 96 open holes: a multiple storage device for performing chemical reactions, especially clinical analysis. Describes a type of reaction vessel. The risk of contamination in this case is also very high.

【0007】

All of the above proposals for containers that automatically process some reactions include that the reaction vessel cannot be effectively sealed or that the reaction vessel has a closing lid that is not suitable for automation. There are drawbacks. As such, these devices can only be used for high volume routine applications with significant contamination potential.

【0008】

[Means for solving problems]

The present invention is contamination-free for automatically performing reaction sequences in a reaction vessel. An operation) device, the device being a number of reaction vessels, several reagent storage vessels, several sample vessels, individual sealing devices for some of the vessels, and the above. It comprises a device for automatically opening the sealing device, and the opening device is for removing the sealing device withdrawing means for pulling out the sealing device from the container and removing the sealing device from the sealing device withdrawing means. It is characterized in that the sealing device taking-out means including a component is a removal plug having a removing ring, and the component for removing the sealing device from the sealing device removing means is a removing jacket. Further, the lid according to claim 2 is a lid for a container having a portion extending into the opening of the container, and the extending portion is for removing the lid as a means for removing the lid from the container. The space between the lid and the container is closed by the sealing ring, and the space between the lid removing means and the sealing ring is provided. It is characterized by becoming. The lid preferably has a partition wall into which a pipetting needle can be inserted.

【0009】

The technique can also be used to handle sample and reagent storage containers. That is, the present invention is also a non-contamination treatment method of a reaction chain for automatically processing a chemical reaction in several containers, a step of supplying a reaction mixture into the reaction vessel, and individual sealing. The reaction vessel is sealed by the device, the reaction mixture is thermally treated, and the individual sealing devices are automatically opened by the device for removing the sealing device from the reaction vessel. The device is characterized in that it has a portion extending into the opening of the container, the extending portion having means for removing the sealing device from the container.

【0010】

Further, the present invention is a reaction chain non-contamination treatment method for automatically processing a chemical reaction using several reagent storage containers, in which the reagent storage containers are sealed by individual sealing devices. The step of supplying the reagent solution, the step of automatically opening each sealing device by the device for removing the sealing device from the reagent storage container, the step of taking out the reagent solution, and the step of automatically reopening the reagent storage container by the individual sealing device. It comprises a step of sealing, wherein the sealing device has a portion extending into the opening of the container, and the extending portion has a means for removing the sealing device from the container. ..

【0011】

Further, the present invention is a reaction chain non-contamination treatment method for automatically processing nucleic acid analysis in a sample solution from several sample containers, in which the sample containers are sealed by individual sealing devices. By a step of supplying a sample solution containing nucleic acid, a step of automatically opening each sealing device by a device for removing the sealing device from the sample container, a step of taking out the sample solution, and an individual sealing device. It comprises a step of automatically resealing the sample container, wherein the sealing device has a portion extending into the opening of the container, and the extending portion has a means for removing the sealing device from the container. It is characterized by being a container.

【0012】

The present invention also relates to a reaction vessel and a sealing device suitable for the above-mentioned device.

【0013】

The device of the present invention is suitable for automatically performing a reaction. In this regard, automatic means that the manual processing step from the introduction of the sample into the apparatus until the measurement result is obtained is omitted. The processing step proceeds in a predetermined manner in a device preferably divided by a computer program. The device is also suitable for performing several consecutive reactions in different reagent storage vessels.

【0014】

In principle, any type of chemical reaction can be carried out in the apparatus of the present invention. These chemical reactions involve steps that the components of the sample must pass through, especially during the analytical process. The reaction that occurs depends on the type of sample to be determined. Thus, for example, an immunologically active mixture is usually determined by an immunological determination method. In this case, the interaction of antigens and adherents on the one hand and the antibodies directed at them on the other are utilized. In the case of nucleic acid testing, relatively special interactions between nucleic acids that are essentially complementary to each other are used.

【0015】

Nucleic acid analysis methods can be essentially divided into the steps of sample preparation, nucleic acid processing and detection of processed nucleic acids. The apparatus of the present invention is specifically configured to carry out a nucleic acid processing reaction. These reactions specifically involve amplification for nucleic acids. There are many ways to do this. Some amplifications are performed at an isothermal temperature, such as the method described in European Patent No. 0 329 822, but many other amplifications are described, for example, in European Patent No. 0 200 362. It is carried out using a temperature change cycle (thermal cycle).

【0016】

The apparatus of the present invention includes several reaction vessels suitable for taking out the reaction solution. The amount of the reaction solution is preferably between 5 μl and 5 ml, particularly preferably between 10 μl and 200 μl. The reaction vessel is made of a plastic-like material such as polystyrene or polypropylene that does not chemically react with the reaction solution. These reaction vessels also need to be heat resistant if they are intended to carry out a thermal cycle. For example, a commonly used PCR (polymerase chain reaction) test tube manufactured by Perkin Elmer is suitable as the reaction vessel of the present invention.

【0017】

An essential feature of the present invention is the use of individual sealing devices for these reaction vessels. Individual lidging of reaction vessels has never been considered before and has helped designers of combined test tube plates by worrying about the variety of lids to be particularly suitable. It may have been. Moreover, the use of individual sealing devices is a technical challenge achieved by the individual sealing devices of the present invention. Like the container, the sealing device is preferably manufactured from a heat-resistant material such as polystyrene or polypropylene, and is particularly preferably manufactured by a known injection molding method.

【0018】

The reactions that can be performed in the reaction vessel may be of physical type (eg, absorption) or biochemical type (eg, enzymatic reaction, amplification, etc.). A more important feature of the device of the present invention is a device for automatically opening a sealing device. This device is technically in harmony with the individual sealing devices used in each case.

【0019】

To carry out the reaction automatically, the liquid is dispensed into the reaction vessel. The use of an automatic pipetting device, preferably a device with a disposable tip, has been found to be particularly advantageous for this. In this process, the components of the reaction to be carried out can be pipetted in any suitable order. For example, the sample solution containing the sample can be added first to the reagent storage container. If desired, this process can be performed earlier, if desired, outside the apparatus of the present invention without the use of an automatic pipetting apparatus. In subsequent steps, which can optionally be performed using an automatic pipetting apparatus, the reagents required for the reaction can be added to the sample solution in a reagent storage vessel, and as a result, the methods of the invention can also be used. it can.

【0020】

In another embodiment, the required reagents are pre-added to the reaction vessel in solid or liquid form, to which the sample solution is pipetted.

【0021】

[Example]

Next, an apparatus and a method for automatically carrying out the reaction of the present invention will be described with reference to the accompanying drawings.

【0022】

FIG. 1 is an elevation sectional view showing a composite plate having a lid of the present invention, FIG. 2 is an elevation sectional view showing an embodiment of the lid of the present invention, a removing device and a reaction vessel, and FIG. 3 is an elevation sectional view of the lid of the present invention. Another Example, an elevation cross-section showing the removal device and the reaction vessel, FIG. 4 is yet another embodiment of the lid of the invention, an elevation cross-section showing the removal device and the reaction vessel, FIG. 5 shows the reaction. Schematic explanatory view of the apparatus of the present invention for carrying out, FIG. 6 is an elevation sectional view showing an embodiment of the screw lid of the present invention, and FIG. 7 is an elevation sectional view showing an improved lid of the present invention. 8 is a cross-sectional view showing a stop unit for the removal device of the present invention, FIG. 9 is a cross-sectional view showing the outer casing of the present invention including a stop element, and FIG. 10 is a partial cross-sectional view showing the manual remover of the present invention. 11 is a cross-sectional explanatory view illustrating the process of the method of the present invention for removing the lid from the container.

【0023】

A device for automatically performing the reaction in a holding plate having 96 pores in the microtiter plate will be described below, which is a particularly preferred embodiment for arranging several reaction vessels. The device is particularly suitable for performing polymer chain reactions using thermal cycles. A holding plate for the Perkin Elmer cycler 9600 device is shown in Figure 1. Reaction vessel 1 is drawn to be located in one of the holes 3 of the holding plate 2 for 96 test tubes. The reaction vessel is secured in the holding plate by a bard 102. When the holding plate with the reaction vessel is inserted into the PCR cycler, the tip of the reaction vessel reaches the heat block 4 of the cycler. A lid 5 suitable for the present invention is schematically depicted in FIG. To seal the reaction vessel, the lid 5 is pushed into the reaction vessel up to the stopper 501.

【0024】

Further, the present invention relates to a lid for a reaction vessel having a portion extending into the opening of the reaction vessel, which portion comprises a device for removing the lid from the reaction vessel.

【0025】

The lid according to the present invention has at least two parts. One portion, 550, is configured to extend into the opening of each reaction vessel when the reaction vessel is closed by a lid. The other part 500 is then still protruding from the reaction vessel.

【0026】

The portion 550 includes a device 551 that removes the lid from the reaction vessel. Further, this portion has a means for sealing the reaction vessel with the lid, ie, a closed rim or a closed ring 552.

【0027】

The portion 500 preferably includes a stopper 501 so that the lid does not disappear completely into the reaction vessel when the reaction vessel is closed. This also guarantees the accurate installation of the lid surface 503. The lid portion 500 preferably projects only slightly from the opening of the reaction vessel. The portion of the lid protruding from the reaction vessel is essentially flush with surface 503. This is advantageous if it is intended to use cap heating to prevent the water vapor from the reaction from condensing on the lid. Thereby, the cap heating can be placed just on the surface 503 of some reaction vessels.

【0028】

All configurations that ensure that the resistance between the lid and the device that automatically removes the lid is greater than the resistance between the lid and the inner wall of the reaction vessel are as devices 551 for removing the lid. It may be used. A technically feasible configuration is, for example, the use of a conical surface or ring to engage the device with portion 550.

【0029】

A first embodiment of the lid of the present invention (with the reaction vessel closed) and a suitable opening device 7 are shown in FIG. In this case, the device for removing the lid is configured as a conical recess 551 that is closed to the reaction vessel but open to the surrounding environment to remove the lid. Can be adapted to the tapered removal plug 700 of the device. In this process, a component with an annular sealing ring 552 pushes the inner wall 101 of the reaction vessel. A particularly advantageous structural detail of the lid of the present invention is that when the tapered removable plug 700 is inserted into the portion 550, the pressure acting on the device does not result in a substantial increase in the pressure acting on the closed edge. The device 551 is structurally separated from the closed edge 552. Structural separation is achieved by providing an intervening space 553 between the device 551 and the closed edge 552, as shown in FIG. In FIG. 2, this intervening space is open to the inside of the reaction vessel. Since the lid surface 503 is relatively large in this case, the contact surface with the cap heater and the heat transfer from the heater to the lid are particularly large.

【0030】

A particularly advantageous lid is shown in Figure 3. The essential difference from the lid shown in FIG. 2 is that the intervening space 553 is configured to be open to the reaction vessel rather than to the surrounding environment. In this example, since the contact surface of the lid with the reaction solution or the space of the gas covering it is small, only a small surface is effective for the condensation of the liquid on the lid.

【0031】

A lid without the intervening space 553 is shown in FIG. This lid can also be used in the apparatus of the present invention, but when the tapered removal plug is inserted, the pressure acting on the inner wall of the reagent storage vessel increases, resulting in the aforementioned lid for removing the lid from the reaction vessel. Greater power is needed than Ai.

【0032】

The device for automatically opening the sealing device is in harmony with the shape of the mechanism inside the lid for removing the lid. A tapered plug with a removal ring 701 that must be pushed into portion 550 beyond the resistance of the lid has been found to be particularly advantageous. When the tapered plug is pulled in, the ring acts as a resistor. The pressure exerted by the lid on the inner wall of the reaction vessel is less than the resistance of the ring to the lid, so the lid is removed from the reaction vessel.

【0033】

It is also preferred that the device include components for removing the lid removed from the reaction vessel from the tapered removal plug so that the tapered removal plug 700 can be used multiple times. For example, this can be a removable jacket 710.

【0034】

The equipment is arranged in the same manner as the reaction vessel arrangement for processing the sample in the reaction vessel. If a composite plate is used, the device can be placed on top of the XY moving unit and moved over each reaction vessel. The tip of the tapered removal plug is inserted into the lid by transferring the tapered removal plug 700 in the direction of the closed reaction vessel (Z direction). This movement will generally be perpendicular to the transfer plane. Both transfer steps can in principle be performed by a computer-controlled robot, but by replacing the rack gears commonly used for pipetting with rack gears with device 7, for example from Tecan. It is also possible to modify traditional automated pipettors. In addition to the pipetting unit, it is also possible to attach a device for automatically opening the lid to the rack gears of the automated pipettor. Therefore, the automated pipettor is programmed.

【0035】

Several devices 7 can be mounted on top of the transfer unit so that several reaction vessels can be opened simultaneously, i.e., several individual lids can be opened simultaneously to process a row of microtiter plates. .. However, this example would obviously be undesirable when it comes to avoiding contamination.

【0036】

The present invention, in particular the auto-operated lid, can be used not only for reaction vessels, but for all types of vessels in devices that carry out some reactions. These vessels include a vessel in which the reaction between several reaction partners proceeds internally, in this case a vessel called a reaction vessel, as well as a vessel for storing individual reaction partners for the desired reaction. In the case of analysis of a liquid sample, the sample container and the reagent storage container can be separated. Liquids intended to determine the presence or amount of one or several specimens are stored in a sample container. After sampling (eg, blood sampling of the patient to be examined) and feasible processing (separation of plasma and serum), the sample fluid is transferred into a sample container and, if necessary, an automated lid according to the invention. Sealed by. The sample container is then inserted individually or with additional sample containers into a device such as an automated analyzer. As the process according to the invention proceeds, the lid is removed from the sample container and the desired amount of sample solution is transferred from the sample container to the reaction vessel before the sample solution is transferred. The advantage of the device of the present invention is that the sample container is also closed by the automatic operation lid, but the sample container is easily closed again by the same lid or a new lid. In the preferred case of re-covering with a new lid, the old lid is transferred from the tapered removal plug to the waste container by the removal jacket and the new lid is pulled out of the lid storage container onto the tapered removal plug. There, the new lid is placed on the sample container and the tapered removal plug is removed, but the lid remains on the sample container. The lid is removed from the same sample again for further sample determination, this time a certain amount of sample solution is transferred. If the lid has a septum, the sample can be transferred from the sample container using an insertion pipette without removing the lid. Once the reaction is carried out, additional reagents may need to be transferred to the reaction vessel. unknown. Reagents are stored in reagent storage containers that may be opened and closed according to the present invention. Therefore, the reagent storage vessel is understood to mean a storage vessel for reagents, and these reagents are used to carry out the reaction in the reaction vessel. These reagent storage vessels are often in the form of bottles with capacities between 20 ml and 500 ml, as reagents generally have to be supplied in large quantities, for example to carry out some reactions. Such reagent storage containers may also be closed by the automatic operating lid of the present invention. This is commonly done by manufacturers. The reagent storage container is placed in a suitable position on the device. When an attempt is made to perform a reaction in a reaction vessel that requires the use of reagents stored in a particular reagent storage vessel, the automated operating lid is immediately removed from the reagent storage vessel and the required amount of reagent is withdrawn. Transferred into a reaction vessel. Subsequently, the reagent storage container can be reclosed with a removed lid or a new lid (see sample container).

【0037】

In the case of a large-capacity reagent storage container, it is possible in principle to use a conventionally used reagent storage container by improving a normal screw lid by using the automatic operation lid of the present invention. Therefore, it is preferable that a small opening closed by the lid of the present invention is provided on the upper surface of the screw lid. The advantage of this device is that the reagent storage container is filled with a relatively large opening closed by a screw cap, while removal of the reagent solution can be performed by a relatively small opening closed by the present invention.

【0038】

Furthermore, the present invention is a method for automatically carrying out a chemical reaction in a reaction vessel, (1) preparing a reagent and / or / or a reaction mixture in the reaction vessel, and (2) individually. It is characterized by comprising a step of closing the reaction vessel by the sealing device of (3) and a step of automatically opening each sealing device by a device for removing the lid from the container.

【0039】

The method proceeds in principle in the same manner as a conventional known manual method, except that an auto-operated lid is used and performed in the apparatus. Of course, the automatic operation lid can also be opened and closed manually. The main feature is the preparation of a reaction vessel containing the reaction mixture and closed by an automatic operation lid. This applies especially to reactions that proceed in the thermal cycle. After the reaction is complete, the lid is removed from the reaction vessel with the help of a device for removing the lid. This is achieved by inserting the device into the means provided in the lid and pulling out the device with the lid. The lid can then be removed from the tapered removal plug by pulling back the tapered removal plug inside the removal jacket. The reaction mixture can then be removed from the reaction vessel, but can also be further processed in it and, if necessary, reclosed with a (new) lid.

【0040】

The advantage of the lid of the present invention is that a cap heater can be used because a particularly simple thing that a flat surface can be provided can be realized. Also, by moving the means for opening the lid into the space below the container, material can be saved and the manufacturing method can be simplified. That is, the movement of the means into the space below the container reduces the space of the gas above the reaction solution. Therefore, it is not necessary to use a large amount of the reaction solution to reduce the space, so that the assembly method can be performed by saving the amount of expensive reagents and reaction solutions used and by using a relatively large reaction vessel that is easy to handle. Can be simplified. A further advantage is that the portion of the lid protruding into the reaction vessel allows the gas space above the reaction solution to be as small as possible. As a result of this advantage, there is no need to reduce the wall height of the reaction vessel, which would be disadvantageous in terms of the possibility of flying splashes during pipette operation. Since the lid of the present invention is suitable for automatic operation, it facilitates full automation of the device for detecting a sample in a liquid.

【0041】

The present invention can be utilized for component analysis of body fluids, particularly according to the principles of immunoassay or nucleic acid diagnosis. Due to the advantages described above, the application of nucleic acid detection is particularly preferred. The method for detecting nucleic acid can also include a step or another apparatus for performing amplification of nucleic acid in a liquid, for example a polymer chain reaction (European Patent No. 0 200 362). Subsequent mixing reactions with detection needles can also be performed in the reaction vessel of the present invention. In this case, it is possible to open the reaction vessel many times with the help of the lid of the present invention.

【0042】

FIG. 1 shows a composite plate with a reaction vessel 1, a heat block 4, and a lid of the present invention.

【0043】

FIGS. 2-4 show an overall view of the lid performing the method of the invention, along with a preferred remover and reaction vessel.

【0044】

FIG. 5 shows the apparatus of the present invention for carrying out the reaction. In this case, the composite plate with the reaction vessel and lid is indicated by reference numeral 010. Reagent storage vessels 011, 012 and 013 and some basic sample vessels 014, 015 and 016 may be provided, from which the amount required for the reaction is removed by the pipetting unit 017 and contained in the reaction vessel 1. Moved to. Sealing of the reaction vessel, which can be performed by the removal device 7 if necessary, but after the reaction and opening of the lid by the removal device, additional reagents can be added by pipette. If necessary, the signal measurement is then carried out in the reaction vessel itself or in the measurement unit after the reaction mixture has been transferred. The rack gear 018 of the XYZ transfer unit of a conventional automated pipettor (eg, manufactured by Tecan) is shown in the figure.

【0045】

The above-mentioned fear of contamination can also be attributed to the reagents exposed on the device. FIG. 6 shows an embodiment of a screw lid according to the present invention that can be processed using a conventional manufacturing machine and then used in an apparatus. The screw lid 8 has a thread 801 and an opening 802 for sampling that can be opened and closed by the lid of the present invention. In one embodiment with a septum, it is applied to a sample that can be transferred by syringe into a special sample tube immediately after blood collection. FIG. 7 shows a lid of the present invention improved for this purpose. It has a bulkhead 555 that can be inserted by a general pipetting needle. The advantage of this embodiment is that the sample can be taken out many times without removing the lid of the present invention. The partition wall is preferably arranged in a portion extending into the sample container of the lid.

【0046】

FIG. 8 shows a stop unit for the removal device. It is operated by the pressing piece 2101, which, along with the stopping element 2103, holds the pressing spring 2106 around the pressing pin 2105 at the starting point. The stop element 2103 moves inside the stop jacket 2102, and the pressing pin 2105 moves inside the open casing 710.

【0047】

This stop unit can be used manually as well as in the device for removing the lid. The stopping element is preferably held in the outer casing 2002 shown in FIG. The stop unit is designated by reference numeral 2100.

【0048】

The manual remover is shown in Figure 10.

【0049】

FIGS. 11 to 16 show steps A to F for removing the lid from the container. Step A shows the initial position of the removal element and the container sealed by the lid of the present invention. In step B, the removal element is placed on top of the element, including the container. In step C, a tapered removal plug is inserted into the lid portion 550. By pulling back the tapered removal plug slightly, the tapered removal plug engages in a pre-arranged narrowed position (step D). The lid is further removed from the container by removing the tapered removal plug and pulling it back with the jacket (step E). The removal unit is lifted from the element along with the reaction vessel in step F. In the next step G (not shown), the lid is removed by removing the removal jacket and pushing it onto the removal ring. The removal unit can then be repositioned or returned to its original position (step A).

【0050】

[Example 1] The procedure for processing an amplification reaction is used as an example of a possible procedure for interaction between a laboratory robot and an object of the present invention as follows. The reaction chain can be carried out by the interaction between all the elements.

【0051】

The pre-step of collecting a sample, including a sample, from a fluid (eg, blood) in the body is performed according to conventional procedures, but this is not mentioned here. A sample in a sample solution (in this case, purified nucleic acid from a blood sample) present in a reaction vessel having an individual sealing device according to the present invention is the first starting point for a series of automated amplification processes. Is.

【0052】

The equipment for the contamination-free processing of the reaction chain is further (1) a Tecan Company laboratory robot (RS9652SLD), the first arm containing the automatic pipetting unit 017 and the second arm. Is a PerkinElmer thermocycler that includes an opening and removal device 7 and (2) a composite plate 2 with a reaction vessel 1 and also includes an individual sealing device 5 for the composite plate to hold the reaction vessel. It is composed of a holding plate having a recess and a thermal block 4 having a corresponding recess located below the holding plate.

【0053】

A pipette tip support with the required reagent container, closed sample with sample for amplification reaction, measurement cell and storage container, waste container and pipette is placed on the laboratory robot platform. All containers are sealed by individual sealing devices and opened before use.

【0054】

The following processing sequence assumes that all units, vessels, elements and materials have been prepared for processing and that the amplification reaction is about to begin anew. The "standby" position of the arm indicates a predetermined position on the laboratory robot platform outside the container zone, allowing another arm to approach the same container. Each of the two arms has its own "standby" position.

【0055】

[Processing procedure] Process operation Start of processing procedure 1 The first arm with the pipetting unit is the tip of the pipette from the starting position. Move to the cage and attach a new pipette tip to the unit.

【0056】

2 The first arm with the opening and removal device 7 is the sample 014 from the starting position. Transfer to a sample container containing.

【0057】

3 Processing procedure for opening and removing device (second arm) The device moves into the starting position and Place the removal element on the surface of the lid and Insert the tapered removal plug 700 inside the lid and Tapered removal plug Remover Push by lifting device 7 Put in, The lid is removed from the container A second arm with a lid is in the "standby" position next to the vessel from the area of the reaction vessel opening. Move into the room.

【0058】

4 The first arm with the pipetting unit and the tip of the pipette are pipette Move from the tip cage to the sample container of the sample.

【0059】

5 Sample removal processing procedure (1st arm) Lower the pipetting unit with the tip of the pipette, Inhale the sample solution into the tip and Lift the pipetting unit with the tip of the pipette and the sample solution To.

【0060】

6 First with pipetting unit with pipette tip and sample solution The arm moves into the "standby" position next to the opening of the amplification vessel.

【0061】

7 The second arm with the lid moves to the opening of the sample container with the sample.

【0062】

8 Covering and sealing procedure (2nd arm) A second arm with a lid moves over the container opening and The lid is lowered onto the container and The lid is pulled away from the tapered removal plug of the opening device, The container is sealed with a lid, The second arm moves to the "standby" position.

【0063】

9 The second arm, which has an opening and a removal device, is a composite pre from the "standby" position. Move to reaction vessel 1 (amplification vessel) at a predetermined position on the container.

【0064】

10 Aperture and removal device processing procedure (2nd arm) Device 7 moves into the moving position and The withdrawal element is placed on the surface of the lid of reaction vessel 1 and A tapered removal plug is inserted inside the lid, The taper removal plug is pushed in, The lid is removed from the container The opening and removal device is removed from the container along with the lid, A second arm with a lid moves from the "standby" position to the area of the container opening.

【0065】

11 First with pipette tip and pipetting unit with sample solution The arm moves from the "standby" position to the opening of vessel 1 (amplification vessel).

【0066】

12 Sample delivery procedure (1st arm) Lower the pipette unit with the pipette tip and sample, The sample liquid is transferred from the tip, Pull up and remove the pipetting unit with the tip of the pipette, The first arm moves to the waste container, The used tip is released, The first arm moves into the starting position.

【0067】

13 The second arm with the lid moves to the opening of the amplification vessel.

【0068】

14 Processing procedure to cover and seal (2nd arm) The second arm with the lid moves over the container opening and The lid is lowered onto the container and The lid is pulled out of the taper removal plug of the opening device, The container is sealed with a lid, The second arm moves to the "standby" position.

【0069】

15 The first arm with the pipetting unit is the pipette tip from the starting position. Move to the end cage and attach a new pipette tip to the unit.

【0070】

16 The second arm, which has an opening and removal device, is plugged from the "standby" position. Transfer to the reagent container on the tofoam.

【0071】

17 Processing procedure for opening and removing device (2nd arm) Device 7 moves into the starting position Detachable elements are placed on the surface of the lid, A tapered removal plug is guided to the surface of the lid, A tapered removal plug is inserted inside the lid, The taper removal plug is pushed in, The lid is removed from the container The opening with lid and the removal unit are removed from the container Second arm with lid moves from open area of container to "standby" position next to container Move.

【0072】

18 First arm with pipetting unit and pipette tip is pipette Move from the tip cage to reagent vessel 011 on the platform.

【0073】

19 Reagent delivery procedure (1st arm) Pipetting unit with pipette tip (penetration through septum if necessary) Will be) lowered, Suck the required amount of liquid into the tip, The pipette tip and the pipetting unit with the liquid are pulled up and removed To.

【0074】

20 First arm with pipette tip and pipetting unit with liquid Moves into the "standby" position next to the opening of the amplification vessel.

【0075】

21 The second arm with the lid moves to the opening of the reagent container.

【0076】

22 Processing procedure to cover and seal (2nd arm) The second arm with the lid moves over the container opening and The lid is lowered onto the container and For the lid, use the removal jacket 710 to remove the opening device with a taper. Pulled out of the rug The container is sealed with a lid The second arm moves to the "standby" position.

【0077】

23 The second arm, which has an opening and a removal device, is a composite pre from the "standby" position. Move to the amplification vessel in place on the tongue.

【0078】

24 Processing procedure for opening and removing device (2nd arm) The device moves into the starting position and Detachable elements are placed on the surface of the lid, A tapered removal plug is inserted inside the lid, The taper removal plug is pushed in and The lid is removed from the container The opening with lid and the removal unit are removed from the container Second arm with lid moves from area of container opening to "standby" position next to container Move.

【0079】

25 Second arm with pipette tip and pipetting unit with liquid Moves from the "standby" position to the opening of the amplification vessel.

【0080】

26 Reagent delivery procedure (1st arm) Lower the pipette unit with the pipette tip and sample, The reagent solution is transferred from the tip, The pipetting unit with the pipette tip is pulled up, The first arm moves to the waste container, The used tip is released, The first arm moves into the starting position.

【0081】

27 The second arm with lid moves to the opening of the amplification vessel.

【0082】

28 Processing procedure to cover and seal (2nd arm) The second arm with the lid moves over the container opening and The lid is lowered onto the container and Is the lid a taper removal plug for the opening device using a removal jacket? Is torn off The container is sealed with a lid The second arm moves to the "standby" position.

【0083】

Steps 15-28 represent the cycle to be performed each time a new reagent is added to the amplification vessel.

【0084】

If you have a commercial kit from Hoffmann-La Roche Company (PCR buffer, Taq-polymerase (trade name, DNA polymer), and primer), 3 cycles of reagent addition If necessary, 28 more steps must be performed.

【0085】

68 The second arm with the pipetting unit moves into the starting position. 69 Amplified solution in solution is recommended for the kit or thermal cycler mentioned above Go through a heat program that is done.

【0086】

70 end 71 The second arm with opening and removal device is a composite plate from the starting position Move to the amplification vessel in place above.

【0087】

72 Processing procedure for opening and removing device (2nd arm) The device moves into the starting position and Detachable elements are placed on the surface of the lid, A tapered removal plug is inserted inside the lid, The taper removal plug is pushed in and The lid is removed from the container The opening with lid and the removal unit are removed from the container Second arm with lid moves from area of container opening to "standby" position next to container Move.

【0088】

73 The second arm with a pipetting unit with a pipette tip opens. Move from the starting position to the opening of the amplification vessel.

【0089】

74 Amplified mixing (1st arm) Lower the pipetting unit with the tip of the pipette, Inhale the required amount of liquid into the tip and The pipette unit with the tip of the pipette and the liquid is lifted. 75 Increased first arm with pipetting unit with pipette tip Move to the "standby" position next to the width container opening.

【0090】

76 The second arm with lid moves to the amplification opening.

【0091】

77 Processing procedure to cover and seal (2nd arm) The second arm with the lid moves over the container opening and The lid is lowered onto the container and Is the lid a taper removal plug for the opening device using a removal jacket? Is torn off The container is sealed with a lid The second arm moves to the "standby" position.

【0092】

78 The second arm, which has an opening and a removal device, is a measuring cell from the standby position. Or move to a storage container in place on the platform.

【0093】

79 Processing procedure for opening and removing device (2nd arm) The device moves into the starting position and The removal element is placed on the surface of the lid of the measuring cell or storage container, A tapered removal plug is inserted inside the lid, The taper removal plug is pushed in and The lid is removed from the container The opening with lid and the removal unit are removed from the container Second arm with lid moves from area of container opening to "standby" position next to container Move.

【0094】

80 Second with pipette tip and pipetting unit with sample solution The arm moves from the "standby" position to the measurement cell or storage container opening.

【0095】

81 Reagent delivery procedure (1st arm) Lower the pipette unit with the pipette tip and sample, The sample liquid is transferred from the tip, Lifted the pipetting unit with the pipette tip, The first arm moves to the waste container, The used tip is released, The first arm moves into the starting position.

【0096】

82 The second arm with the lid moves to the opening of the measuring cell or storage container.

【0097】

83 Processing procedure to cover and seal (2nd arm) A second arm with a lid moves over the container opening and The lid is lowered onto the container and The lid is pulled out of the taper removal plug of the opening device, The container is sealed with a lid, The second arm moves to the "standby" position.

【0098】

End of procedure Here, the amplification mixture is ready to directly detect the amplification being formed or to carry out a further reaction.

【0099】

[Effect of the invention]

According to the present invention, the individual lids for sealing the vessel can be automatically removed or attached so that the reaction can be carried out continuously, automatically and unaffected by contamination in the reaction vessel. can do.

[Simple explanation of drawings]

[Figure 1]

It is an elevation sectional view which shows the composite plate which has the lid of this invention.

[Figure 2]

FIG. 5 is an elevation sectional view showing an embodiment of the lid of the present invention, a removing device, and a reaction vessel.

[Fig. 3]

FIG. 5 is an elevation sectional view showing another embodiment of the lid of the present invention, a removing device and a reaction vessel.

[Fig. 4]

FIG. 5 is an elevational cross-sectional view showing still another embodiment of the lid of the present invention, a removal device and a reaction vessel.

[Fig. 5]

It is the schematic explanatory drawing of the apparatus of this invention for carrying out a reaction.

[Fig. 6]

It is an elevation sectional view which shows one Example of the screw lid of this invention.

[Fig. 7]

It is an elevation sectional view which shows the improved lid of this invention.

[Fig. 8]

It is sectional drawing which shows the stop unit for the removal device of this invention.

[Fig. 9]

It is sectional drawing which shows the outer casing of this invention including a stop element.

[Fig. 10]

It is a partial cross-sectional view which shows the manual removal tool of this invention.

[Fig. 11]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Fig. 12]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Fig. 13]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Fig. 14]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Fig. 15]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Fig. 16]

It is sectional drawing explaining the process of the method of this invention which removes a lid from a container.

[Explanation of symbols]

1 Reaction vessel 101 Inner wall of reaction vessel 102 Engagement part 2 Holding plate 3 Retention plate holes 4 heat block 5 lid 500 The part of the reaction vessel protruding from the reaction vessel 501 stopper 503 Lid surface 550 The part that extends into the opening of the reaction vessel 551 Lid removal device 552 Sealed ring 553 Intervening space 554 Inner wall 101 The outer surface of the lid facing towards 555 bulkhead 7 Aperture device and removal device 700 Tapered removable plug 701 Removal ring 710 Removable jacket 2100 Stop unit 2101 Press piece 2102 Stop jacket 2103 Stop element 2105 Press pin 2106 Pressing spring 2001 mounting 2002 external casing 2003 Counter-support 2004 Stop spring 2005 cross slot screw 010 Composite plate with reaction vessel and lid 011 Reagent storage container 012 Reagent storage container 013 Reagent storage container 014 Sample container 015 Sample container 016 Sample container 017 Pipetting unit 018 Rack gear 8 screw lid 801 thread 802 Screw lid opening for sampling