JPH06225752A - Device for automatically measuring dna probe and method for measuring nucleic acid in sample - Google Patents

Abstract

PURPOSE:To obtain a DNA probe automatically measuring device labeling the DNA with an enzyme by disposing a sample/reagent unit, a sample/reagent- injection unit, a reaction container-carrying unit, a hybridization unit, a B/F separation unit, and a light-measuring unit. CONSTITUTION:A hydridization unit D, a B/F separation unit E and a light- measuring unit F are disposed so as to be used on a circulative line, and a sample/reagent-injection unit B and a reaction container carrying unit C are arranged on an arm capable of being transferred in the directions of the X-Y-Z axes. The sample/reagent unit A comprises sample tubes 2, reagent cassettes 5 and an alkali solution-receiving container 6, and loaded on a prescribed position of a measurement section 1.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention uses a nucleic acid hybridization method to hybridize nucleic acid in a sample with a DNA probe reagent labeled with an enzyme,
The present invention relates to a DNA probe automatic measuring device and a method for measuring nucleic acid in a sample, which are capable of automatically and continuously measuring an optically detectable amount of change in a substrate under the action of an enzyme.

[0002]

2. Description of the Related Art Nucleic acid hybridization methods are used in a wide range of fields as highly specific measuring methods utilizing the complementarity of nucleic acids. As the hybridization method, dot hybridization for fixing nucleic acid directly to a membrane, colony hybridization performed by culturing bacteria in a membrane, nucleic acid as it is or cleaved with a restriction enzyme,
Southern blotting, in which fractions are obtained by electrophoresis and transferred to a nitrocellulose membrane for hybridization, and the above treatment is performed on slide glass.
In situ hybridization method and the like are known. As a support for immobilizing nucleic acids, attempts have been made to immobilize conventional membranes to microplates, beads, latex particles and the like. The DNA probe reagent used in the above hybridization also varies depending on the label, and for example, a probe reagent using a radioactive substance as a label, a probe reagent using an enzyme, a probe reagent using a chemiluminescent substance, a chemiluminescent substance. A probe reagent and the like using is known.

[0003]

DISCLOSURE OF THE INVENTION Problems to be solved by the present invention, such as infectious disease test and genetic disease test using a DNA probe, have been attracting much attention in recent years and various attempts have been made. A diagnostic kit for infectious diseases has also been released. In the above-mentioned inspections and the like, particularly in the DNA probe measurement work, it is necessary to handle a small amount of sample with high accuracy for a long time, and it becomes a monotonous and easy to get tired work. Measurement errors are likely to occur due to problems such as variations and contamination due to trace amounts of analytes, and it is earnestly desired in the market to mechanize this DNA probe measurement automatically, but it is still desired. At present, the device is not completed.

The object of the present invention is to solve the above-mentioned problems, and is suitable for a DNA probe hybridization method in which an enzyme is used as a label and an optically detectable amount of change produced in a substrate under the action of the enzyme is measured. There is a new automatic measuring device. Another object of the present invention is to provide a method for measuring a nucleic acid in a sample, which allows a large number of analytes to be continuously and efficiently measured.

[0005]

[Means for Solving the Problems] The inventors of the present invention have successively performed a sample single-strand step, a sample / reagent dispensing step, a hybridization step, a B / F separation step and a photometric step of a DNA probe hybridization method. As a result of repeated research on equipment that can be performed, the necessary equipment is arranged so that each process of hybridization, B / F separation and photometry can be performed on a line that can be circulated, and sample / reagent dispensing and reaction container transportation The present invention has been accomplished successfully by using an apparatus using an arm portion movable in the X, Y, Z axis directions and combining them to achieve the above object.

[0006] That is, the automatic DNA probe measuring apparatus of the present invention, by allowing a nucleic acid in a sample to hybridize with a DNA probe reagent labeled with an enzyme,
An automatic DNA probe measuring device for measuring nucleic acid in a sample, comprising at least (A): sample / reagent unit, (B): sample / reagent dispensing unit, (C): reaction container transport unit, (D): Hybridization unit, (E): B / F separation unit and (F):
The sample / reagent unit (A) is preferably provided with a sample tube containing a sample, a reaction container having a capture DNA probe fixed thereto, and an enzyme-labeled DNA probe reagent. And a reagent cassette including an enzyme substrate, a reagent pack unit, a mixing tank, and a storage unit that detachably stores the reaction container, and an alkali denaturing liquid storage container,
The cleaning liquid storage container is installed at a predetermined position, and the sample / reagent dispensing unit (B) has an arm portion movable in the XYZ axis directions, a tip nozzle provided in the arm portion, and A chip rack that holds a chip attached to a nozzle and is installed at a predetermined position, a mechanism that moves the arm portion, and the arm portion is moved to attach the chip to the tip of the tip nozzle. It is composed of a mechanism for sucking and injecting the sample in the sample tube and the reagent in the reagent pack, respectively, and the reaction container transport unit (C) is provided with the container holding device on the arm part, and the hybridization unit (D) is provided. However, the measurement rack containing the reaction container is circulated from the stocker section to the stocker section via the incubator section equipped with the heating device and shaking mechanism. And the circulation mechanism for, sample
A single-stranded sample is continuously sucked from the mixing tank of the reagent cassette and the enzyme-labeled DNA probe reagent is continuously sucked from the reagent pack portion with the tip nozzle equipped with the tip of the reagent dispensing unit, and then from the stocker portion to the incubator portion. The B / F separation unit (E) comprises a mechanism for dispensing to a conveyed reaction container, and a discharge nozzle and a cleaning liquid nozzle installed in the arm portion and a hybridization nozzle at the discharge nozzle. The photometric unit (E) is composed of a mechanism for sucking the content from the reaction container and discharging it to the outside, sucking the cleaning liquid from the cleaning liquid storage container with the cleaning liquid nozzle and injecting the cleaning liquid into the reaction container, and the circulation mechanism of the hybridization unit. Does not absorb the enzyme substrate from the reagent pack part of the reagent cassette with the tip nozzle equipped with the tip of the sample / reagent dispensing unit. And a mechanism for injecting into a reaction vessel after B / F separation, and the circulation mechanism of the hybridization unit is made more photometric device for measuring the optically detectable change that occurs in the reaction vessel.

Further, the method for measuring nucleic acid in a sample of the present invention uses the above-mentioned automatic DNA probe measuring apparatus,
With the tip nozzle equipped with the tip of the sample / reagent dispensing unit, the sample is continuously sucked from the sample tube and the alkaline denaturing solution is continuously sucked from the container for denaturing the alkaline solution, and simultaneously dispensed and mixed into the mixing tank of the reagent cassette. hand,
In the step of making the nucleic acid in the sample single-stranded, in the reaction container transport unit, the reaction container in which the capture probe is fixed in advance is transported from the reagent cassette to the storage unit of the measurement rack in the stocker unit of the hybridization unit,
With the tip nozzle equipped with the tip of the sample / reagent dispensing unit, the single-stranded sample is sucked from the above mixing tank and dispensed into the above reaction container, and the tip nozzle equipped with the tip is further equipped with the reagent in the reagent cassette After the enzyme-labeled DNA probe reagent is sucked from the pack part and dispensed into the reaction container, the measurement rack is passed through an incubator part by a circulation mechanism, heated and shaken, and hybridization is carried out in the reaction container. Hybridization step, with the discharge nozzle of the sample / reagent dispensing unit, the contents are sucked and discharged from the reaction container after the hybridization, and then with the washing liquid nozzle of the dispensing unit, from the washing liquid storage container Suction the cleaning solution and pour it into the above reaction vessel. B and F separation is performed in the reaction container by passing through the heating section, and then the cleaning liquid is sucked and discharged from the reaction container after the B / F separation by the discharging nozzle. / F separation process, with the tip nozzle equipped with the tip of the sample / reagent dispensing unit,
After sucking the enzyme substrate from the reagent pack part of the reagent cassette and injecting it into the reaction vessel, the measurement rack is heated by passing through the incubator part by the circulation mechanism of the hybridization unit, and then the measurement rack is measured by the circulation mechanism. It is characterized in that it is carried to a section and a photometric step is carried out to measure the amount of change in the light generated in the reaction vessel by a photometric device.

[0008]

According to the apparatus having the above construction, the nucleic acid in the sample can be automatically measured. Therefore, the measurement operator is freed from monotonous and tired work, and measurement errors due to problems such as variations in temperature management based on individual differences among operators and contamination due to a small amount of analyte are generated. It can be resolved. Further, according to the method using the apparatus having the above-mentioned configuration, it becomes possible to measure the change in light due to the sample in the reaction container automatically and continuously with high accuracy. Therefore, the nucleic acid in the sample can be measured with high efficiency and reliability.

[0009]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A DNA probe automatic measuring apparatus according to an embodiment of the present invention will be described below in more detail with reference to the drawings. FIG. 1 is a schematic diagram showing an internal configuration of a measuring unit of a DNA probe automatic measuring device used in a hybridization processing system using the DNA probe reagent. In the figure, 1 is a measurement unit, which includes a sample / reagent unit (A), a sample / reagent dispensing unit (B) provided with a reaction container transport unit (C), and a hybridization unit (D). B / F separation unit (E)
And a photometric unit (F). In addition,
The operation of the device can be controlled semi-automatically to fully automatically by various methods, but it is preferable to control the instrumental operation of the unit by a program so that the nucleic acid in the sample can be fully automatically measured.

The sample / reagent unit (A) comprises a sample tube 2 for containing a sample, a reagent cassette 5, an alkali liquid container 7 for denaturing the sample into a single chain, and a washing liquid container 6. , The measuring unit 1
Is placed at a predetermined position.

The reagent cassette has the structure shown in the side view of FIG. 2, in which reference numeral 5 is a reagent cassette, which contains a reagent pack portion 8 for containing a plurality of reagents, a mixing tank 10 and a reaction container 11. The storage part 5'to be formed is integrally formed. The reagent pack section 8 contains reagents necessary for each measurement item, such as enzyme-labeled DNA probe reagents, buffers, substrates and the like, and the opening thereof is usually sealed with a laminate seal 9 or the like. A sample and an alkaline denaturing solution are dispensed into the mixing tank 10, and the double-stranded nucleic acid in the sample is denatured into a single strand by sufficiently mixing the samples. In the storage part 5 ', the captured DNA is previously attached to the inner surface thereof
A reaction container 11 having a probe fixed therein is detachably housed.

Sample / reagent dispensing unit (B)
Is an arm portion arranged above the measuring portion of the automatic DNA probe measuring device and movable in the XYZ axis directions;
A tip nozzle provided on the arm section, a tip rack that holds a tip mounted on the nozzle and is installed at a predetermined position of the measuring section, a mechanism for moving the arm section, and a moving section for the arm section. Thus, the chip is attached to the tip of the tip nozzle, and a mechanism for aspirating and injecting the sample in the sample tube and the reagent in the reagent pack is configured.

FIG. 3 is a partial perspective view showing an arm portion of the above dispensing device. In the figure, B is a dispensing device which is movable in the XYZ axis directions, and the arm portion 21 has a tip nozzle 22 on which a tip can be attached and detached, and the tip nozzle 22 holds samples and reagents. It is connected to a fixed-volume dispenser (not shown) for quantitative suction and dispensing. Further, the arm portion 23 installed in parallel with the arm portion 21 has a B / F
It has a discharge nozzle 24 for discharging a sample, a reagent, etc. in a reaction vessel for a separation unit, and a cleaning nozzle 25 for injecting a cleaning liquid. The discharge nozzle 24 uses a dedicated discharge pump (not shown) for cleaning. The nozzle 25 is connected to a constant-volume dispenser (not shown) for aspirating and dispensing the cleaning liquid.

The reaction container transport unit (C) is the above X-
A container holding device for holding a reaction container is provided side by side with an arm portion that is movable in the YZ axis directions. As the container holding device 27, a known system such as a vacuum system or a chuck system can be used. A known mechanism can be used as it is as a method of moving the arm portion of the dispensing device in the X-axis, Y-axis, and Z-axis directions.

Hybridization unit (D)
Is a reagent cassette with a circulation mechanism that circulates the measurement rack containing the reaction container from the stocker section to the stocker section through the incubator section equipped with a heating device and a shaking mechanism, and a chip nozzle equipped with the chip of the sample / reagent dispensing unit. The single-stranded sample is continuously sucked from the mixing tank, the enzyme-labeled DNA probe reagent is sucked from the reagent pack part, and dispensed into the reaction container transported from the stocker part to the incubator part. is there.

FIG. 4 is a schematic diagram showing the structure of the hybridization unit (D). As shown in FIG.
A stocker section 14 for storing a measurement rack 16 having a storage section 16a capable of storing a plurality of reaction vessels 11, and a reaction stored in the measurement rack 16 at a predetermined position 18 while carrying the measurement rack 16 to an incubator section 15. An incubator for heating the dispensing line 17 for dispensing the single-stranded sample and the reagent and the measurement rack 16 into the container 11 and shaking them as necessary to perform hybridization in the reaction container. A circulation mechanism including a section 15 and a transfer line 20 that transfers the measurement rack 16 after hybridization to the stocker section 14 is provided.

As described above, the hybridization unit is basically composed of a circulation mechanism. In this embodiment, the stocker section 14 and the incubator section 15 are used.
8 measurement racks 16 each capable of accommodating 5 reaction vessels 11 such as microplates are installed, and by operating the motor M connected to the dispensing line 17, the measurement racks 16 are provided at the outlet of the stocker unit 14. There is one measuring rack
They are individually conveyed to the inlet of the incubator unit 15 and transferred into the incubator unit 15 by the pushing tool 19. Due to this transfer, the empty measurement rack at the exit of the incubator unit 15 is pushed out to the transfer line 20, and is moved to the inlet of the stocker unit 14 by operating the motor M connected to this transfer line 20 to push out the pusher tool 19. 'To transfer to the stocker section 14. Due to this transfer, the measurement rack at the exit of the stocker unit 14 is moved to the dispensing line 1
7 is extruded. Hereinafter, the circulation of the measurement rack described above is repeated, and the stocker section 14 and the incubator section 15 are repeated.
There are always eight measurement racks in the.

FIG. 5 is a schematic cross-sectional view for explaining the incubator section. The incubator section 15 shakes the heater 32 for heating the reaction vessel 11, the sensor 33 for controlling the temperature, and the entire incubator section. A shaking mechanism 34 for fixing and a fixture 35 for fixing the measurement rack 16 at the time of shaking are provided. This incubator section 15
Then, the reaction container 11 housed in the measurement rack 16 is heated to an appropriate temperature for a certain period of time. It should be noted that the shaking mechanism may be omitted, but it is preferable to install it side by side because the hybridization reaction in the reaction container is made uniform and the reaction time is shortened.

The B / F separation unit (E) is attached to the sample / reagent unit (A), the sample / reagent dispensing unit (B), and the hybridization unit (D). That is, the discharge nozzle and the cleaning liquid nozzle installed in the arm that is movable in the X, Y, and Z axis directions, and the discharge nozzle sucks the contents from the reaction container after hybridization and discharges them to the outside. A mechanism for sucking the cleaning liquid from the cleaning liquid storage container by the cleaning liquid nozzle and injecting the cleaning liquid into the reaction container, and a circulation mechanism for the hybridization unit.

The photometric unit (F) is a mechanism for sucking the enzyme substrate from the reagent pack portion of the reagent cassette with the chip nozzle equipped with the chip of the sample / reagent dispensing unit and injecting it into the reaction container after B / F separation. And a circulation mechanism of the hybridization unit, and a photometric device for measuring an optically detectable change occurring in the reaction container. FIG. 4 shows an example in which the photometric device is provided on the transfer line 20 of the circulation mechanism of the hybridization unit. In the present invention, the photometric device used in a normal DNA probe hybridization method can be preferably used. When performing luminescence detection as a specific example of the photometric device, as shown in the schematic cross-sectional view of FIG. 6, for example, a gap 52 between the light receiving section 51 of the luminescence detection section 50 and the opening 11a of the reaction container 11 is formed.
Is used, the stray light 53 can be covered with the luminescence detection device S, so that stray light from outside reaction chambers and other reaction containers accommodated in the measurement rack can be blocked, and even minute changes in light can be prevented. It is preferable because the measurement can be performed with high accuracy.

The automatic DNA probe measuring device having the above-mentioned structure is constructed by using the enzyme labeled NBT:
Nitro blue tetrazolium and BCIP: 5-BROMO-4-chlor
A colorimetric measurement method using o-3-indolylphosphate or the like at an absorption wavelength of 570 nm, a fluorescent measurement method in which a lanthanide or a europium chelating agent which is a fluorescent dye is labeled, and a fluorescence excited by ultraviolet irradiation is measured, and an enzyme is labeled. It can be suitably used for a luminescence measuring method for measuring luminescence generated by reacting a probe with a luminescent substrate such as luminol or a dioxycetane derivative. As the detector used in the photometric device, a light source, an interference filter, and
The light receiving element or the like may be selectively used.

The method for measuring nucleic acid in a sample of the present invention is characterized by using the DNA probe automatic measuring apparatus having the above-mentioned constitution. The processing system for hybridization with a DNA probe reagent, which is the object of the present invention, utilizes hybridization of a known nucleic acid in a sample with a DNA probe reagent, and a hybridization method is used in the measurement of the nucleic acid in the sample. 1
In the case of step method and 2-step method (sandwich method),
Although there are some differences between the DNA sample measurement and the RNA sample measurement, in the following description, FIG.
The description will be made based on the general sandwich hybridization method for a DNA sample shown in the flowchart of FIG. The other measuring methods are essentially the same.

Usually, in the above-mentioned sandwich hybridization method for a DNA sample, since the DNA in the sample is double-stranded, a single-stranded step of making it single-stranded by acid, alkali, or heat and the above-mentioned single-stranded formation. Synthetic DNA complementary to the sample DNA on a solid support such as a microplate
Is added to a fixed reaction container, and a first hybridization step is performed in which hybridization is carried out at an appropriate constant temperature. Then, a DNA probe reagent labeled with an enzyme is injected into the reaction container, the same as the first hybridization step. A second hybridization step of sandwich hybridization of sample DNA captured at a constant temperature appropriate for the above, followed by a sample that was not captured in the reaction vessel and a free labeled DNA probe in the same manner as in a known immunoassay. The reagent and the one immobilized on the solid phase carrier by sandwich hybridization are
/ F separation, and a photometry step in which the substrate is added to the reaction vessel and an enzyme activity is applied to the substrate at the same time, or a change appearing in the substrate is optically measured.

In the sample single-strand step, the arm 21 of the dispensing apparatus B shown in FIG. 3 is moved downward, and the tip 3a held in the tip rack 3 shown in FIG. After this, sample tube 2
A predetermined amount of the alkaline denaturing liquid and the sample stored in
Are simultaneously poured into the mixing tank 10. In this mixing tank 10, the tip nozzle 22 sucks and injects the mixed liquid several times to mix them sufficiently. By this operation, the sample DNA is made single-stranded. After that, the reaction container 11 in which the capture DNA probe that matches the set measurement item is stored in the reagent cassette shown in FIG. 2 is held by the container holding device 27 of the dispensing device B, and the hybridization unit It is conveyed to the storage section 16a of the measurement rack 16 in the stocker section 14 of (D).

The hybridization step is performed twice. In the first hybridization step, first, the tip nozzle 22 equipped with the tip 3a of the dispensing apparatus B is used.
The single-stranded sample is sucked from the mixing tank 10 of the reagent cassette and dispensed into the reaction container 11 housed in the measurement rack at the predetermined position 18 on the dispensing line 17 in FIG. Then, the measurement rack 16 is passed through the incubator section 15 by the circulation mechanism of the hybridization unit, heated at about 150 ° C. for 15 to 30 minutes while keeping the reaction vessel 11 under a heating atmosphere, and heated in the reaction vessel 11. Allow hybridization to occur.

In the second hybridization step, the tip nozzle 22 equipped with the above-mentioned chip is attached with the labeled probe reagent of the item set from the reagent pack section 8 of the reagent cassette 5.
And then dispensed into the reaction vessel 11 at the predetermined position 18 of the dispensing line 17 after the completion of the first hybridization step, and then the measurement rack 16 is passed through the incubator section 15 by the circulation mechanism in the same manner as above. And allow hybridization to occur. In the one-step method, the above-described hybridization step may be performed after the single-stranded sample and the labeled probe reagent are dispensed into the reaction container 11, respectively.

In the B / F separation step, after the hybridization step is completed, the content of the reaction container 11 is sucked out by the discharge nozzle 24 of the pipetting apparatus B and discharged to the outside, and then the sample is washed by the cleaning nozzle 25. The cleaning liquid is sucked from the cleaning liquid storage container 6 of the reagent unit and injected into the reaction container 11, and then the measurement rack 16 is heated in the incubator section 15 for a certain period of time (50 ° C. × 10 minutes) by the circulation mechanism as in the hybridization step. ) Do. By this operation, B / F separation is performed in the reaction container 11. After the completion of the incubator, the cleaning liquid is sucked in by the discharge nozzle 24 and discharged to the outside to remove the free labeled probe reagent and the sample remaining in the reaction container. The discharge nozzle 24 is connected to a diaphragm pump or the like, and the cleaning nozzle 25 is connected to a fixed-volume dispenser that can suck and inject a cleaning liquid in a fixed amount.

In the photometric step, the tip nozzle 22 equipped with the tip of the dispensing device B dispenses the enzyme substrate from the reagent pack section 8 into the reaction container 11 which has been subjected to the B / F separation, and dispenses the hybridization. Similar to the process, after heating in the incubator unit 15 for a certain period of time (37 ° C. × 15 minutes), the change in light generated in the reaction vessel 11 is measured by the photometric device F on the transfer line 20 of the circulation mechanism.

[0029]

As described in detail above, according to the DNA probe automatic measuring apparatus of the present invention, the nucleic acid in the sample is automatically measured, which frees the measuring operator from the tedious and tiresome work. At the same time, it is possible to eliminate the occurrence of measurement errors due to variations in temperature management based on individual differences among workers and problems such as contamination due to a small amount of analyte. Further, according to the method for measuring nucleic acid in a sample using the above-mentioned DNA probe automatic measuring device, it becomes possible to measure the change of light due to the sample in the reaction container automatically and continuously with high accuracy. Highly reliable and highly efficient measurement becomes possible.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing an internal configuration of a measuring unit of a DNA probe automatic measuring apparatus showing an embodiment of the present invention.

FIG. 2 is a side view showing the configuration of a reagent cassette.

FIG. 3 is a perspective view showing an arm portion provided with a nozzle of a dispensing unit.

FIG. 4 is a schematic diagram showing a configuration of a hybridization unit.

FIG. 5 is a schematic cross-sectional view showing a configuration of an incubator section.

FIG. 6 is a schematic cross-sectional view showing an example of a photometric unit.

FIG. 7 is a flowchart showing a measurement outline of the method of the present invention.

[Explanation of symbols]

 1 Measuring Section (A) Sample / Reagent Unit (B) Sample / Reagent Dispensing Unit (C) Reaction Container Transport Unit (D) Hybridization Unit (E) B / F Separation Unit (F) Photometric Unit

Claims (4)

[Claims] 1. An automatic DNA probe measuring device for measuring nucleic acid in a sample by hybridizing nucleic acid in the sample with a DNA probe reagent labeled with an enzyme, comprising at least the following (A), (B),
An automatic DNA probe measuring device comprising the units (C), (D), (E) and (F). (A): Sample / reagent unit. (B): Sample / reagent dispensing unit. (C): Reaction container transport unit. (D): Hybridization unit. (E): B / F separation unit. (F): Photometric unit. 2. An automatic DNA probe measuring device for measuring nucleic acid in a sample by hybridizing nucleic acid in the sample with a DNA probe reagent labeled with an enzyme, comprising at least the following (A): sample / reagent unit And (B): sample / reagent dispensing unit, (C): reaction container transport unit, (D): hybridization unit, (E): B / F separation unit, (F): photometric unit DNA comprising: and each unit having the following structure
Automatic probe measuring device. (A): A sample / reagent unit includes a sample tube containing a sample, a reaction container having a capture DNA probe fixed thereto, a reagent pack unit containing an enzyme-labeled DNA probe reagent and an enzyme substrate, a mixing tank, and the reaction container. The reagent cassette, which is integrally formed with a storage unit that detachably stores, an alkali denaturing liquid storage container, and a cleaning liquid storage container are installed at predetermined positions. (B): The sample / reagent dispensing unit holds an arm part movable in the XYZ axis directions, a chip nozzle provided in the arm part, and a chip attached to the nozzle, and holds the chip at a predetermined position. A chip rack to be installed, a mechanism for moving the arm part, and the arm part is moved so that the chip is attached to the tip of the tip nozzle, whereby the sample in the sample tube and the reagent in the reagent pack are attached. Each is composed of a mechanism for sucking and injecting. (C): The reaction container transport unit includes a container holding device installed on an arm unit that is movable in the X, Y, and Z axis directions, a mechanism for moving the arm unit, and a unit for moving the arm unit. It comprises a mechanism for transporting the reaction container held by the container holding device to the storage portion of the measurement rack. (D): The hybridization unit is equipped with a circulation mechanism that circulates the measurement rack containing the reaction container from the stocker section to the stocker section through the incubator section that includes the heating device and the shaking mechanism, and the chip of the sample / reagent dispensing unit is mounted. The single-stranded sample is continuously sucked from the mixing tank of the reagent cassette with the tip nozzle, and the enzyme-labeled DNA probe reagent is continuously sucked from the reagent pack section, and dispensed into the reaction container transported from the stocker section to the incubator section. It consists of a mechanism to (E): The B / F separation unit is installed in the arm that is movable in the XYZ axis directions, and the discharge nozzle and the washing liquid nozzle, and the discharge nozzle from the reaction container after hybridization. The contents are sucked and discharged to the outside,
It comprises a mechanism for sucking the cleaning liquid from the cleaning liquid container by the cleaning liquid nozzle and injecting the cleaning liquid into the reaction container, and a circulation mechanism for the hybridization unit. (F): a mechanism in which the photometric unit sucks the enzyme substrate from the reagent pack portion of the reagent cassette with the tip nozzle equipped with the tip of the sample / reagent dispensing unit and injects it into the reaction container after B / F separation, It comprises a circulation mechanism of the hybridization unit and a photometric device for measuring an optically detectable change occurring in the reaction container. 3. A method for measuring nucleic acid in a sample by performing the following steps using the automatic DNA probe measuring device according to claim 2. 1. Sample single-strand process: (1) Sample from the sample tube with the tip nozzle equipped with the tip of the sample / reagent dispensing unit.
The alkaline denaturing liquid is continuously sucked from the container for denaturing the alkaline denaturing liquid, and simultaneously dispensed and mixed into the mixing tank of the reagent cassette to make the nucleic acid in the sample into a single strand. 2. Hybridization step: (2) In the reaction container transport unit, the reaction container in which the capture probe is fixed in advance is transported from the reagent cassette to the storage unit of the measurement rack in the stocker unit of the hybridization unit. (3) With the tip nozzle equipped with the tip of the sample / reagent dispensing unit, the single-stranded sample is sucked from the mixing tank and dispensed into the reaction container, and the tip nozzle equipped with the tip is used to carry out the reagent After the enzyme-labeled DNA probe reagent is sucked from the reagent pack part of the cassette and dispensed into the above reaction container, the measurement rack is passed through an incubator part by a circulation mechanism, and heating and shaking are applied to carry out hybridization in the above reaction container. Let's do it in. 3. B / F separation and washing process: (4) The sample / reagent dispensing unit discharge nozzle sucks and discharges the contents from the reaction container after hybridization, and then the washing liquid nozzle of the dispensing unit. Then, the cleaning liquid is sucked from the cleaning liquid storage container and injected into the reaction container, and the measurement rack is passed through the incubator part by the circulation mechanism to be heated and shaken in the same manner as above to perform B / F separation in the reaction container. Let (5) Then, the cleaning liquid is sucked and discharged from the reaction container after the B / F separation by the discharging nozzle. 4. Photometric process: (6) After the enzyme substrate is sucked from the reagent pack portion of the reagent cassette and injected into the reaction container with the tip nozzle equipped with the tip of the sample / reagent dispensing unit, the measurement rack is attached to the hybridization unit. The circulation mechanism heats the incubator through the incubator. (7) Next, the measurement rack is conveyed to the photometry unit by the circulation mechanism, and the amount of change in light generated in the reaction container is measured by the photometry device. 4. A method for measuring nucleic acid in a sample by carrying out the following steps using the automatic DNA probe measuring device according to claim 2. 1. Sample single-strand process: (1) Sample from the sample tube with the tip nozzle equipped with the tip of the sample / reagent dispensing unit.
The alkaline denaturing liquid is continuously sucked from the container for denaturing the alkaline denaturing liquid, and simultaneously dispensed and mixed into the mixing tank of the reagent cassette to make the nucleic acid in the sample into a single strand. 2. Hybridization step: (2) In the reaction container transport unit, the reaction container in which the capture probe of the reagent cassette is fixed in advance is transported to the storage unit of the measurement rack in the stocker unit of the hybridization unit. (3) With the tip nozzle equipped with the tip of the sample / reagent dispensing unit, the single-stranded sample is sucked from the mixing tank and dispensed into the reaction container of the measurement rack, and the measurement rack is circulated by an incubator section. Heating and shaking are applied to allow hybridization to occur in the reaction vessel. (4) Then, after the DNA probe reagent labeled with the enzyme is sucked from the reagent pack portion of the reagent cassette and injected into the reaction container with the chip nozzle equipped with the chip of the dispensing unit, the measurement rack is connected to the circulation mechanism. To allow hybridization to proceed in the reaction vessel by passing through the incubator section. 3. B / F separation and washing step (5) The discharge nozzle of the sample / reagent dispensing unit sucks and discharges the contents from the reaction container after the hybridization, and then the washing liquid nozzle of the dispensing unit. The cleaning liquid is sucked from the cleaning liquid storage container and injected into the reaction container, and the measurement rack is passed through the incubator section by the circulation mechanism, heated and shaken, and the B /
F separation is performed in the reaction vessel. (6) Next, the cleaning liquid is sucked and discharged from the reaction container after the B / F separation by the discharge nozzle. 4. Photometric process (7) The enzyme nozzle is sucked from the reagent pack part of the reagent cassette with the tip nozzle equipped with the tip of the sample / reagent dispensing unit and injected into the reaction container, and then the measurement rack is circulated through the hybridization unit. It is heated by passing through the incubator by the mechanism. (8) Next, the measurement rack is transported to the photometry unit by the circulation mechanism, and the amount of change in light generated in the reaction container is measured by the photometry device.