WO2021176561A1

WO2021176561A1 - Detection device, dispensing device, and dispensing method

Info

Publication number: WO2021176561A1
Application number: PCT/JP2020/008923
Authority: WO
Inventors: 雪夫小野; 稔章平塚; 隼司石塚; 貴之野田; 曽根原　剛志; 庄司　智広
Original assignee: 株式会社日立ハイテク
Priority date: 2020-03-03
Filing date: 2020-03-03
Publication date: 2021-09-10

Abstract

A detection device of the present disclosure detects light from an analyte in a liquid, wherein the detection device is characterized: by comprising a light source that projects light to the liquid, and a detection unit that detects light from the analyte in the liquid; and in that at least the detection unit is disposed in a dispensing tip by which the liquid is suctioned.

Description

Detection device, dispensing device and dispensing method

This disclosure relates to a detection device, a dispensing device and a dispensing method.

Genome medicine has started in earnest, and three oncogene panel diagnostic agents have been covered by insurance. The gene profiling test used in oncogene panel diagnosis is a comprehensive next-generation DNA sequencer (NGS) that extracts nucleic acids from blood and pathological specimens collected from cancer patients and uses gene mutation test kit reagents. It is to be analyzed. Based on the analyzed results, after the treatment policy is decided by a group of specialists such as doctors, pathologists, bioinformaticians, pharmacists, etc., anti-cancer drugs, molecular-targeted drugs, or immune checkpoints suitable for the patient Inhibitors are provided. However, in addition to the high cost of genetic panel testing, all tests are required to be successful because one patient can only use it once within the insurance coverage. On the other hand, at present, 10 to 20% of the samples have poor analysis results.

There are various pretreatment steps for nucleic acid samples, but in the nucleic acid sample amplification step, the concentration adjustment of the double-stranded nucleic acid solution after library preparation keeps the initial nucleic acid concentration to some extent in order to carry out the amplification reaction in a reproducible manner. Need to be aligned with. Also, when loading the amplified nucleic acid sample into the flow cell for sequencing, it is necessary to adjust the nucleic acid concentration to be suitable for the sequencing.

Conventionally, the quantification and concentration adjustment of the nucleic acid extracted by the nucleic acid extractor has been performed manually. That is, in order to adjust the nucleic acid concentration, the nucleic acid concentration was quantified, and the amount of dispensing or dilution was manually determined according to the concentration. These series of processes have a large number of steps and may have specialized knowledge, and there is an increasing need for automation so as not to depend on the proficiency level of the technician.

Since sample quality confirmation (QC) with a pretreatment automation device already on the market is performed by a dedicated measuring device provided in the device, the pretreatment automation device is generally large. In addition, since the number of dispensings increases dramatically as the number of processed samples increases, the overall processing time tends to increase.

In Patent Document 1, in the nucleic acid detection device, "the incubator 6 of the reaction vessel 1 is arranged, and the fluorescence detection unit 7 for performing fluorescence measurement in the reaction vessel 1 is arranged below the incubator 6, and each of these container racks. , The enzyme reagent dispensing mechanism 8 and the lid catching mechanism 9 are installed on the transfer device 10 arranged so as to be horizontally movable above the chip disposal unit 5 and the incubator 6. ” Is described (see the abstract of the same document).

Japanese Unexamined Patent Publication No. 2009-77369

The nucleic acid detection device of Patent Document 1 has a dispensing device and a fluorescence detection unit, but since the nucleic acid is only detected by fluorescence, the presence or absence of nucleic acid can be detected, but the desired nucleic acid concentration can be obtained. To adjust, it is necessary to perform a separate operation such as measuring the concentration. Even if the nucleic acid detection device of Patent Document 1 is provided with a measuring device for measuring the nucleic acid concentration, it is considered that the entire device becomes large. As a result, it becomes difficult to install the device in a laboratory such as a hospital where the sample is collected, and it becomes impossible to measure the concentration of the nucleic acid of the sample on the spot, so that the total processing time of the sample increases. there is a possibility.

Therefore, the present disclosure provides a technique that enables rapid measurement of the concentration of an analysis target in a liquid.

The detection device of the present disclosure is a detection device that detects light from an analysis target in a liquid, and is a detection device that detects a light source that irradiates the liquid with light and light from the analysis target in the liquid. And, at least, the detection unit is arranged in a dispensing chip to which the liquid is sucked.

Further features relating to this disclosure will become apparent from the description herein and the accompanying drawings. In addition, the aspects of the present disclosure are achieved and realized by the combination of elements and various elements and the following detailed description and the aspect of the appended claims.
The description of the present specification is merely a typical example, and does not limit the scope of claims or application examples of the present disclosure in any sense.

According to the detection device of the present disclosure, it is possible to quickly measure the light from the analysis target in the liquid sucked into the dispensing chip.
Issues, configurations and effects other than the above will be clarified by the following description of the embodiments.

It is a schematic diagram which shows the dispensing apparatus which concerns on 1st Embodiment. It is schematic cross-sectional view which shows the internal structure of the chip which concerns on 1st Embodiment. It is a B arrow view of FIG. 1B. It is a flowchart which shows the dispensing method using the dispensing apparatus which concerns on 1st Embodiment. It is a flowchart which shows the dispensing method using the dispensing apparatus which concerns on 1st Embodiment. It is a schematic diagram which shows the dispensing apparatus which concerns on 2nd Embodiment. It is the schematic sectional drawing which shows the internal structure of the chip which concerns on 2nd Embodiment. It is a B arrow view of FIG. 4B. It is a schematic diagram which shows the dispensing apparatus which concerns on 3rd Embodiment. It is a schematic cross-sectional view which shows the structure inside and outside of the chip which concerns on 3rd Embodiment. It is a schematic diagram which shows the automatic dispensing apparatus which concerns on 4th Embodiment.

An embodiment of the present disclosure will be described below. The steps and components for forming the components of each embodiment are not limited to the following descriptions. In addition, when the shape and function of a component are referred to, unless otherwise specified, those having a shape or function similar to the shape and arrangement of the component are included. In addition, even if some of the components are changed, if the main functions are not lost, the components are considered to be the same as before the change.

In the present disclosure, as an example, a nucleic acid is used as a concentration analysis target, and a liquid (detected liquid) dispensed by the dispensing device of the present disclosure is a mixture of a solution containing nucleic acid and a fluorescent dye that binds to nucleic acid. Make it a liquid. However, in the application of the technique of the present disclosure, the analysis target is not limited to nucleic acid, and the dye that binds to the analysis target is not limited to the fluorescent dye.

Examples of fluorescent dyes for nucleic acid staining include 3,6-bis (dimethylamino) acridines hydrochloride, ethidium bromide, propidium iodide, ethidium homodimer, SYTOX Green, POPO-1, BOBO-1, YOYO-1, TOTO. -1, SYBR (registered trademark) Gold, SYBR Green I, SYBR Green II and the like. The above fluorescent dye may be used as the standard fluorescent dye used when determining the calibration curve used for measuring the concentration of nucleic acid. For example, tetramethylrhodamine isothiocyanate, Texas Red, Cy3, Cy5, HNPP (2-hydroxy- 3-naphthoic acid-2'-phenylanilidephosphate), Alexa Fluor (registered trademark) 488, Alexa Fluor 546 and the like may be used. A mixed solution of these fluorescent dyes and a nucleic acid solution of known concentration can be used in the preparation of the calibration curve.

[First Embodiment]
<Configuration of dispensing device>
FIG. 1A is a schematic view showing a dispensing device 100 according to the first embodiment. As shown in FIG. 1A, the dispensing device 100 includes a chip 1 and a main body 2.

The dispensing device 100 can be, for example, a cylinder-driven pneumatic electric pipette. Although not shown, the main body 2 has a built-in dispensing mechanism necessary for the dispensing operation (suction operation and discharge operation) by the dispensing device 100, and the dispensing mechanism is a cylinder in which a plunger is built. , Has a motor for reciprocating the plunger.

The chip 1 can be attached to and detached from the main body 2, and may be disposable. The liquid to be detected 101 (liquid) is sucked into the tip of the chip 1 by the suction operation of the dispensing device 100, and is discharged from the chip 1 by the discharging operation of the dispensing device 100.

Inside the chip 1, a fluorescence detection device 10 (detection device) is provided above the region where the liquid to be detected 101 is sucked. The fluorescence detection device 10 is connected to the main body 2 by a cable 3. The details will be described later, but to outline the operation of the fluorescence detection device 10, the light to be detected 101 is irradiated to detect fluorescence from the fluorescence-labeled nucleic acid (analysis target) present in the detection liquid 101. , The detection result is output to the main body 2.

The main body 2 has a calculation unit 21, a power supply unit 22, and a switch 23. As the hardware configuration of the arithmetic unit 21, for example, a small microcomputer system such as Arduino (registered trademark) or Raspberry Pi (registered trademark) can be used.

The calculation unit 21 transmits an instruction value according to a predetermined suction amount or discharge amount to the above-mentioned dispensing mechanism (not shown), and drives the plunger based on the instruction value to obtain a predetermined amount. The liquid to be detected 101 can be sucked and discharged. A storage device (not shown) such as a memory built in the main body 2 stores upper and lower limits of the suction amount and the discharge amount of the liquid by the dispensing device 100. Further, an input unit (not shown) for setting the suction amount can be provided in the main body 2 so that the user can appropriately set the suction amount and the discharge amount.

The power supply unit 22 supplies power to each component (calculation unit 21, dispensing mechanism, etc.) of the main body 2 to drive them. The power supply unit 22 also supplies power to the fluorescence detection device 10 via the cable 3. The power supply unit 22 may be, for example, a small battery, or may be a mechanism that receives power from an external power supply line and supplies power to each component of the main body 2 and the fluorescence detection device 10. By providing the power supply unit 22 in the main body 2 in this way, it is not necessary to provide the power supply unit in the fluorescence detection device 10, so that the fluorescence detection device 10 has a simple configuration and can be miniaturized.

The switch 23 is provided for the user to instruct the timing of suction and discharge of the liquid. For example, when the switch 23 is pressed by the user, the calculation unit 21 detects that the switch 23 is turned on and starts the suction operation by the dispensing mechanism, and when the switch 23 is pressed again by the user, the dispensing is performed. The discharge operation by the mechanism is started.

FIG. 1B is a schematic cross-sectional view showing the internal configuration of the chip 1. In FIG. 1B, only the region of the chip 1 in which the fluorescence detection device 10 exists is shown. As shown in FIG. 1B, the fluorescence detection device 10 is supported by the chip 1 by the support 4. The fluorescence detection device 10 can be attached to and detached from the chip 1. As the material of the support 4, a flexible material such as rubber or resin can be used. A vent 5 is provided between the support 4 and the fluorescence detection device 10.

The fluorescence detection device 10 includes a substrate 11, a light receiving element 12 (detection unit), a fluorescence filter 13, a light emitting element 14 (light source), a convex lens 15 (condensing lens), a current / voltage conversion circuit 16 (detection unit), and an AD conversion circuit 17. It has a (detection unit) and a light source drive circuit 18.

The light emitting element 14 and the light receiving element 12 are provided on the substrate 11 so as to face the liquid to be detected 101. The light emitting element 14 and the light receiving element 12 may be independently provided on the substrate 11, or an element in which they are integrated may be provided on the substrate 11.

The light emitting element 14 irradiates the liquid to be detected 101 with light (excitation light). As the light emitting element 14, an element whose central wavelength of the emitted light is close to the excitation wavelength of the fluorescent dye contained in the liquid to be detected 101 can be used. As a specific example of the light emitting element 14, for example, a light emitting diode (LED) or the like which can be easily miniaturized, is inexpensive, has low power consumption, and has a long life can be used. Examples of the material of the light emitting diode include indium gallium nitride, gallium nitride, zinc selenide, zinc oxide, perovskite semiconductor, gallium aluminum gallium arsenide, and gallium arsenide.

A convex lens 15 is provided after the light emitting element 14 (on the side of the liquid to be detected 101). As a result, the light from the light emitting element 14 can be focused, and the excitation light can be irradiated to an arbitrary place at a predetermined distance with respect to the liquid to be detected 101.

The light receiving element 12 detects the light emitted from the liquid to be detected 101 (the fluorescence emitted by the fluorescent dye in the liquid to be detected 101 by the irradiation of the light from the light emitting element 14), and outputs a current signal. As the light receiving element 12, an element capable of detecting the wavelength of fluorescence emitted by the fluorescent dye can be used. As a specific example of the light receiving element 12, an element that converts light into an electric current, such as a photomultiplier tube (PMT), a photodiode (PD), and a phototransistor, can be used.

The fluorescence filter 13 is provided in front of the light receiving element 12 (on the side of the liquid to be detected 101), and causes light of a specific wavelength to enter the light receiving element 12. For example, as the fluorescence filter 13, one that transmits the fluorescence wavelength of the fluorescent dye and cuts the excitation wavelength of the light emitting element 14 can be used.

The number of light receiving elements 12 and light emitting elements 14 may be one or a plurality, respectively, as shown in the figure.

The current-voltage conversion circuit 16 converts the current signal output from the light receiving element 12 into a voltage signal, and the AD conversion circuit 17 converts the voltage signal into a digital signal, and the calculation unit 21 (FIG. 1A) via the cable 3. A digital signal (detection result) is transmitted to.

The light source drive circuit 18 controls the irradiation of light by the light emitting element 14. As the drive method of the light source, for example, a constant current drive method, a current limiting resistance method, a high frequency drive method, or the like can be adopted. The light source drive circuit 18 can control the lighting or extinguishing of the light emitting element 14 by an external trigger, for example, an on / off operation by a user of the switch 23 provided in the main body 2.

The calculation unit 21 reads the digital signal received from the AD conversion circuit 17 and digitizes it as a voltage value. After that, the calculation unit 21 uses a calibration curve prepared based on the fluorescence measurement value (voltage value) of the fluorescence standard solution having a known nucleic acid concentration measured in advance, and corresponds to the quantified voltage value of the liquid to be detected 101. Calculate the concentration of nucleic acid in. Further, the calculation unit 21 determines the discharge amount of the liquid to be detected 101 based on the calculated nucleic acid concentration and the suction amount of the dispensing device 100, and transmits an indicated value to the dispensing mechanism according to the determined discharge amount. ..

Although the example in which the calculation unit 21 is mounted on the main body 2 is shown, the calculation unit 21 may be mounted on the substrate 11 of the fluorescence detection device 10.

The calculation result (voltage value, nucleic acid concentration, suction amount, discharge amount, etc.) by the calculation unit 21 may be displayed on a display unit (not shown) provided in the main body unit 2.

FIG. 1C is a view taken along the line B of FIG. 1B. Note that FIG. 1B is a cross-sectional view taken along the line AA shown in FIG. 1C. As shown in FIG. 1C, the support 4 is an annular member, and the four corners of the rectangular plate-shaped substrate 11 are supported by the support 4, and a part of each side of the substrate 11 and the support 4 are supported. The gap is a vent 5. Since the vent 5 is formed, the liquid to be detected 101 can be sucked and discharged by the cylinder drive. The vent 5 does not have to be a gap provided between the substrate 11 and the support 4, and may be in the form of a through hole provided in the substrate 11 itself. Further, the support 4 may be formed of a breathable material (for example, a porous material such as sponge or urethane) to serve as a vent.

As shown in FIG. 1C, the support 4 can be one annular member, but a plurality of arcuate members may be used in combination. Further, the shape of the support 4 is not limited to the annular shape, and other shapes may be adopted depending on the dimensions and shape of the substrate 11. As described above, the number, material, and shape of the support 4 are not limited as long as the substrate 11 can be supported in the chip 1.

<Analysis method>
FIG. 2 is a flowchart showing a dispensing method using the dispensing device 100 according to the present embodiment. In this embodiment, the user manually performs the dispensing operation using the dispensing device 100. The method of FIG. 2 is performed to prepare a calibration curve using a standard solution containing nucleic acid of known concentration and a fluorescent dye.

In step S11, the user mounts the support 4 and the fluorescence detection device 10 inside the chip 1, connects the fluorescence detection device 10 and the main body 2 with a cable 3, and mounts the chip 1 on the main body 2. .. Alternatively, the main body 2 and the fluorescence detection device 10 may be previously connected by a cable 3 and a chip 1 provided with a support 4 inside may be supplied to the user. In this case, the user only needs to insert the fluorescence detection device 10 into the chip 1 and attach the chip 1 to the main body 2, so that the operation is simple. Of course, the dispensing device 100 in the state shown in FIG. 1A may be provided to the user. As described above, there is no particular limitation on to what stage the dispensing device 100 is provided to the user in an assembled state.

In step S12, the user immerses the tip of the chip 1 in a fluorescent standard solution having a known nucleic acid concentration, and turns on the switch 23 for starting the suction operation. When the calculation unit 21 detects that the switch 23 is turned on, it transmits an instruction value to the dispensing mechanism to drive the plunger, thereby sucking a predetermined amount of the fluorescent standard solution into the chip 1.

In step S13, the calculation unit 21 transmits an instruction to the light source drive circuit 18 to irradiate the fluorescence standard solution with excitation light from the light emitting element 14. When the light receiving element 12 detects fluorescence from the fluorescence standard solution in the chip 1, it outputs a detection signal to the current-voltage conversion circuit 16.

In step S14, the current-voltage conversion circuit 16 converts the detection signal into a voltage signal, the AD conversion circuit 17 converts the voltage signal (analog signal) into a digital signal, and transmits the digital signal to the arithmetic unit 21 via the cable 3. do. When the calculation unit 21 receives the digital signal, the calculation unit 21 calculates the voltage value based on the digital signal. The calculated voltage value is stored in, for example, a storage device (not shown) built in the main body 2.

In order to prepare a calibration curve, the user performs the above steps S12 to S14 for a plurality of fluorescent standard solutions having different concentrations.

In step S15, the calculation unit 21 creates a calibration curve from the voltage value calculated for the fluorescence standard solution of each concentration and the concentration of the fluorescence standard solution at that time. The state inside the chip 1 without a solution is used as the background. The created calibration curve is stored in a storage device (not shown).

FIG. 3 is a flowchart showing a dispensing method using the dispensing device 100 according to the present embodiment. The method of FIG. 3 is carried out to dispense the nucleic acid to be detected 101 containing the nucleic acid of unknown concentration and measure the nucleic acid concentration.

In step S21, the user prepares the dispensing device 100 in the same manner as in step S11 of FIG.

In step S22, the user immerses the tip of the chip 1 in the liquid to be detected 101 having an unknown nucleic acid concentration, and turns on the switch 23 for starting the suction operation. When the calculation unit 21 detects that the switch 23 is turned on, it transmits an instruction value to the dispensing mechanism to drive the plunger, thereby detecting a nucleic acid having an unknown concentration and a fluorescent dye in the chip 1. A predetermined amount of the liquid 101 is sucked. Let A be the suction amount of the liquid to be detected 101. Further, the upper limit of the suction amount of the dispensing device 100 is set to B.

In step S23, the calculation unit 21 transmits an instruction to the light source drive circuit 18 to irradiate the liquid to be detected 101 with excitation light from the light emitting element 14. When the light receiving element 12 detects fluorescence from the liquid to be detected 101 in the chip 1, it outputs a detection signal to the current-voltage conversion circuit 16.

In step S24, the voltage value (V) is calculated by the calculation unit 21 in the same manner as in step S14 of FIG.

In step S25, the calculation unit 21 reads the calibration curve from the storage device and obtains the nucleic acid concentration (C1) corresponding to the voltage value (V) calculated in step S24 based on the calibration curve.

In step S26, the calculation unit 21 calculates the required discharge amount (D) of the liquid to be detected 101 from the obtained nucleic acid concentration (C1) and the required concentration (C2). The required discharge amount (D) is calculated by D = C2 × (1 / C1).

In step S27, the calculation unit 21 determines the discharge amount (F) of the liquid to be detected 101 based on the required discharge amount (D). Here, for example, when the required discharge amount (D) is equal to or less than the lower limit (E) of the discharge amount of the dispensing device 100 (D ≦ E), the lower limit of the discharge amount (E) is set as the discharge amount (F). Further, for example, when the required discharge amount (D) is equal to or higher than the upper limit (B) of the suction amount (D ≧ B), the total amount of the liquid to be detected 101 sucked into the chip 1 is referred to as the discharge amount (F). do. When the required discharge amount (D) is larger than the discharge amount lower limit (E) of the dispensing device 100 and less than the suction amount upper limit (B) (E <D <B), the required discharge amount (D) = discharge amount. Let it be (F).

In step S28, the user turns on the switch 23 for instructing the discharge. When the calculation unit 21 detects that the switch 23 is turned on, it transmits an indicated value according to the discharge amount (F) to the dispensing mechanism, and drives the plunger to discharge the liquid to be detected 101.

<Technical effect>
As described above, the fluorescence detection device 10 (detection device) according to the first embodiment is provided inside the chip 1 and receives light from the liquid to be detected 101 (liquid) sucked into the chip 1 as a light receiving element. Detect by 12. By having such a configuration, the distance between the light receiving element 12 and the liquid surface can be shortened, and only air exists between them, so that the fluorescence from the liquid to be detected 101 can be emitted. It can be detected with high sensitivity. Further, since the device for detecting fluorescence is housed in the chip 1, fluorescence can be detected only by sucking the liquid to be detected 101. Therefore, the time for performing the process for detecting fluorescence can be shortened. Further, it is not necessary to provide the fluorescence detection unit in the apparatus as in Patent Document 1, and it is possible to suppress the increase in size of the apparatus.

Since the light emitting element 14 is also provided in the chip 1, the distance between the liquid surface of the liquid to be detected 101 and the light emitting element 14 can be made as close as possible, so that it is not necessary to increase the light emitting intensity of the light source. As a result, a large light source such as a laser light source or a shading means becomes unnecessary.

In the dispensing device 100 according to the first embodiment, the calculation unit 21 measures the concentration of nucleic acid (analysis target) in the detected liquid 101 based on the fluorescence detection result of the fluorescence detection device 10, and the detected liquid The discharge amount of 101 is determined. With such a configuration, it is possible to measure the concentration of nucleic acid in the sucked liquid 101 to be detected and discharge a desired amount of the liquid 101 to be detected by one dispensing operation. As described above, the user only presses the switch 23 during suction and discharge of the liquid to be detected 101. Therefore, it is not necessary to perform a plurality of operations such as measuring the nucleic acid concentration and re-dispensing the required amount after dispensing the liquid to be detected 101 containing the analysis target, so that the quality of the liquid to be detected 101 can be confirmed easily and quickly. Can be carried out. Further, as described above, since the light receiving element 12 can detect the fluorescence from the liquid to be detected 101 with high sensitivity, the calculation unit 21 of the dispensing device 100 can also calculate the nucleic acid concentration with high accuracy.

[Second Embodiment]
In the first embodiment, the fluorescence detection device 10 in which both the light emitting element 14 and the light receiving element 12 are mounted on the substrate 11 has been described. If the fluorescence from the liquid to be detected 101 can be detected by the light receiving element 12 inside the chip 1, the arrangement of each element of the fluorescence detection device 10 can be appropriately changed. Therefore, in the second embodiment, we propose an example in which the arrangement of the light receiving element 12 and the light emitting element 14 is changed.

<Configuration example of dispensing device>
FIG. 4A is a schematic view showing the configuration of the dispensing device 200 according to the second embodiment. As shown in FIG. 4A, the appearance configuration of the dispensing device 200 is the same as that of the dispensing device 100 of the first embodiment, and thus the description thereof will be omitted.

FIG. 4B is a schematic cross-sectional view showing the internal configuration of the chip 1 of the dispensing device 200. As shown in FIG. 4B, a fluorescence detection device 20 is provided inside the chip 1. The components of the fluorescence detection device 20 are the same as those of the fluorescence detection device 10 according to the first embodiment, but in this embodiment, the arrangement of the light receiving element 12 and the light emitting element 14 is different. Specifically, only the light receiving element 12 and the fluorescence filter 13 are provided at the center of the surface of the substrate 11 facing the liquid to be detected 101. A plurality of light emitting elements 14 and convex lenses 15 are provided on the inner peripheral surface of the fluorescence detection device 20. Further, the light emitting element 14 is supported by the support 6 on the inner peripheral surface of the chip 1. The light emitting element 14 and the substrate 11 are connected by, for example, wiring (not shown). For example, the circuit of the substrate 11 and the light emitting element 14 is a printed circuit board circuit, or a single-sided substrate, a double-sided substrate, a multilayer substrate, or a single-sided substrate, a double-sided substrate, or a multilayer board in which a circuit pattern arrangement is constructed by vapor deposition, gravure printing, or inkjet printing of a conductive thin film or a conductor. It may be connected by a build-up board.

FIG. 4C is a view taken along the line B of FIG. 4B. Note that FIG. 4B is a cross-sectional view taken along the line AA shown in FIG. 4C. As shown in FIG. 4C, four light emitting elements 14 and four convex lenses 15 are provided at intervals of 90 degrees along the circumferential direction of the chip 1, and the light receiving element 12 is the chip 1 when viewed from below the dispensing device 200. It is located on the central axis, and the light emitting element 14 is located around the light receiving element 12.

The number and arrangement intervals of the light emitting element 14 and the convex lens 15 are not limited to those shown in FIG. 4C, and can be set arbitrarily.

<Technical effect>
As described above, in the dispensing device 200 according to the second embodiment, the light receiving element 12 is arranged on the central axis in the chip 1, and the light emitting element 14 is arranged along the inner circumference of the chip 1. .. By adopting such a structure, the light of the light emitting element 14 can be more efficiently irradiated to the liquid to be detected 101 as compared with the first embodiment, so that the detection efficiency of the low-concentration fluorescent sample is improved. be able to.

[Third Embodiment]
In the first embodiment, it has been described that the light detection result (digital signal) by the fluorescence detection device 10 is transmitted to the calculation unit 21 of the main body 2 via the cable 3, that is, by wire. On the other hand, in the third embodiment, an example of wirelessly transmitting the detection result of the fluorescence detection device 10 to the outside is proposed.

<Configuration of dispensing device>
FIG. 5A is a schematic view showing the dispensing device 300 according to the third embodiment. As shown in FIG. 5A, the dispensing device 300 is different from the dispensing device 200 of the first embodiment in that the main body 2 further includes a receiving unit 24 and a transmitting unit 25 (wireless communication circuit). The receiving unit 24 receives the light detection result by the fluorescence detection device 30. The transmission unit 25 wirelessly transmits, for example, the calculation result of the calculation unit 21 to an external computer terminal or the like.

FIG. 5B is a schematic cross-sectional view showing the internal configuration of the chip 1 of the dispensing device 300. As shown in FIG. 4B, the components of the fluorescence detection device 30 are the same as those of the fluorescence detection device 10 according to the first embodiment, but in the fluorescence detection device 30, the wireless communication circuit 31 and the battery 32 are further mounted on the substrate 11. It is provided in. The battery 32 supplies power to each component provided on the substrate 11.

The wireless communication circuit 31 wirelessly transmits the digital signal generated by the AD conversion circuit 17 to the receiving unit 24 of the main unit 2. As the wireless communication method, known methods such as Bluetooth (registered trademark), Wi-Fi, and infrared rays can be adopted. When the receiving unit 24 receives the digital signal from the wireless communication circuit 31, it outputs the digital signal to the arithmetic unit 21.

The receiving unit 24, the transmitting unit 25, and the processing unit that executes the arithmetic processing in the arithmetic unit 21 do not necessarily have to be provided in the main body 2, but may be provided in an external device. For example, a computer terminal such as a smartphone 110, a personal computer 120, a tablet, or a mobile phone may execute the processing of the receiving unit 24, the transmitting unit 25, and the arithmetic unit 21. Further, data on the concentration of nucleic acid and the discharge amount of the liquid to be detected 101 may be displayed on the screen of the computer terminal. In this way, when the processing of the arithmetic unit 21 is executed on the computer terminal, an application or the like for executing the necessary processing on the computer terminal may be installed.

<Dispensing method>
Since the dispensing method using the dispensing device 300 according to the third embodiment is almost the same as that of the first embodiment (FIGS. 2 and 3), the description thereof will be omitted.

<Technical effect>
As described above, the dispensing device 300 according to the third embodiment adopts a configuration in which the detection result by the fluorescence detecting device 30 provided in the chip 1 is wirelessly transmitted to the main body or an external device. .. As a result, as compared with the dispensing device 100 of the first embodiment, it is not necessary to connect the fluorescence detection device 30 in the chip 1 and the main body 2 with a cable, which facilitates assembly.

[Fourth Embodiment]
In the first to third embodiments, the pipette type dispensing devices 100 to 300 operated by the user have been described. The dispensing devices 100 to 300 of each embodiment can also be mounted on a device that automatically dispenses and analyzes a liquid. Therefore, in the present embodiment, we propose an automatic dispensing device that automatically performs the dispensing operation. Here, the automatic dispensing device of the present embodiment can be incorporated into an analyzer such as a preprocessing automation device of a next-generation sequencer, for example. The automatic dispensing device incorporated in such a pretreatment automation device can be used, for example, for quality confirmation (measurement of nucleic acid concentration) of a nucleic acid solution having an unknown concentration prepared by the pretreatment automation device.

<Configuration of automatic analyzer>
FIG. 6 is a schematic view showing the automatic dispensing device 400 according to the fourth embodiment. As shown in FIG. 6, the automatic dispensing device 400 includes a dispensing device 300, a moving mechanism 401, a control unit 402, a display unit 403, and a reaction vessel 102 containing the liquid to be detected 101, which are the same as those in the third embodiment. An empty container 103 for discharging the liquid to be detected 101 is provided. Instead of the dispensing device 300, the dispensing device 100 of the first embodiment or the dispensing device 200 of the second embodiment may be used.

The moving mechanism 401 has, for example, a holding portion for holding the dispensing device 300 and an actuator for moving the holding portion in the horizontal and vertical directions, and moves the dispensing device 300 in the horizontal and vertical directions. In the moving mechanism 401 shown in FIG. 6, a rail for moving the dispensing device 300 in the vertical direction and the left-right direction on the paper surface is shown, but a rail for moving the dispensing device in the front-rear direction is further provided. May be.

The control unit 402 controls the drive of the moving mechanism 401. Further, the control unit 402 instructs the start of the suction operation and the discharge operation by the dispensing device 300. Further, the control unit 402 may execute the calculation process of the calculation unit 21 (not shown in FIG. 6) described in the first embodiment (FIGS. 2 and 3).

The display unit 403 displays the data processed by the calculation unit 21 or the control unit 402 of the dispensing device 300.

<Dispensing method>
The dispensing method using the automatic dispensing device 400 of the present embodiment will be outlined. Since the basic procedure for measuring the nucleic acid concentration and determining the discharge amount is the same as that in the first embodiment (FIGS. 2 and 3), description thereof will be omitted here.

First, the control unit 402 drives the moving mechanism 401 to immerse the tip of the chip 1 of the dispensing device 300 in the reaction vessel 102 containing the liquid to be detected 101, and instructs the dispensing device 300 to start the suction operation. To send. When the instruction to start the suction operation is input, the calculation unit 21 of the dispensing device 300 drives the plunger to suck the liquid to be detected 101 into the chip 1. When the suction of the liquid to be detected 101 is completed, the control unit 402 drives the moving mechanism 401 to pull up the dispensing device 300. After that, the calculation unit 21 of the dispensing device 300 causes the fluorescence detection device 30 to irradiate the light and detect the light from the liquid to be detected 101. The fluorescence detection device 30 wirelessly transmits a detection result (digital signal) to the calculation unit 21 of the main body unit 2. At this time, the fluorescence detection device 30 may transmit a detection result (digital signal) to the control unit 402.

The calculation unit 21 or the control unit 402 calculates the concentration of nucleic acid in the liquid to be detected 101 and determines the discharge amount of the liquid to be detected 101. After that, the control unit 402 drives the moving mechanism 401 to move the dispensing device 300 so that the tip portion of the tip 1 is located inside the container 103, and then starts the dispensing operation to the dispensing device 300. Send instructions. When the instruction to start the discharge operation is input, the calculation unit 21 drives the plunger to discharge the liquid to be detected 101 into the container 103 according to the determined discharge amount.

Since the amount of the liquid to be detected 101 discharged into the container 103 and the concentration of nucleic acid have been calculated, it can be used for subsequent processing in an analyzer such as a pretreatment automation device. For example, the analyzer may be configured to further supply the required amount of diluent to the container 103.

<Technical effect>
As described above, the automatic dispensing device according to the fourth embodiment includes the dispensing device 300 provided with the fluorescence detection device 30 in the chip 1, and automatically executes the dispensing operation by the dispensing device 300. .. As a result, the concentration of nucleic acid in the liquid to be detected 101 prepared by the analyzer can be measured simply by incorporating the automatic dispensing device into the analyzer, so that the size of the analyzer can be suppressed. Since such a small analyzer can be installed in one laboratory (for example, a place where a sample is collected such as a hospital), the pretreatment of the nucleic acid sequencer can be rapidly performed on the spot. Further, since the concentration of nucleic acid can be calculated and the required amount can be discharged only by one dispensing operation, an increase in processing time is suppressed.

[Modification example]
<Modification example 1>
In the first to fourth embodiments, it has been described that the light emitting element 14 of the fluorescence detection device provided in the chip 1 irradiates the liquid to be detected 101 with light. Alternatively, the light emitting element 14 may irradiate light from the outside of the chip 1. For example, when the automatic dispensing device 400 of the fourth embodiment is provided with a light source unit for irradiating light from below or laterally of the dispensing device 300, the nucleic acid concentration in the liquid to be detected 101 is measured. The dispensing device 300 is moved above or laterally to the light source unit to irradiate the chip 1 with light.

By adopting such a configuration, the fluorescence detection device can be downsized by the amount of the light emitting element 14 and the light source drive circuit 18. As a result, the fluorescence detection device can be attached to the chip 1 having a smaller capacity, so that the concentration of nucleic acid in the liquid to be detected 101 can be accurately measured even when the amount of the liquid to be detected 101 is very small. be able to.

<Modification 2>
In the above, it has been described that a fluorescent dye is bound to the nucleic acid in the liquid to be detected 101, and the concentration of the nucleic acid is measured by detecting the fluorescence. However, the target for measuring the concentration is not limited to nucleic acid, and the light to be detected is not limited to fluorescence. For example, a liquid containing an analysis target and a dye that can be bound to the analysis target may be used as the liquid to be detected 101, and the wavelength of the color emitted by the dye may be detected.

<Others>
The present disclosure is not limited to the embodiments described above, but includes various modifications. For example, the above-described embodiment has been described in detail in order to explain the present disclosure in an easy-to-understand manner, and does not necessarily have all the configurations described. In addition, a part of one embodiment can be replaced with the configuration of another embodiment. It is also possible to add the configuration of another embodiment to the configuration of one embodiment. It is also possible to add, delete, or replace a part of the configuration of another embodiment with respect to a part of the configuration of each embodiment.

100, 200, 300 ... Dispensing device 400 ... Automatic dispensing device 101 ... Liquid 102 to be detected ... Reaction container 103 ... Container 1 ... Chip 2 ... Main body 3 ...

Cable

4, 6 ... Support 5 ...

Vents

10, 20 , 30 ... Fluorescence detection device 11 ... Substrate 12 ... Light receiving element 13 ... Fluorescent filter 14 ... Light emitting element 15 ... Convex lens 16 ... Current / voltage conversion circuit 17 ... AD conversion circuit 18 ... Light source drive circuit 21 ... Calculation unit 22 ... Power supply unit 23 ... Switch 24 ... Receiver 25 ... Transmitter 31 ... Wireless communication circuit 32 ... Battery

Claims

A detector that detects light from an analysis target in a liquid.
A light source that irradiates the liquid with light,
A detection unit for detecting light from the analysis target in the liquid is provided.
A detection device characterized in that at least the detection unit is arranged in a dispensing chip to which the liquid is sucked.
The detection device according to claim 1.
A detection device characterized in that the light source is arranged in the dispensing chip.
The detection device according to claim 1.
A detection device further comprising a wireless communication unit that wirelessly transmits the light detection result to the outside of the detection device.
The detection device according to claim 1.
A detection device further comprising a condenser lens arranged after the light source.
The detection device according to claim 1.
A detection device that is arranged in front of the detection unit and further includes a filter that transmits light of a specific wavelength.
The detection device according to claim 1.
A substrate having at least the detection unit and further provided in the dispensing chip.
A detection device characterized in that the substrate is provided with a vent.
A dispensing device that dispenses the liquid containing the analysis target.
The detection device according to claim 1 and
With the dispensing tip
A support that supports the detection device in the dispensing chip, and
The main body to which the dispensing tip is mounted and having a dispensing mechanism,
A calculation unit that receives the light detection result from the detection device and performs a calculation is provided.
The calculation unit is a dispensing device, which calculates the concentration of the analysis target in the liquid based on the detection result.
The dispensing device according to claim 7.
A dispensing device, wherein the calculation unit determines a discharge amount of the liquid based on the concentration of the analysis target.
The dispensing device according to claim 8.
A dispensing device, wherein the calculation unit determines a discharge amount of the liquid based on the concentration and an upper limit suction amount or a lower limit discharge amount of the dispensing device.
The dispensing device according to claim 7.
A dispensing device in which the calculation unit is arranged in the main body unit.
The dispensing device according to claim 7.
A dispensing device, characterized in that the support supports the detection device so as to form a vent in the dispensing chip.
The dispensing device according to claim 7.
The detection device further includes a wireless communication unit that wirelessly transmits the detection result to a computer terminal outside the detection device.
A dispensing device characterized in that the computer terminal executes the processing of the arithmetic unit.
The dispensing device according to claim 7.
A moving mechanism for moving the dispensing device and
A dispensing device further comprising a control unit for controlling the drive of the moving mechanism.
A method for dispensing a liquid using the dispensing device according to claim 7.
Placing the detection device in the dispensing chip and
By sucking the liquid into the dispensing tip by the operation of the dispensing mechanism,
Irradiating the liquid with light by the light source
The detection unit detects the light from the liquid and
A dispensing method comprising calculating the concentration of the analysis target in the liquid by the calculation unit based on the detection result of the light of the detection unit.
The dispensing method according to claim 14.
A dispensing method characterized in that the calculation unit determines the discharge amount of the liquid based on the concentration of the analysis target.