TWM597295U - Nucleic acid analysis apparatus - Google Patents

Abstract

The nucleic acid analysis device includes a main frame, a fluid delivery unit, a first temperature control unit, a second temperature control unit, an optical unit, a rotary drive unit, and a linear drive unit. The main frame body has a cavity to install a cassette, and the cassette includes a plurality of detection slots. The fluid delivery unit performs nucleic acid extraction and nucleic acid amplification, and has a fluid adapter. The first temperature control unit provides a first temperature for nucleic acid amplification. The second temperature control unit provides a second temperature higher than the first temperature for nucleic acid denaturation before nucleic acid amplification. The optical unit performs nucleic acid detection. The rotation drive unit rotates the cassette during nucleic acid detection to align the detection slot with the optical unit. The linear drive unit lifts the fluid adapter and the second temperature control unit upward during nucleic acid extraction to closely contact the bottom surface of the cassette.

Description

Nucleic acid analysis device

This case relates to a nucleic acid analysis device, especially an all-in-one nucleic acid analysis device.

The Point of Care (POC) test is an analysis method performed outside of a central medical institution or laboratory, and is performed using a device that can interpret the results in real time. Globalization trends have caused outbreaks of new or recurring infectious diseases, and accelerated their speed from epidemic to pandemic (epidemic-to-pandemic), and in the face of such threats, decentralization of clinical treatment at the frontline Diagnostic tests will help to implement the response to public health events early to reduce the severity of the event. In developing countries, due to the lack of clinical laboratory infrastructure and funding constraints, the diagnosis is not easy to increase the burden of highly infectious diseases, and the use of POC tests can help improve this situation.

In recent years, some POC devices have been developed for isothermal molecular diagnosis of nucleic acid amplification. The disposable cartridges used in these devices are installed in the device for fluid processing, and then amplified and optically detected. However, current devices only provide a single heating zone for amplification and detection (eg, 60-65°C), lacking a higher heating temperature zone (eg, 95°C), and failing to make nucleic acids more efficiently after nucleic acid extraction Denaturation to generate single-stranded nucleic acids for amplification. Therefore, in order to improve the lack of the prior art, it is necessary to provide an improved POC device.

One of the purposes of this case is to provide a fully functional integrated nucleic acid analysis device, so that sample purification, nucleic acid extraction, nucleic acid amplification, and nucleic acid detection processes can be performed on this fully functional integrated device to achieve real-time nucleic acid analysis.

Another object of this case is to provide a nucleic acid analysis device having a second temperature control unit that can provide a higher heating temperature zone to denature nucleic acids, thereby improving nucleic acid amplification efficiency.

Another object of the present case is to provide a nucleic acid analysis device having a linear drive unit, which can ensure non-leakage fluid delivery and reliable high-temperature heating.

To achieve the above purpose, the present case provides a nucleic acid analysis device, which includes a housing, a main frame, a fluid delivery unit, a first temperature control unit, a second temperature control unit, at least one optical unit, and a rotation drive unit And a linear drive unit. The casing includes an upper casing and a lower casing. The main frame is arranged in the lower casing, and has a chamber for installing a cassette therein, the cassette includes a plurality of detection slots. The fluid delivery unit is disposed in the lower housing and is constructed on the sample and/or reagent in the delivery cassette for nucleic acid extraction and/or nucleic acid amplification. The fluid delivery unit has a fluid adapter and the fluid is mated The device is arranged below the main frame. The first temperature control unit is disposed on the main frame and is configured to provide a first temperature for nucleic acid amplification. The second temperature control unit is disposed below the main frame and is configured to provide a second temperature higher than the first temperature before nucleic acid amplification to denature the nucleic acid. The optical unit is disposed on the main frame and includes a plurality of optical elements for nucleic acid detection. The rotation driving unit is disposed in the lower casing and is configured to rotate the cassette according to a preset program during nucleic acid detection, so as to align the detection slot of the cassette with the optical unit. The linear driving unit is arranged in the lower casing and is constructed to lift up the fluid adapter and the second temperature control unit during nucleic acid extraction, so that the bottom surface of one of the fluid adapter, the second temperature control unit and the cassette Close contact.

In one embodiment, during nucleic acid amplification and detection, the linear drive unit drives the fluid adapter and the second temperature control unit to move downward to maintain a gap with the bottom surface of the cassette.

In one embodiment, after the nucleic acid detection is completed, the linear drive unit drives the fluid adapter and the second temperature control unit to move upward, and pushes the cassette upward so that the top surface of the cassette is significantly higher than the top surface of the main frame To remove the Eli Cartridge.

In an embodiment, the linear drive unit includes a stepping motor, a screw, a slider, and a cantilever, wherein the slider is connected to the stepping motor through the screw, the cantilever is fixed to the slider, and the fluid adapter and The second temperature control unit is carried on the cantilever.

In an embodiment, the linear driving unit includes at least one position sensing unit.

In an embodiment, the upper housing includes a plurality of blocking portions. When the cassette is pushed upward by the linear driving unit, the top surface of the cassette will abut against the blocking portion.

In one embodiment, the cassette includes at least one guide groove, and the rotary drive unit includes at least one protrusion, the protrusion can slide in the guide groove, and the rotary drive unit clamps the cassette through the protrusion to drive the rotation of the cassette .

In one embodiment, the rotary driving unit includes a stepping motor, a rotating shaft, a driving gear, and a driven gear, wherein the driving gear is connected to the stepping motor through the rotating shaft, and the driving gear meshes with the driven gear.

In one embodiment, the protrusion is provided on the driven gear.

In one embodiment, the optical unit includes a light source and a light detector.

In one embodiment, the nucleic acid analysis device includes a plurality of optical units, each of which provides light of a unique wavelength to detect multiple targets.

In one embodiment, the cassette includes a cassette body, a reaction wafer, and a heat-treated wafer. The reaction wafer is disposed on the top of the cassette body, and the heat-treated wafer is disposed on the bottom of the cassette body.

In one embodiment, the first temperature control unit surrounds the cassette body and carries the reaction wafer thereon.

In one embodiment, the second temperature control unit performs contact heating on the heat-treated wafer.

In one embodiment, the bottom surface of the heat-treated wafer includes a plurality of openings, and the top surface of the fluid adapter includes a plurality of openings, which can be correspondingly connected to the plurality of openings of the heat-treated wafer to form a fluid delivery pipe.

In an embodiment, the reaction wafer includes at least two fluid networks, wherein each fluid network includes a plurality of detection tanks, a main fluid flow channel, and a gas release flow channel, the main fluid flow channel is connected to the detection tank and is constructed in Distribute the sample or control liquid into the detection tank. The gas release flow channel is connected to the detection tank and is configured to release gas from the detection tank. One of the structures of the fluid network is used for quality control.

In one embodiment, the gas release channel is significantly narrower than the main fluid channel.

In one embodiment, the shape of the reaction wafer is substantially a regular polygon, and each detection slot has at least one plane.

In one embodiment, the reaction wafer further includes at least one sample loading hole provided on the top surface of the reaction wafer for adding the sample to the cassette.

In one embodiment, the nucleic acid analysis device further includes an embedded lens for scanning a bar code or a QR code on a sample tube or cartridge to obtain relevant information.

Some embodiments embodying the characteristics and advantages of this case will be described in detail in the following paragraphs. It should be understood that this case can have various changes in different forms, and all of them do not deviate from the scope of this case, and the descriptions and drawings therein are essentially for illustration purposes, rather than to limit this case.

This case provides an all-in-one nucleic acid analysis device, which integrates a fluid delivery unit, a temperature control unit, a driving unit, and an optical unit on a single device to enable sample purification, nucleic acid extraction, and nucleic acid expansion Processes such as augmentation and nucleic acid detection can be performed on this fully functional integrated device to achieve real-time nucleic acid analysis.

FIG. 1 shows a schematic diagram of a nucleic acid analysis device according to an embodiment of the present invention, in which the nucleic acid analysis device is turned on. Figure 2 shows a schematic diagram of the main internal structure of the nucleic acid analysis device, in which the connection of wires and fittings is not shown, to more clearly show the internal structure of the nucleic acid analysis device. Figure 3 shows an exploded view of the nucleic acid analysis device of Figure 2. Figures 4 and 5 show schematic diagrams of cassettes at different angles. The nucleic acid analysis device in this case includes a housing 1, a main frame 2, a fluid delivery unit 3, a first temperature control unit 4, a second temperature control unit 5, at least one optical unit 6, and a rotary drive unit 7和和一个一个线驱动单元8。 And a linear drive unit 8. The housing 1 includes an upper housing 11 and a lower housing 12, and the main housing 2, the fluid delivery unit 3, the first temperature control unit 4, the second temperature control unit 5, the part of the optical unit 6, the rotation drive unit 7 Both the linear drive unit 8 and the linear drive unit 8 are disposed in the lower housing 12.

The main frame 2 has a chamber 21 specially designed for installing a cassette 9 therein, wherein the cassette 9 includes a plurality of detection slots 921. The fluid delivery unit 3 is configured to deliver samples and/or reagents in the cassette 9 for nucleic acid extraction and/or nucleic acid amplification. The fluid delivery unit 3 has a fluid adapter 31, and the fluid adapter 31 is disposed at Below the main frame 2. The first temperature control unit 4 is disposed on the main frame 2 and is configured to provide a first temperature for nucleic acid amplification. The second temperature control unit 5 is disposed below the main frame 2 and is configured to provide a second temperature higher than the first temperature before nucleic acid amplification to denature the nucleic acid. The optical unit 6 is provided on the main frame 2 and includes a plurality of optical elements for nucleic acid detection. The rotation driving unit 7 is disposed in the lower casing 12 and is configured to rotate the cassette 9 according to a preset program during nucleic acid detection, so that the detection slot 921 of the cassette 9 is aligned with the optical unit 6. The linear drive unit 8 is disposed in the lower housing 12 and is configured to lift the fluid adapter 31 and the second temperature control unit 5 upward during nucleic acid extraction, so that the fluid adapter 31 and the second temperature control unit 5 are One of the bottom surfaces of the cassette 9 is in close contact.

In one embodiment, the nucleic acid analysis device further includes a controller, such as a micro control unit (MCU), to control the fluid delivery unit 3, the first temperature control unit 4, the second temperature control unit 5, and the optical unit 6. Operation of rotary drive unit 7 and linear drive unit 8.

In one embodiment, the upper casing 11 and the lower casing 12 are connected by a hinge, but not limited to this. The upper casing 11 can be opened so that the cassette 9 can be placed in the chamber 21 of the main frame 2. When the upper casing 11 is closed, a closed space is formed in the casing 1.

As shown in FIGS. 4 and 5, the cassette 9 includes a cassette body 91, a reaction wafer 92, and a heat treatment wafer 93, wherein the reaction wafer 92 is disposed on top of the cassette body 91, and the heat treatment wafer 93 is disposed At the bottom of the cassette body 91. The reaction wafer 92 can be used for at least one of nucleic acid amplification reagent storage, reaction solution processing, nucleic acid amplification, and detection. The cartridge body 91 is also called an extraction wafer, and can be used for at least one of sample storage, sample preparation reagent storage, sample cell lysis, nucleic acid extraction, and purification. The heat-treated wafer 93 can be used for nucleic acid denaturation.

In one embodiment, the reaction wafer 92 is a planar fluid wafer, including at least two fluid networks 9A and 9B, which are configured to perform nucleic acid amplification and detection, wherein one of the fluid networks 9A and 9B is used to construct Quality control. For example, the longer detection fluid network 9A is a sample fluid network for detecting nucleic acids extracted from the sample, and the shorter detection fluid network 9B is a control fluid network for detecting positive control fluid and negative control fluid . In some embodiments, each of the fluid networks 9A and 9B includes a plurality of detection tanks 921, a main fluid flow channel 922, and at least one gas release flow channel 923, wherein the detection tank 921 accommodates a sample or a control liquid, The main fluid flow channel 922 is connected to the detection tank 921 and is configured to distribute the sample or control liquid into the detection tank 921, and the gas release flow channel 923 is connected to the detection tank 921 and is configured to release gas from the detection tank 921, and the gas The release channel 923 is significantly narrower than the main fluid channel 922.

In one embodiment, the detection tank 921 contains reagents for nucleic acid amplification and detection. For example, the detection tank 921 may be coated with reagents for nucleic acid amplification and detection, such as reagents including different fluorescent dyes. The number of detection slots 921 is not limited, and can be as many as 40 or more, so the device of the present invention can perform multiplexing nucleic acid analysis. In an embodiment, the shape of the reaction wafer 92 is substantially a regular polygon, so that the reaction wafer 92 has a plurality of flat sides, which can be arranged in a line with the optical unit 6 to facilitate light focusing. Of course, the shape of the reaction wafer 92 is not limited to a regular polygon, it may be a circle or other shapes, because the arrangement of the optical elements through the optical unit 6 can also focus the light on the sample in the detection tank 921. In one embodiment, each detection slot 921 has at least one plane. For example, the shape of the detection slot 921 may be rectangular, and during the nucleic acid detection process, the detection slot 921 has a plane aligned with the photodetector 62 of the optical unit 6 in a straight line.

In one embodiment, the cartridge body 91 includes a plurality of tanks that store reagents for sample purification and nucleic acid extraction. The cassette body 91 also includes a plurality of flow channels, which are connected to the tank for fluid delivery. In an embodiment, the cassette body 91 may be, but not limited to, a cylindrical body.

In one embodiment, the reaction wafer 92 further includes at least one sample loading hole 924, which is opened on the top surface of the reaction wafer 92, and the sample loading hole 924 is aligned and communicated with at least one slot of the cassette body 91, It is used to add a sample to the cassette 9 and perform sample purification and nucleic acid extraction in the cassette body 91.

In some embodiments, the reaction wafer 92 further includes a plurality of grooves 925 and flow channels, which can be used to perform fluid functions of various fluid processing processes, including but not limited to metering, mixing, debubbling, And at least one of dispensing.

In one embodiment, the heat-treated wafer 93 includes a plurality of openings 931 and a heating tank 932 provided on the bottom surface of the heat-treated wafer 93.

In one embodiment, the cassette 9 further includes at least one guide groove 94 disposed on the outer surface of the cassette body 91 and the heat-treated wafer 93, which is a vertical groove. The guide groove 94 can work together with the rotary driving unit 7 to control the movement of the cassette 9.

Figure 6 shows a schematic diagram of a part of the structure of the nucleic acid analysis device. The fluid adapter 31 includes a plurality of openings 311 and a plurality of joints 312. A plurality of openings 311 are provided on the top surface of the fluid adapter 31 and corresponding to the plurality of openings 931 connected to the bottom surface of the heat-treated wafer 93 to form a pipeline for fluid transportation. A plurality of connectors 312 are provided on the side of the fluid adapter 31, and are connected to the fluid delivery unit 3 through a plurality of hoses (not shown). During operation, once the sample is loaded into the cassette 9, the sample loading hole 924 is sealed, and the cassette 9 is placed in the chamber 21 of the nucleic acid analysis device, the linear drive unit 8 drives the fluid delivery unit 3 upward So that the cassette 9 is in close contact with the fluid adapter 31 of the fluid delivery unit 3 to avoid leakage. Next, the fluid transport unit 3 performs fluid processing. The fluid delivery unit 3 operates simultaneously with the cassette 9 to perform sample purification, nucleic acid extraction, and fluid delivery, thereby achieving a fully automatic device. Fluid delivery can be via pneumatic, vacuum, plunger, chamber deformation, thermal-induced expansion, acoustics, centrifugal force or other available The method of completing sample processing in the cassette 9 is implemented.

In one embodiment, the flow system is driven pneumatically via micro-pipes and holes. For example, the fluid delivery unit 3 is similar to the fluid integration module described in Taiwan Patent Application No. 105123156 (Patent Announcement No. I641823) filed by the applicant on July 22, 2016, and the entire contents of the aforementioned application are combined in This is for disclosure and will not be repeated here. Briefly, the fluid delivery unit 3 of the present embodiment includes the fluid manifold part, rotary valve stator, rotary valve rotor, rotary valve housing, and fluid source (such as a pump) described in Taiwan Patent Publication No. I641823. The fluid manifold portion has a plurality of micro-pipes, and is connected to the groove in the cassette 9 through the plurality of hoses, the fluid adapter 31, and the opening 931 at the bottom of the cassette 9. Since the perforations and/or grooves on the rotary valve stator and rotary valve rotor have a corresponding alignment relationship when the rotary valve rotor rotates, when the rotary valve rotor rotates to a different position, multiple fluid paths can be switched, and further Regulate the fluid operation of cassette 9 Therefore, the samples or reagents stored in the cassette 9 can be transported to the position to be transported by the pneumatic force provided by the pump of the fluid transport unit 3, and then the processes of sample purification, nucleic acid extraction, and nucleic acid amplification can be automatically performed. . Of course, the fluid conveying unit is not limited to the above-mentioned design. Any other type of fluid conveying unit, as long as it can achieve multiple fluid conveying and path switching functions in the cassette 9, will not deviate from the scope disclosed in this case.

Fig. 7 shows a schematic diagram of a part of the structure of the nucleic acid analysis device, and Fig. 8 shows the structure shown in Fig. 7 and the cassette mounted thereon. The first temperature control unit 4 is disposed on the main frame 2 and includes a heater 41 and a radiator 42. In one embodiment, the heater 41 includes a heating element 411 with a temperature control algorithm, and the heat sink 42 includes a plurality of fins, which are integrated on the heater 41 and arranged in a ring shape to accelerate heat energy distribution. The heater 41 has a central hole, and the diameter of the central hole is slightly larger than the diameter of the cassette body 91 of the cassette 9 so that the cassette 9 can be easily installed therein. When the cassette 9 is installed in the chamber 21, the first temperature control unit 4 carries the reaction wafer 92 thereon, and the reaction wafer 92 contacts the heater 41 and is surrounded by the heat sink 42.

In one embodiment, the nucleic acid analysis device in this case is designed for nucleic acid amplification performed in an isothermal manner, so only a single heating interval (for example, 60-65°C) is required in the nucleic acid amplification stage, and no multiple The thermal cycle control in different temperature ranges, therefore, the first temperature control unit 4 can be significantly simplified, and thus the nucleic acid analysis device can be designed in a compact, portable, and suitable for POC diagnosis. In addition, the first temperature control unit 4 further includes a thermal insulation layer 43, which can provide good thermal insulation, so that the temperature inside the chamber 21 of the nucleic acid analysis device can be easily maintained. Once the chamber 21 is in a uniform temperature environment, the heat loss from the detection tank 921 and the sample to the environment can be minimized, and during the nucleic acid amplification and detection process, regardless of whether the cassette 9 is rotating or stationary, the entire The samples in the closed chamber 21 and each detection tank 921 have the same temperature.

The first temperature control unit 4 provides the temperature required inside the chamber 21 during the operation, wherein the temperature control is not affected by the number and shape of the detection grooves 921. In an embodiment, the first temperature control unit 4 further includes at least one temperature sensor for controlling the temperature accuracy.

In an embodiment, the nucleic acid analysis device includes a plurality of optical units 6. The optical unit 6 includes optical elements such as a light source, a lens, a filter, and a light detector to realize optical detection, so that the sample can be detected in real time during nucleic acid amplification. As shown in FIG. 1, the optical unit 6 may include a light source 61 and a light detector 62. In an embodiment, the light source 61 may be, but not limited to, a light emitting diode (LED), and the light detector 62 may be, but not limited to, a photodiode. During the operation, each light source 61 will be aligned with one of the detection slots 921 to provide an effective light source for detection, and each light detector 62 will be aligned with the detection slot 921 for detection and acquisition Analyze the results. The rotation of the cassette 9 allows each detection slot 921 to pass through different optical units 6 in sequence. In one embodiment, each optical unit 6 can provide a unique wavelength of light, thereby providing different colors of light for fluorescent detection, so that the nucleic acid analysis device can simultaneously detect multiple targets and achieve multiplexing detection (multiplexing detection) ).

The rotary drive unit 7 can be actuated by a motor through a gear, belt, chain, rack, worm, or other mechanical transmission mechanism, and can drive the rotation of the cassette 9 as required. Figure 9 shows a schematic diagram of the rotary drive unit of the embodiment of the present invention. In this embodiment, the rotation driving unit 7 is a gear transmission mechanism, including a stepping motor 71, a rotating shaft 72, a driving gear 73, a driven gear 74, and a rotation sensing unit 75. The driven gear 74 has a central hole, and the diameter of the central hole is slightly larger than the diameter of the cassette body 91, so that the cassette 9 can be easily installed therein, and the driven gear 74 further includes at least one protrusion 741, such as a positioning pin or other A similar structure can be correspondingly inserted into the guide groove 94 on the cassette 9 to clamp and control the movement of the cassette 9.

Figure 10 shows a partial schematic diagram of a nucleic acid analysis device. As shown in FIGS. 9 and 10, the driving gear 73 is connected to the stepping motor 71 through the rotating shaft 72, and the driving gear 73 meshes with the driven gear 74, so when the stepping motor 71 drives the driving gear 73 to rotate through the rotating shaft 72 , The driven gear 74 and the cassette 9 clamped by the driven gear 74 can be driven to rotate at the same time, and then the cassette 9 is controlled to rotate according to a predetermined program, so that the detection slot 921 and each optical unit 6 are sequentially aligned to Nucleic acid detection is performed, and the rotation sensing unit 75 can monitor whether the detection slot 921 is aligned with the optical unit 6. In one embodiment, the rotary drive unit 7 further includes a bearing 76 disposed inside the driven gear 74, which can limit the absolute position of the driven gear 74 to achieve a smoother and more reliable rotation.

Figure 11 shows a partial cross-sectional view of the cassette when installation is complete. According to the design of this case, the guide groove 94 of the cassette 9 has a certain length. When the cassette 9 is installed in the chamber 21 by the user, the guide groove 94 cooperates with the protrusion 741 of the driven gear 74 to make the cassette 9 You can slide down to the end position, at this time, the detection slot 921 of the cassette 9 and the photodetector 62 of the optical unit 6 are perfectly located on the same horizontal plane, and the horizontal axis of the two is controlled by the rotary drive unit 7 Can be accurately aligned. Therefore, the design of this case defines the absolute initial position of the cassette 9 when it is installed, which is beneficial to the initial optical correction and background signal measurement. The design of this case also defines the absolute terminal position of the cassette 9 to maintain the consistency of rotation and alignment during nucleic acid amplification and detection.

In one embodiment, the cassette 9 includes three guide grooves 94, and accordingly, the driven gear 74 also includes three protrusions 741. Of course, the number of guide grooves 94 and protrusions 741 is not limited to three.

Figure 12 shows a schematic diagram of a part of the structure of a nucleic acid analysis device. The second temperature control unit 5 includes a heater 51, a radiator 52 and a fan 53. The heater 51 includes a heating element with a temperature control algorithm, the radiator 52 includes a plurality of fins and is installed below the heater 51, and the fan 53 is installed below the radiator 52. The top surface of the heater 51 may be in contact with the bottom surface of the cassette 9 so that the heating groove 932 on the bottom surface of the heat-treated wafer 93 is in contact with the heater 51 to perform contact heating. The purpose of the setting of the second temperature control unit 5 is to provide a higher heating temperature zone (for example, 95°C) to heat-treat the nucleic acid extracted from the sample to denature the nucleic acid to produce single-stranded nucleic acid to improve the subsequent The efficiency of nucleic acid amplification. Therefore, after the nucleic acid extraction is completed in the cassette body 91, the sample with the extracted nucleic acid will be transported downward to the heat-treated wafer 93 for denaturation, and then transported upward to the reaction wafer 92 for nucleic acid amplification and detection. After the nucleic acid is denatured, the radiator 52 and the fan 53 can first cool the sample to reduce the temperature to the temperature required for isothermal amplification (for example, 60-65°C), and then transport the sample upward.

In the nucleic acid extraction stage, the bottom surface of the cassette 9 must be in close contact with the fluid adapter 31 of the fluid delivery unit 3 to achieve a sealed state for fluid treatment and avoid leakage, and in close contact with the second temperature control unit 5 for heat treatment. In the nucleic acid amplification and detection stage, the cassette 9 must be rotated so that each detection slot 921 is sequentially aligned with a different optical unit 6, and in order to reduce the friction force when the cassette 9 rotates, the cassette 9 rotates smoothly The bottom surface of the cassette 9 must have a gap with the fluid adapter 31 and the second temperature control unit 5. Therefore, in addition to the rotation drive unit 7 to control the rotation and alignment of the cassette 9, the nucleic acid analysis device of the present case is further provided with a linear drive unit 8 to control the fluid adapter 31 and the second temperature control unit 5 in the vertical direction The movement, in turn, meets the sealing requirements of the nucleic acid extraction stage and the rotation requirements of the nucleic acid amplification and detection stage, respectively.

Please refer to Figure 6 and Figure 12. In an embodiment, the linear driving unit 8 includes a stepping motor 81, a screw 82, a slider 83, and a cantilever 84, wherein the slider 83 is connected to the stepping motor 81 through the screw 82, and the cantilever 84 is fixed to Slider 83. Since the fluid adapter 31 and the second temperature control unit 5 of the fluid delivery unit 3 are carried on the cantilever 84, when the stepper motor 81 controls the slider 83 to slide up and down via the screw 82, the fluid on the cantilever 84 can be driven The movement of the adapter 31 and the second temperature control unit 5 in the vertical direction. Therefore, in the nucleic acid extraction stage, the linear driving unit 8 can drive the fluid adapter 31 and the second temperature control unit 5 to move upward and cling to the bottom surface of the cassette 9 to perform nucleic acid extraction by the fluid delivery unit 3, and extract the nucleic acid Then, the second temperature control unit 5 performs heat treatment to denature the nucleic acid. In the nucleic acid amplification and detection stage, the linear drive unit 8 can drive the fluid adapter 31 and the second temperature control unit 5 to move downward to maintain a gap with the bottom surface of the cassette 9, and then the rotary drive unit 7 drives the cassette The rotation of 9 causes each detection slot 921 to be sequentially aligned with a different optical unit 6 for detection.

In an embodiment, after the nucleic acid detection is completed, the linear driving unit 8 can further drive the fluid adapter 31 and the second temperature control unit 5 to move upward and cling to the bottom surface of the cassette 9 and further push the cassette 9 upward The top surface of the cassette 9 is significantly higher than the top surface of the main frame 2 to facilitate the user to take out the cassette 9.

In one embodiment, the linear driving unit 8 further includes at least one position sensing unit 85, which can monitor the position of the cantilever 84 and accurately control the fluid adapter 31 and the second temperature control by a preset logic control algorithm Linear movement of unit 5. For example, the linear drive unit 8 includes three sets of position sensing units 85, which can monitor the three positions of the fluid adapter 31 and the second temperature control unit 5, including the initial position from low to high, and close contact with the cassette 9 And the position where the cassette 9 is ejected. Or, the three groups of position sensing units 85 can be monitored by logical arrangement and combination, and up to 8 (2 ³ ) different positions can be controlled to realize more functional position control.

In an embodiment, the nucleic acid analysis device further includes a pressure sensor. When the fluid adapter 31 and the second temperature control unit 5 reach a position in close contact with the cassette 9, the fluid inside the cassette 9 can be monitored in real time The pressure further adjusts the positions of the fluid adapter 31 and the second temperature control unit 5 more accurately, ensuring non-leaking fluid delivery and reliable high-temperature heating.

In an embodiment, the upper housing 11 of the nucleic acid analysis device includes a plurality of stop portions 13 such as check-valve plungers, and a spring 131 is provided inside. When the cassette 9 is pushed upward by the linear drive unit 8, it will abut against the stop portion 13 of the upper housing 11, so the stop portion 13 can limit the maximum lifting height of the linear drive unit 8, and can absorb the action on the upper housing 11 pressure.

Figures 13A to 13F show the workflow of the nucleic acid analysis device for full-function integration detection. For ease of explanation, only part of the structure of the main components is shown in the figure. First, the user adds the sample to the cassette 9 and installs the cassette in the chamber 21. At this time, the guide groove 94 of the cassette 9 is aligned with the protrusion 741 of the driven gear 74, so that the cassette 9 can slide downward (as shown in FIG. 13A). When the cassette 9 slides down to the end position, that is, the protrusion 741 reaches the top of the guide groove 94, then the detection slot 921 of the cassette 9 and the photodetector 62 of the optical unit 6 are perfectly located on the same horizontal plane. Next, the upper casing 11 of the nucleic acid analysis device is closed, and the cassette 9 is clamped on a predetermined horizontal surface by the protrusion 741 and the stopper 13 (as shown in FIG. 13B ).

When the upper casing 11 of the nucleic acid analysis device is closed, the linear drive unit 8 starts to move, so that the fluid adapter 31 and the second temperature control unit 5 move upward and cling to the bottom surface of the cassette 9 so that the fluid delivery unit 3 Nucleic acid extraction is performed, and after the nucleic acid extraction, heat treatment is performed by the second temperature control unit 5 to denature the nucleic acid (as shown in FIG. 13C). The position sensing unit 85 can simultaneously monitor the positions of the fluid adapter 31 and the second temperature control unit 5 to ensure non-leakage fluid delivery and reliable high-temperature heating.

When all the fluid processing and heating/cooling processes are completed according to a predetermined program, the linear drive unit 8 will drive the fluid adapter 31 and the second temperature control unit 5 to move down to separate from the cassette 9 (as shown in FIG. 13D ). At this time, the cassette 9 will return to the initial position, so that the detection slot 921 of the cassette 9 and the photodetector 62 of the optical unit 6 are on the same horizontal plane. At the same time, the sample will be transported to the reaction wafer 92, and after measurement, mixing, and/or defoaming, etc., the sample will be distributed to the detection tank 921 for nucleic acid amplification and detection. Then, the rotation drive unit 7 drives the rotation of the cassette 9 so that each detection slot 921 is sequentially aligned with the different optical unit 6 and is detected (as shown in FIG. 13E).

After the nucleic acid detection is completed, the user can open the upper casing 11, and the linear drive unit 8 will again drive the fluid adapter 31 and the second temperature control unit 5 to move upward and cling to the bottom surface of the cassette 9, and The cassette 9 is further pushed up so that the top surface of the cassette 9 is significantly higher than the top surface of the main frame 2 to facilitate the user to take out the cassette 9 (as shown in FIG. 13F). This function may be accomplished in another way. For example, at least one raised structure such as a pin may be fixed on the fluid adapter 31. The convex structure and the guide groove 94 of the cassette 9 have appropriate dimensions so that they can be accommodated in the guide groove 94 of the cassette 9 and are vertically aligned with the guide groove 94 without affecting the fluid adapter 31 and The movement of the second temperature control unit 5 in the vertical direction. When the cassette 9 needs to be jacked up, the convex structure and the guide groove 94 of the cassette 9 can be misaligned by rotating the cassette 9 at a certain angle, and the linear drive unit 8 will drive the fluid adapter 31 and The second temperature control unit 5 moves upward to lift the cassette 9. This method can adjust the height of the jack 9 more flexibly. The jacking height can be optimized by adjusting the length of the raised structure.

In one embodiment, the lower case 12 of the nucleic acid analysis device of this case further includes a power switch 14, an embedded lens 15 and a touch screen 16. The embedded lens 15 can scan the barcode or QR code on the sample tube and cassette to obtain relevant information, and the touch screen 16 can be used for user operation and result display.

In one embodiment, the nucleic acid analysis device in this case is designed for isothermal amplification and can be used to perform all isothermal amplification methods, such as nucleic acid sequence-dependent amplification technology (nucleic acid sequence-based amplification, NASBA), Strand displacement amplification (SDA), helicase-dependent amplification (HDA), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification Recombinase polymerase amplification (RPA) and nicking enzyme amplification reaction (NEAR).

In summary, the embodiments of the present invention provide a fully functional integrated nucleic acid analysis device, which integrates a fluid delivery unit, a temperature control unit, a driving unit, and an optical unit on a single device to enable sample purification, nucleic acid extraction, and nucleic acid expansion Processes such as augmentation and nucleic acid detection can be performed on this fully functional integrated device to achieve real-time nucleic acid analysis, so the nucleic acid analysis device in this case provides simple and fast nucleic acid analysis. In particular, in addition to the first temperature control unit that provides a single temperature interval required for isothermal amplification, the temperature control unit in this case also includes a second temperature control unit that can provide a higher heating temperature region to denature nucleic acids. Can improve the efficiency of nucleic acid amplification. Furthermore, the nucleic acid analysis device in this case includes two sets of drive units, one of which is a rotary drive unit, which can drive the rotation of the cassette, so that each detection slot is sequentially aligned with a different optical unit and detected. The other is a linear drive unit, which can drive the part of the fluid delivery unit and the second temperature control unit to move upward and closely contact the cassette for fluid processing such as nucleic acid extraction; and drive the part of the fluid delivery unit during nucleic acid amplification and detection And the second temperature control unit moves downward to separate from the cassette to facilitate subsequent cassette rotation; it can also eject the cassette after nucleic acid detection is completed, so that the user can take out the cassette. Therefore, the nucleic acid analysis device of the present case can complete automated full-function integrated detection, which is helpful for POC diagnosis and has extremely high industrial utilization value.

Even though the new model has been described in detail in the above embodiments and can be modified by any person skilled in the art, it is not as easy to protect as the scope of the patent application.

1: shell 11: Upper case 12: Lower case 13: Stop 131: Spring 14: Power switch 15: Embedded lens 16: Touch screen 2: main frame 21: chamber 3: Fluid delivery unit 31: Fluid adapter 311: opening 312: Connector 4: The first temperature control unit 41: Heater 411: Heating element 42: Radiator 43: Thermal insulation 5: Second temperature control unit 51: Heater 52: Radiator 53: Fan 6: Optical unit 61: Light source 62: light detector 7: rotary drive unit 71: Stepper motor 72: shaft 73: Drive gear 74: driven gear 741: Protrusion 75: rotation sensing unit 76: Bearing 8: Linear drive unit 81: stepper motor 82: screw 83: slider 84: cantilever 85: Position sensing unit 9: Cassette 91: Cassette body 92: Reaction chip 921: detection slot 922: Main fluid channel 923: Gas release channel 924: Sample loading hole 925: slot 93: Heat-treated wafer 931: opening 932: Heating tank 94: guide groove 9A, 9B: fluid network

Figure 1 shows a schematic diagram of the nucleic acid analysis device of the embodiment of the present invention. Figure 2 shows a schematic diagram of the main internal structure of the nucleic acid analysis device. Figure 3 shows an exploded view of the nucleic acid analysis device of Figure 2. Figures 4 and 5 show schematic diagrams of cassettes at different angles. Figure 6 shows a schematic diagram of a part of the structure of the nucleic acid analysis device. Figure 7 shows a schematic diagram of a part of the structure of a nucleic acid analysis device. Figure 8 shows the structure shown in Figure 7 and the cassette mounted on it. Figure 9 shows a schematic diagram of the rotary drive unit of the embodiment of the present invention. Figure 10 shows a partial schematic diagram of a nucleic acid analysis device. Figure 11 shows a partial cross-sectional view of the cassette when installation is complete. Figure 12 shows a schematic diagram of a part of the structure of a nucleic acid analysis device. Figures 13A to 13F show the workflow of the nucleic acid analysis device for full-function integration detection.

2: main frame

3: Fluid delivery unit

31: Fluid adapter

4: The first temperature control unit

5: Second temperature control unit

6: Optical unit

7: rotary drive unit

8: Linear drive unit

9: Cassette

Claims (20)

A nucleic acid analysis device, including: A shell, including an upper shell and a lower shell; A main frame body, which is arranged in the lower casing and has a chamber for installing a cassette therein, the cassette including a plurality of detection slots; A fluid delivery unit is provided in the lower housing and is configured to deliver samples and/or reagents in the cassette for nucleic acid extraction and/or nucleic acid amplification, wherein the fluid delivery unit has a fluid adapter , And the fluid adapter is arranged below the main frame; A first temperature control unit is disposed on the main frame and is configured to provide a first temperature for nucleic acid amplification; A second temperature control unit is arranged below the main frame and is configured to provide a second temperature higher than the first temperature for nucleic acid denaturation before nucleic acid amplification; At least one optical unit, disposed on the main frame, and including a plurality of optical elements for nucleic acid detection; A rotary drive unit, disposed in the lower housing, and configured to rotate the cassette with a preset program during nucleic acid detection to align the detection slot of the cassette with the optical unit; and A linear drive unit is provided in the lower housing and is structured to lift the fluid adapter and the second temperature control unit upward during nucleic acid extraction so that the fluid adapter and the second temperature control unit Close contact with the bottom of one of the cassettes. The nucleic acid analysis device according to claim 1, wherein during nucleic acid amplification and detection, the linear drive unit drives the fluid adapter and the second temperature control unit to move downward to maintain a gap with the bottom surface of the cassette . The nucleic acid analysis device according to claim 1, wherein after the nucleic acid detection is completed, the linear drive unit drives the fluid adapter and the second temperature control unit to move upward, and pushes the cassette upward so that the cassette The top surface is obviously higher than the top surface of the main frame to facilitate the removal of the cassette. The nucleic acid analysis device according to claim 1, wherein the linear driving unit includes a stepping motor, a screw, a slider, and a cantilever, wherein the slider is connected to the stepping motor through the screw, and the cantilever is fixed On the slider, and the fluid adapter and the second temperature control unit are carried on the cantilever. The nucleic acid analysis device according to claim 1, wherein the linear driving unit includes at least one position sensing unit. The nucleic acid analysis device according to claim 1, wherein the upper casing includes a plurality of blocking portions, and when the cassette is pushed upward by the linear driving unit, the top surface of the cassette will abut against the blocking portion . The nucleic acid analysis device according to claim 1, wherein the cassette includes at least one guide groove, the rotary drive unit includes at least one protrusion, the protrusion can slide in the guide groove, and the rotary drive unit passes through the protrusion Part clamps the cassette to drive the rotation of the cassette. The nucleic acid analysis device according to claim 7, wherein the rotary driving unit includes a stepping motor, a rotating shaft, a driving gear, and a driven gear, wherein the driving gear is connected to the stepping motor through the rotating shaft, and the The driving gear meshes with the driven gear. The nucleic acid analysis device according to claim 8, wherein the protrusion is provided on the driven gear. The nucleic acid analysis device according to claim 1, wherein the optical unit includes a light source and a light detector. The nucleic acid analysis device according to claim 1, wherein the nucleic acid analysis device includes a plurality of optical units, each of which provides light of a unique wavelength to detect a plurality of targets. The nucleic acid analysis device according to claim 1, wherein the cassette includes a cassette body, a reaction wafer, and a heat treatment wafer, the reaction wafer is disposed on top of the cassette body, and the heat treatment wafer is disposed on the card The bottom of the box body. The nucleic acid analysis device according to claim 12, wherein the first temperature control unit surrounds the cassette body and carries the reaction wafer thereon. The nucleic acid analysis device according to claim 12, wherein the second temperature control unit performs contact heating on the heat-treated wafer. The nucleic acid analysis device according to claim 12, wherein the bottom surface of the heat-treated wafer includes a plurality of openings, and the top surface of the fluid adapter includes a plurality of openings, which can correspond to the plurality of openings of the heat-treated wafer, to Form a pipeline for fluid transport. The nucleic acid analysis device according to claim 12, wherein the reaction wafer includes at least two fluid networks, wherein each fluid network includes the plurality of detection tanks, a main fluid flow path, and a gas release flow path, the main The fluid flow channel is connected to the detection tank and is configured to distribute the sample or control liquid into the detection tank, and the gas release flow channel is connected to the detection tank and is configured to release gas from the detection tank, wherein the fluid network One of these frameworks is used for quality control. The nucleic acid analysis device according to claim 16, wherein the gas release flow path is significantly narrower than the main fluid flow path. The nucleic acid analysis device according to claim 12, wherein the shape of the reaction wafer is substantially a regular polygon, and each of the detection slots has at least one plane. The nucleic acid analysis device according to claim 12, wherein the reaction wafer further includes at least one sample loading hole provided on the top surface thereof for adding the sample to the cassette. The nucleic acid analysis device according to claim 1, further comprising an embedded lens for scanning a code or a QR code on a sample tube or the cassette to obtain relevant information.

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