Nucleic acid detection device

Abstract

A nucleic acid detection device comprises a pipe fitting placing groove, a light emitting module, a detection module and a driving module. The pipe fitting containing groove is used for containing a pipe fitting and comprises a reagent and a magnetisable element. The light-emitting module comprises a light source, a magnet and a shell, and the light source and the magnet are installed on the shell. The driving module comprises a motor, a driving screw and a sliding block, the driving screw is coupled with the motor and driven by the motor to rotate, and the sliding block is arranged on the driving screw in a sleeving mode and moves along with rotation of the driving screw. The light-emitting module is coupled with the sliding block and linked with the sliding block, and performs reciprocating rectilinear motion in the direction parallel to the guide screw. When the magnet is driven to deviate from the normal level of the pipe fitting, the magnetizable element is maintained at a first site, and when the magnet is driven to align with the normal level of the pipe fitting, the magnetizable element moves from the first site to a second site under the magnetic attraction action of the magnet, so that the reagent in the pipe fitting is mixed.

Description

Technical Field

[0001] This case is about a nucleic acid detection device, in particular, a nucleic acid detection device with magnetic mixing function. Background Art

[0002] The outbreak of severe acute respiratory syndrome (COVID-19) in 2019 has made the field of molecular diagnosis a global focus, prompting a rapid increase in the demand for in vitro diagnostic (IVD) test kits and testing equipment. In order to meet the rapidly growing demand for IVD testing and to achieve accurate and precise testing and analysis, how to preserve the test reagents (such as enzymes and antibodies) filled in the test kit is an important issue. If the test reagents become ineffective due to improper storage, it will lead to false positive or false negative results.

[0003] In the past, if molecular detection reagents are stored and transported in wet form, they are usually supplemented with glycerin, mineral oil or other related additives to ensure the stability of the chemical structure of the reagents in wet form. In addition, wet-form molecular detection reagents must be stored in a low-temperature environment (e.g., -20 degrees Celsius) and must be transported by cold chain transportation. However, the necessity of low-temperature storage and cold chain transportation in wet reagent form not only increases the cost of the detection kit, but also causes inconvenience to users in storing the detection kit. In addition, the refrigeration equipment, refrigerants and coolants required for cold chain storage and transportation also cause energy loss and environmental pollution.

[0004] The above inconvenience can now be solved by the mature freeze-drying (lyophilization) technology. By placing the frozen molecular detection reagents in a vacuum environment for dehydration and drying, a stable solid form is formed, which improves the stability of the reagents and greatly extends the shelf life. In addition, freeze-dried reagents have greatly reduced weight and volume, and no longer require cold chain transportation specifically to maintain the stability of the reagents, reducing costs and reducing the impact on the environment. During the freeze-dried storage process of molecular detection reagents, lyoprotectants (also known as excipients) are often used to prevent denaturation of the components. Lyoprotectants can be divided into carbohydrates, polyols, polymers, surfactants, amino acids, and salts according to their ingredients.

[0005] However, each lyophilized reagent and its lyophilized protective agent have fixed and optimized components and ratios, that is, the added relevant lyophilized protective agent has a certain volume concentration, which directly causes the lyophilized reagent to have a certain degree of concentration gradient distribution problem after being rehydrated with water, buffer or reaction reagent. Therefore, it is necessary to give a period of time to allow the molecules in the solution to diffuse evenly, or an active disturbance method (such as using an oscillator or using a micropipette to pipette up and down) must be provided to evenly distribute the components of the reagent reaction. If the uneven concentration distribution generated after the lyophilized reagent is rehydrated and dissolved is not resolved, it is easy to reduce the reaction efficiency between the reagent and the reactant, and reduce the consistency of the reaction results, thereby affecting the interpretation of the test results.

[0006] Therefore, how to improve the above-mentioned shortcomings of the prior art is a problem to be overcome urgently. Summary of the invention

[0007] The purpose of this case is to provide a nucleic acid detection device, which mainly proposes an improved design for reagent mixing to solve the problem of uneven concentration distribution after reconstitution of freeze-dried reagents.

[0008] To achieve the above-mentioned purpose, the present invention provides a nucleic acid detection device, including: a plurality of tube placement slots for accommodating a plurality of tubes, wherein each tube includes a reagent and a magnetizable element; a light-emitting module, which is arranged on one side of the plurality of tube placement slots and includes a plurality of light sources, a magnet and a housing, wherein the plurality of light sources and the magnet are mounted on the housing; a detection module, which is arranged on the opposite side of the plurality of tube placement slots relative to the light-emitting module; and a driving module, which includes a motor, a lead screw and a slider, wherein the lead screw is coupled to the motor and driven by the motor to rotate, and the slider is sleeved on the lead screw and performs reciprocating linear motion in response to the rotation of the lead screw. The light-emitting module is coupled to the slider and is linked to the slider, so that the plurality of light sources and the magnet are synchronously driven by the motor to perform reciprocating linear motion in a direction parallel to the lead screw. When the magnet is driven to deviate from the normal position of the tube, the magnetizable element remains at the first position. When the magnet is driven to align with the normal position of the tube, the magnetizable element is attracted by the magnet and moves from the first position to the second position, thereby mixing the reagents in the tube.

[0009] In one embodiment, the first point is approximately located at the bottom of the tube and the second point is approximately located near the magnet.

[0010] In one embodiment, the reagent is a lyophilized reagent.

[0011] In one embodiment, the magnetizable element is made of metal material.

[0012] In one embodiment, the magnetizable element is spherical, and its diameter is between 0.5 mm and 2 mm.

[0013] In one embodiment, the magnet is made of neodymium iron boron material.

[0014] In one embodiment, the maximum magnetic energy product of the magnet is between 45 and 55 MGOe, the cross-sectional area is between 1.5 and 3.5 mm, and the length is between 1 and 8 mm.

[0015] In one embodiment, the vertical distance between the magnet and the top of the housing is between 2.5 and 3.5 mm.

[0016] In one embodiment, the number of the magnet is single.

[0017] In one embodiment, the installation position of the magnet is offset from the installation position of the light source.

[0018] In one embodiment, the driving module further includes a carrier, which is coupled to and linked with the slider.

[0019] In one embodiment, the light emitting module is carried on a carrier.

[0020] In one embodiment, the driving module further includes a photoelectric sensor, and the carrier includes an alignment fin having at least one slit thereon for determining whether the light source is aligned with the tube.

[0021] In one embodiment, the driving module further includes a linear slide rail, which is arranged in parallel with the lead screw, and the carrier is slidably arranged on the linear slide rail.

[0022] In one embodiment, the slider includes a nut that is threaded onto the lead screw.

[0023] In one embodiment, the arrangement direction of the plurality of tube placement grooves is parallel to the lead screw.

[0024] In one embodiment, the tube further comprises an annular paraffin layer disposed on the top of the reagent.

[0025] In one embodiment, the light emitting module further includes a cover plate for accommodating a plurality of filters.

[0026] In one embodiment, the detection module includes a plurality of lens groups, a plurality of filters, and a plurality of light detectors.

[0027] In one embodiment, the nucleic acid detection device also includes a temperature control module, which includes a thermoelectric cooling chip. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A schematic diagram showing the nucleic acid detection device in this case;

[0029] Figure 2 show Figure 1 Schematic diagram of the partial structure of the nucleic acid detection device;

[0030] Figure 3 show Figure 2 A-A' section structure diagram;

[0031] Figure 4A A schematic diagram showing the light-emitting module of the present invention;

[0032] Figure 4B show Figure 4A B-B' cross-sectional structure diagram;

[0033] Figure 4C show Figure 4A Exploded structure diagram;

[0034] Figure 5A A schematic diagram showing a light emitting module according to another embodiment of the present invention;

[0035] Figure 5B show Figure 5A C'-C' cross-sectional structure diagram;

[0036] Figure 5C show Figure 5A Exploded structure diagram;

[0037] Figure 6 and Figure 7 A schematic diagram showing the light-emitting module and the driving module of the present invention;

[0038] Figure 8 A schematic diagram showing the carrier of this case;

[0039] Fig. 9A and Fig. 9B Schematic diagram showing the magnet not being aligned with the pipe;

[0040] Fig. 10A and Fig. 10B A schematic diagram showing the alignment of the magnet and the pipe;

[0041] FIG. 11A to FIG. 11F A schematic diagram showing the sequential alignment of magnets and pipes;

[0042] FIG. 12A to FIG. 12F A schematic diagram showing the alignment of light sources and pipe fittings in sequence;

[0043] FIG. 13A to FIG. 13C Demonstrating the effect of a magnetic mixing mechanism in promoting reagent mixing in one embodiment;

[0044] FIG. 14A to FIG. 14C Another embodiment shows the effect of a magnetic mixing mechanism in promoting reagent mixing.

[0045]

Explanation of symbols

[0046] 1: Pipe placement slot

[0047] 11: Light entrance

[0048] 12: Light exit

[0049] 2: Light emitting module

[0050] 21: Light Source

[0051] 22: Filter

[0052] 23: Magnet

[0053] 24: Shell

[0054] 241: Slot

[0055] 242: Slot

[0056] 243: Light Exit

[0057] 244: Opening

[0058] 25: Cover

[0059] 251: Slot

[0060] 252: Light Exit

[0061] 253: Opening

[0062] 3: Detection module

[0063] 31: Lens group

[0064] 32: Filter

[0065] 33: Light Detector

[0066] 34: Signal acquisition circuit board

[0067] 4: Driver module

[0068] 41: Motor

[0069] 42: Lead screw

[0070] 43: Slider

[0071] 431: Nut

[0072] 44: Carrier

[0073] 441: First pair of fins

[0074] 442: Slit

[0075] 443: Second pair of fins

[0076] 444: Slit

[0077] 45: Linear guide rail

[0078] 461: First photoelectric sensor

[0079] 462: Two photoelectric sensors

[0080] 5: Pipe fittings

[0081] 51: Reagents

[0082] 52: Magnetizable components

[0083] 53: Annular paraffin layer

[0084] 6: Temperature control module

[0085] 61: Thermal conductive silicone sheet

[0086] 62: Thermoelectric cooling chip

[0087] 63: Heat sink

[0088] X, Y, Z: Axis

[0089] A-A', B-B', C-C', D-D', E-E': hatching DETAILED DESCRIPTION

[0090] Some embodiments that embody the features and advantages of the present invention will be described in detail in the following description. It should be understood that the present invention can have various variations in different aspects without departing from the scope of the present invention, and the description and drawings are essentially for illustrative purposes rather than for limiting the present invention.

[0091] This case provides a nucleic acid detection device, which proposes an improved design for reagent mixing, which is used to solve the problem of uneven concentration distribution of freeze-dried reagents after reconstitution, especially to solve the problem of fluorescence baseline drift and inconsistent detection results caused by uneven concentration distribution during the real-time polymerase chain reaction (qPCR) process. In view of the fact that the conventional magnetic mixing technology causes a significant increase in equipment volume and cost due to the mechanism setting, this case integrates the magnet into the light-emitting module required for nucleic acid detection, so that the magnet can be driven together with the light-emitting module to simplify the mechanism setting. According to this concept, this case mainly drives the magnetizable element in the reaction tube to move through the magnetic field contributed by a single magnet and moving along a single axis, thereby providing a driving force for active disturbance of the reagents in the reaction tube, so as to achieve the purpose of uniform reagent concentration distribution in the reaction tube.

[0092] Figure 1 A schematic diagram showing the nucleic acid detection device in this case, Figure 2 show Figure 1 Schematic diagram of the partial structure of the nucleic acid detection device. Figure 3 show Figure 2 A-A' cross-sectional structure diagram. Figures 1 to 3As shown, the nucleic acid detection device of the present case mainly includes a plurality of tube placement slots 1, a light emitting module 2, a detection module 3, and a driving module 4. The plurality of tube placement slots 1 are made of metal material and are arranged in a straight line along the X-axis to accommodate a plurality of tubes (such as PCR reaction tubes) 5, so the plurality of tubes 5 are also arranged in a straight line. The light emitting module 2 and the detection module 3 are respectively arranged on two opposite sides of the plurality of tube placement slots 1, that is, respectively arranged on two opposite sides of the plurality of tubes 5. The light emitting module 2 can provide light of a specific wavelength to excite the fluorescent substance in the tube 5, and then the detection module 3 receives the fluorescence emitted by the fluorescent substance to achieve the purpose of nucleic acid detection. The driving module 4 drives the movement of the light emitting module 2 to align with the tube 5.

[0093] In one embodiment, the light emitting module 2 includes a plurality of light channels, the spacing between two adjacent light channels is substantially equal to the spacing between two adjacent tubes 5, and each light channel includes a light source 21 and a filter 22. The detection module 3 also includes a plurality of light channels, the spacing between two adjacent light channels is substantially equal to the spacing between two adjacent tubes 5, and each light channel includes a lens group 31, a filter 32, and a light detector 33, and the light detector 33 is further coupled to a signal acquisition circuit board 34. The light source 21 can be a light emitting diode, but is not limited thereto. A light emitting diode with a specific wavelength provides a detection light source, and is incident on the tube 5 through the filter 22 and the light inlet 11 of the tube placement slot 1, thereby exciting the fluorescent substance in the tube 5; the fluorescence generated by the amplification reaction can be emitted through the light outlet 12 of the tube placement slot 1, and is collected by the lens group 31, the filter 32, and the light detector 33, and is sensed by the rear signal acquisition circuit board 34 and converted into a readable value.

[0094] In one embodiment, the nucleic acid detection device of the present case further includes a temperature control module 6 for applying different temperatures for regulation, driving the nucleic acid molecules and reaction reagents to continuously cycle through the steps of denaturation, annealing and extension to achieve the purpose of nucleic acid molecule amplification. Figure 3 As shown, the temperature control module 6 includes a thermal conductive silicone sheet 61, a thermoelectric cooling (TEC) chip 62, a heat sink 63 and a cooling fan (not shown). In addition, the tube placement slot 1 is made of metal material, such as copper with excellent thermal conductivity. Therefore, the tube placement slot 1 is used not only to place the tube 5, but also to conduct the heat energy generated by the thermoelectric cooling chip 62 to facilitate the PCR reaction.

[0095] Figure 4A A schematic diagram showing the light-emitting module of the present invention is shown. Figure 4B show Figure 4A The B-B' cross-sectional structure diagram, Figure 4C show Figure 4AExploded structure diagram. FIG. 4A to FIG. 4C As shown, in addition to the aforementioned light source 21 and filter 22, the light emitting module 2 of the present invention further includes a magnet 23, a housing 24, and a cover plate 25. The housing 24 includes a slot 241 (shown in FIG. 24 ) that can accommodate a plurality of light sources 21. Figure 3 ), and a slot 242 for accommodating the magnet 23, so the light source 21 and the magnet 23 are both mounted and fixed on the housing 24, wherein the mounting position of the magnet 23 is offset from the mounting position of the light source 21, and is approximately located between two adjacent light sources 21 in the X-axis direction. The housing 24 also includes a plurality of light outlets 243 corresponding to the plurality of light sources 21, and an opening 244 corresponding to the magnet 23. The cover plate 25 includes a slot 251 (shown in FIG. 25 ) for accommodating a plurality of filters 22. Figure 3 ), and a plurality of light outlets 252 corresponding to the plurality of filters 22, wherein the light outlet 243 of the housing 24 and the light outlet 252 of the cover plate 25 are aligned with each other in each light channel.

[0096] In the aforementioned embodiment, the magnet 23 is shielded by the cover plate 25 . However, in another embodiment, the magnet 23 may be exposed and not shielded by the cover plate 25 . Figure 5A A schematic diagram showing a light emitting module according to another embodiment of the present invention is shown. Figure 5B show Figure 5A The C-C' cross-sectional structure diagram, Figure 5C show Figure 5A Exploded structure diagram. FIG. 5A to FIG. 5C As shown, the cover 25 further includes an opening 253 corresponding to the magnet 23 , which is aligned with the opening 244 of the housing 24 , so that the magnet 23 on the light-emitting module 2 is exposed without being shielded by the cover 25 , and can be closer to the tube 5 for magnetic mixing.

[0097] Figure 6 and Figure 7 A schematic diagram showing the light-emitting module and the driving module of the present invention, wherein the light-emitting module is driven by the driving module and is located at different positions. Figure 6 and Figure 7As shown, the driving module 4 is a linear slide assembly, which mainly includes a motor 41, a lead screw 42 and a slider 43. The lead screw 42 is coupled to the motor 41 and driven by the motor 41 to rotate. The slider 43 is sleeved on the lead screw 42 and performs reciprocating linear motion on the lead screw 42 in response to the rotation of the lead screw 42. The light-emitting module 2 is coupled to the slider 43 and linked with the slider 43, so that the light source 21 and the magnet 23 on the light-emitting module 2 are synchronously driven by the motor 41 to perform reciprocating linear motion in a direction parallel to the lead screw 42 (X-axis direction). In other words, the arrangement direction of the plurality of pipe placement slots 1, the arrangement direction of the plurality of pipes 5, and the axial direction of the lead screw are all parallel to the X-axis direction, and the light source 21 and the magnet 23 on the light-emitting module 2 move in this single axial direction. Of course, the design of the driving module 4 driving the light-emitting module 2 can also be adjusted according to different needs, and is not limited to the embodiment described in this case.

[0098] In one embodiment, the motor 41 is a stepping motor, which can programmatically control the movement mode of the light emitting module 2 , but is not limited thereto.

[0099] In one embodiment, the slider 43 includes a nut 431 which is screwed onto the lead screw 42 , thereby driving the slider 43 to perform reciprocating linear motion on the lead screw 42 .

[0100] In one embodiment, the driving module 4 further includes a carrier 44 and a linear slide 45, and the linear slide 45 is arranged in parallel with the lead screw 42. The carrier 44 is coupled and linked with the slider 43, and is slidably arranged on the linear slide 45. The light emitting module 2 is carried on the carrier 44, so it can be coupled and linked with the slider 43, so that the light source 21 and the magnet 23 on the light emitting module 2 are synchronously driven by the motor 41 to perform reciprocating linear motion in a direction parallel to the lead screw 42 (X-axis direction).

[0101] In one embodiment, the driving module 4 further includes a design of an alignment structure for determining whether the light source 21 is aligned with the tube 5. Figures 6 to 8 ,in Figure 8 A schematic diagram showing the carrier for this case. Figures 6 to 8As shown, the driving module 4 includes a first photoelectric sensor 461, which is a photointerrupter, having a light-emitting element and a light-receiving element arranged relatively. When the light-shielding part or the light-transmitting part of the object passes through, the current changes due to whether the light path of the photointerrupter is blocked or not. The carrier 44 includes a first alignment fin 441, on which a plurality of slits 442 are provided, and the spacing between two adjacent slits 442 is roughly equal to the spacing between two adjacent tubes 5. When the slits 442 pass through the first photoelectric sensor 461, the first photoelectric sensor 461 detects that the light path is not blocked, indicating that at least one light source 21 is aligned with the tube 5, and the magnet 23 is not aligned with the tube 5, and the light source 21 switch can be turned on to emit the detection light source. Through the design of the alignment structure, the magnetic mixing and optical detection paths can be separated to avoid shielding the detection light source path due to magnetic mixing during optical detection, which affects the acquisition of detection data.

[0102] In one embodiment, the driving module 4 further includes a second photoelectric sensor 462, and the stage 44 includes a second alignment fin 443, on which a slit 444 is provided. The slit 444 is provided for zero point positioning. When the slit 444 passes through the second photoelectric sensor 462, the second photoelectric sensor 462 detects that the light path is not blocked, indicating that the stage 44 has returned to the origin of the reciprocating linear motion (the position shown in FIG. 5 ).

[0103] The following will further cooperate Fig. 9A , Fig. 9B , Fig. 10A and Fig. 10B The magneto-hybrid mechanism of this case is described, where Fig. 9A and Fig. 9B Schematic diagram showing the magnet not aligned with the pipe. Fig. 9B for Fig. 9A The D-D' local cross-sectional structure diagram, and Fig. 10A and Fig. 10B Schematic diagram showing the alignment of magnets and pipe fittings, Fig. 10B for Fig. 10A E-E' local cross-sectional structure diagram.

[0104] First of all, it is explained that the pipe 5 is made of a polymer material and is a conical container that is open upward and wide at the top and narrow at the bottom. The pipe placement groove 1 is also an upwardly open and wide at the top and narrow at the bottom structure, and its inner diameter is designed to conform to and fit closely to the pipe 5. The pipe 5 mainly contains reagents 51 and magnetizable elements 52. Reagents 51 include but are not limited to PCR reaction reagents, especially PCR reaction reagents in freeze-dried form. The reagent package includes nucleotides, primers, probes, enzymes, carbohydrates and stabilizers. The pipe 5 also includes an annular paraffin layer 53 on the top of the reagent 51. This annular structure allows the solution to be tested, such as nucleic acid extract, to be added from the center, and then melted by the temperature control module 6, and covered on the surface of the reaction solution to form an isolation layer to prevent the evaporation of the reaction solution during the temperature control cycle and prevent the amplification product from escaping into the environment and causing pollution.

[0105] In one embodiment, the magnetizable element 52 is made of metal, and its surface is usually coated with a layer of corrosion-resistant material (such as nickel, chromium, molybdenum or polymer material) to avoid oxidation reaction with the reagent 51. The magnetizable element 52 will naturally settle to the bottom of the tube 5 due to gravity. The shape of the magnetizable element 52 can be, but not limited to, spherical, square, or cylindrical, wherein the spherical shape is preferred because it can conform to and fit closely to the bottom of the tube 5, reducing the generation of bubbles, and the diameter of the sphere is between 0.5 and 2 mm, for example, 0.5, 0.58, 0.8 or 2 mm. For example, the magnetizable element 52 can be a steel ball.

[0106] When the light-emitting module 2 embedded with the magnet 23 is driven by the driving module 4 so that the center of the magnet 23 deviates from the normal position of the tube 5 (i.e., it is not aligned with the tube 5), the magnetizable element 52 placed in the tube 5 is not actuated by the magnetic field and naturally maintains the first position due to its own mass, i.e., it is approximately located at the bottom of the tube 5 (e.g., Fig. 9A and Fig. 9B When the light-emitting module 2 embedded with the magnet 23 is driven by the driving module 4 so that the center of the magnet 23 is aligned with the normal position of the respective tube 5 (i.e. aligned with the tube 5), the magnetic field contributed by the magnet 23 will drive the magnetizable element 52 in the tube 5 to move from the first position to the second position, i.e., move upward from the bottom of the tube 5 to a position close to the magnet 23 (e.g. Fig. 10A and Fig. 10B As shown in FIG. 4 ). When the driving module 4 drives the magnet 23 to move to the next tube 5, the center of the magnet 23 deviates from the normal position of the original tube 5, and the magnetizable element 52 that was originally attracted to the second position is no longer attracted to the magnet and naturally settles back to the first position due to its own mass. Therefore, through programmable repeated movement and alignment, the magnetizable element 52 in the tube 5 is driven to move, thereby providing a driving force for active disturbance of the reagent 51 in the tube 5, so as to achieve the purpose of making the concentration distribution of the reagent 51 in the tube 5 uniform.

[0107] The above embodiment is described by taking the light-emitting module 2 inlaid with a single magnet 23 as an example, but the invention is not limited thereto. The magnetic mixing of the reagent 51 in the tube 5 can be adjusted by programmatically controlling the stop time and the number of cycles of the magnet 23 moving along the multiple tubes 5. For example, the stop time can be set to 0 seconds, 200 milliseconds, 300 milliseconds, 400 milliseconds, 500 milliseconds, 600 milliseconds, 700 milliseconds, 800 milliseconds, 900 milliseconds or 1 second, and the number of cycles can be set to 10 times, 20 times or 30 times. These parameters can be adjusted according to different needs and are not limited to those described in the embodiments of the present invention.

[0108] According to the design of the nucleic acid detection device of the present case, when the tube 5 injected with the solution to be tested is placed in the tube placement slot 1, the light emitting module 2 can be driven by the driving module 4 in a programmable manner, and the alignment structure can be designed to determine whether the light source 21 is aligned with the tube 5 to control the light source 21 switch, so as to avoid the magnetizable element 52 being attracted by the magnet in the tube 5 and moving during the optical detection, thereby blocking the detection light source path and affecting the detection data acquisition. For example, the nucleic acid detection device first performs a magnetic mixing operation, such as FIG. 11A to FIG. 11F As shown, the magnet 23 is driven to align with each tube 5 in sequence to mix the reagents, and can be circulated to mix them fully. Then, the temperature control module 6 is used to amplify the nucleic acid molecules in the tube 5. After the amplification is completed, the driving module 4 drives the light source 21 to align with the tube 5, and the light source 21 is turned on in the aligned state to perform optical detection, and as shown in FIG. FIG. 12A to FIG. 12F As shown, the light source 21 is driven to align with the pipe 5 in sequence. During the optical detection operation, the light source 21 is aligned with the normal position of the pipe 5, and during the magnetic mixing operation, the center of the magnet 23 is aligned with the normal position of each pipe 5, thereby creating a light source 21 and a magnet 23 that can be set on the same mechanism component and their functions do not interfere with each other, greatly reducing the complexity of the mechanism.

[0109] In one embodiment, the magnet 23 is made of neodymium iron boron material, the maximum magnetic energy product ((BH)max) is between 45 and 55 MGOe, the cross-sectional area of ​​the magnet 23 is between 1.5 and 3.5 mm, the length of the magnet 23 is between 1 and 8 mm, and the vertical distance between the magnet 23 and the top of the shell 24 is between 2.5 and 3.5 mm.

[0110] In one embodiment, the number of the magnet 23 is only one, and it is embedded in the center of the housing 24. Because it is driven by the driving module 4 to move and align with each tube 5, only one magnet 23 is needed to achieve magnetic mixing of all tubes 5. Of course, the number of the magnet 23 is not limited to one, and can be more than two, thereby shortening the moving distance of the light-emitting module 2.

[0111] The following will use a specific embodiment to illustrate the magnetic mixing effect of the nucleic acid detection device of this case. In this specific embodiment, the nucleic acid detection device is provided with a single magnet 23, which is made of neodymium iron boron, has a maximum magnetic energy product ((BH)max) of 50-53MGOe, a cross-sectional area of ​​2mm and a length of 6mm. The magnet 23 is located in the center of the shell 24 and is 2.5mm vertically away from the top of the shell 24, so that when the magnetizable element 52 actuated by the magnetic field moves to the second position, it is just adjacent to and located at the bottom of the paraffin isolation layer. The magnetizable material 52 contained in the pipe 5 is spherical in shape, with a diameter of 0.58mm, and its material is 440 series stainless steel. When the magnet 23 is driven to move cyclically along multiple pipes 5, the waiting time of individual pipes 5 is set to 200 milliseconds, and the number of mixing cycles is set to 10 times, 20 times and 30 times.

[0112] In this specific embodiment, the reagent used for detection is 25μL SARS-CoV-2 lyophilized-cake, and the nucleic acid detection device of this case is used with the nucleic acid extraction cartridge to perform the full process test of extraction and RT-PCR on the SARS-CoV-2 standard, and the detection target is the RdRp and N genes of the virus. Using viral transport medium (VTM) as the matrix, 750cp / mL SARS-CoV-2 standard and 3x 10^4 human cells are prepared, and 0.4mL is taken as a sample. When the extraction is completed, the magnetic mixing mechanism is set to: no mixing, mixing 10 times, mixing 20 times and mixing 30 times, and subsequent nucleic acid amplification and optical detection are performed. FIG. 13A to FIG. 13C The effect of the magnetic mixing mechanism in this embodiment to promote the mixing of reagents is shown, wherein Fig.13A Display the RT-PCR fluorescence curve signal of RdRp gene, Fig. 13B Displays the RT-PCR fluorescence curve signal of N gene. Fig. 13C Display the Cq value and amplification fluorescence value (ΔRFU). As shown in the figure, as the number of mixing increases, the fluorescence signal gradually increases and forms a typical S-shaped amplification curve. Then calculate the Cq value and amplification fluorescence value (ΔRFU) using the regression model. The results are as follows Fig. 13C As shown, the magnetically induced mixing mechanism of the nucleic acid detection device in this case does help to improve the reaction performance.

[0113] To verify the feasibility again, in another specific embodiment, a magnetic mixing test was performed with 10 μL of SARS-CoV-2 lyophilized-cake, wherein the sample included 1,000 cp / mL SARS-CoV-2 standard and 3x 10^4 human cells, and 0.4 mL of the sample was taken for full-process testing. FIG. 14A to FIG. 14CThe effect of the magnetic mixing mechanism in this embodiment to promote the mixing of reagents is shown, wherein Fig.14A Display the RT-PCR fluorescence curve signal of RdRp gene, Fig. 14B Displays the RT-PCR fluorescence curve signal of N gene. Fig. 14C Display Cq value and amplification fluorescence value (ΔRFU). As shown in the figure, when 10μL of drying reagent is not mixed, the RT-PCR fluorescence curve is flat and there is no amplification, and neither the RdRp gene nor the N gene can be detected. As the number of mixing increases to 10 and 20 times, the fluorescence signal is significantly enhanced.

[0114] The above verification results also show that when the volume of the dried reagent is reduced, the shape is more dense, and it may be more difficult to homogenize after re-dissolving due to the larger concentration gradient. If combined with the magnetic mixing mechanism of this case, it can indeed effectively accelerate the mixing of reagents to meet the subsequent PCR results. In other words, the magnetic mixing mechanism of this case has the effect of promoting the mixing of PCR solutions.

[0115] In summary, the present case provides a nucleic acid detection device, including a plurality of tube placement slots, a light-emitting module, a detection module and a driving module, and further integrates a magnet into the light-emitting module so that the magnet can be driven together with the light-emitting module to simplify the mechanism setting. That is, in addition to providing the excitation light source required for PCR fluorescence detection, the light-emitting module is also embedded with a magnet to magnetically mix the reagents in the tube. With the actuation of the driving module, the magnet moves in a single axis parallel to the arrangement direction of the tube, and through programmable repeated movement and alignment, the magnetizable element in the tube is driven to move, thereby providing a driving force for active disturbance of the reagent in the tube, promoting uniform distribution of reagent concentration in the tube, so as to improve reaction efficiency and consistency of reaction results, and avoid a substantial increase in equipment size and cost.

[0116] Although the present invention has been described in detail by the above embodiments, it can be modified in various ways by those skilled in the art without departing from the scope of protection as defined in the appended claims.

Claims

1. A nucleic acid detection device, characterized in that: include: A plurality of tube placement slots for accommodating a plurality of tubes, wherein each of the tubes comprises a reagent and a magnetizable element; A light emitting module is disposed on one side of the plurality of tube placement slots and comprises a plurality of light sources, a magnet and a shell, wherein the plurality of light sources and the magnet are mounted on the shell; a detection module, disposed on the opposite side of the plurality of tube placement slots relative to the light emitting module; and A driving module includes a motor, a lead screw and a slider, wherein the lead screw is coupled to the motor and driven by the motor to rotate, and the slider is sleeved on the lead screw and performs reciprocating linear motion according to the rotation of the lead screw. The light-emitting module is coupled to the slider and moves in conjunction with the slider, so that the plurality of light sources and the magnet are synchronously driven by the motor to perform reciprocating linear motion in a direction parallel to the lead screw. When the magnet is driven to deviate from a normal position of the tube, the magnetizable element is maintained at a first position. When the magnet is driven to align with the normal position of the tube, the magnetizable element is attracted by the magnet and moves from the first position to a second position, thereby mixing the reagent in the tube.

2. The nucleic acid detection device according to claim 1, characterized in that: The first point is approximately located at the bottom of the tube, and the second point is approximately located near the magnet.

3. The nucleic acid detection device according to claim 1, characterized in that: This reagent is a freeze-dried reagent.

4. The nucleic acid detection device according to claim 1, characterized in that: The magnetizable element is made of a metal material.

5. The nucleic acid detection device according to claim 1, characterized in that: The magnetizable element is spherical, and its diameter is between 0.5 and 2 mm.

6. The nucleic acid detection device according to claim 1, characterized in that: The magnet is made of neodymium iron boron material.

7. The nucleic acid detection device according to claim 1, characterized in that: The maximum magnetic energy product of the magnet is between 45 and 55 MGOe, the cross-sectional area is between 1.5 and 3.5 mm, and the length is between 1 and 8 mm.

8. The nucleic acid detection device according to claim 1, characterized in that: The vertical distance between the magnet and the top of the shell is between 2.5 and 3.5 mm.

9. The nucleic acid detection device according to claim 1, characterized in that: The number of the magnet is single.

10. The nucleic acid detection device according to claim 1, characterized in that: The installation position of the magnet is misaligned with the installation position of the light source.

11. The nucleic acid detection device according to claim 1, characterized in that: The driving module also includes a carrier, which is coupled and linked to the slider.

12. The nucleic acid detection device according to claim 11, characterized in that: The light emitting module is carried on the carrier.

13. The nucleic acid detection device according to claim 11, characterized in that: The driving module further comprises a photoelectric sensor, and the carrier comprises an alignment fin, on which at least one slit is provided for judging whether the light source is aligned with the tube.

14. The nucleic acid detection device according to claim 11, characterized in that: The driving module further comprises a linear slide rail which is arranged in parallel with the lead screw, and the carrier is slidably arranged on the linear slide rail.

15. The nucleic acid detection device according to claim 1, characterized in that: The sliding block includes a nut which is threadedly mounted on the lead screw.

16. The nucleic acid detection device according to claim 1, characterized in that: The arrangement direction of the plurality of pipe placement grooves is parallel to the lead screw.

17. The nucleic acid detection device according to claim 1, characterized in that: The tube also includes an annular paraffin layer, which is arranged on the top of the reagent.

18. The nucleic acid detection device according to claim 1, characterized in that: The light emitting module further comprises a cover plate for accommodating a plurality of filters.

19. The nucleic acid detection device according to claim 1, characterized in that: The detection module includes a plurality of lens groups, a plurality of filters, and a plurality of light detectors.

20. The nucleic acid detection device according to claim 1, characterized in that: The invention also comprises a temperature control module, which comprises a thermoelectric cooling chip.

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