CN210916022U

CN210916022U - Nucleic acid extraction and amplification system and molecular detection device

Info

Publication number: CN210916022U
Application number: CN201920884347.5U
Authority: CN
Inventors: 不公告发明人
Original assignee: Guangzhou Zhixin Technology Co ltd
Current assignee: Du Yikong
Priority date: 2019-06-12
Filing date: 2019-06-12
Publication date: 2020-07-03
Anticipated expiration: 2029-06-12

Abstract

The utility model discloses a nucleic acid extraction and amplification system, which comprises a temperature control mechanism, a pushing mechanism, a magnetic attraction mechanism, a carrying unit and a control unit; one side of the carrying unit is provided with a temperature control mechanism for respectively controlling the temperature of the carrying bins, and the other side of the carrying unit is provided with a pushing mechanism; the pushing mechanism comprises a pushing extrusion mechanism and a pushing separation mechanism; the pushing and extruding mechanism is used for opening the breakable sealing layer to communicate the adjacent carrying bins; the pushing and blocking mechanism is used for blocking the fluid of the adjacent loading bins from moving in a mutual series manner; the pushing and extruding mechanism comprises a plurality of baffle plates; the pushing and blocking mechanism comprises a plurality of push plates. And a molecular detection device comprising a housing, an optical module and the nucleic acid extraction and amplification system; an accommodating chamber is arranged in the shell and used for the nucleic acid extraction and amplification system; the optical module is used for optically detecting the substances in the loading bin. The utility model has the advantages of totally closed, portable, integrated detection.

Description

Nucleic acid extraction and amplification system and molecular detection device

Technical Field

The utility model relates to a biological detection field, more specifically relates to a nucleic acid draws amplification system and detection device.

Background

Generally, molecular diagnostic techniques involve three steps of nucleic acid extraction, amplification and detection. The prior art tends to perform these three steps in steps. Firstly, the extraction process of nucleic acid has extremely high requirements on the collection, storage and transportation environments of samples in order to obtain accurate detection results. Then, nucleic acid is extracted by conventional extraction techniques including an ultrasonic method, a freeze-thaw method, a magnetic bead method, and the like. The magnetic bead method is commonly used at present and is generally carried out by a nucleic acid extractor. However, since the process required for nucleic acid extraction is complicated, a series of processes including lysis and purification of a sample takes a long time, and the extracted nucleic acid is easily cross-contaminated during the transfer to PCR. Finally, molecular detection is performed. The existing molecular diagnosis technology mainly depends on Polymerase Chain Reaction (PCR) to amplify nucleic acid molecules, and is mostly completed by a fluorescent quantitative PCR instrument. In the amplification process, various required data are obtained by a fluorescence quantitative method, qualitative or quantitative results are obtained by algorithm analysis, and the whole amplification and analysis process can be completed in hours. Meanwhile, the method has extremely high requirements on laboratories and operators. If the steps are to be completed, on one hand, the required site space is very large, the number of related devices is large, the possibility of cross contamination is increased, on the other hand, the requirements on operators are very high due to the fact that the devices are multiple, the steps are multiple, and the operation is complicated, and the actual molecular diagnosis work can be really performed after a large amount of time is spent on training and early practice before the post.

Such a detection process has a long period, is complex to operate, has high requirements on operators, greatly influences the detection efficiency, and has a lot of labor cost. Some prior arts have improved the aspects of simplified flow, reduced floor space, reduced requirements, etc. of molecular diagnosis, for example, patent document with application No. 201611199449.0, which is a mechanical device integrating nucleic acid extraction, amplification and detection, and adopts technical means such as sample cell, reagent bin, hybridization tank, motor, bar code reader, sampling needle, etc. to realize the technical scheme of integrating nucleic acid extraction, amplification and detection. Although the similar prior art realizes integration of molecular diagnosis, there still remains a problem of cross contamination such as contamination of reagents by moving reaction liquid using a sampling needle, waste liquid tank, and the like. Therefore, further improvement is still required in terms of integration of nucleic acid extraction, amplification and detection.

SUMMERY OF THE UTILITY MODEL

The utility model discloses it is complicated to extract nucleic acid and PCR amplification process among the prior art, requires high to operating personnel, and the treatment effeciency is low, and the easy technical problem who pollutes of sample provides a nucleic acid extraction amplification system and device.

The utility model discloses a main objective does: a nucleic acid extraction and amplification system is provided.

The utility model discloses another purpose is: provided is a molecular detection device for realizing integrated portable nucleic acid extraction and amplification and detecting amplified nucleic acid.

In order to solve the technical problems that the process of extracting nucleic acid and PCR amplification is complex in the prior art, the requirement on operators is high, the treatment efficiency is low, and the sample is easy to pollute the utility model discloses:

provides a nucleic acid extraction and amplification system, which comprises a temperature control mechanism, a pushing mechanism, a magnetic attraction mechanism, a carrying unit and a control unit. The carrying unit comprises a carrying cavity, wherein a plurality of breakable sealing layers are arranged in the carrying cavity so as to divide the carrying cavity into a plurality of mutually independent carrying bins which are connected in sequence. And a sealable sample injection port is arranged at the top end of the object carrying chamber. One side of the loading unit is provided with a temperature control mechanism for respectively controlling the temperature of the plurality of loading bins, and the other side of the loading unit is provided with a pushing mechanism. The pushing mechanism comprises a pushing extrusion mechanism and a pushing separation mechanism; the pushing and extruding mechanism is used for opening the breakable sealing layer to communicate the adjacent carrying bins; the pushing and blocking mechanism is used for blocking the fluid of the adjacent loading bins from moving in a mutual series mode. The pushing and extruding mechanism comprises a plurality of baffle plates; the pushing and blocking mechanism comprises a plurality of push plates; the baffle and the push plate are respectively connected with a first driving device. The baffle plate is close to the breakable sealing layer, so that fluids can be prevented from flowing in a mutual serial mode, and the push plate is close to the carrying bin and can break the breakable sealing layer in an extrusion mode to realize the flowing of reagents between the adjacent carrying bins; the extruded surface of the loading bin is of a flexible structure, and the extrusion of the push plate is facilitated. The magnetic attraction mechanism is used for attracting the magnetic beads in the object carrying bins and driving the magnetic beads to move to the corresponding positions of the object carrying bins. The temperature control mechanism, the pushing mechanism and the magnetic attraction mechanism are respectively and electrically connected with the control unit.

The general principle of the utility model is that different reaction reagents are divided into different reaction bins in advance through the destructible sealing layer, then reagent mixing of adjacent loading bins is realized through breaking the destructible sealing layer, the waste liquid after reaction is separated by the baffle plate, cross contamination caused by series flow is prevented, and meanwhile, the extraction of nucleic acid is carried out by utilizing a magnetic bead method; then, the nucleic acid is amplified. The whole system is completely closed, and the risk of cross contamination does not exist.

At first, the utility model discloses use and carry out processes such as sample treatment, nucleic acid extraction, amplification in year storehouse, with required reagent encapsulation in advance inside carrying the storehouse, combine pushing mechanism to get through the interval between the adjacent year storehouse, utilize temperature control mechanism to control each temperature that carries the storehouse respectively simultaneously, whole reaction sequence once only integrates and accomplishes, has reached the sample and has advanced, the effect that the result goes out. In addition, need not detect step by step like traditional equipment to save the intermediate process, very big improvement detection efficiency, whole process is 15 ~ 30 minutes.

The utility model discloses pushing mechanism, accuse temperature mechanism, magnetism inhale the mechanism and carry the thing unit and all can accomplish than little volume, so take up an area of very little. Compared with the traditional molecular diagnosis equipment, the device has the advantage of portability.

The utility model discloses well year storehouse can adopt the form of sealed lid to seal, and sealed lid can set up in the entrance, and its connected mode with the main part who carries the storehouse can be for screw thread locking, buckle connection, cap formula cooperation, adopts screw thread locking best. The structure not only plays a role in sealing the pre-loaded reagent in the storage and transportation processes, but also ensures that the whole reaction process is finished in the loading bin, thereby achieving the fully-closed effect. The main structure of the loading bin is made of flexible high polymer materials, the surface of the loading bin can resist the temperature of not less than 100 ℃, and meanwhile, the loading bin has certain elasticity and toughness, can have certain expansibility under specific conditions and cannot crack. The structure of the carrying unit comprises a bag or a long cylinder, and a bracket can be arranged outside the carrying unit and used for supporting the flexible carrying unit. The bracket can also be matched with a sheath for use, so that the leakage-proof performance of the carrying unit is further improved. The shape of the sheath comprises a polygonal sleeve shape, the wall thickness is more than or equal to 0.2mm, the sheath can be directly sleeved on the carrying unit, and the relative position of the sheath and the carrying unit is fixed in a tight fit or clamping manner. The carrier unit comprises a plurality of carrier bins, and the volume of each carrier bin is not more than 2 ml. The object carrying unit comprises a sample inlet made of high polymer materials, and is tightly matched with the sealing cover to prevent liquid leakage and air leakage. The sample inlet is communicated with the loading bin, and the connection form of the sample inlet comprises glue bonding and ultrasonic welding.

As a preferred scheme of the present invention, the magnetic attraction mechanism includes a magnetic attraction portion and a second driving device for driving the magnetic beads to move to each position of the loading bin; the magnetic part is close to one side of the object carrying bin, and the second driving device is used for realizing the movement of the magnetic beads in the processes of nucleic acid extraction and amplification.

As a more preferable scheme of the utility model, be equipped with the slider on the second drive arrangement, the slider is connected the portion is inhaled to magnetism.

The magnetic attraction part is used for fixing magnetic beads in the carrying bin and realizing the repeated movement of the reagent in the carrying bin by pushing the pushing and extruding mechanism; the magnetic part can be any object capable of generating magnetic force, preferably a magnet, and the shape of the magnet can be any polygon, preferably a hexahedron or a cylinder. The magnetic part is fixed on the sliding block and can be fixed on the sliding block in a buckling or bonding mode, and the magnetic part is not limited to the buckling or bonding fixing mode and can be fixed on the sliding block as long as the effect of the magnetic part can be achieved. When reagent is processed in the loading bins at different positions, the driving device drives the sliding block to move, and the fixation of magnetic beads in the loading bins at different positions is realized.

As the utility model discloses an optimized scheme, magnetism is inhaled the mechanism and is still including magnetism suction threaded rod, magnetism suction threaded rod threaded sleeve connects the slider, second drive arrangement is the motor, the motor is connected the threaded rod is inhaled to magnetism. A specific connection mode of the magnetic attraction mechanism is given, but the magnetic attraction mechanism is not limited to the combination of a motor and a threaded rod, and the movement and the positioning of the magnetic attraction part can be realized.

As a preferred embodiment of the present invention, the first driving device includes a motor, the baffle is connected to the motor, the push plate is connected to the motor, and the motor can separately control the baffle and the push plate; the baffle plate props against the breakable sealing layer, and the push plate props against the loading bin; the number of the baffle plates, the push plates and the motors corresponds to the number of the loading bins and the number of the breakable sealing layers. A conventional pushing means is shown here, but it is obvious that any means for achieving a connection to open adjacent bins, such as a crank connection or a threaded rod pushing means, is possible as long as the squeezed bins are achieved. The motor in the scheme can also be embodied as a direct current motor, a stepping motor and a servo motor, different motors can be installed according to needs, obviously, the effects of thrust, acceleration, deceleration and the like can be generated on the pushing mechanism, and the motors of positions or corners can be controlled according to needs. For the control effect, the number of motors is a plurality, preferably 21, which can be adapted to the use of a carrier unit with 11 carrier bins. The number of the baffle plates, the number of the push plates and the number of the loading bins are also corresponding. Baffle and push pedal drive mode, not limited to the threaded rod drive, can realize that the promotion of baffle makes it can block that reagent flows through from this position, can realize that the push pedal promotes the structure of the axial repetitive motion's of reagent in the reagent storehouse effect all can, for example connecting rod connection or guide bar connection or spring combination guide bar or threaded rod.

As a preferred aspect of the present invention, the baffle includes a first end and a bracket; the end part is an elastic end part, and the support is a hard support. The hard support may be made of a polymer material or a metal material, and obviously, a material having sufficient support strength is all possible. The soft end part can be made of high molecular elastic materials, including TPU and silica gel. In addition, the length of the end part can be manufactured according to the width of the loading bin, and preferably, the length range is 1-50 mm. Also, the ends need not all be soft, for example, in a configuration that wraps around the ends where the outer layer is an elastic material.

As the preferred scheme of the utility model, the push plate comprises a second end part, a push plate heating film and a push plate seat; the second end part is fixedly connected with a push plate heating film; the push plate heating film is fixedly connected with the push plate seat. The structure of the push plate is not limited to the connection mode that the second end is fixed on the push plate seat, and any connection mode that the second end can be fixed, so that the push plate can be pushed and has enough strength can be adopted. The shape of the second end portion comprises a hexahedron, a cylinder, preferably a hexahedron. Since the second end portion also has a function of conducting heat, the material thereof needs to be selected from a high temperature resistant or metal material, preferably a metal material.

As the preferred scheme of the utility model, the temperature control mechanism comprises a heating support plate, a heating element and a temperature sensor; the heating element and the temperature sensor are respectively and fixedly connected with the heating plate; the heating plate is fixedly connected to the heating support plate.

As the preferable scheme of the utility model, the heating element and the heating plate are fixedly connected in a clamping or bonding way; the temperature sensor and the heating plate are fixedly connected in a clamping or bonding mode. The heating plate can be made of stainless steel, aluminum alloy or copper with good heat conductivity, preferably brass. The number of heating plates can be customized according to requirements, and the best number is 10 or 11.

As a preferred embodiment of the present invention, the heating element is a heating film or a semiconductor or a combination of a heating film and a semiconductor. The number of the heating elements is at least 1, preferably 11, so that the better temperature control effect can be achieved. The shape of the heating element can be circular or any polygon, preferably quadrilateral.

As a preferred embodiment of the present invention, the semiconductor is a TEC semiconductor.

In order to achieve integrated portable nucleic acid extraction and amplification and detection of amplified nucleic acids, another object of the present invention is:

providing a molecular detection device comprising a housing, an optical module and a molecular detection system; an accommodating chamber is arranged in the shell and used for bearing the nucleic acid extraction and amplification system; the optical module is used for optically detecting the substances in the loading bin.

The utility model discloses a carry the thing storehouse and keep totally closed state in the processing procedure always, with external environment pollution-free, whole reaction sequence has realized totally closed. When in actual use, the whole reaction system is inside the equipment shell, no matter between the shell and the loading bin or between the shell and the external environment, no pollution is caused, and a good environment is provided for the accuracy of a sample detection result.

The utility model discloses for the complicated operation form and the system of traditional molecular diagnosis to reagent, equipment and each flow, only need 2 to 3 minutes's artifical application of sample, all the other processes are accomplished by the system. The requirement on the operators is reduced, and long-term complex training on the operators is not needed.

The utility model discloses have the advantage of no place restriction, the concrete performance does: because the system is small and flexible and is convenient to carry, compared with the traditional molecular diagnosis which can only be carried out in a relevant laboratory, the system allows an operator to work in other places except the laboratory according to the requirements of test projects.

The utility model discloses have the advantage that expansibility is good, the concrete manifestation is in: the flexibility of the system determines that the same equipment can correspond to different detection items, so that the system has great advantages in openness. The platform is not only suitable for quick detection in clinical departments and inspection centers of hospitals, but also suitable for scientific research institutions or related personnel in the field of molecular detection, and self detection projects are developed on the platform.

As a preferred embodiment of the present invention, the optical module is disposed on the peripheral side of the loading bin.

As the preferred scheme of the utility model, the heating region with correspond the position be equipped with the through-hole on the heating element, one side of through-hole is equipped with optical module.

As the utility model discloses a preferred scheme, the holding chamber is for can sealing the cavity, the further leakproofness that has improved the molecule processing environment, and the main objective is in order to prevent that the sample from receiving external interference, improves and detects the success rate.

As a preferred scheme of the utility model, the accommodating chamber is sealed by a sealing cover; the seal cover is arranged on the shell.

As a preferred aspect of the present invention, the cover is hinged or slidably connected to the housing.

As a preferable embodiment of the present invention, the through hole has a circular shape or a polygonal shape.

As a preferred embodiment of the present invention, the number of the through holes is at least 1.

As a preferred embodiment of the present invention, the optical module includes an excitation light device and an optical detection device. The excitation light device comprises a light-emitting element, a lens, an excitation filter and an excitation dichroic mirror. The lens is an optional element for converging the exciting light, the quantity of the lens is determined by the light intensity of the exciting light source, and if the light intensity is sufficiently converged, the lens can be selected not to be used. The excitation filter is used for filtering light waves in unnecessary wave bands in the excitation light, the number of the excitation filter is at least 1, and the specific number is determined according to the excitation light source. The wave bands of the excitation filters correspond to the wave bands of the excitation light sources one to one. The shape of the excitation filter comprises a cylinder and a hexahedron, and the sizes of the excitation filter can be different according to the size of the light path. The excitation dichroic mirror is used for exciting the transmission of light with a specific waveband in a light path according to different reflection and transmission wavebands of the excitation light sources so as to irradiate an object to be detected; the shape of the optical fiber comprises a cylinder and a hexahedron, and the size of the optical fiber is set according to the size of an optical path; types include mirrors, dichroic trichroic mirrors, dichroic polychromatic mirrors. The optical detection device comprises a detection element, a lens, a detection optical filter and a detection dichroic mirror.

As a preferable scheme of the utility model, the exciting light device is an LED lamp, a black light or an incandescent lamp. Wherein the number of LED lamps is at least 1, preferably 2 LED lamps corresponding to different wave bands.

As a preferable scheme of the present invention, the wavelength bands of the LED lamp include FAX, HEX, ROX, and CY 5.

As a preferred embodiment of the present invention, the optical detection device includes a photodiode PD or a photomultiplier tube PMT.

As a preferred embodiment of the present invention, the molecule detecting device further includes a display and a power supply disposed on the housing. The display, which may be a touch screen. The connection form of the power supply and the equipment comprises the built-in machine or the independent external connection.

As the utility model discloses an optimal scheme, the molecule detection device still includes the bar code scanner, the bar code scanner set up in on the lateral wall of casing, carry be equipped with on the thing unit with bar code scanner assorted sweeps a yard region. The thing unit of carrying can add and establish the protective sheath, if add and establish the protective sheath, sweep the corresponding setting in code region and be in on the protective sheath.

As the utility model discloses a preferred scheme, sweep the code area and set up two-dimensional code or bar code.

The control unit in the utility model comprises a main control module, a temperature control module, a liquid processing module, an optical module, a display module and a power supply module; the temperature control module, the liquid processing module, the optical module, the display module and the power supply module are respectively and electrically connected with the main control module; the temperature control module is electrically connected with the temperature control mechanism; the optical module comprises an exciting light device and an optical detection device which are electrically connected; the liquid treatment module is electrically connected with the pushing mechanism; the liquid processing module is electrically connected with the magnetic attraction mechanism; the display module is electrically connected with the display; the power supply module is electrically connected with a power supply; the code scanning module is electrically connected with the code scanner.

As the preferred scheme of the utility model, the nucleic acid detection control system also comprises a code scanning module; sweep a yard module electric connection master control module. The communication module comprises any one or more combined common communication modes such as WiFi, Bluetooth, internet access, GPS and the like, and can be selected according to requirements.

As a preferred embodiment of the present invention, the nucleic acid detection control system further comprises a communication module; the communication module is electrically connected with the main control module.

The molecular detection device of the utility model can be used for the molecular diagnosis of the field including clinical departments or ICU or emergency treatment or disease control center or food safety or scientific research or criminal investigation or family or military affairs or drugs or customs inspection and quarantine or pets or aquatic products or water quality or forestry or animal husbandry.

The applicable reagent of the nucleic acid extraction and amplification system of the utility model comprises a dry or liquid reagent. The kit specifically comprises lysis solution, proteinase k, buffer solution and PCR premix solution.

The reagent of the utility model is suitable for preservation or transportation at the temperature of-20 ℃ to 30 ℃.

Compared with the prior art, the beneficial effects of the utility model are that:

the utility model discloses use and carry out processes such as sample treatment, nucleic acid extraction, amplification in year storehouse, with required reagent encapsulation in advance inside carrying the thing storehouse, combine pushing mechanism to get through the interval between the adjacent thing storehouse of carrying, utilize temperature control mechanism to control each temperature that carries the thing storehouse respectively simultaneously, whole reaction sequence once only integrates and accomplishes, has reached the sample and has advanced, the effect that the result goes out. In addition, need not detect step by step like traditional equipment to save the intermediate process, very big improvement detection efficiency, whole process is 15 ~ 30 minutes.

The utility model discloses pushing mechanism, accuse temperature mechanism, magnetism inhale mechanism and carry the thing unit and all can accomplish than little volume, so, take up an area of very little, for example be 170mm × 200mm × 230mm in the geometric dimensions, weight is less than or equal to 4kg the casing in can accomplish the whole processes that detect.

Drawings

FIG. 1 is a schematic diagram of the front view structure of the nucleic acid extraction and amplification system of the present invention.

FIG. 2 is a schematic diagram of a side view of the nucleic acid amplification system of the present invention.

FIG. 3 is a schematic diagram of a side view cross-sectional structure of the nucleic acid extraction and amplification system of the present invention.

Fig. 4 is a schematic view of a part a of the enlarged structure in fig. 3.

FIG. 5 is a schematic top view of the nucleic acid amplification system of the present invention.

FIG. 6 is a perspective view of the nucleic acid amplification system of the present invention.

FIG. 7 is a schematic view of the protective sheath of the nucleic acid amplification system of the present invention.

Fig. 8 is a schematic structural view of the totally-enclosed integrated portable molecular detection device of the present invention.

Fig. 9 is a schematic longitudinal sectional structure view of the totally-enclosed integrated portable molecular detection device of the present invention.

Fig. 10 is a schematic perspective view of the totally-enclosed integrated portable molecular detection device of the present invention.

Fig. 11 is a schematic structural diagram of an optical module.

Fig. 12 is a schematic diagram of an optical module.

Fig. 13 is a schematic structural view of the molecular detection system of the present invention.

In the figure: 1. a housing; 11. a display; 12. a power source; 13. a code scanner; 2. an accommodating chamber; 21. sealing the cover; 3. a carrying unit; 31. a carrier bin 32, a breakable seal; 33. scanning a code area; 34. a support; 35. a protective sleeve; 4. a temperature control mechanism; 41. heating the support plate; 42. heating plates; 43. a heating element; 44. A temperature sensor; 45. a through hole; 5. a pushing mechanism; 51. a baffle plate; 511. a first end portion; 512. a support; 52. pushing the plate; 521. a second end portion; 522. pushing the plate to heat the membrane; 523. a push plate seat; 53. a first driving device; 54. pushing the threaded rod; 6. a magnetic attraction mechanism; 61. a magnetic part; 62. a slider; 63. a second driving device; 64. magnetically attracting the threaded rod; 7. an optical module; 71. an excitation light device; 72. an optical detection device; 7111. a light emitting element; 7112. a lens; 7113. exciting the optical filter; 7114. exciting a dichroic mirror; 7221. a detection element; 7222. a lens; 7223. detecting the optical filter; 7224. and detecting the dichroic mirror.

Detailed Description

The present invention will be further described with reference to the following embodiments.

The same or similar reference numerals in the drawings of the embodiments of the present invention correspond to the same or similar parts; in the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "left", "right", "top", "bottom", "inner", "outer", etc., indicating orientations or positional relationships based on the orientations or positional relationships shown in the drawings, the description is merely for convenience and simplicity of description, and it is not intended to indicate or imply that the devices or elements being referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore, the terms describing the positional relationships in the drawings are for illustrative purposes only and are not to be construed as limiting the present patent.

Furthermore, if the terms "first," "second," and the like are used for descriptive purposes only, they are used for mainly distinguishing different devices, elements or components (the specific types and configurations may be the same or different), and they are not used for indicating or implying relative importance or quantity among the devices, elements or components, but are not to be construed as indicating or implying relative importance.

Example 1

As shown in FIGS. 1-6, the present embodiment provides a nucleic acid extraction and amplification system, which includes a temperature control mechanism 4, a pushing mechanism 5, a magnetic attraction mechanism 6, a carrying unit 3, and a control unit. The loading unit 3 comprises a loading chamber, and a plurality of breakable sealing layers 32 are arranged in the loading chamber so as to divide the loading chamber into a plurality of loading bins 31 which are connected in sequence and are mutually independent. The top end of the object carrying chamber is provided with a sealable sample injection port. One side of the loading unit 3 is a temperature control mechanism 4 for respectively controlling the temperature of the plurality of loading bins 31, and the other side is a pushing mechanism 5. The pushing mechanism 5 comprises a pushing extrusion mechanism and a pushing separation mechanism; the pushing extrusion mechanism is used for opening the breakable sealing layer to communicate the adjacent carrying bin 31; the pushing blocking mechanism is used for blocking the fluid of the adjacent loading bins 31 from moving in series. The pushing and extruding mechanism comprises a plurality of baffle plates 51; the pushing barrier mechanism comprises a plurality of push plates 52; the baffle 51 and the push plate 52 are respectively connected with a first driving device 53. The baffle plate 51 is positioned close to the breakable sealing layer 32 to prevent the fluids from flowing in series, and the push plate 52 is positioned close to the carrier bins 31 to break the breakable sealing layer 32 in an extrusion manner to realize the flow of the reagent between the adjacent carrier bins 31; the pressed surface of the loading bin 31 is of a flexible structure, so that the pressing of the push plate 52 is facilitated. The magnetic attraction mechanism 6 is used for attracting magnetic beads in the loading bins 31 and driving the magnetic beads to move to corresponding positions of the loading bins 31. The temperature control mechanism 4, the pushing mechanism 5 and the magnetic attraction mechanism 6 are respectively electrically connected with the control unit.

As a preferred embodiment of the present invention, the magnetic attraction mechanism 6 includes a magnetic attraction portion 61 and a second driving device 63 for driving the magnetic beads to move to the positions of the loading bins 31; the magnetic part 61 is close to one side of the loading chamber 31, and the second driving device 63 is used for moving the magnetic beads in the process of extracting and amplifying the nucleic acid. The second driving device 63 is provided with a slide block 62, and the slide block 62 is connected with the magnetic suction part 61. The magnetic part 61 is used for fixing magnetic beads in the loading bin 31 and realizing the reciprocating motion of the reagent in the loading bin 31 by the pushing of the pushing and squeezing mechanism; the magnetic attraction portion 61 is an object capable of generating a magnetic force, such as a magnet. The shape of the magnet can be any polygon, and preferably, the magnet can be a hexahedron or a cylinder. The magnetic part 61 is fixed on the slider 62, and can be fixed on the slider 62 by a snap or adhesive manner, and is not limited to the snap or adhesive manner, as long as the effect of fixing the magnetic part 61 on the slider 62 can be achieved. When the reagent is processed in the loading bins 31 at different positions, the second driving device 63 drives the sliding block 62 to move, so that the magnetic beads in the loading bins 31 at different positions are fixed. The magnetic attraction mechanism 6 further comprises a magnetic attraction threaded rod 64, the magnetic attraction threaded rod 64 is in threaded sleeve connection with the sliding block 62, the second driving device 63 is a motor, and the motor is connected with the magnetic attraction threaded rod 64. A specific connection mode of the magnetic attraction mechanism 6 is given here, but the magnetic attraction mechanism is not limited to a combination of a motor and a threaded rod, and the movement and positioning of the magnetic attraction part can be realized.

The first driving means 53 includes a motor which is connected to the shutter 51 and the push plate 52 and is capable of individually controlling the shutter 51 and the push plate 52, respectively. The baffle plate 51 is pressed against the breakable sealing layer 32, and the push plate 52 is pressed against the loading bin 31; the number of baffles 51, push plates 52 and motors corresponds to the number of bins 31 and rupturable seal 32. A conventional pushing means is shown here, but it is obvious that any means for achieving a connection opening through adjacent magazines 31 is possible, such as a crank connection or a threaded rod pushing means, as long as the squeezed magazines are reached. The motor in the scheme can also be embodied as a direct current motor, a stepping motor and a servo motor, different motors can be installed according to needs, obviously, the effects of thrust, acceleration, deceleration and the like can be generated on the pushing mechanism, and the motors of positions or corners can be controlled according to needs. For the control effect, the number of motors is a plurality, preferably 21, which can be adapted to the use of a carrier unit with 11 carrier bins. The number of the baffle plates 51 and the number of the push plates 52 correspond to the number of the carrier bins 31. The driving mode of the baffle 51 and the push plate 52 is not limited to the driving by a threaded rod, and the structure capable of pushing the baffle 51 to prevent the reagent from flowing through the position and pushing the push plate 52 to push the reagent in the reagent bin to move axially and repeatedly is all possible, for example, a connecting rod connection or a guide rod connection or a spring combination guide rod or a threaded rod.

The baffle 51 includes a first end 511 and a bracket 512; the first end 511 is an elastic end and the bracket 512 is a rigid bracket. The hard support may be made of a polymer material or a metal material, and obviously, a material having sufficient support strength is all possible. The elastic end part can be made of high polymer elastic materials including TPU and silica gel. In addition, the length of the elastic end part can be made according to the width of the loading bin, and preferably, the length range is 1-50 mm. Also, the first end portion need not be entirely soft, such as by being wrapped around the end portion where the outer layer is an elastomeric material. The push plate 52 includes a second end 521, a push plate heating film 522, and a push plate seat 523; the second end 521 is fixedly connected with the push plate heating film 522; the push plate heating film 522 is fixedly connected to the push plate holder 523. The structure of the push plate 52 is not limited to the connection mode that the second end 521 is fixed on the push plate seat 523, and any connection mode that can fix the second end 521 so that it can be pushed and has sufficient strength may be used. The shape of the second end 521 includes a hexahedron, a cylinder, and preferably a hexahedron. Since the second end portion also has a function of conducting heat, the material thereof needs to be selected from a high temperature resistant or metal material, preferably a metal material.

The temperature control mechanism 4 comprises a heating support plate 41, a heating plate 42, a heating element 43 and a temperature sensor 44; the heating element 43 and the temperature sensor 44 are respectively fixedly connected with the heating plate 42; the heating plate 42 is fixedly attached to the heating support plate 41. The heating element 43 is fixedly connected with the heating plate 42 in a clamping or bonding mode; the temperature sensor 44 is fixedly connected with the heating plate 42 by clamping or bonding. The heating plate 42 may be made of stainless steel, aluminum alloy or copper, preferably brass. The number of heating plates 42 can be customized according to the requirements, and the best number is 10 or 11. The heating element 43 is a heating film or a semiconductor or a combination of a heating film and a semiconductor. The number of the heating elements 43 is at least 1, preferably 11, which can achieve better temperature control effect. The shape of the heating element 43 may be circular or any polygon, preferably a quadrangle. The semiconductor is a TEC semiconductor.

The control unit of the embodiment shown in fig. 10-12 includes a main control module, a temperature control module, a liquid processing module, an optical module, a display module, and a power module; the temperature control module, the liquid processing module, the optical module, the display module and the power supply module are respectively and electrically connected with the main control module; the temperature control module is electrically connected with the temperature control mechanism; the optical module comprises an exciting light device and an optical detection device which are electrically connected; the liquid processing module is electrically connected with the pushing mechanism; the liquid processing module is electrically connected with the magnetic attraction mechanism; the display module is electrically connected with the display; the power module is electrically connected with the power supply. The code scanning module is electrically connected with the main control module and can read two-dimensional codes or bar codes, and the two-dimensional codes or the bar codes record corresponding detection item information. The communication module is electrically connected with the main control module, and the communication module comprises any one or more combined common communication modes such as WiFi, Bluetooth, a network port and GPS, and can be selected according to requirements.

Example 2

As shown in fig. 7 to 10, the molecular detection device in the present embodiment includes a housing 1, an optical module 7, and a molecular detection system; the housing 1 is internally provided with an accommodating chamber 2, and the accommodating chamber 2 is used for bearing the nucleic acid extraction and amplification system; the optical module 7 is used for optically detecting the substance in the loading bin 31.

The holding chamber 2 is a sealable chamber, further improves the sealing property of the molecular processing environment, and the main purpose is to prevent the sample from being interfered by the outside world and improve the detection success rate. The accommodating chamber 2 is sealed by a sealing cover 21; the cover 21 is provided on the housing 1. Preferably, the cover 21 is hingedly or slidably connected to the housing. Preferably, the housing 1 is further provided with a display 11, which may be a touch screen. The form of connection of the power source 12 to the device may include an internal connection to the machine or a separate external connection. Furthermore, a code scanner 13 is arranged on the housing 1, a code scanning area 33 matched with the code scanner is arranged on the carrier unit 3, the code scanning area 33 can also be arranged on the protective sleeve 35, and as a preferred embodiment, the code scanning area 33 is arranged on the protective sleeve 35. The code scanning area 33 is provided with a two-dimensional code or a bar code. In this embodiment, the code scanning area 33 is provided with a two-dimensional code. The shape of the through hole 45 is any one of circular or polygonal; the number of through holes 45 is at least 1. The heating element 43 is a heating film or a semiconductor or a combination of a heating film and a semiconductor. In this embodiment, the semiconductor is a TEC semiconductor.

The optical module 7 comprises an excitation light device 71 and an optical detection device 72. The excitation light device 71 includes a light emitting element 7111, a lens 7112, an excitation filter 7113, and an excitation dichroic mirror 7114. The lens 7112 is an optional element for converging the exciting light, the amount of which is determined by the intensity of the exciting light source, and if the intensity is sufficiently converged, the lens is not used. The excitation filter 7113 is used to filter the light wave of unnecessary wavelength band in the excitation light, the number is at least 1, and the specific number is determined according to the excitation light source. The wave bands of the excitation filters correspond to the wave bands of the excitation light sources one to one. The shape of the excitation filter comprises a cylinder and a hexahedron, and the sizes of the excitation filter can be different according to the size of the light path. An excitation dichroic mirror 7114, whose reflection and transmission wave bands are different according to the wave bands of each excitation light source, is used for transmitting the light with specific wave bands in the excitation light path, so that the light irradiates the object to be measured; the shape of the optical fiber comprises a cylinder and a hexahedron, and the size of the optical fiber is set according to the size of an optical path; types include mirrors, dichroic trichroic mirrors, dichroic polychromatic mirrors. The optical detection device includes a detection element 7221, a lens 7222, a detection filter 7223, and a detection dichroic mirror 7224. According to the totally-enclosed integrated portable molecular detection device, the exciting light device is an LED lamp or a black light or an incandescent lamp. Wherein the number of LED lamps is at least 1, preferably 2 LED lamps corresponding to different wave bands. The wavelength bands of the LED lamp include FAX, HEX, ROX and CY 5. The optical detection means comprise a photodiode PD or a photomultiplier tube PMT. Both of these elements are preferably photodiodes PD.

The utility model discloses a concrete working method does:

taking the extraction of nucleic acid from a blood sample and performing PCR detection as an example:

various reaction reagents pre-packaged in the loading bin; the carrying unit 3 comprises 10 sections of carrying bins, and each section is represented by the number 31- (1-10); the length of the loading bin 31-1 is longer and is about 2 times of the length of each other loading bin, and the lengths of the other loading bins are equal; the bins are separated by breakable seals 32.

Each carrying bin contains a pre-packaged reagent: 31-1 is a sample receiving area which can receive a blood sample; 31-2, encapsulating 80 mu L of buffer solution; 31-3, encapsulating 80 mu L of internal control liquid; 31-4 encapsulating 80 μ L proteinase k; 31-5, encapsulating 80 mu L of magnetic particles; 31-6, packaging 80 mu L of lysate; 31-7, packaging 80 mu L of washing solution; packing eluent 80 μ L in 31-8; 31-9 packaging 50 mu L of PCR premix 1; 31-10, 50. mu.L of PCR premix 2 was packed.

The utility model discloses contained extraction, purification and PCR reaction and the analysis to nucleic acid to integrate all steps in the nucleic acid extraction amplification system of integration, complete closure processing is realized to whole process, makes the molecular diagnosis environment can not be contaminated.

The method comprises the following specific steps:

the samples were loaded manually. The sealing cap of the loading unit is unscrewed, and 50. mu.L of blood is added to the reagent reservoir 31-1 by using a pipette or a pipette tip. The sealing cover is screwed to ensure the sealing performance of the chip;

and starting the computer. Pressing the power source 12 of the housing 1 to start the device;

and scanning the code and inputting information. The code scanning area 33 on the carrier unit 3 is aligned with the code scanner 13 on the shell 1, and after code scanning, the detection system records the detection item information corresponding to the chip. At this time, the baffle 51 and the push plate 52 in the liquid processing system are both reset to the initial position, and a space is left for allowing the carrying unit 3 to be completely inserted from the accommodating chamber 2 to the bottom of the inner side of the device;

inserted into the carrier unit. Opening the sealing cover 21, inserting the object carrying unit 3 into the accommodating chamber 2, and pressing a start key of a screen to start a full-automatic processing and analyzing process after closing the sealing cover 21;

and (4) cracking the sample. After the chip is inserted, the baffles 31- (1-2) positioned at the first two positions are positioned at the initial position, and the other baffles move leftwards to press the corresponding breakable sealing layer 32 to close the reagent bins 31- (2-10) at all sections. Then, (the shutter 51 and the push plate 52 both move closer to or away from the carrier unit 3 along the x-axis, and in this embodiment, the shutter 51 or the push plate 52 moves closer to the carrier unit 3 along the x-axis when the shutter 51 or the push plate 52 is described to be closed, and the shutter 51 or the push plate 52 moves away from the carrier unit 3 along the x-axis when the shutter 51 or the push plate 52 is described to be opened), the shutter 51-1 in the first position and the push plate 52-1 in the first position are closed, and the shutter 51-3 in the third position is opened. The push plate 52-2 in the second position is then closed, causing the rupturable seal 32 in the first position to open under pressure, thereby allowing the sample in the carrier 31-1 to flow into the carrier 31-2. Then, the second and third push plates 52- (2-3) are alternately pushed in and pulled out, so that the sample and the buffer solution are mixed back and forth in the loading chamber 31-1 and the loading chamber 31-2. After mixing for a plurality of times, pulling out the second push plate 52- (2-3), opening the baffle 51-4 at the fourth position, pushing the fourth push plate 52-4, destroying the breakable sealing layer 32 between the loading bin 31-3 and the loading bin 31-2, and leading the internal control liquid in the loading bin 31-3 to flow into the loading bin 31-2. Then the second, third and fourth push plates 52- (2, 3, 4) are alternately extruded to fully and evenly mix the sample, the buffer solution and the internal control solution. The heating area from the loading bin 31-1 to the loading bin 31-7 is heated to 56 ℃. And opening the loading bin 31-4, the loading bin 31-5 and the loading bin 31-6 in the same way to fully and uniformly mix the sample, the buffer solution, the internal control solution, the proteinase k, the magnetic particles and the lysis solution to form a mixed solution. Extruding the mixed solution to a loading bin 31-4, a loading bin 31-5 and a loading bin 31-6, pushing a baffle plate 51-5 at a fifth position, and alternately extruding push plates 52- (5, 6 and 7) at fifth, sixth and seventh positions to ensure that the mixed solution can only flow in the loading bin 31-4, the loading bin 31-5 and the loading bin 31-6, and culturing for 3 minutes at the temperature of 56 ℃;

and (4) collecting nucleic acid. The magnet is moved to the area corresponding to the loading bin 31-5, the push plates 52- (5, 6, 7) are alternately extruded to enable the mixed liquid to alternately flow in the area of the loading bin 31- (4, 5, 6), and when the mixed liquid passes through the loading bin 31-6, the magnetic particles are gradually gathered in the area after multiple cycles due to the action of the magnetic field generated by the magnet. At this time, the fifth baffle 51-5 is opened, the push plates 52- (7, 6, 5, 4, 3) at the (7, 6, 5, 4, 3) th positions are sequentially pressed, waste liquid is squeezed into the loading bin 31-1, and the baffle 51-3 at the third position is closed, so that the waste liquid does not flow out downwards. At this point, the magnetic particles remain in the loading chamber 31-5 and the nucleic acid has bound to the magnetic particles;

and (3) washing the nucleic acid. The fifth shutter 51-5 is closed, and the magnet 61 is moved to the region other than the loading chamber 31- (4, 5, 6), thereby canceling the magnetic field in the above region. Then, the eighth baffle 51-8 is opened, and the eighth push plate 52-8 is squeezed, so that the washing liquid in the loading bin 31-7 is squeezed into the loading bin 31-6. Then the eight baffles 51 are closed, and the push plates 52- (7, 6, 5) at the (7, 6, 5) th positions are sequentially pressed, so that the magnetic particles are washed back and forth by the washing liquid. Magnet 61 is then moved to the region of carrier 31-5, as described above, to collect the magnetic particles in the region of carrier 31-5 during the washing process. The waste liquid is then transferred to the loading chamber 31-1 and the shutter 51-3 in the third position is closed, as described above, so that the waste liquid does not flow downwards. At this time, the residues and inhibitors, etc. other than the nucleic acid, on the magnetic particles have been removed;

and (4) eluting the nucleic acid. The fifth shutter 51-5 is closed to hold the magnet 61 in the position of the loading compartment 31-5. Then, the ninth baffle 51-9 is opened, the ninth pushing plate 52-9 is pressed, and the eluent in the area of the loading bin 31-8 is delivered to the area of the loading bin 31-5. Then, pushing an eighth baffle plate 51-8, adjusting the temperature of a corresponding heating plate in the loading bin 31-5 to 70 ℃, repeatedly extruding a push plate 52- (7, 6, 5) in sequence, and enabling the eluent to repeatedly wash the magnetic particles in the loading bin 31-5 from the loading bin 31-4 to the loading bin 31-6 so as to elute the nucleic acid from the magnetic particles;

and (3) amplifying nucleic acid and analyzing results. The baffles 51- (8, 9, 10) at the (8, 9, 10) th positions are opened in sequence, and after the eluent is squeezed to the loading bin 31-8, the baffles 51-9 at the ninth position are closed. Then, the shutter 51-11 at the eleventh position is opened, the pusher 52- (9, 10, 11) at the (9, 10, 11) position is alternately pressed to mix the eluent with the

PCR premix

1,2, and the mixing operation is repeated for 2 min. Then, the temperature of the heating area corresponding to the loading bin 31-8 is adjusted to 95 ℃, the temperature of the heating area corresponding to the loading bin 31-9 is adjusted to 72 ℃, and the temperature of the heating area corresponding to the loading bin 31-10 is adjusted to 55 ℃. And extruding the final mixed liquor to a loading bin 31-8 for 5 seconds, extruding the final mixed liquor to a loading bin 31-10 for 8 seconds, and extruding the final mixed liquor to a loading bin 31-9 for 11 seconds. The above 3 temperature zones and holding times were used as one cycle, i.e., 95 ℃ for 5 seconds, 55 ℃ for 8 seconds, and 72 ℃ for 11 seconds. And fluorescence is detected by the optical module corresponding to the optical detection device 72 during the last 5s of each cycle. The excitation light is emitted from the optical module 14 40 times under the control of the main control module for a total of 40 cycles, so as to obtain 40 sets of fluorescence data, and the schematic diagrams are shown in fig. 11 and 12. And performing curve fitting and algorithm operation on the obtained data to obtain results including but not limited to an amplification curve, a dissolution curve, negative and positive judgment, sample concentration quantification and the like.

And (4) waste treatment. The cover 21 is opened, the reacted carrier unit 3 is taken out, and the used disposable carrier unit 3 is disposed according to the disposal clause of the wastes in the relevant industry.

It is obvious that the above embodiments of the present invention are only examples for clearly illustrating the present invention, and are not limitations to the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A nucleic acid extraction and amplification system is characterized by comprising a temperature control mechanism, a pushing mechanism, a magnetic attraction mechanism, a carrying unit and a control unit;

the carrying unit comprises a carrying cavity, and a plurality of breakable sealing layers are arranged in the carrying cavity so as to divide the carrying cavity into a plurality of carrying bins which are connected in sequence and are mutually independent; the top end of the object carrying chamber is provided with a sealable sample injection port;

one side of the carrying unit is provided with a temperature control mechanism for respectively controlling the temperature of the carrying bins, and the other side of the carrying unit is provided with a pushing mechanism; the pushing mechanism comprises a pushing extrusion mechanism and a pushing separation mechanism; the pushing and extruding mechanism is used for opening the breakable sealing layer to communicate the adjacent carrying bins; the pushing and blocking mechanism is used for blocking the fluid of the adjacent loading bins from moving in a mutual series manner;

the pushing and extruding mechanism comprises a plurality of baffle plates; the pushing and blocking mechanism comprises a plurality of push plates; the baffle and the push plate are respectively connected with a first driving device; the baffle plate is close to the breakable sealing layer, so that fluids can be prevented from flowing in a mutual serial mode, and the push plate is close to the carrying bin and can break the breakable sealing layer in an extrusion mode to realize the flowing of reagents between the adjacent carrying bins; the extruded surface of the loading bin is of a flexible structure, so that the extrusion of the push plate is facilitated;

the magnetic attraction mechanism is used for attracting the magnetic beads in the carrying bins and driving the magnetic beads to move to corresponding positions of the carrying bins;

the temperature control mechanism, the pushing mechanism and the magnetic attraction mechanism are respectively and electrically connected with the control unit.

2. The nucleic acid extraction and amplification system of claim 1, wherein the magnetic attraction mechanism comprises a magnetic attraction portion and a second driving device for driving the magnetic beads to move to each of the bin positions; the magnetic part is close to one side of the object carrying bin, and the second driving device is used for realizing the movement of the magnetic beads in the processes of nucleic acid extraction and amplification.

3. The nucleic acid extraction and amplification system of claim 2, wherein the second driving device is provided with a slider, and the slider is connected to the magnetic attraction part.

4. The nucleic acid extraction and amplification system of claim 3, wherein the magnetic attraction mechanism further comprises a magnetic attraction threaded rod in threaded connection with the slider, and the second driving device comprises a plurality of motors connected to the magnetic attraction threaded rod.

5. The nucleic acid extraction and amplification system of claim 1, wherein the first driving device comprises a plurality of motors, the baffle is connected to the motors, and the push plate is connected to the motors, wherein the motors are capable of controlling the baffle and the push plate separately; the number of the baffle plates, the push plates and the motors corresponds to the number of the loading bins and the number of the breakable sealing layers.

6. The nucleic acid extraction amplification system of claim 1, wherein the baffle comprises a first end and a bracket.

7. The nucleic acid extraction and amplification system of claim 6, wherein the first end is a flexible end.

8. The nucleic acid extraction and amplification system of claim 1, wherein the push plate comprises a second end, a push plate heating membrane, a temperature sensor, and a push plate seat; the second end part is fixedly connected with a push plate heating film; the push plate heating film is fixedly connected with the push plate seat; the temperature sensor is fixedly connected with the push plate seat.

9. The nucleic acid extraction and amplification system of claim 1, wherein the temperature control mechanism comprises a heating support plate, a heating plate, a plurality of heating elements, and a plurality of temperature sensors; the heating element and the temperature sensor are respectively and fixedly connected with the heating plate; the heating plate is fixedly connected to the heating support plate; the heating element is positioned on one side of the loading bin.

10. The nucleic acid extraction and amplification system of claim 9, wherein the heating element is fixedly connected to the heating plate by clamping or bonding; the temperature sensor and the heating plate are fixedly connected in a clamping or bonding mode.

11. The nucleic acid extraction and amplification system of claim 9, wherein the heating element is a heating film or a semiconductor or a combination of a heating film and a semiconductor.

12. The nucleic acid extraction amplification system of claim 11, wherein the semiconductor is a TEC semiconductor.

13. A molecular detection device comprising a housing, an optical module, and the nucleic acid extraction and amplification system according to any one of claims 9 to 12;

an accommodating chamber is arranged in the shell and used for the nucleic acid extraction and amplification system;

the optical module is used for optically detecting the substances in the loading bin.

14. The molecular detection device of claim 13, wherein the optical module is disposed on a peripheral side of the carrier.

15. The molecular detection device of claim 14, wherein the heating element is provided with a through hole, and one side of the through hole is provided with the optical module.

16. The molecular testing device of claim 13, wherein the containment chamber is sealed by a lid; the seal cover is arranged on the shell.

17. The molecular testing device of claim 16, wherein the cover is hingedly or slidably coupled to the housing.

18. The molecular detection device according to claim 15, wherein the shape of the through-hole is any one of circular and polygonal.

19. The molecular testing device of claim 15, wherein the number of through-holes is at least 1.

20. The molecular detection device of claim 13, wherein the optical module comprises an excitation light device and an optical detection device.

21. The molecular detection device of claim 13, further comprising a code scanner disposed on the sidewall of the housing, wherein the carrier unit has a code scanning area matching the code scanner.

22. The molecular detection device of claim 21, wherein the code scanning area is provided with a two-dimensional code or a bar code.