CN114774273A - Portable nucleic acid detector and nucleic acid detection method - Google Patents
Portable nucleic acid detector and nucleic acid detection method Download PDFInfo
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- CN114774273A CN114774273A CN202210431366.9A CN202210431366A CN114774273A CN 114774273 A CN114774273 A CN 114774273A CN 202210431366 A CN202210431366 A CN 202210431366A CN 114774273 A CN114774273 A CN 114774273A
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Abstract
The present invention provides a portable nucleic acid detecting instrument, comprising: the card box comprises a kit, a slide block and a reaction tube for amplification reaction; the reagent kit is provided with a plurality of cavities, a plurality of channels and a liquid outlet; the bottom of a cavity is communicated with a channel; the top end of the reaction tube is communicated with the liquid outlet; the slide block is connected with the reagent box in a sliding way; the sliding block is provided with a transition cavity and an injector structure; the injector structure comprises an outer cylinder and a piston; the displacement mechanism is used for driving the sliding block to slide relative to the reagent box; the piston driving mechanism comprises a first driving piece and a push-pull rod which are connected; the push-pull rod is inserted into the outer cylinder and clamped on the piston; the first driving piece is used for driving the push-pull rod to drive the piston to move in the outer cylinder so as to draw or push the transition cavity, so that the card box can extract nucleic acid. The invention also provides a nucleic acid detection method. The detector body omits a mechanical arm liquid-transferring structure, a series of nucleic acid extraction operations are completed inside the card box, and the whole structure is simple, small and convenient to carry.
Description
Technical Field
The invention relates to the field of nucleic acid detection equipment, in particular to a portable nucleic acid detector and a nucleic acid detection method.
Background
With the development of science and technology, nucleic acid detection has been increasingly applied to the fields of medical diagnosis, food safety detection and the like. The nucleic acid detector is an automatic device integrating nucleic acid extraction, amplification and detection, and the existing nucleic acid detector has the defects of high cost, large volume, complex operation and inconvenient transportation.
Disclosure of Invention
In order to achieve the above object, the present invention is achieved by the following technical solutions.
The present invention provides a portable nucleic acid detecting instrument, comprising:
the card box comprises a kit, a slide block and a reaction tube for amplification reaction; the kit is provided with a plurality of containing cavities, a plurality of channels and a liquid outlet; the bottom of the cavity is communicated with the channel; the top end of the reaction tube is communicated with the liquid outlet; the slide block is connected with the reagent box in a sliding way; the sliding block is provided with a transition cavity and an injector structure; the syringe structure comprises an outer barrel and a piston;
the displacement mechanism is used for driving the sliding block to slide relative to the reagent box so as to switch the channel or the liquid outlet to be communicated with the transition cavity;
the piston driving mechanism comprises a first driving piece and a push-pull rod which are connected; the push-pull rod is inserted into the outer cylinder and clamped on the piston; the first driving piece is used for driving the push-pull rod to drive the piston to move in the outer cylinder so as to draw or push the transition cavity, and the card box is used for extracting nucleic acid.
Preferably, the displacement mechanism comprises:
the clamping assembly comprises two ejector rods which are used for respectively abutting against the end surfaces of two sides of the sliding block;
and the translation assembly is used for driving the clamping assembly to move along the sliding direction of the sliding block.
Preferably, the clamping assembly comprises a positive and negative tooth screw rod structure for driving the two ejector rods to move oppositely or oppositely.
Preferably, one of the top rods is of a fixed rod structure; the clamping assembly comprises a screw nut structure and is used for driving the other ejector rod to reciprocate.
Preferably, the amplification mechanism comprises:
the heating aluminum seat is provided with a slot and a detection hole; the detection hole is communicated with one side of the slot;
the heat preservation cover is provided with a jack and a yielding hole; the heat insulation cover surrounds the heating aluminum seat, and the jack is communicated with the slot; the reaction tube penetrates through the jack and then is inserted into the slot; (ii) a The abdicating hole is used for exposing the detection hole;
the heat dissipation element is fixed on the bottom wall of the heat preservation cover and forms a space for accommodating the heating aluminum seat together with the heat preservation cover; the bottom wall of the heating aluminum seat is abutted against the upper surface of the heat dissipation element;
a fan connected with the heat dissipation element.
Preferably, the reading mechanism comprises:
the reading head is provided with a plurality of LED lamps with different colors, a plurality of detectors and a plurality of optical holes;
and the driving component is used for driving the reading head to move so as to switch the LED lamp matched with the reaction tube.
Preferably, a plurality of the LED lamps are matched with a plurality of the detectors so as to carry out FAM, HEX, ROX and CY5 four-color detection.
Preferably, the reading head is provided with a convex part, and a plurality of optical holes are arranged on the convex part; the amplification mechanism is provided with a first groove, and a detection hole of the amplification mechanism is arranged in the first groove; the end face of the bulge part is abutted against the inner surface of the first groove so as to be matched with the detection hole.
Preferably, also include magnetic attraction mechanism, it includes:
a magnet for attracting magnetic beads in the reagent cartridge;
and the second driving piece is used for driving the magnet to be close to or far away from the magnetic bead.
Preferably, the displacement mechanism and the magnetic attraction mechanism are respectively positioned at two sides of the card box.
The invention also provides a nucleic acid detection method, which adopts the portable nucleic acid detector for detection.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a portable nucleic acid detector.A displacement mechanism is used for driving a sliding block to slide relative to a kit so as to adjust the position of the sliding block, further to enable a transition cavity to be communicated with a target containing cavity, and further to suck, spit or transfer liquid in the corresponding containing cavity so as to finish nucleic acid extraction. The product of nucleic acid extraction is finally discharged into the reaction tube through the transition cavity and the liquid outlet so as to be amplified by the amplification mechanism. The structure design of the kit, the slide block and the reaction tube forms a closed nucleic acid extraction cavity, the structure is simple and small, the nucleic acid extraction operation is simple and convenient, and aerosol pollution is prevented. The detector body omits a mechanical arm liquid-transferring structure, a series of nucleic acid extraction operations are completed inside the card box, and the whole structure is simple, small and convenient to carry.
The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical means of the present invention more clearly understood and to be implemented according to the content of the description, the following detailed description is given with reference to the preferred embodiments of the present invention and the accompanying drawings. The detailed description of the present invention is given in detail by the following examples and the accompanying drawings.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the invention without limiting the invention. In the drawings:
fig. 1 is a schematic perspective view of a detector body according to an embodiment of the present invention;
fig. 2 is a schematic perspective view of a detector body according to an embodiment of the invention;
FIG. 3 is a schematic view showing an assembled structure of the cartridge, the displacement mechanism and the piston driving mechanism according to still another embodiment of the present invention;
FIG. 4 is a schematic perspective view of an amplification mechanism according to the present invention;
FIG. 5 is a sectional view showing the structure of an amplification mechanism of the present invention;
FIG. 6 is a schematic diagram of an explosive structure of the amplification mechanism of the present invention;
FIG. 7 is a schematic perspective view of a reading mechanism according to the present invention;
FIG. 8 is a first perspective view of the cartridge of the present invention;
FIG. 9 is a structural cross-sectional view of the cartridge of the present invention;
FIG. 10 is a schematic perspective view of a kit according to the present invention;
FIG. 11 is a schematic perspective view of the cartridge of the present invention;
fig. 12 is a schematic view of the detection flow of the detector body according to the present invention.
In the figure: 100. a detector body;
10. a card box; 11. a kit; 111. a channel; 1111. a passage port; 112. a liquid outlet; 113. a chute; 1141. a sample position; 1142. the cracking liquid level; 1143. combining the liquid levels; 1144. a first cleaning solution; 1145. magnetic bead position; 1146. a second cleaning solution; 1147. a water level; 1148. the elution liquid level; 1149. i-MIX reaction level; 115. a second groove; 12. a slider; 121. a transition chamber; 122. an injector structure; 1221. an outer cylinder; 1222. a piston; 1223. a stopper; 1224. a seal ring; 123. a limiting bulge; 124. a concave cavity; 13. a reaction tube; 14. a silica gel pad; 15. pressing a plate; 151. a through groove;
20. a displacement mechanism; 21. a clamping assembly; 211. a first motor; 212. a screw rod with positive and negative teeth; 213. a first screw block; 2141. a first ejector rod; 2142. a second ejector rod; 215. an adaptor; 2161. a third ejector rod; 2162. a fourth ejector rod; 217. a second motor; 2181. a first lead screw; 2182. a second screw block; 22. a translation assembly;
30. a piston drive mechanism; 31. a first driving member; 32. a push-pull rod;
40. an amplification mechanism; 41. heating the aluminum base; 411. a slot; 412. a detection hole; 42. a heat-preserving cover; 421. a jack; 422. a hole of abdication; 423. a first groove; 43. a heat dissipating element; 44. a fan; 45. a high and low temperature control element;
50. a reading mechanism; 51. a reading head; 511. an optical aperture; 512. a boss portion; 52. a drive assembly;
60. a magnetic attraction mechanism; 61. a magnet; 62. a second driving member; 621. a magnetic motor; 622. a second lead screw; 623. a third screw block;
70. a power supply module; 71. a switch; 72. a temperature controller; 73. a PCB board; 74. a power source;
80. a base plate;
91. a base.
Detailed Description
The foregoing and other objects, features, aspects and advantages of the present invention will become more apparent from the following detailed description of the present invention when taken in conjunction with the accompanying drawings, which will enable those skilled in the art to practice the present invention with reference to the accompanying specification. In the drawings, the shape and size may be exaggerated for clarity, and the same reference numerals will be used throughout the drawings to designate the same or similar components. In the following description, terms such as center, thickness, height, length, front, back, rear, left, right, top, bottom, upper, lower, and the like are used based on the orientation or positional relationship shown in the drawings. In particular, "height" corresponds to the dimension from top to bottom, "width" corresponds to the dimension from left to right, and "depth" corresponds to the dimension from front to back. These relative terms are for convenience of description and generally are not intended to require a particular orientation. Terms concerning attachments, coupling and the like (e.g., "connected" and "attached") refer to a relationship wherein structures are secured or attached to one another either directly or indirectly through intervening structures, as well as both movable or rigid attachments or relationships, unless expressly described otherwise.
The present invention is further described with reference to the accompanying drawings and the detailed description, and it should be noted that, in the case of no conflict, any combination between the embodiments or technical features described below may form a new embodiment.
Example 1
The present invention provides a portable nucleic acid detecting instrument, as shown in fig. 1 to 11, including a detecting instrument body 100, the detecting instrument body 100 including:
a cartridge 10 including a reagent cartridge 11, a slide 12, and a reaction tube 13 for amplification reaction; the reagent box 11 is provided with a plurality of cavities, a plurality of channels 111 and a liquid outlet 112; a bottom of said chamber communicating with a said channel 111; the top end of the reaction tube 13 is communicated with the liquid outlet 112; the slide block 12 is connected with the reagent box 11 in a sliding way; the sliding block 12 is provided with a transition cavity 121 and an injector structure 122; the syringe structure 122 includes an outer cylinder 1221, a piston 1222;
a displacement mechanism 20, configured to drive the slide block 12 to slide relative to the reagent cartridge 11, so as to switch the communication between the channel 111 or the liquid outlet 112 and the transition cavity 121;
a piston driving mechanism 30 comprising a first driving member 31 and a push-pull rod 32 connected with each other; the push-pull rod 32 is inserted into the outer cylinder 1221 and is clamped to the piston 1222; the first driving member 31 is used to drive the push-pull rod 32 to drive the piston 1222 to move in the outer cylinder 1221, so as to draw or push the transition chamber 121, so that the cartridge 10 can perform nucleic acid extraction.
In this embodiment, the displacement mechanism 20 is used to drive the slide block 12 to slide relative to the reagent kit 11, so as to adjust the position of the slide block 12, further enable the transition cavity 121 to communicate with the target containing cavity, and further suck, spit or transfer the liquid in the corresponding containing cavity, so as to complete the nucleic acid extraction. The product of nucleic acid extraction is finally discharged into the reaction tube 13 through the transition chamber 121 and the liquid outlet 112 for amplification by the amplification mechanism 40. The structure design of the reagent kit 11, the slide block 12 and the reaction tube 13 forms a closed nucleic acid extraction cavity, the structure is simple and small, the nucleic acid extraction operation is simple and convenient, and aerosol pollution is prevented. The detecting instrument body 100 omits a mechanical arm liquid transfer structure, completes a series of nucleic acid extraction operations inside the cartridge 10, and has a simple and small overall structure and convenient transportation.
In one embodiment, the cartridge 10 is a consumable structure for performing nucleic acid extraction on a single sample, and the cleaning operation for different sample detection is omitted.
In one embodiment, as shown in fig. 1-3, the displacement mechanism 20 includes:
the clamping assembly 21 comprises two ejector rods which are used for respectively abutting against the end surfaces of two sides of the sliding block 12;
a translation assembly 22 for driving the clamping assembly 21 to move along the sliding direction of the sliding block 12. Specifically, the two ejector rods respectively abut against end surfaces of two sides of the sliding block 12 to clamp the sliding block 12, and the two ejector rods drive the sliding block 12 to slide relative to the reagent kit 11 under the driving of the translation assembly 22, so that the transition cavity 121 is communicated with the target accommodating cavity or the liquid outlet 112.
In one embodiment, as shown in fig. 1 and 2, the clamping assembly 21 includes a positive and negative lead screw structure for driving the two rods to move toward or away from each other. Specifically, the clamping assembly 21 includes a first motor 211, a positive and negative tooth screw rod 212, two first screw blocks 213, a first ejector rod 2141, and a second ejector rod 2142; the two first screw blocks 213 are respectively sleeved on the positive tooth rod and the negative tooth rod of the positive and negative tooth screw rod 212; the first top rod 2141 and the second top rod 2142 are connected to the two first screw blocks 213 through a connecting piece 215. The first motor 211 rotates, the positive and negative teeth screw rod 212 and the two first screw blocks 213 are matched, so that the two first screw blocks 213 drive the first ejector rod 2141 and the second ejector rod 2142 to move in opposite directions or in opposite directions, and the clamping or loosening operation on the end surfaces of the two sides of the sliding block 12 is further realized. The first ejector rod 2141 and the second ejector rod 2142 have a large range of distance, and can clamp a large-size slide block 12, and the clamping assembly 21 has a simple overall structure, and can be driven by one motor.
In another embodiment, as shown in fig. 3, one of the carrier rods is a fixed rod structure; the clamping assembly 21 comprises a screw nut structure for driving the other ejector rod to reciprocate. Specifically, the clamping assembly 21 includes a third push rod 2161, a fourth push rod 2162, a second motor 217, a first lead screw 2181, and a second screw block 2182 sleeved on the periphery of the first lead screw 2181; the third ejector rod 2161 is fixed, and the driving end of the second motor 217 is coaxially connected with one end of the first screw rod 2181; the fourth push rod 2162 is connected to the second screw block 2182. The second motor 217 rotates to drive the first lead screw 2181 to rotate, the second screw 2182 moves linearly along the first lead screw 2181, and then the fourth push rod 2162 is driven to move close to or away from the third push rod 2161, so that the end surfaces of the two sides of the sliding block 12 are clamped or loosened. Because the third top bar 2161 is fixed, the clamping of the slide block 12 by the clamping component 21 is more stable.
In one embodiment, the translation assembly 22 is a lead screw nut assembly to achieve linear drive.
In one embodiment, an amplification mechanism 40 is further included to amplify the extracted product in the reaction tube 13.
Further, as shown in fig. 4 to 6, the amplification mechanism 40 includes:
a heating aluminum base 41 having a slot 411 and a detection hole 412; the detection hole 412 is communicated with one side of the slot 411;
the heat preservation cover 42 is provided with a jack 421 and a abdicating hole 422; the heat-insulating cover 42 surrounds the heating aluminum seat 41, and the insertion holes 421 are communicated with the insertion slots 411; the reaction tube 13 passes through the insertion hole 421 and then is inserted into the insertion slot 411; the relief hole 422 is used for exposing the detection hole 412;
a heat dissipation element 43 fixed to the bottom wall of the heat insulation cover 42 and forming a space for accommodating the heating aluminum base 42 together with the heat insulation cover 42; the bottom wall of the heating aluminum seat 41 is abutted against the upper surface of the heat dissipation element 43;
a fan 44 connected to the heat dissipating member 43. Specifically, the heating aluminum base 41 is used to heat the reaction tube 13 to form a temperature environment required for amplification. The heat-insulating cover 41 is used to prevent or reduce the temperature loss of the heating aluminum base 41 and the reaction tube 13, so as to ensure the stability of the heating temperature. The heat dissipation element 43 is used to cool the heating aluminum base 41 to form the high/low temperature cycling condition required for amplification. The fan 44 is used to accelerate the heat dissipation speed of the heat dissipation element 43. After the cartridge 10 is installed in the work station, the reaction tube 13 is directly inserted into the slot 411 at a portion where the cartridge 10 is exposed. When the cartridge 10 completes the nucleic acid extraction, the amplification mechanism 40 starts amplifying the nucleic acid extraction product in the reaction tube. The extraction and amplification process saves a mechanical transfer structure of the reaction tube, is beneficial to the miniaturization design of the detector body 100, and improves the detection speed.
Further, a fan 44 is provided at one side of the heating element 43 to avoid increasing the overall height of the amplification mechanism 40.
Further, the amplification mechanism 40 further includes a high-temperature and low-temperature control element 45 disposed between the heating aluminum base 41 and the heat dissipation element 43.
In one embodiment, a reading mechanism 50 is further included to read the amplification product in the reaction tube 13.
Further, as shown in fig. 7, the reading mechanism 50 includes:
a reading head 51 provided with a plurality of LED lamps of different colors, a plurality of detectors, and a plurality of optical holes 511;
and the driving assembly 52 is used for driving the reading head 51 to move so as to switch the LED lamp matched with the reaction tube 13. Specifically, different LEDs emit light of different colors, and the light enters the reaction tube 13 after passing through the optical hole 511 and the detection hole 412, so as to excite the amplification product to emit fluorescence, and the generated fluorescence is fed back to the detector, so as to obtain a detection result through analysis of a fluorescence signal.
Further, a plurality of the LED lamps are matched with a plurality of the detectors to carry out FAM, HEX, ROX and CY5 four-color detection.
Further, in order to simplify the movement track of the reading mechanism 50, a plurality of LED lamps are arranged in a row along the sliding direction of the sliding block 12, the reading head 51 is abutted against the amplification mechanism 40 towards the end surface of the amplification mechanism 40, and the driving assembly 52 is configured to drive the reading head 51 to move along the sliding direction of the sliding block 12.
Furthermore, the reading head 51 is provided with a protruding portion 512, and a plurality of optical holes 511 are arranged on the protruding portion 512; the amplification mechanism 40 is provided with a first groove 423, and the abdication hole 422 of the amplification mechanism 40 is arranged in the first groove 423; the end surface of the protrusion 512 abuts against the inner surface of the first groove 423 to match the detection hole 412. The first groove 423 partially surrounds the outer end face of the protrusion 512, so as to reduce the interference detected by the external light detector, avoid fluorescence crosstalk, and ensure the stability of the reading mechanism 50.
In one embodiment, as shown in fig. 1 and 2, the monitor body 100 further includes a magnetic attraction mechanism 60, which includes:
a magnet 61 for attracting magnetic beads in the cartridge 11;
a second driving member 62 for driving the magnet 61 to approach or separate from the magnetic bead.
Further, the second driving member 62 includes a magnetic attraction motor 621, a second screw rod 622 and a third screw block 623 sleeved on the periphery of the second screw rod 622, and the magnet 61 is fixed to the third screw block 623.
Further, the displacement mechanism 20 and the magnetic attraction mechanism 60 are respectively located on two sides of the cartridge 10, and the arrangement of the mechanisms is compact.
In one embodiment, the monitor body 100 further includes a power module 70; the power module 70 comprises a switch 71, a temperature controller 72, a PCB 73 and a power supply 74.
In one embodiment, the meter body 100 includes a base plate 80, and the cartridge 10 is secured to the base plate 80 by a base 91.
Further, the amplification mechanism 40 and the reading mechanism 50 are located below the base 91.
In one embodiment, as shown in fig. 8 to 11, the reagent cartridge 11 is U-shaped to form a sliding slot 113 for slidably connecting the sliding block 12, so as to improve the wrapping performance of the sliding slot 113 to the sliding block 12.
In one embodiment, the number of chambers is nine, including sample level 1141, lysis level 1142, binding level 1143, first wash level 1144, bead level 1145, second wash level 1146, water level 1147, elution level 1148, and i-MIX reaction level 1149, in that order.
Furthermore, the cavity is opened above and covered with a gas-permeable and nucleic acid-impermeable membrane structure or a sealing cover. Specifically, the upper opening of the cavity is covered by a breathable nucleic acid-impermeable membrane structure, so that the air pressure inside and outside the cavity is balanced, the nucleic acid is prevented from leaking, and in addition, the reagent can be prevented from leaking in the transportation process. Or, the sealing cover covers the openings above the plurality of cavities to avoid nucleic acid leakage, and a cavity for containing gas is formed between the sealing cover and the kit 11 to balance the gas pressure inside and outside the cavities when liquid is transferred.
In an embodiment, the liquid outlet 112 and the plurality of channels 111 are respectively disposed on the bottom wall of the sliding chute 113, so as to reduce the distance between the liquid outlet 112 and the plurality of channels 111 and the first through hole at the bottom of the transition cavity 121, and reduce the length of the channel 111, so as to increase the flowing efficiency of the liquid during pumping; the passage ports 1111 at one ends of the passages 111 and the liquid outlet 112 are arranged in a line along the moving direction of the slider 12 to match the sliding direction of the slider 12, and the slider 12 can slide back and forth along one direction, so that different passage ports 1111 communicated with the bottom of the transition cavity 121 or the liquid outlet 112 can be switched, and the structure is simple and the operation is convenient.
Further, a plurality of the channel ports 1111 are disposed close to each other, so that the path of the slider 12 during the nucleic acid extraction process is shortened and the length of the channel 111 is shortened.
Further, a plurality of the channels 111 extend obliquely toward the transition chamber 121, respectively, such that a plurality of the channel ports 1111 are disposed closely, and the length of the channels 111 is reduced and the length of the channel 111 is reduced while the path of the slider 12 is shortened during the nucleic acid extraction process.
Further, a second groove 115 is formed in the bottom wall of the sliding chute 113, and the channel port 1111 and the liquid outlet 112 are respectively disposed in the second groove 115;
a silica gel pad 14 is arranged in the second groove 115; the silica gel pad 14 is provided with a plurality of second through holes to be respectively communicated with the plurality of passage ports 1111 and the liquid outlet 112. Specifically, through the matching of the second groove 115 and the silicone rubber pad 14, when the first through hole is communicated with the channel port 1111, the communicating position of the first through hole and the channel port 1111 is sealed, and when the first through hole is communicated with the liquid outlet 112, the communicating position of the first through hole and the liquid outlet 112 is sealed, so that the sealing performance of the formed whole pumping and releasing cavity is improved.
Further, the cartridge 10 further includes a pressing plate 15 fixed to the upper surface of the reagent cartridge 11 and pressing the slider 12 against the upper surface of the silicone pad 14. Specifically, the slider 12 is pressed by the pressing plate 15 to increase the sealability between the slider 12 and the silicone pad 14.
Further, the pressing plate 15 is provided with a through groove 151, and the sliding block 12 is provided with a limiting protrusion 123; the through groove 151 is matched with the limiting protrusion 123 to limit the movement position of the sliding block 12 and limit the maximum stroke of the sliding block 12, so that stroke control is matched, and a control algorithm is simplified.
In one embodiment, the channel 111 is a groove structure, which is easy to machine, and the lower opening of the channel 111 is sealed by a plastic film.
In one embodiment, the syringe structure 122 further includes a stopper 1223 disposed at the opening of the outer cylinder 1221 for preventing the piston 1222 from being removed from the outer cylinder 1221 along with the movement of the push-pull rod 32, and for assisting in the removal of the push-pull rod 32 from the piston 1222.
In one embodiment, the slider 12 is provided with a cavity 124, and the outer cylinder 1221 is disposed within the cavity 124.
Further, a sealing ring 1224 is disposed at a position where the head of the outer cylinder 1221 is communicated with the transition cavity 121.
Example 2
The invention provides a nucleic acid detection method, which adopts the portable nucleic acid detector for detection.
When the nucleic acid is detected by the detection apparatus body 100, the cartridge 10 containing the sample and each reagent is mounted on the base 91, and the reaction tube 13 of the cartridge 10 is inserted into the slot 411 of the amplification mechanism 40 to start the detection.
The cartridge 10 is clamped by the two rods of the displacement mechanism 20, and the push-pull rod 32 of the piston driving mechanism 30 is inserted into the outer cylinder 1221 and engaged with the piston 1222 to start nucleic acid extraction. As shown in FIG. 12, the nucleic acid extraction specifically comprises the steps of: reagent cracking, magnetic bead combination, first cleaning liquid cleaning, second cleaning liquid cleaning, magnetic bead drying and magnetic bead elution. After the nucleic acid is extracted, PCR amplification and detection are carried out, and the card box is released after the detection is finished.
The reagent vessel 11 comprises nine channels 111, one outlet port 112. When the first through hole of the transition chamber 121 communicates with the sample site 1141, the slider is located at position 1; when the first through hole of the transition chamber 121 is in communication with the lysis level 1142, the slider is in position 2; when the first through hole of the transition chamber 121 communicates with the bonding level 1143, the slider is located at position 3; when the first through-hole of the transition chamber 121 is in communication with the first cleaning fluid level 1144, the slide is in position 4; when the first through hole of the transition cavity 121 is communicated with the magnetic bead position 1145, the slider is located at position 5; when the first through-hole of the transition chamber 121 is in communication with the second cleaning fluid level 1146, the slide is in position 6; when the first through hole of the transition chamber 121 is in communication with the water level 1147, the slider is in position 7; when the first through hole of the transition chamber 121 is in communication with the elution level 1148, the slide is in position 8; when the first through-hole of the transition cavity 121 is in communication with the i-MIX reactant level 1149, the slider is in position 9; when the first through hole of the transition cavity 121 is communicated with the liquid outlet 112, the slide block is located at the position 10. The method specifically comprises the following steps:
(1) and (3) cracking of a reagent: the slide 12 moves to position 1 and the injector structure 122 aspirates liquid from the sample site 1141; the slide block 12 moves to the position 2, the injector structure 122 spits the sample liquid, and the sample liquid is put into the lysis liquid level 1142, and cell lysis is completed after the process is finished;
(2) and (3) magnetic bead combination: the syringe structure 122 imbibes from within the lysis level 1142; the slide block 12 moves to the position 3, the injector structure 122 spits liquid, and the liquid after cracking is put into a cracking liquid level 1142; the injector structure 122 sucks, spits and uniformly mixes for 5 times; the injector structure 122 is aspirated, and the slide 12 is moved to position 5; the syringe structure 122 discharges liquid; the injector structure 122 sucks, spits and mixes uniformly for 50 times; feeding a magnetic suction motor, approaching the magnetic bead position 1145, sucking the magnetic beads for 2min, slowly sucking and spitting the injector structure 122 for 2 times, waiting for 1min, slowly sucking and spitting the injector structure 122 for 2 times, and waiting for 1 min; the injector structure 122 aspirates; the slide block 12 moves to the position 1, the injector structure discharges liquid, and the process is finished;
(3) cleaning with a first cleaning solution: the slide 12 moves to position 4 and the syringe structure 122 aspirates a small amount; the slider 12 moves to position 5 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and uniformly mixes for 10 times; the slide 12 moves to position 4 and the injector structure 122 is primed; the slider 12 moves to position 5 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and mixes uniformly for 30 times; feeding a magnetic suction motor, approaching the magnetic bead position 1145, sucking the magnetic beads for 2min, slowly sucking and spitting the injector structure 122 for 2 times, waiting for 1min, slowly sucking and spitting the injector structure 122 for 2 times, and waiting for 1 min; the injector structure 122 aspirates; the slide block 12 moves to the position 2, the injector structure discharges liquid, and the process is finished;
(4) cleaning with a second cleaning solution: the slide 12 moves to position 6 and the injector structure 122 aspirates a small amount; the slider 12 moves to position 5 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and mixes uniformly for 10 times; the slide 12 moves to position 6 and the syringe structure 122 aspirates; the slider 12 moves to position 5 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and mixes uniformly for 30 times; feeding a magnetic suction motor, approaching the magnetic bead position 1145, sucking the magnetic beads for 2min, slowly sucking and spitting the injector structure 122 for 2 times, waiting for 1min, slowly sucking and spitting the injector structure 122 for 2 times, and waiting for 1 min; the injector structure 122 aspirates; the slide block 12 moves to the position 3, and the injector structure discharges liquid; finishing the two-time cleaning;
(5) and (3) drying magnetic beads: the slide 12 moves to position 7 and the syringe structure 122 is primed; the slider 12 moves to position 6 and the syringe structure 122 spits; the injector structure 122 sucks, spits and mixes uniformly for 10 times; the injector structure 122 aspirates; the slider 12 moves to position 4 and the injector structure 122 dispenses the fluid; the slide 12 moves to position 5 and the injector structure 122 aspirates; the slider 12 moves to position 4 and the injector structure 122 dispenses the fluid; the slider 12 moves to position 7 and the injector structure 122 dispenses the fluid; the slider 12 moves to position 6 and the syringe structure 122 spits; the injector structure 122 sucks, spits and mixes uniformly for 10 times; the injector structure 122 aspirates; the slider 12 moves to position 4 and the injector structure 122 dispenses the fluid; waiting for 30 seconds, the injector structure 122 dispenses the liquid; feeding by a magnetic motor; the slider 12 moves to position 5, the injector structure 122 lengthens to aspirate liquid, and the injector structure 122 ejects 1/3; waiting for 3min, the injector structure 122 spits 1/3; waiting for 3min, the injector structure 122 spits 1/3; withdrawing the magnet motor, and ending;
(6) and (3) magnetic bead elution: the slide 12 moves to position 8 and the syringe structure 122 aspirates; the slider 12 moves to position 5 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and uniformly mixes for 30 times; waiting for 5 min; the syringe structure 122 aspirates 1 time; feeding the magnet for 3min, and sucking, spitting and uniformly mixing the injector structure 122 for 2 times; waiting for 1 min; the injector structure 122 is sucked, discharged and uniformly mixed for 2 times; waiting for 1 min; the slide block 12 moves to the position 7, and the injector structure 122 sucks, discharges and uniformly mixes for 10 times; the slide 12 moves to position 5, waiting for 30 s; the injector structure 122 meters pipetting; the slider 12 moves to position 9 and the injector structure 122 dispenses the fluid; the injector structure 122 sucks, spits and mixes uniformly for 3 times, and waits for 1 min; the injector structure 122 aspirates; the slide 12 moves to position 10 and the syringe structure 122 doses; withdrawing the magnet; and resetting all motors and ending.
The foregoing is merely a preferred embodiment of this invention and is not intended to limit the invention in any manner; one of ordinary skill in the art can readily practice the present invention as illustrated and described herein with reference to the accompanying drawings; however, those skilled in the art should appreciate that they can readily use the disclosed conception and specific embodiments as a basis for designing or modifying other structures for carrying out the same purposes of the present invention without departing from the scope of the invention as defined by the appended claims; meanwhile, any equivalent changes, modifications and evolutions made to the above embodiments according to the substantial technology of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (13)
1. A portable nucleic acid detecting instrument, comprising:
a cartridge (10) including a reagent cartridge (11), a slide (12), and a reaction tube (13) for amplification reaction; the kit (11) is provided with a plurality of cavities, a plurality of channels (111) and a liquid outlet (112); a bottom of said chamber communicating with a said passage (111); the top end of the reaction tube (13) is communicated with the liquid outlet (112); the sliding block (12) is connected with the reagent box (11) in a sliding way; the sliding block (12) is provided with a transition cavity (121) and an injector structure (122); the syringe structure (122) comprises an outer cylinder (1221), a piston (1222);
the displacement mechanism (20) is used for driving the sliding block (12) to slide relative to the reagent box (11) so as to switch the communication between the channel (111) or the liquid outlet (112) and the transition cavity (121);
the piston driving mechanism (30) comprises a first driving piece (31) and a push-pull rod (32) which are connected; the push-pull rod (32) is inserted into the outer cylinder (1221) and clamped to the piston (1222); the first driving part (31) is used for driving the push-pull rod (32) to drive the piston (1222) to move in the outer cylinder (1221) so as to draw or push the transition cavity (121) to extract or push the nucleic acid from the cartridge (10).
2. The portable nucleic acid detecting apparatus according to claim 1, wherein the displacement mechanism (20) includes:
the clamping component (21) comprises two ejector rods which are used for respectively abutting against the end surfaces of two sides of the sliding block (12);
the translation assembly (22) is used for driving the clamping assembly (21) to move along the sliding direction of the sliding block (12).
3. The portable nucleic acid detecting apparatus according to claim 2, wherein the holding member (21) includes a screw mechanism with positive and negative teeth for driving the two rods to move toward or away from each other.
4. The portable nucleic acid detecting instrument according to claim 2, wherein the rod is a rod-fixed structure; the clamping assembly (21) comprises a screw nut structure and is used for driving the other ejector rod to reciprocate.
5. The portable nucleic acid detecting apparatus according to any one of claims 1 to 4, further comprising an amplification mechanism (40) for amplifying the extracted product in the reaction tube (13).
6. The portable nucleic acid detecting apparatus according to claim 5, wherein the amplification means (40) includes:
a heating aluminum base (41) provided with a slot (411) and a detection hole (412); the detection hole (412) is communicated with one side of the slot (411);
the heat preservation cover (42) is provided with an insertion hole (421) and a yielding hole (422); the heat preservation cover (42) surrounds the heating aluminum seat (41), and the insertion hole (421) is communicated with the insertion groove (411); the reaction tube (13) passes through the jack (421) and then is inserted into the slot (411); the abdicating hole (422) is used for exposing the detection hole (412);
the heat dissipation element (43) is fixed on the bottom wall of the heat preservation cover (42) and forms a space for accommodating the heating aluminum seat (42) together with the heat preservation cover (42); the bottom wall of the heating aluminum seat (41) is abutted against the upper surface of the heat dissipation element (43);
a fan (44) connected to the heat-dissipating element (43).
7. The portable nucleic acid detecting apparatus according to any one of claims 1 to 4, further comprising a reading mechanism (50) for reading an amplification product in the reaction tube (13).
8. The portable nucleic acid detecting instrument according to claim 7, wherein the reading mechanism (50) includes:
the reading head (51) is provided with a plurality of LED lamps with different colors, a plurality of detectors and a plurality of optical holes (511);
and the driving component (52) is used for driving the reading head (51) to move so as to switch the LED lamp matched with the reaction tube (13).
9. The portable nucleic acid detecting instrument of claim 8, wherein a plurality of the LED lamps are matched with a plurality of the detectors to perform FAM, HEX, ROX, CY5 four-color detection.
10. The portable nucleic acid detecting instrument according to claim 8, wherein the reading head (51) is provided with a boss (512), and the plurality of optical holes (511) are provided on the boss (512); the amplification mechanism (40) is provided with a first groove (41), and a detection hole (412) of the amplification mechanism (40) is arranged in the first groove (41); the end face of the bulge (513) is abutted against the inner surface of the first groove (41) to match with the detection hole (412).
11. The portable nucleic acid detecting instrument according to any one of claims 1 to 4, further comprising a magnetic attraction mechanism (60) including:
a magnet (61) for attracting magnetic beads in the cartridge (11);
a second driving member (62) for driving the magnet (61) to approach or separate from the magnetic bead.
12. The portable nucleic acid detecting apparatus according to claim 11, wherein the displacement mechanism (20) and the magnetic attraction mechanism (60) are respectively disposed on two sides of the cartridge (10).
13. A method for detecting nucleic acid, characterized in that the detection is carried out using the portable nucleic acid detecting instrument according to any one of claims 1 to 12.
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