CN212955086U - Totally enclosed nucleic acid draws and purification card box - Google Patents
Totally enclosed nucleic acid draws and purification card box Download PDFInfo
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- CN212955086U CN212955086U CN202021453799.7U CN202021453799U CN212955086U CN 212955086 U CN212955086 U CN 212955086U CN 202021453799 U CN202021453799 U CN 202021453799U CN 212955086 U CN212955086 U CN 212955086U
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- 238000000746 purification Methods 0.000 title claims abstract description 40
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 28
- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 28
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 87
- 238000000605 extraction Methods 0.000 claims abstract description 40
- 238000012545 processing Methods 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000011324 bead Substances 0.000 claims abstract description 29
- 239000006166 lysate Substances 0.000 claims abstract description 25
- 102000004190 Enzymes Human genes 0.000 claims abstract description 20
- 108090000790 Enzymes Proteins 0.000 claims abstract description 20
- 239000003480 eluent Substances 0.000 claims abstract description 18
- 238000005406 washing Methods 0.000 claims abstract description 16
- 238000002347 injection Methods 0.000 claims abstract description 13
- 239000007924 injection Substances 0.000 claims abstract description 13
- 238000004108 freeze drying Methods 0.000 claims abstract description 11
- 238000003860 storage Methods 0.000 claims description 85
- 239000000243 solution Substances 0.000 claims description 34
- 238000011144 upstream manufacturing Methods 0.000 claims description 31
- 238000004140 cleaning Methods 0.000 claims description 20
- 238000007789 sealing Methods 0.000 claims description 17
- 230000000903 blocking effect Effects 0.000 claims description 9
- 239000005060 rubber Substances 0.000 claims description 4
- 238000004026 adhesive bonding Methods 0.000 claims description 3
- 239000002131 composite material Substances 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 2
- 238000006243 chemical reaction Methods 0.000 claims description 2
- 239000000499 gel Substances 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 125000001153 fluoro group Chemical group F* 0.000 claims 1
- 238000002955 isolation Methods 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 12
- 238000013461 design Methods 0.000 abstract description 7
- 238000012840 feeding operation Methods 0.000 abstract description 2
- 238000004321 preservation Methods 0.000 abstract 6
- 239000000047 product Substances 0.000 description 22
- 238000000034 method Methods 0.000 description 8
- 239000012530 fluid Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 4
- 239000002699 waste material Substances 0.000 description 4
- 229920000742 Cotton Polymers 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 2
- 230000009089 cytolysis Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
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- 238000003912 environmental pollution Methods 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 1
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Abstract
The utility model discloses a totally enclosed nucleic acid extraction and purification card box, the card box at least comprises a main box body, a piston rod and 9 two-way valves, wherein an injection tube, a sample processing cavity, a PK freeze-drying enzyme preservation cavity, a magnetic bead preservation cavity, a lysate preservation cavity, a first washing liquid preservation cavity, a second washing liquid preservation cavity, an eluent preservation cavity, a purification cavity, an extraction product cavity and a water cut cavity are sequentially arranged in the main box body side by side; the two-way valve set up in the top of main box body is equipped with the series connection of runner completion each valve body under the valve between every adjacent valve body in 9 two-way valves, the injection tube links to each other with the two-way valve that sets up in the head end, the two-way valve of intercepting water chamber and tail end is linked together, be equipped with the hole of hindering water on the wall of intercepting water chamber, be equipped with ventilative stagnant water post in the hole of hindering water to be linked together through ventilative stagnant water post and exterior space. Through the structural design of this card box, can realize the presetting to various reagents to need not to carry out loaded down with trivial details reagent feeding operation in the course of handling.
Description
Technical Field
The utility model belongs to nucleic acid draws and purification unit field, especially relates to a totally enclosed nucleic acid draws and purification card box.
Background
Various DNA/RNA researches and applications require extraction and purification of nucleic acids by certain methods.
The current nucleic acid extraction and purification instrument basically adopts a magnetic bead method, but an open-cover purification device is basically adopted no matter a magnetic bar method or a liquid transfer workstation. The uncapping purifier has the risks of cross contamination, laboratory pollution and operator infection.
And most of nucleic acid extractors cannot directly treat samples such as cotton sticks, blood strips, gauze, swabs and the like, and require the samples to be pretreated to obtain liquid, so that the operation is troublesome.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide a: in order to overcome the prior art problem, provide a totally enclosed nucleic acid and draw and purification card box, make through the structural design of this card box and realized reagent in nucleic acid extraction and purification process and preset, need not to carry on loaded down with trivial details reagent feeding operation, also realized operating space's sealed design, stopped the pollution problem.
The purpose of the utility model is realized through the following technical scheme:
a totally enclosed nucleic acid extraction and purification card box, the said card box includes the main cartridge, piston rod and 9 two-way valves at least, wherein, there are injection cylinder, sample processing chamber, PK freeze-drying enzyme that preserves the chamber, magnetic bead that preserve the chamber, lysate preserve the chamber, first cleaning solution that preserve the chamber, second cleaning solution that preserve the chamber, eluant preserve the chamber, purify the chamber, extract product chamber and cut the water chamber side by side sequentially in the said main cartridge; the piston rod is assembled in the injection cylinder, and the top end of the piston rod is provided with a piston; the two-way valve is arranged at the top end of the main box body, a lower valve flow channel is arranged between every two adjacent valve bodies in the 9 two-way valves to complete the series connection of the valve bodies, the injection tube is connected with the two-way valve arranged at the head end, the water intercepting cavity is communicated with the two-way valve at the tail end, a water blocking hole is arranged on the side wall of the water intercepting cavity, and an air-permeable water-blocking column is arranged in the water blocking hole and is communicated with the external space through the air-permeable water-blocking column; and each valve body of the two-way valve from the head end to the tail end is also respectively communicated with the top and the bottom of the sample processing cavity, the PK freeze-drying enzyme storage cavity, the magnetic bead storage cavity, the lysate storage cavity, the first washing liquid storage cavity, the second washing liquid storage cavity, the eluent storage cavity, the purification cavity and the product extraction cavity, and the conversion of each cavity into the lower valve flow channel in series or out of the lower valve flow channel is completed through the valve rotation switching of the two-way valve.
According to a preferred embodiment, the two-way valve at least comprises a valve lower flow channel, a sealing gasket and a rotary valve, wherein a positioning hole is formed in the center of the bottom end of the rotary valve, two inwards-concave valve gating flow channels are symmetrically arranged on two sides with the positioning hole as a circle center, inlet and outlet ports of each cavity, which correspond to the liquid flow channel and each section of the valve lower flow channel, are arranged at the top end of the box body, a positioning column is further arranged at the top end of the box body, the rotary valve is in matched connection with the positioning column through the positioning hole, and the inlet and outlet ports of the liquid flow channel and the valve lower flow channel, which correspond to each cavity, are respectively communicated with four ports of the valve gating flow channel of; the sealing gasket is arranged between the rotary valve and the top end of the box body, and at least one valve through hole matched with the size of the inlet and outlet port is formed in the sealing gasket.
According to a preferred embodiment, the 9 two-way valves are respectively a first two-way valve, a second two-way valve, a third two-way valve, a fourth two-way valve, a fifth two-way valve, a sixth two-way valve, a seventh two-way valve, an eighth two-way valve and a ninth two-way valve, and a valve lower flow passage is arranged between every two adjacent valve bodies in the first two-way valve, the second two-way valve, the third two-way valve, the fourth two-way valve, the fifth two-way valve, the sixth two-way valve, the seventh two-way valve, the eighth two-way valve and the ninth two-way valve to complete the series connection of the valve bodies; the bottom end of the injection tube is provided with a liquid flow passage which is communicated with a valve gating flow passage on the upstream side of the first two-way valve; the top end of the sample processing cavity is communicated with a valve gating flow channel on the downstream side of the first two-way valve through a liquid flow channel, and the bottom end of the sample processing cavity is communicated with a valve gating flow channel on the upstream side of the first two-way valve through a liquid flow channel; the top end of the PK freeze-dried enzyme storage cavity is communicated with a valve gating flow channel on the downstream side of the second two-way valve through a liquid flow channel, and the bottom end of the PK freeze-dried enzyme storage cavity is communicated with a valve gating flow channel on the upstream side of the second two-way valve through a liquid flow channel; the top end of the magnetic bead storage cavity is communicated with a valve gating flow channel on the upstream side of the third two-way valve through a liquid flow channel, and the bottom end of the magnetic bead storage cavity is communicated with a valve gating flow channel on the downstream side of the third two-way valve through a liquid flow channel; the top end of the lysate storage cavity is communicated with a valve gating flow channel on the downstream side of the fourth two-way valve through a liquid flow channel, and the bottom end of the lysate storage cavity is communicated with a valve gating flow channel on the upstream side of the fourth two-way valve through a liquid flow channel; the top end of the first cleaning solution storage cavity is communicated with a valve gating flow channel on the upstream side of the fifth two-way valve through a liquid flow channel, and the bottom end of the first cleaning solution storage cavity is communicated with a valve gating flow channel on the downstream side of the fifth two-way valve through a liquid flow channel; the top end of the second cleaning solution storage cavity is communicated with a valve gating flow channel on the upstream side of the sixth two-way valve through a liquid flow channel, and the bottom end of the second cleaning solution storage cavity is communicated with a valve gating flow channel on the downstream side of the sixth two-way valve through a liquid flow channel; the top end of the eluent storage cavity is communicated with a valve gating flow channel on the upstream side of the seventh two-way valve through a liquid flow channel, and the bottom end of the eluent storage cavity is communicated with a valve gating flow channel on the downstream side of the seventh two-way valve through a liquid flow channel; the top end of the purification cavity is communicated with a valve gating flow channel on the downstream side of the eighth two-way valve through a liquid flow channel, and the bottom end of the purification cavity is communicated with a valve gating flow channel on the upstream side of the eighth two-way valve through a liquid flow channel; the top end of the extraction product cavity is communicated with a valve gating flow channel on the downstream side of the ninth two-way valve through a liquid flow channel, and the bottom end of the extraction product cavity is communicated with a valve gating flow channel on the upstream side of the ninth two-way valve through a liquid flow channel.
According to a preferred embodiment, the sealing pad is further provided with a through hole structure for passing through the positioning column.
According to a preferred embodiment, the seal is made of rubber or silicone or fluorine.
According to a preferred embodiment, the cartridge further comprises a valve cover which is fixedly attached to the top end of the main case body through a rivet post, and the sealing gasket and the rotary valve are disposed between the valve cover and the main case body.
According to a preferred embodiment, the main case is made of PP, and the rotary valve and the valve cover are made of PP or PC.
According to a preferred embodiment, the top end of the sample processing cavity is also provided with a sample processing cavity cover; and an extraction product cavity cover is also arranged at the top end of the extraction product cavity.
According to a preferred embodiment, the bottom end of the sample processing chamber is further provided with a filter column.
According to a preferred embodiment, a bonding film is arranged in the direction of the side wall of the main box body, the bonding film is a PET/CPP composite film, and the bonding film and the main box body are bonded in a gluing or heat sealing mode.
The main scheme and the further selection schemes of the utility model can be freely combined to form a plurality of schemes, which are the schemes that can be adopted and claimed by the utility model; and the utility model discloses also can the independent assortment between (each non-conflict selection) selection and between other choices. The technical solutions to be protected by the present invention, which are various combinations that can be known to those skilled in the art based on the prior art and the common general knowledge after understanding the present invention, are not exhaustive herein.
The utility model has the advantages that: through the structural design of this card box, can realize presetting various reagents, put into corresponding cavity in advance with it. Thus, the complicated operation of adding reagent in the treatment process is not needed. In addition, the card box can directly process samples such as cotton sticks, blood strips, gauze, swabs and the like. The sealed design of card box can not cause environmental pollution, can realize high automation. The card box can be injection molded, and the cost is low. After the product is taken out, all the liquid in the box body is in a sealed state. No special treatment is needed, and the environment is not polluted.
Drawings
FIG. 1 is a schematic diagram of the front structure of the cartridge of the present invention;
FIG. 2 is a schematic view of the back structure of the cartridge of the present invention;
FIG. 3 is a schematic diagram of the cartridge explosion structure of the present invention;
FIG. 4 is a schematic structural view of the main body of the cartridge of the present invention;
FIG. 5 is a schematic structural view of a two-way valve of the cartridge of the present invention;
FIG. 6 is a schematic view of the structure of the water blocking hole of the cartridge of the present invention;
FIG. 7 is a schematic view of the position of the magnet outside the cartridge of the present invention;
FIG. 8 is a schematic view of the piping structure of the cartridge of the present invention;
wherein, the kit comprises a main box body 10, a syringe 100, a liquid flow channel 110, a valve lower flow channel 120, a sample processing cavity 131, a PK freeze-drying enzyme storage cavity 132, a magnetic bead storage cavity 133, a lysate storage cavity 134, a first washing solution storage cavity 135, a second washing solution storage cavity 136, an eluent storage cavity 137, a purification cavity 138, a water cut-off cavity 139, an extraction product cavity 140, a sample processing cavity cover 141, an extraction product cavity cover 142, a water blocking hole 151, an air-permeable water blocking column 152, a filter column 155, a magnet 157, a two-way valve 160, a first two-way valve 161, a second two-way valve 162, a third two-way valve 163, a fourth two-way valve 164, a fifth two-way valve 165, a sixth two-way valve 166, 167-a seventh two-way valve, 168-an eighth two-way valve, 169-a ninth two-way valve, 20-a sealing gasket, 201-a hot riveting column, 202-a positioning column, 203-an inlet and outlet port, 204-a valve through hole, 205-a positioning hole, 206-a valve gating flow channel, 30-a rotary valve, 40-a valve cover, 50-a piston rod and 60-a bonding film.
Detailed Description
The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. The present invention can also be implemented or applied through other different specific embodiments, and various details in the present specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict.
It should be noted that, in order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, not all embodiments. The components of embodiments of the present invention, as generally described and illustrated in the figures herein, may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the accompanying drawings, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.
It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate the position or positional relationship based on the position or positional relationship shown in the drawings, or the position or positional relationship which is usually placed when the product of the present invention is used, and are only for convenience of description and simplification of the description, but do not indicate or imply that the device or element referred to must have a specific position, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance.
Furthermore, the terms "horizontal", "vertical", "overhang" and the like do not imply that the components are required to be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.
In the description of the present invention, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Additionally, the utility model discloses it is pointed out that, in the utility model, if do not write out structure, connection relation, positional relationship, power source relation etc. that concretely relates to very much, then the utility model relates to a structure, connection relation, positional relationship, power source relation etc. are technical personnel in the field on prior art's basis, can not learn through creative work.
Example 1:
referring to FIGS. 1 to 8, there is shown a hermetically sealed nucleic acid extraction and purification cartridge comprising a main cartridge body 10, a sealing gasket 20, a rotary valve 30, a valve cover 40, a piston rod 50 and a bonding film 60.
Preferably, the main case 10 is made of PP, and the rotary valve 30 and the valve cover 40 are made of PP or PC. The sealing gasket 20 is made of rubber, silica gel or fluorine gel. The piston rod 50 is made of PP materials, and the piston is made of rubber or TPE materials. The bonding film 60 is a PET/CPP composite film. The bonding film 60 is located in the direction of the side wall of the main box 10 and is bonded to the main box 10 by gluing or heat sealing.
Preferably, the main cartridge body 10 is provided with a syringe 100, a sample processing chamber 131, a PK freeze-drying enzyme storage chamber 132, a magnetic bead storage chamber 133, a lysate storage chamber 134, a first washing solution storage chamber 135, a second washing solution storage chamber 136, an eluent storage chamber 137, a purification chamber 138, an extraction product chamber 140 and a water cut-off chamber 139 in sequence.
Preferably, the piston rod 50 is assembled in the syringe 100, and the top end of the piston rod 50 is provided with a piston.
Preferably, the main case 10 is intermittently provided at the top thereof with a multi-stage valve lower flow passage 120. And the under-valve flow passage 120, the packing 20 and the rotary valve 30 constitute a two-way valve 160.
Preferably, the two-way valve 160 is provided at the top end of the main case 10. The cartridge includes 9 two-way valves 160. And a valve lower flow passage 120 is arranged between every two adjacent valve bodies in the 9 two-way valves 160 to complete the series connection of the valve bodies.
Preferably, the syringe 100 is connected to a two-way valve 160 disposed at the head end, and the shut-off chamber 139 is communicated to the two-way valve 160 at the tail end. The water stopping cavity 139 is provided with a water stopping hole 151 on the side wall, and a ventilating water stopping column 152 is arranged in the water stopping hole 151 and communicated with the external space through the ventilating water stopping column 152. Preferably, the air-permeable and water-blocking column 152 may be made of air-permeable and water-blocking material.
That is, the air-permeable water-blocking column 152 is the only channel between the inner cavity of the card box and the flow channel and the outside, and since the air-permeable water-blocking column 152 has a small aperture and cannot pass through molecules with a diameter larger than that of water molecules, microbial molecules and reagents in all reagents can be blocked in the box by the air-permeable water-blocking column, and the outside cannot be polluted.
Simultaneously, set up the water-stop chamber 139 at the tail end of card box and have two effects: first, in case moisture permeates the chamber, the water trap chamber 139 allows the moisture to drop into the water trap chamber 139 without directly blocking the air-permeable and water-blocking column 152. Secondly, the air pressure buffering is performed at the tail end, so that the air pressure in the box is not changed violently due to the slow ventilation speed of the ventilation and water blocking column 152.
Preferably, the valve bodies of the two-way valve 160 from the head end to the tail end are further respectively communicated with the top and the bottom of the sample processing chamber 131, the PK freeze-drying enzyme storage chamber 132, the magnetic bead storage chamber 133, the lysate storage chamber 134, the first washing solution storage chamber 135, the second washing solution storage chamber 136, the eluent storage chamber 137, the purification chamber 138 and the extraction product chamber 140.
The switching of each cavity into the valve lower flow passage 120 in series or out of the valve lower flow passage 120 is completed by the valve rotation switching of the two-way valve 160. That is, the two-way valve 160 can selectively connect each chamber in series to the lower valve flow channel 120 to form a pipeline or close the connection pipeline between each chamber and the lower valve flow channel 120.
Preferably, the top end of the sample processing cavity 131 is further provided with a sample processing cavity cover 141. The bottom end of the sample processing chamber 131 is further provided with a filter column 155. The filter column 155 is used to complete the filtration process of the liquid flowing through. The top end of the extraction product cavity 140 is also provided with an extraction product cavity cover 142.
Preferably, a positioning hole 205 is formed in the center of the bottom end of the rotary valve 30, and two inward-recessed valve gate flow passages 206 are symmetrically formed on both sides of the positioning hole 205.
The sample processing cavity 131, the PK freeze-drying enzyme storage cavity 132, the magnetic bead storage cavity 133, the lysate storage cavity 134, the first washing solution storage cavity 135, the second washing solution storage cavity 136, the eluent storage cavity 137, the purification cavity 138, and the inlet and outlet ports 203 of each cavity in the extraction product cavity 140, which correspond to the liquid flow channel 110 and each section of the valve lower flow channel 120, are arranged at the top end of the box body.
The top end of the box body is further provided with a positioning column 202, the rotary valve 30 is in fit connection with the positioning column 202 through a positioning hole 205, and the inlet and outlet ports 203 of the liquid flow passage 110 and the valve lower flow passage 120 corresponding to each cavity are respectively communicated with four ports of the valve gating flow passage 206 of the corresponding rotary valve 30.
The sealing gasket 20 is arranged between the rotary valve 30 and the top end of the box body, and at least a valve through hole 204 matched with the size of the inlet and outlet port 203 is arranged on the sealing gasket 20. The sealing gasket 20 is further provided with a through hole structure for passing through the positioning column 202.
Therefore, after the rotary valve 30 rotates by 90 degrees, the communication state between the liquid flow passage 110 of the corresponding chamber and the under-valve flow passage 120 can be switched once. I.e. from on to off, or from off to on.
Preferably, the valve cover 40 is fixed to the top end of the main case 10 via a rivet 201. And the packing 20 and the rotary valve 30 are disposed between the valve cover 40 and the main case 10.
Specifically, as shown in fig. 8, the 9 two-way valves 160 are a first two-way valve 161, a second two-way valve 162, a third two-way valve 163, a fourth two-way valve 164, a fifth two-way valve 165, a sixth two-way valve 166, a seventh two-way valve 167, an eighth two-way valve 168, and a ninth two-way valve 169, respectively.
A valve lower flow passage 120 is arranged between each adjacent valve body of the first two-way valve 161, the second two-way valve 162, the third two-way valve 163, the fourth two-way valve 164, the fifth two-way valve 165, the sixth two-way valve 166, the seventh two-way valve 167, the eighth two-way valve 168 and the ninth two-way valve 169 to complete the series connection of the valve bodies. The direction of the main box body pipeline close to the injection cylinder 100 is the upstream side, and the side close to the water-stopping cavity 139 is the downstream side.
Preferably, the bottom end of the syringe barrel 100 is provided with a liquid flow passage 110 communicating with a valve gate flow passage 206 on the upstream side of the first two-way valve 161.
Preferably, the top end of the sample processing chamber 131 is communicated with the valve gating channel 206 on the downstream side of the first two-way valve 161 via the liquid channel 110, and the bottom end of the sample processing chamber 131 is communicated with the valve gating channel 206 on the upstream side of the first two-way valve 161 via the liquid channel 110.
Preferably, the top end of the PK holding chamber 132 communicates with the gate flow path 206 on the downstream side of the second two-way valve 162160 via the liquid flow path 110, and the bottom end of the PK holding chamber 132 communicates with the gate flow path 206 on the upstream side of the second two-way valve 162 via the liquid flow path 110.
Preferably, the top end of the magnetic bead storage chamber 133 is connected to the valve gating channel 206 on the upstream side of the third two-way valve 163 via the liquid channel 110, and the bottom end of the magnetic bead storage chamber 133 is connected to the valve gating channel 206 on the downstream side of the third two-way valve 163 via the liquid channel 110.
Preferably, the top end of the lysate holding chamber 134 communicates with the valve gating flow path 206 on the downstream side of the fourth two-way valve 164 through the liquid flow path 110, and the bottom end of the lysate holding chamber 134 communicates with the valve gating flow path 206 on the upstream side of the fourth two-way valve 164 through the liquid flow path 110.
Preferably, the top end of the first cleaning solution holding chamber 135 communicates with the gate flow path 206 on the upstream side of the fifth two-way valve 165 through the liquid flow path 110, and the bottom end of the first cleaning solution holding chamber 135 communicates with the gate flow path 206 on the downstream side of the fifth two-way valve 165 through the liquid flow path 110.
Preferably, the top end of the second cleaning solution holding chamber 136 communicates with the gate flow path 206 on the upstream side of the sixth two-way valve 166 via the liquid flow path 110, and the bottom end of the second cleaning solution holding chamber 136 communicates with the gate flow path 206 on the downstream side of the sixth two-way valve 166 via the liquid flow path 110.
Preferably, the top end of the eluent holding cavity 137 is communicated with the valve gating flow channel 206 on the upstream side of the seventh two-way valve 167 through the liquid flow channel 110, and the bottom end of the eluent holding cavity 137 is communicated with the valve gating flow channel 206 on the downstream side of the seventh two-way valve 167 through the liquid flow channel 110.
Preferably, the top end of the purifying chamber 138 communicates with the valve gating flow path 206 on the downstream side of the eighth two-way valve 168 via the liquid flow path 110, and the bottom end of the purifying chamber 138 communicates with the valve gating flow path 206 on the upstream side of the eighth two-way valve 168 via the liquid flow path 110.
Preferably, the top end of the extraction product chamber 140 communicates with the valve gating flow path 206 on the downstream side of the ninth two-way valve 169 via the liquid flow path 110, and the bottom end of the extraction product chamber 140 communicates with the valve gating flow path 206 on the upstream side of the ninth two-way valve 169 via the liquid flow path 110.
The working principle or the working method flow of the card box is as follows:
in the initial state of the cartridge valve, as shown in FIG. 8, the syringe and the water shut-off chamber 139 formed by the syringe cartridge 100 and the piston rod are directly connected through the two-way valve 160. That is, the two-way valve 160 does not connect the chambers into the valve down flow passage 120.
The first step is as follows: the sample processing chamber cover 141 is opened and a sample such as a swab is added to the sample processing chamber 131. Sample chamber cover 141 is closed.
The second step is that: the valves of the second two-way valve 162 and the fourth two-way valve 164 corresponding to the freeze-dried PK enzyme storage cavity 132 and the lysate storage cavity 134 are rotated, so that the freeze-dried PK enzyme storage cavity 132 and the lysate storage cavity 134 are connected to the valve lower flow channel 120 and the solution is extracted by a syringe. Because the freeze-dried PK enzyme storage cavity 132 and the lysate storage cavity 134 are reversed, the lysate liquid will first pass through the freeze-dried PK enzyme storage cavity 132, and all the lysate liquid is sucked by the syringe after the freeze-dried PK enzyme is dissolved.
Because the other end of the cavity is connected with the breathable water-blocking column 152, gas can freely enter and exit from the box, so that the pressure in the box can not change due to the movement of the injector, and the problem that the gas cannot be sucked or cannot be discharged due to uneven air pressure can not be caused. This problem is not described in detail in the following steps.
The third step: and closing the valves of the two-way valves 160 corresponding to the freeze-drying PK enzyme storage cavity 132 and the lysate storage cavity 134, and cutting the valve lower flow channel 120 of the freeze-drying PK enzyme storage cavity 132 and the lysate storage cavity 134. The valve of the sample cavity 131 corresponding to the two-way valve 160 is opened, the piston of the injector is pushed, the liquid is injected into the sample cavity 131, and the piston moves up and down to blow and mix the liquid. The filter column 155 of the sample processing chamber 131 has a large pore size and allows the reagents to pass through. If heating is required, a heating device may be added to the sample processing chamber 131 externally to heat the reagent.
The fourth step: after lysis is complete, the fluid in the sample processing chamber 131 is aspirated by a syringe. Due to the presence of the filter column 155, the swab will be retained and after lysis, the liquid containing the nucleic acids and other cellular impurities will be sucked into the syringe in its entirety.
The fifth step: the sample processing chamber 131 is cut out by closing the corresponding valve of the first two-way valve 161 of the sample processing chamber 131. The valves of the third two-way valve 163 and the eighth two-way valve 168 corresponding to the magnetic bead storage chamber 133 and the purification chamber 138 are opened, the syringe is pushed, and the liquid in the fourth step enters the purification chamber 138 through the magnetic bead storage chamber 133 together with the magnetic beads.
And a sixth step: and closing a valve of the third two-way valve 163 corresponding to the magnetic bead storage cavity 133, moving the piston up and down to blow and pump the liquid in the mixing and purifying cavity 138, so that the nucleic acid is adsorbed on the magnetic beads.
The seventh step: an external magnet 157 is loaded outside the purification chamber 138, as shown in fig. 7, to adsorb magnetic beads. After the magnetic beads were fully adsorbed, the waste liquid was aspirated into the syringe. The fourth two-way valve 164 corresponding to the lysate holding cavity 134 is opened to inject the waste fluid into the lysate holding cavity 134.
Eighth step: the valve of the fourth two-way valve 164 corresponding to the lysate holding chamber 134 is closed. The syringe is pulled and no liquid is drawn into the syringe at this point because only the purification chamber valve is open and the magnetic beads are attracted by the magnet. Then the magnet 157 is removed, the valve of the fifth two-way valve 165 corresponding to the first cleaning solution storage cavity 135 is opened, the injector is pushed, the first cleaning solution is pushed into the purifying cavity 138, and the piston is moved up and down to blow and mix the cleaning solution. Then, the magnet 157 is loaded to adsorb the magnetic beads.
The ninth step: by pulling the syringe plunger, all waste fluid will enter the first cleaning fluid chamber 135 and not the syringe, since the first cleaning fluid holding chamber 135 is upright. The valve of the first cleaning solution holding chamber 135 corresponding to the fifth two-way valve 165 is closed.
The tenth step: in the same manner as the first wash solution, the beads are washed a second time with a second wash solution, and the waste solution is returned to the second wash solution holding chamber 136.
The eleventh step: and opening a valve of a seventh two-way valve 167 corresponding to the eluent storage cavity 137, pushing the injector, pushing the eluent into the purification cavity 138, and moving the injector up and down to blow and mix the magnetic beads. Then, the magnet 157 is loaded to adsorb the magnetic beads, and after the magnetic beads are sufficiently adsorbed, the valve of the seventh two-way valve 167 corresponding to the eluent storage chamber 137 is closed, and the eluent containing nucleic acids is sucked into the syringe.
The twelfth step: the valve of the eighth two-way valve 168 corresponding to the purification chamber 138 is closed, and the valve of the ninth two-way valve 169 corresponding to the extraction product chamber 140 is opened, and the nucleic acid eluate is injected into the extraction product chamber 140. And closes the valve of the ninth two-way valve 169 corresponding to the extraction product chamber 140.
The thirteenth step: the extraction product chamber cover 142 is opened and the final nucleic acid eluate is aspirated. The nucleic acid extraction and purification process is complete.
Through the structural design of this card box, can realize presetting various reagents, put into corresponding cavity in advance with it. Thus, the complicated operation of adding reagent in the treatment process is not needed. In addition, the card box can directly process samples such as cotton sticks, blood strips, gauze, swabs and the like. The sealed design of card box can not cause environmental pollution, can realize high automation. The card box can be injection molded, and the cost is low. After the product is taken out, all the liquid in the box body is in a sealed state. No special treatment is needed, and the environment is not polluted.
The aforesaid the utility model discloses basic embodiment and each further alternative can the independent assortment in order to form a plurality of embodiments, is the utility model discloses can adopt and claim the embodiment of protection. In the scheme of the utility model, each selection example can be combined with any other basic examples and selection examples at will.
The above description is only exemplary of the present invention and should not be taken as limiting the scope of the present invention, as any modifications, equivalents, improvements and the like made within the spirit and principles of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A totally enclosed cassette for nucleic acid extraction and purification, characterized in that the cassette comprises at least a main cassette body, a piston rod and 9 two-way valves,
wherein, an injection tube, a sample processing cavity, a PK freeze-drying enzyme storage cavity, a magnetic bead storage cavity, a lysate storage cavity, a first washing liquid storage cavity, a second washing liquid storage cavity, an eluent storage cavity, a purification cavity, an extraction product cavity and a water interception cavity are sequentially arranged in the main box body side by side;
the piston rod is assembled in the injection cylinder, and the top end of the piston rod is provided with a piston;
the two-way valve is arranged at the top end of the main box body, a lower valve flow channel is arranged between every two adjacent valve bodies in the 9 two-way valves to complete the series connection of the valve bodies, the injection tube is connected with the two-way valve arranged at the head end, the water intercepting cavity is communicated with the two-way valve at the tail end, a water blocking hole is arranged on the side wall of the water intercepting cavity, and an air-permeable water-blocking column is arranged in the water blocking hole and is communicated with the external space through the air-permeable water-blocking column;
and each valve body of the two-way valve from the head end to the tail end is also respectively communicated with the top and the bottom of the sample processing cavity, the PK freeze-drying enzyme storage cavity, the magnetic bead storage cavity, the lysate storage cavity, the first washing liquid storage cavity, the second washing liquid storage cavity, the eluent storage cavity, the purification cavity and the product extraction cavity, and the conversion of each cavity into the lower valve flow channel in series or out of the lower valve flow channel is completed through the valve rotation switching of the two-way valve.
2. The totally enclosed nucleic acid extraction and purification cartridge of claim 1, wherein the two-way valve comprises at least a valve-lower flow channel, a gasket, and a rotary valve,
a positioning hole is arranged at the center of the bottom end of the rotating valve, two inwards concave valve gating flow passages are symmetrically arranged at two sides taking the positioning hole as the circle center,
the inlet and outlet ports of each cavity corresponding to the liquid flow passage and each section of valve lower flow passage are arranged at the top end of the box body, the top end of the box body is also provided with a positioning column,
the rotary valve is matched and connected with the positioning column through a positioning hole,
the inlet and outlet ports of the liquid flow passage and the valve lower flow passage corresponding to each cavity are respectively communicated with the four ports of the valve gating flow passage of the corresponding rotating valve;
the sealing gasket is arranged between the rotary valve and the top end of the box body, and at least one valve through hole matched with the size of the inlet and outlet port is formed in the sealing gasket.
3. The hermetically sealed nucleic acid extracting and purifying cartridge of claim 2, wherein the 9 two-way valves are a first two-way valve, a second two-way valve, a third two-way valve, a fourth two-way valve, a fifth two-way valve, a sixth two-way valve, a seventh two-way valve, an eighth two-way valve, and a ninth two-way valve, respectively,
a valve lower flow channel is arranged between every two adjacent valve bodies in the first two-way valve, the second two-way valve, the third two-way valve, the fourth two-way valve, the fifth two-way valve, the sixth two-way valve, the seventh two-way valve, the eighth two-way valve and the ninth two-way valve to complete the series connection of the valve bodies;
the bottom end of the injection tube is provided with a liquid flow passage which is communicated with a valve gating flow passage on the upstream side of the first two-way valve;
the top end of the sample processing cavity is communicated with a valve gating flow channel on the downstream side of the first two-way valve through a liquid flow channel, and the bottom end of the sample processing cavity is communicated with a valve gating flow channel on the upstream side of the first two-way valve through a liquid flow channel;
the top end of the PK freeze-dried enzyme storage cavity is communicated with a valve gating flow channel on the downstream side of the second two-way valve through a liquid flow channel, and the bottom end of the PK freeze-dried enzyme storage cavity is communicated with a valve gating flow channel on the upstream side of the second two-way valve through a liquid flow channel;
the top end of the magnetic bead storage cavity is communicated with a valve gating flow channel on the upstream side of the third two-way valve through a liquid flow channel, and the bottom end of the magnetic bead storage cavity is communicated with a valve gating flow channel on the downstream side of the third two-way valve through a liquid flow channel;
the top end of the lysate storage cavity is communicated with a valve gating flow channel on the downstream side of the fourth two-way valve through a liquid flow channel, and the bottom end of the lysate storage cavity is communicated with a valve gating flow channel on the upstream side of the fourth two-way valve through a liquid flow channel;
the top end of the first cleaning solution storage cavity is communicated with a valve gating flow channel on the upstream side of the fifth two-way valve through a liquid flow channel, and the bottom end of the first cleaning solution storage cavity is communicated with a valve gating flow channel on the downstream side of the fifth two-way valve through a liquid flow channel;
the top end of the second cleaning solution storage cavity is communicated with a valve gating flow channel on the upstream side of the sixth two-way valve through a liquid flow channel, and the bottom end of the second cleaning solution storage cavity is communicated with a valve gating flow channel on the downstream side of the sixth two-way valve through a liquid flow channel;
the top end of the eluent storage cavity is communicated with a valve gating flow channel on the upstream side of the seventh two-way valve through a liquid flow channel, and the bottom end of the eluent storage cavity is communicated with a valve gating flow channel on the downstream side of the seventh two-way valve through a liquid flow channel;
the top end of the purification cavity is communicated with a valve gating flow channel on the downstream side of the eighth two-way valve through a liquid flow channel, and the bottom end of the purification cavity is communicated with a valve gating flow channel on the upstream side of the eighth two-way valve through a liquid flow channel;
the top end of the extraction product cavity is communicated with a valve gating flow channel on the downstream side of the ninth two-way valve through a liquid flow channel, and the bottom end of the extraction product cavity is communicated with a valve gating flow channel on the upstream side of the ninth two-way valve through a liquid flow channel.
4. The hermetically sealed nucleic acid extraction and purification cartridge of claim 3, wherein the gasket further comprises a through hole structure for passing the positioning post.
5. The hermetically sealed nucleic acid extraction and purification cartridge of claim 4, wherein the gasket is made of rubber or silica gel or fluoro gel.
6. The hermetically sealed nucleic acid extraction and purification cartridge of claim 5, further comprising a valve cover, wherein the valve cover is fixed to the top of the main cartridge body via a rivet stem, and the sealing gasket and the rotary valve are disposed between the valve cover and the main cartridge body.
7. The hermetically sealed nucleic acid extraction and purification cartridge of claim 6, wherein the main cartridge body is made of PP, and the rotary valve and the valve cover are made of PP or PC.
8. The hermetically sealed nucleic acid extraction and purification cartridge of claim 1, wherein a sample processing chamber cover is further disposed at a top end of the sample processing chamber;
and an extraction product cavity cover is also arranged at the top end of the extraction product cavity.
9. The hermetically sealed nucleic acid extraction and purification cartridge of claim 1, wherein the bottom end of the sample processing chamber further comprises a filter column.
10. The hermetically sealed cartridge for nucleic acid isolation and purification of claim 1, wherein a bonding film is disposed on the main cartridge in the direction of the sidewall, the bonding film is a PET/CPP composite film, and the bonding film is bonded to the main cartridge by gluing or heat sealing.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202021453799.7U CN212955086U (en) | 2020-07-22 | 2020-07-22 | Totally enclosed nucleic acid draws and purification card box |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202021453799.7U CN212955086U (en) | 2020-07-22 | 2020-07-22 | Totally enclosed nucleic acid draws and purification card box |
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| CN212955086U true CN212955086U (en) | 2021-04-13 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111826273A (en) * | 2020-07-22 | 2020-10-27 | 赵毅 | Automatic totally-enclosed micro-fluidic chip for nucleic acid detection |
| CN114558632A (en) * | 2022-04-26 | 2022-05-31 | 广州国家实验室 | Liquid transfer device, multi-channel liquid transfer device and method |
| CN114574324A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Sample extraction device |
| CN114570449A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Liquid transfer device and multi-path parallel liquid transfer device |
| CN114574349A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Equipment that adaptation card box used |
| WO2023206093A1 (en) * | 2022-04-26 | 2023-11-02 | 广州国家实验室 | Sample extraction device |
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2020
- 2020-07-22 CN CN202021453799.7U patent/CN212955086U/en active Active
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111826273A (en) * | 2020-07-22 | 2020-10-27 | 赵毅 | Automatic totally-enclosed micro-fluidic chip for nucleic acid detection |
| CN114558632A (en) * | 2022-04-26 | 2022-05-31 | 广州国家实验室 | Liquid transfer device, multi-channel liquid transfer device and method |
| CN114574324A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Sample extraction device |
| CN114570449A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Liquid transfer device and multi-path parallel liquid transfer device |
| CN114574349A (en) * | 2022-04-26 | 2022-06-03 | 广州国家实验室 | Equipment that adaptation card box used |
| CN114574349B (en) * | 2022-04-26 | 2022-10-14 | 广州国家实验室 | Equipment for adapting to card box |
| WO2023206093A1 (en) * | 2022-04-26 | 2023-11-02 | 广州国家实验室 | Sample extraction device |
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Inventor after: Zhao Yi Inventor before: Zhao Yi Inventor before: Zhang Jianyi |
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Effective date of registration: 20211104 Address after: 201499 3rd floor, building 1, 368 Xiaonan Road, Fengxian District, Shanghai Patentee after: Shanghai fenghuotai Enterprise Management Co.,Ltd. Address before: No.46 Zhengdong street, Huilong Town, Yingshan County, Nanchong City, Sichuan Province Patentee before: Zhao Yi |
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Effective date of registration: 20231228 Address after: Unit 202, Building 1, Phase II, Suzhou Biomedical Industrial Park, No. 218, Sangtian Street, Suzhou Area, China (Jiangsu) Pilot Free Trade Zone, Suzhou City, Jiangsu Province, 215000 Patentee after: SUZHOU PRECIGENOME Co.,Ltd. Address before: 201499 3rd floor, building 1, 368 Xiaonan Road, Fengxian District, Shanghai Patentee before: Shanghai fenghuotai Enterprise Management Co.,Ltd. |