CN113667598A - Nucleic acid detection device and nucleic acid detection method - Google Patents
Nucleic acid detection device and nucleic acid detection method Download PDFInfo
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- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 78
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 78
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 78
- 238000001514 detection method Methods 0.000 title abstract description 18
- 239000007788 liquid Substances 0.000 claims abstract description 175
- 238000006243 chemical reaction Methods 0.000 claims abstract description 131
- 238000004891 communication Methods 0.000 claims abstract description 31
- 238000007789 sealing Methods 0.000 claims description 44
- 239000003153 chemical reaction reagent Substances 0.000 claims description 31
- 238000000034 method Methods 0.000 claims description 7
- 239000011148 porous material Substances 0.000 claims description 5
- 230000003321 amplification Effects 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 4
- 238000010521 absorption reaction Methods 0.000 claims description 3
- 230000000717 retained effect Effects 0.000 claims description 3
- 239000011324 bead Substances 0.000 description 22
- 238000005406 washing Methods 0.000 description 21
- 239000000203 mixture Substances 0.000 description 20
- 102000004190 Enzymes Human genes 0.000 description 15
- 108090000790 Enzymes Proteins 0.000 description 15
- 108010067770 Endopeptidase K Proteins 0.000 description 14
- 239000000243 solution Substances 0.000 description 11
- 239000011550 stock solution Substances 0.000 description 8
- 239000003480 eluent Substances 0.000 description 6
- 238000000605 extraction Methods 0.000 description 6
- 239000000443 aerosol Substances 0.000 description 4
- 238000009434 installation Methods 0.000 description 4
- 238000003752 polymerase chain reaction Methods 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 230000009089 cytolysis Effects 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
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Abstract
The present invention relates to a nucleic acid detection device and a nucleic acid detection method, wherein the nucleic acid detection device comprises: the liquid storage assembly comprises a fixing frame, a piston bin and a plurality of liquid storage bins, and the fixing frame is provided with a fixing frame piston hole and a plurality of fixing frame liquid storage holes; the reaction bin is connected with the fixing frame in a matching mode, and the fixing frame is provided with a reaction bin liquid inlet channel communicated with the reaction bin; the runner base is arranged on one side of the liquid storage assembly and is provided with a base piston hole and a plurality of runners; the rotary valve is arranged on one side of the flow channel base, which is far away from the liquid storage assembly, a rotary valve piston hole and a rotary valve communication hole which are mutually communicated are formed in the rotary valve, and the rotary valve piston hole is correspondingly communicated with the base piston hole; the rotary valve can rotate relative to the flow channel base by using the central axis of the rotary valve piston hole as a rotating shaft, so that the rotary valve communication hole can alternatively communicate with the flow channel.
Description
Technical Field
The invention relates to the field of biological detection, in particular to a nucleic acid detection device and a nucleic acid detection method.
Background
Nucleic acids are carriers of genetic information of living bodies and are essential constituent materials for all known life forms. Nucleic acids, DNA and RNA, are mainly present in the nucleus and are present in a state of being bound to proteins. With the rapid development of molecular biology, research and analysis of nucleic acid are being promoted and applied in clinical diagnosis, food safety, environmental detection, agriculture, forestry, animal husbandry and other fields.
Conventional nucleic acid extraction needs to be performed in a PCR (Polymerase Chain Reaction, abbreviation) laboratory. However, the extraction of nucleic acid requires more matched instruments, and aerosol pollution is easily caused by carelessness, so that the requirements on operating environment and operators are high, and timely detection under emergency is difficult to deal with.
Disclosure of Invention
Therefore, it is necessary to provide a nucleic acid detecting apparatus and a nucleic acid detecting method, which can achieve the technical effects of reducing the requirements of the experimental environment and preventing the pollution of the aerosol sample, in order to solve the problem that the requirements of the nucleic acid detection on the experimental environment are high.
According to one aspect of the present application, there is provided a nucleic acid detecting apparatus comprising:
the liquid storage assembly comprises a fixing frame, a piston bin and a plurality of liquid storage bins, wherein the piston bin and the plurality of liquid storage bins are arranged on one side of the fixing frame;
the reaction bin is connected to the fixing frame in a matching mode, and a reaction bin liquid inlet channel communicated with the reaction bin is formed in the fixing frame;
the runner base is arranged on one side of the liquid storage assembly, a base piston hole and a plurality of runners are formed in the runner base, the base piston hole is communicated with the fixing frame piston hole, and one end of each runner is communicated with one fixing frame liquid storage hole or the reaction bin liquid inlet channel; and
the rotary valve is arranged on one side of the flow channel base, which is far away from the liquid storage assembly, a rotary valve piston hole and a rotary valve communication hole which are mutually communicated are formed in the rotary valve, and the rotary valve piston hole is correspondingly communicated with the base piston hole;
the rotary valve can rotate relative to the flow channel base by taking the central axis of the piston hole of the rotary valve as a rotating shaft so as to enable the rotary valve communication hole to be alternatively communicated with the flow channel, and the piston bin can generate negative pressure for absorbing the reagent in the liquid storage bin or the reaction bin or positive pressure for injecting the reagent into the liquid storage bin or the reaction bin.
In one embodiment, the plurality of liquid storage bins include a liquid discharge bin, the fixing frame is provided with a reaction bin liquid discharge channel communicated with the reaction bin, the plurality of flow channels include a reaction bin liquid discharge flow channel and a liquid discharge bin flow channel, the reaction bin liquid discharge flow channel is communicated with the reaction bin liquid discharge channel, and the liquid discharge bin flow channel is communicated with the liquid discharge bin;
the rotary valve is provided with a rotary valve communicating groove which is selectively communicated with the reaction bin liquid drainage flow channel and the liquid drainage bin flow channel.
In one embodiment, the plurality of flow channels further comprise a reaction bin liquid inlet flow channel, and the reaction bin liquid inlet flow channel is communicated with the reaction bin liquid inlet channel;
when the rotary valve intercommunicating pore passes through the reaction bin liquid inlet flow channel and is communicated with the reaction bin liquid inlet flow channel, the rotary valve intercommunicating pore is communicated with the reaction bin liquid discharge flow channel and the liquid discharge bin flow channel.
In one embodiment, the liquid storage bin comprises an open end and a closed end which are oppositely arranged, the closed end is provided with a communication hole communicated with the liquid storage hole of the fixing frame, the nucleic acid detection device further comprises a liquid storage bin sealing film, and at least part of the open end of the liquid storage bin is sealed by the liquid storage bin sealing film.
In one embodiment, the plurality of storage chambers includes a sample chamber, and the nucleic acid detecting device further includes a sealing cover openably and closably connected to an open end of the sample chamber.
In one embodiment, the piston cabin comprises a piston cabin body and a piston, the piston cabin body is installed on the fixing frame, the piston is contained in the piston cabin body and is in interference fit with the inner wall of the piston cabin body, and the piston can reciprocate in the piston cabin body to generate negative pressure or positive pressure.
In one embodiment, the nucleic acid detecting apparatus further comprises a sealing member disposed between the flow path base and the rotary valve, the sealing member being provided with a sealing member communication hole communicating the flow path base and the rotary valve.
In one embodiment, the nucleic acid detecting device further comprises a mounting seat, the mounting seat is coupled to a side of the flow channel assembly away from the liquid storage assembly, one end of the rotary valve is rotatably limited in the mounting seat, and the other end of the rotary valve extends out of the mounting seat.
In one embodiment, the reaction chamber comprises a reaction chamber main body and a reaction chamber sealing film, and the reaction chamber sealing film is coated outside the reaction chamber main body.
In one embodiment, the fixing frame is provided with an installation groove, one end of the reaction chamber is convexly provided with a clamping arm, and the clamping arm is clamped in the installation groove so that the reaction chamber is matched and connected with the fixing frame.
According to another aspect of the present invention, there is provided a nucleic acid detecting method using the above-mentioned nucleic acid detecting apparatus, comprising the steps of:
s1: adding various reagents for extracting nucleic acid and a sample to be detected into the liquid storage bins respectively;
s2: the rotary valve is rotated to sequentially communicate with the flow channel corresponding to the liquid storage bin in an alternative mode, and the piston bin is driven to generate positive pressure or negative pressure so as to suck and/or inject the sample to be detected in the liquid storage bin and a reagent for extracting nucleic acid to mix, and selectively carry out magnetic absorption treatment and/or heating treatment;
s3: sucking the liquid containing the nucleic acid after the treatment and introducing the liquid into the reaction chamber for amplification treatment.
Above-mentioned nucleic acid detection device only needs can make piston storehouse correspond the intercommunication with the stock solution storehouse of difference through rotating the rotary valve, and then realizes the exchange of reagent, has reduced the interference of external factor to the extraction result, is showing the operating environment restriction who has reduced nucleic acid detection, has reduced the requirement to operating personnel.
Drawings
FIG. 1 is a schematic structural view of a nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 2 is an exploded view of the nucleic acid detecting apparatus shown in FIG. 1;
FIG. 3 is a schematic view showing the construction of a rotary valve of the nucleic acid detecting apparatus shown in FIG. 1;
FIG. 4 is a schematic view showing the manner of communication among the reservoir unit, the flow path unit, and the rotary valve of the nucleic acid detecting apparatus shown in FIG. 1.
The reference numbers illustrate:
100. a nucleic acid detecting device; 110. a liquid storage assembly; 112. a fixed mount; 1121. a bottom fixing portion; 1123. a side fixing part; 1123a and a reaction bin liquid inlet channel; 1123b, a reaction chamber liquid discharge channel; 114. a piston chamber; 1141. a piston bin body; 1143. a piston; 116. a liquid storage bin; 1161. a sample bin; 1162. a protease K bin; 1163. a magnetic bead bin; 1164a, a first washing liquid bin; 1164b, a second washing liquid bin; 1164c, a third washing liquid bin; 1165. an eluent bin; 1166. an exhaust bin; 1167. a Taq enzyme compartment; 1168. a Mix enzyme bin; 1169. a liquid discharge bin; 120. a reaction bin; 121. a reaction bin main body; 1212. a liquid inlet hole of the reaction bin; 1214. a liquid discharge hole of the reaction bin; 1216. a chucking arm; 123. a first reaction chamber sealing film; 125. a second reaction chamber sealing film; 130. a flow channel base; 132. a base piston bore; 134. a flow channel; 1341. a reaction bin liquid inlet flow channel; 1342. a liquid discharge flow channel of the reaction bin; 1343. a drainage bin flow channel; 1344. a liquid storage bin flow channel; 140. rotating the valve; 141. rotating the valve body; 1412. rotating a valve piston bore; 1414. a rotary valve communicating hole; 1416. a rotary valve communicating groove; 143. a rotating part; 145a and a first liquid storage bin sealing film; 145b and a second liquid storage bin sealing film; 160. a sealing cover; 170. a seal member; 180. and (7) mounting a seat.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the invention.
Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless otherwise expressly stated or limited, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through an intermediate. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like as used herein are for illustrative purposes only and do not denote a unique embodiment.
Referring to fig. 1 and 2, an embodiment of the present invention provides a nucleic acid detecting apparatus 100, and an operator can extract nucleic acid through the nucleic acid detecting apparatus 100. Because all the transfer and mixing processes of the reagents are carried out in the nucleic acid detecting device 100, the interference of external factors can be conveniently avoided, and aerosol pollution is avoided.
The nucleic acid detecting device 100 includes a reservoir assembly 110, a reaction chamber 120, a flow channel base 130, and a rotary valve 140. The reservoir assembly 110 is used for storing various reagents, the reaction chamber 120 is used for performing a polymerase chain reaction, and the flow channel base 130 is matched with the rotary valve 140 for transferring the reagents in the reservoir assembly 110 and the reaction chamber 120.
The reservoir assembly 110 includes a mounting bracket 112, a piston chamber 114, and a plurality of reservoirs 116. The fixing frame 112 includes a bottom fixing portion 1121 and a side fixing portion 1123. The bottom fixing portion 1121 has an elongated structure, a longitudinal direction of the bottom fixing portion 1121 is a first direction (X direction in fig. 1), a width direction of the bottom fixing portion 1121 is a second direction (Y direction in fig. 1), and a thickness direction of the bottom fixing portion 1121 is a third direction (Z direction in fig. 1). The side fixing portion 1123 is located at one end of the bottom fixing portion 1121 in the first direction, and the side fixing portion 1123 extends upward from the upper surface of the bottom fixing portion 1121 in the third direction. The piston chamber 114 and the liquid storage chamber 116 are both cylindrical hollow tubular structures, one end of the piston chamber 114 and one end of the liquid storage chamber 116 are fixedly connected to the upper surface of the bottom fixing portion 1121, and the other end extends upward along the third direction.
Further, the bottom fixing portion 1121 is provided with a fixing frame piston hole and a plurality of fixing frame liquid storage holes, and the fixing frame piston hole and each fixing frame liquid storage hole penetrate through the upper surface and the lower surface of the bottom fixing portion 1121 along the third direction.
The piston chamber 114 includes a piston chamber main body 1141 and a piston 1143, the piston chamber main body 1141 includes an open end and a closed end which are oppositely arranged in a third direction, the closed end of the piston chamber 114 is located at one end of the piston chamber 114 connected with the bottom fixing portion 1121, and the closed end of the piston 1143 is provided with a communication hole for communicating with a piston hole of the fixing frame. The piston 1143 is accommodated in the piston chamber body 1141 and is in interference fit with the chamber wall of the piston chamber 114, and the piston 1143 can reciprocate in the piston chamber body 1141, so as to generate a negative pressure for sucking the reagent in the liquid storage chamber 116 or the reaction chamber 120, or a positive pressure for injecting the reagent into the liquid storage chamber 116 or the reaction chamber 120.
Thus, the reagent can enter and exit the piston chamber 114 through the piston hole of the fixing frame, and can enter and exit the liquid storage chamber 116 through the liquid storage hole of the fixing frame. It is understood that the types and the number of the liquid storage bins 116 can be set according to different requirements to meet different experimental requirements.
Furthermore, the side fixing portion 1123 is opened with a reaction chamber liquid inlet channel 1123a communicating with the reaction chamber 120 and a reaction chamber liquid discharge channel 1123b communicating with the reaction chamber 120, and the reaction chamber 120 can communicate with the piston chamber 114 through the reaction chamber liquid inlet channel 1123a and communicate with the liquid discharge chamber 1169 through the reaction chamber liquid discharge channel 1123 b.
The reaction chamber 120 is detachably inserted into the side fixing portion 1123 of the fixing frame 112. The reaction chamber 120 is a hollow flat structure, and includes a reaction chamber main body 121 and a reaction chamber sealing film, the reaction chamber main body 121 is provided with a reaction chamber allowing liquid to flow, and the reaction chamber main body 121 is covered with the reaction chamber sealing film to seal the reaction chamber. Specifically, in some embodiments, the reaction chamber 120 includes a first reaction chamber sealing film and a second reaction chamber sealing film, which are respectively attached to two side surfaces of the reaction chamber body 121 and are thermally fused with the reaction chamber body 121. Because the reaction bin 120 is of a flat structure, the reaction bin has a high cooling speed, and the experimental efficiency is improved.
Further, a reaction chamber liquid inlet hole 1212 and a reaction chamber liquid outlet hole 1214 are convexly formed in one end of the reaction chamber main body 121 facing the fixing frame 112, the reaction chamber liquid inlet hole 1212 and the reaction chamber liquid outlet hole 1214 are arranged at intervals in the third direction, when the reaction chamber 120 is inserted into the side fixing portion 1123, the reaction chamber liquid inlet hole 1212 extends into the reaction chamber liquid inlet channel 1123a to be communicated with the reaction chamber liquid inlet channel 1123a, and the reaction chamber liquid outlet hole 1214 extends into the reaction chamber liquid outlet channel 1123b to be communicated with the reaction chamber liquid outlet channel 1123 b.
Preferably, in some embodiments, in order to firmly couple the reaction chamber main body 121 and the fixing frame 112, two retaining arms 1216 are protruded from one end of the reaction chamber main body 121 facing the fixing frame 112, and the two retaining arms 1216 are spaced in the third direction. The side fixing portion 1123 of the fixing frame 112 has two mounting slots spaced apart in the third direction, and each of the retaining arms 1216 is capable of generating a recoverable deformation to be retained in one of the mounting slots. In this manner, the reaction chamber 120 is detachably coupled to the fixing frame 112 by the retaining arm 1216.
Referring to fig. 4, the flow channel base 130 is located on a side of the bottom fixing portion 1121 of the fixing frame 112 away from the liquid storage chamber 116, and a base piston hole and a plurality of flow channels 134 are formed on an upper surface of the flow channel base 130 facing the bottom fixing portion 1121, so as to guide the flow of the reagent and communicate the liquid storage assembly 110 and the rotary valve 140.
The base piston hole penetrates through the upper surface and the lower surface of the flow channel base 130 along the third direction, and the base piston hole and the fixing frame piston hole are correspondingly and coaxially arranged in the third direction and are communicated with each other. The plurality of flow channels 134 include a reaction chamber liquid inlet flow channel 1341, a reaction chamber liquid discharge flow channel 1342, a liquid discharge chamber flow channel 1343, and a plurality of liquid storage chamber flow channels 1344.
One end of the reaction bin liquid inlet passage 1341 is communicated with the reaction bin liquid inlet passage 1341 formed in the fixing frame 112, and the other end of the reaction bin liquid inlet passage 1341 penetrates through the upper surface and the lower surface of the passage base 130 along the third direction. One end of the reaction chamber liquid drainage channel 1342 is communicated with the reaction chamber liquid drainage channel 1342 formed on the fixing frame 112, and the other end of the reaction chamber liquid drainage channel 1342 penetrates through the upper and lower surfaces of the channel base 130 along the third direction. One end of the liquid drainage chamber passage 1343 is communicated with the liquid drainage chamber 1169 through the liquid storage hole of the fixing frame, and the other end of the liquid drainage chamber passage 1343 penetrates through the upper and lower surfaces of the passage base 130 along the third direction. One end of each liquid storage bin flow passage 1344 is communicated with one liquid storage bin 116 through a liquid storage hole of the fixing frame, and the other end of each liquid storage bin flow passage 1344 penetrates through the upper surface and the lower surface of the flow passage base 130 along the third direction.
Referring to fig. 3 and 4, the rotary valve 140 is disposed on a side of the flow channel base 130 away from the reservoir assembly 110, a rotary valve piston hole 1412, a rotary valve communication hole 1414 and a rotary valve communication groove 1416 are formed on a surface of the rotary valve 140 facing the reservoir assembly 110, the rotary valve piston hole 1412 and the rotary valve communication hole 1414 are spaced apart and communicate with each other, and the rotary valve communication groove 1416 is disposed on a side of the rotary valve piston hole 1412.
When the rotary valve 140 is coupled to the flow channel base 130, the rotary valve piston hole 1412 is correspondingly communicated with the base piston hole, and the rotary valve 140 can rotate relative to the flow channel base 130 with the central axis of the rotary valve piston hole 1412 as a rotating axis, so that the rotary valve communication hole 1414 is selectively communicated with the flow channel 134, and the rotary valve communication groove 1416 is selectively communicated with the reaction chamber drainage flow channel 1342 and the drainage chamber flow channel 1343.
Specifically, when the rotary valve communication hole 1414 is in communication with one of the reservoir channels 1344 of the channel 134 assembly, the reagent in the reservoir 116 that the channel 134 is in communication with can flow from the reservoir channel 1344 into the rotary valve communication hole 1414 and then into the piston chamber 114 through the rotary valve piston hole 1412, and conversely, the liquid in the piston chamber 114 can also flow into the corresponding reservoir channel 1344 through the rotary valve piston hole 1412 and then into the corresponding reservoir 116 through the selectively rotary valve communication hole 1414.
Further, when the rotary valve communication hole 1414 is communicated with the reaction bin liquid inlet channel 1341, the rotary valve communication groove 1416 is communicated with the reaction bin liquid outlet channel 1342 and the liquid outlet channel 1343, the reagent in the piston bin 114 can sequentially pass through the rotary valve piston hole 1412, the rotary valve communication hole 1414, the reaction bin liquid inlet channel 1123a and the reaction bin liquid inlet channel 1123a to enter the reaction bin 120, and the reagent in the reaction bin 120 can sequentially pass through the reaction bin liquid outlet channel 1123b, the reaction bin liquid outlet channel 1342 and the liquid outlet channel 1343 to enter the liquid outlet bin 1169.
In some embodiments, the nucleic acid detecting device 100 further comprises a reservoir sealing film that seals at least a portion of the open end of the reservoir 116, thereby preventing the reservoir 116 storing the reagent from communicating with the external environment to form aerosol contamination. Specifically, in one embodiment, the sealing film of the liquid storage compartment encloses the open ends of the proteinase K compartment 1162, the magnetic bead compartment 1163, the first washing solution compartment 1164a, the second washing solution compartment 1164b, the third washing solution compartment 1164c, the eluent compartment 1165, the gas exhaust compartment 1166, the Taq enzyme compartment 1167, the Mix enzyme compartment 1168, and the liquid discharge compartment 1169. Thus, after the reagent is injected into the reservoir 116 from the open end of the reservoir 116, a reservoir sealing film may be covered on the open end of the reservoir 116 to seal the reservoir 116.
It is understood that the nucleic acid detecting apparatus 100 may include different numbers of cartridge sealing films so that the cartridges 116 to be sealed are sealed, depending on the arrangement of the cartridges 116. Specifically, in one embodiment, the liquid discharge chamber 1169 is spaced apart from the other liquid discharge chambers 116, and the nucleic acid detecting apparatus 100 includes a first liquid discharge chamber sealing film 145a and a second liquid discharge chamber sealing film 145b having different shapes, the first liquid discharge chamber sealing film 145a is used to seal the opening end of the liquid discharge chamber 116 except for the liquid discharge chamber 1169, and the second liquid discharge chamber sealing film 145b is used to seal the liquid discharge chamber 1169.
In some embodiments, the nucleic acid detecting device 100 further comprises a sealing cover 160, and the sealing cover 160 is openably and closably connected to the open end of the sample compartment 1161. Thus, after a sample is added to sample compartment 1161, closure flap 160 may be used to close sample compartment 1161. Since sealing lid 160 can be easily switched between the open state and the closed state, the sample can be easily added while ensuring the sealing effect of sample compartment 1161.
In some embodiments, the nucleic acid detecting apparatus 100 further includes a sealing member 170, the sealing member 170 is disposed between the flow path base 130 and the rotary valve 140, and the sealing member 170 is provided with a sealing member communication hole for communicating the flow path base 130 and the rotary valve 140. Thus, the flow channel base 130 and the rotary valve 140 are hermetically connected by the sealing ring, and the leakage of the reagent from between the flow channel base 130 and the rotary valve 140 is effectively prevented. Specifically, in one embodiment, the sealing member 170 is formed of a material that is resiliently deformable under an external force to provide a good seal.
In some embodiments, the nucleic acid detecting device 100 further comprises a mount 180 for limiting the rotary valve 140. Specifically, mount pad 180 includes the mount pad diapire and extends the mount pad lateral wall that forms towards the same direction from the edge of mount pad diapire, and the mount pad lateral wall encircles the mount pad diapire along circumference in order to define jointly with the mount pad diapire and form the installation cavity, and the mounting groove of intercommunication installation cavity is seted up to the mount pad lateral wall. The flow channel base 130 is provided with a mounting buckle matching with the mounting groove, so that the mounting seat 180 is coupled to a side of the flow channel base 130 away from the liquid storage assembly 110 by the buckle.
The rotary valve 140 includes a rotary valve body 141 and a rotary portion 143 connected to the rotary valve body 141, the rotary valve piston hole 1412, the rotary valve communication hole 1414 and the rotary valve communication groove 1416 are all opened in the rotary valve body 141, the rotary valve body 141 is rotatably limited in the mounting seat 180, and the rotary portion 143 extends out of the mounting seat bottom wall of the mounting seat along a third direction.
In this manner, an operator may hold the end of the rotary valve 140 extending beyond the mounting base 180 to rotate the rotary valve 140. In other embodiments, the end of the rotary valve 140 extending beyond the mounting seat 180 may also be drivingly connected to an external drive mechanism for rotation by the external drive mechanism.
The present application also provides a nucleic acid extraction method using the nucleic acid detection device 100, including the steps of:
s1: adding various reagents for extracting nucleic acid and a sample to be detected into each liquid storage bin 116 respectively;
specifically, the operator prepackages each reagent for extracting nucleic acid into each of the reservoirs 116, and adds the sample to be tested in the sample compartment 1161, and then closes the sealing lid 160 of the open end of the sample compartment 1161, and seals the open end of the reservoir 116 with the reservoir sealing film 145.
S2: the rotary valve 140 is rotated to sequentially and alternatively communicate with the flow channel corresponding to the liquid storage bin 116, and the piston bin is driven to generate positive pressure or negative pressure so as to suck and/or inject the sample to be detected in the liquid storage bin 116 and the reagent for extracting nucleic acid for mixing, and selectively carry out magnetic absorption treatment and/or heating treatment;
in some embodiments, step S2 includes the steps of:
s21: the sample in one of the reservoirs 116 is aspirated and injected into the reservoir 116 storing proteinase K to form a mixture of the sample and proteinase K.
Specifically, the rotary valve 140 is rotated to connect the sample chamber 1161 and the piston chamber 114, the piston chamber 114 sucks the sample in the sample chamber 1161, then the rotary valve 140 is rotated to connect the piston chamber 114 and the proteinase K chamber, and then the piston chamber 114 injects the sample into the proteinase K chamber 1162 to mix the sample with the proteinase K.
S22: the mixture of the sample and the proteinase K is aspirated and injected into the reservoir 116 storing the magnetic beads to form a mixture of the sample, the proteinase K, and the magnetic beads.
Specifically, after the piston chamber 114 sucks the mixture of the sample and the proteinase K, the rotary valve 140 is rotated to connect the piston chamber 114 and the magnetic bead chamber 1163, and then the piston chamber 114 injects the mixture into the magnetic bead chamber 1163 to mix the sample and the magnetic beads.
S23: the mixture of the sample, proteinase K and magnetic beads is heated to accelerate the lysis rate of the sample.
Specifically, the magnetic bead bin 1163 is heated by an external heating device to accelerate the lysis speed of the sample. It will be appreciated that in other embodiments, the sample may not be heated.
S24: the washing solution in one of the reservoirs 116 is aspirated and the reservoir 116 storing the mixture of the sample, proteinase K and magnetic beads is filled.
Specifically, the rotary valve 140 is rotated to connect the piston chamber 114 with the first washing liquid chamber 1164a, the piston chamber 114 sucks the first washing liquid in the first washing liquid chamber 1164a, then the rotary valve 140 is rotated again to connect the piston chamber with the magnetic bead chamber 1163, and then the piston chamber 114 is used to inject the first washing liquid into the magnetic bead chamber 1163 to wash the magnetic beads.
S25: the mixture of the sample, proteinase K, magnetic beads, and wash solution is aspirated and injected into the reservoir 116 to wash the magnetic beads.
Specifically, the piston chamber 114 first sucks the liquid containing the magnetic beads washed by the first washing solution, the rotary valve 140 is rotated to connect the piston chamber 114 and the second washing solution chamber 1164b, the piston chamber 114 injects the second washing solution chamber 1164b to continuously wash the magnetic beads, then the magnetic beads washed by the second washing solution are sucked, the rotary valve 140 is rotated to connect the piston chamber 114 and the third washing solution chamber 1164c, and then the piston chamber 114 is injected into the third washing chamber to continuously wash the magnetic beads by using the third washing solution.
S26: the washed mixture is aspirated and injected into a reservoir 116 holding an eluent to separate the nucleic acids from the magnetic beads.
Specifically, the piston chamber 114 sucks the cleaned mixture, the rotary valve 140 is then rotated to connect the piston chamber 114 and the eluent chamber 1165, and then the cleaned mixture is injected into the eluent chamber 1165 to separate the nucleic acids from the magnetic beads and dissolve the nucleic acids in the eluent.
S27: the eluate mixed with the nucleic acid is aspirated and injected into the reservoir 116 storing the Taq enzyme.
Specifically, the piston chamber 114 sucks the washed nucleic acid-containing eluate, and then the rotary valve 140 is rotated to connect the piston chamber 114 with the Taq enzyme chamber 1167, and then the eluate mixed with the nucleic acid is injected into the Taq enzyme chamber 1167.
S28: the mixture containing the Taq enzyme is aspirated and injected into the reservoir 116 where the Mix enzyme is stored.
Specifically, the piston chamber 114 sucks the mixture containing the Taq enzyme, and then the rotary valve 140 is rotated to communicate the piston chamber 114 with the Mix enzyme chamber 1168, and then the mixture containing the Taq enzyme is injected into the Mix enzyme chamber 1168.
In some embodiments, step S2 includes the following: the sample and the nucleic acid extraction reagent in the reservoir 116 are aspirated and mixed.
Specifically, rotary valve 140 is driven to rotate so that exchange hole 212 communicates with sample chamber 1161 and reservoir 116 in turn, and piston 1143 moves in the third direction to sequentially aspirate sample and nucleic acid extraction reagent.
In some embodiments, after step S2, the method further includes the following steps:
s3: the liquid containing the nucleic acid after the treatment is aspirated and introduced into the reaction chamber 120 to perform an amplification treatment.
Specifically, the piston chamber 114 sucks up the liquid containing the nucleic acid, and then the rotary valve 140 is rotated to connect the piston chamber 114 and the reaction chamber 120, and then the liquid containing the nucleic acid is injected into the reaction chamber 120 for the subsequent amplification process.
It is to be understood that the specific steps of the experiment performed by the nucleic acid detecting apparatus 100 are not limited, and different experimental procedures may be established as needed.
Above-mentioned nucleic acid detecting device only needs manual reagent that adds once, and the user only need rotate rotary valve 140 and can make piston storehouse 114 and different stock solution storehouse 116 or reaction storehouse 120 intercommunication each other, and then carries out the transfer of reagent under the closed environment, and then realizes nucleic acid and draws, has higher reagent transfer efficiency when effectively having prevented external factors interference. Different reagents can be selectively configured in the reaction chamber 120, and the operation process can be flexibly adjusted, so as to achieve the detection purpose of different requirements.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (11)
1. A nucleic acid detecting apparatus, comprising:
the liquid storage assembly comprises a fixing frame, a piston bin and a plurality of liquid storage bins, wherein the piston bin and the plurality of liquid storage bins are arranged on one side of the fixing frame;
the reaction bin is connected to the fixing frame in a matching mode, and a reaction bin liquid inlet channel communicated with the reaction bin is formed in the fixing frame;
the runner base is arranged on one side of the liquid storage assembly, a base piston hole and a plurality of runners are formed in the runner base, the base piston hole is communicated with the fixing frame piston hole, and one end of each runner is communicated with one fixing frame liquid storage hole or the reaction bin liquid inlet channel; and
the rotary valve is arranged on one side of the flow channel base, which is far away from the liquid storage assembly, a rotary valve piston hole and a rotary valve communication hole which are mutually communicated are formed in the rotary valve, and the rotary valve piston hole is correspondingly communicated with the base piston hole;
the rotary valve can rotate relative to the flow channel base by taking the central axis of the piston hole of the rotary valve as a rotating shaft so as to enable the rotary valve communication hole to be alternatively communicated with the flow channel, and the piston bin can generate negative pressure for absorbing the reagent in the liquid storage bin or the reaction bin or positive pressure for injecting the reagent into the liquid storage bin or the reaction bin.
2. The apparatus according to claim 1, wherein the plurality of liquid storage compartments include a liquid discharge compartment, the holder defines a liquid discharge channel for the reaction compartment, the plurality of flow channels include a liquid discharge channel for the reaction compartment and a liquid discharge channel for the reaction compartment, the liquid discharge channel for the reaction compartment is in communication with the liquid discharge channel for the reaction compartment, and the liquid discharge channel for the reaction compartment is in communication with the liquid discharge compartment;
the rotary valve is provided with a rotary valve communicating groove which is selectively communicated with the reaction bin liquid drainage flow channel and the liquid drainage bin flow channel.
3. The nucleic acid detecting device according to claim 2, wherein a plurality of the flow channels further include a reaction chamber liquid inlet flow channel, and the reaction chamber liquid inlet flow channel is communicated with the reaction chamber liquid inlet channel;
when the rotary valve intercommunicating pore passes through the reaction bin liquid inlet flow channel and is communicated with the reaction bin liquid inlet flow channel, the rotary valve intercommunicating pore is communicated with the reaction bin liquid discharge flow channel and the liquid discharge bin flow channel.
4. The nucleic acid detecting device according to claim 1, wherein the reservoir includes an open end and a closed end that are disposed opposite to each other, the closed end being opened with a communication hole that communicates with the reservoir hole of the holder, the nucleic acid detecting device further including a reservoir sealing film that seals at least a part of the open end of the reservoir.
5. The apparatus for detecting nucleic acid according to claim 4, wherein the plurality of storage chambers includes a sample chamber, and the apparatus further includes a sealing lid openably and closably connected to an open end of the sample chamber.
6. The nucleic acid detecting device according to claim 1, wherein the piston chamber includes a piston chamber body and a piston, the piston chamber body is mounted on the fixing frame, the piston is accommodated in the piston chamber body and is in interference fit with an inner wall of the piston chamber body, and the piston can reciprocate in the piston chamber body to generate negative pressure or positive pressure.
7. The nucleic acid detecting apparatus according to claim 1, further comprising a sealing member disposed between the flow path base and the rotary valve, the sealing member having a sealing member communication hole communicating the flow path base and the rotary valve.
8. The apparatus according to claim 1, further comprising a mounting base, wherein the mounting base is coupled to a side of the flow channel assembly away from the liquid storage assembly, one end of the rotary valve is rotatably retained in the mounting base, and the other end of the rotary valve extends out of the mounting base.
9. The nucleic acid detecting device according to claim 1, wherein the reaction chamber includes a reaction chamber main body and a reaction chamber sealing film, and the reaction chamber sealing film covers the reaction chamber main body.
10. The apparatus for detecting nucleic acid according to claim 1, wherein the fixing frame has a mounting groove, and a retaining arm is protruded from one end of the reaction chamber, and the retaining arm is retained in the mounting groove to couple the reaction chamber to the fixing frame.
11. A nucleic acid detecting method using the nucleic acid detecting apparatus according to any one of claims 1 to 10, comprising the steps of:
s1: adding various reagents for extracting nucleic acid and a sample to be detected into the liquid storage bins respectively;
s2: the rotary valve is rotated to sequentially communicate with the flow channel corresponding to the liquid storage bin in an alternative mode, and the piston bin is driven to generate positive pressure or negative pressure so as to suck and/or inject the sample to be detected in the liquid storage bin and a reagent for extracting nucleic acid to mix, and selectively carry out magnetic absorption treatment and/or heating treatment;
s3: sucking the liquid containing the nucleic acid after the treatment and introducing the liquid into the reaction chamber for amplification treatment.
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| CN202110943952.7A CN113667598A (en) | 2021-08-17 | 2021-08-17 | Nucleic acid detection device and nucleic acid detection method |
| PCT/CN2022/110369 WO2023011607A1 (en) | 2021-08-04 | 2022-08-04 | Nucleic acid detection device, and method for detecting nucleic acid |
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Cited By (1)
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| WO2023011607A1 (en) * | 2021-08-04 | 2023-02-09 | 圣湘生物科技股份有限公司 | Nucleic acid detection device, and method for detecting nucleic acid |
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