CN116640659A - Sample extraction and detection device - Google Patents
Sample extraction and detection device Download PDFInfo
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- CN116640659A CN116640659A CN202310732909.5A CN202310732909A CN116640659A CN 116640659 A CN116640659 A CN 116640659A CN 202310732909 A CN202310732909 A CN 202310732909A CN 116640659 A CN116640659 A CN 116640659A
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- 238000001514 detection method Methods 0.000 title claims abstract description 56
- 238000000605 extraction Methods 0.000 title claims abstract description 52
- 238000006243 chemical reaction Methods 0.000 claims abstract description 72
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 13
- 238000007789 sealing Methods 0.000 claims description 41
- 238000004108 freeze drying Methods 0.000 claims description 35
- 239000007788 liquid Substances 0.000 claims description 35
- 238000010828 elution Methods 0.000 claims description 29
- 230000009089 cytolysis Effects 0.000 claims description 16
- 238000005406 washing Methods 0.000 claims description 16
- 238000005192 partition Methods 0.000 claims description 9
- 238000004891 communication Methods 0.000 claims description 6
- 239000003480 eluent Substances 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 abstract description 32
- 108020004707 nucleic acids Proteins 0.000 abstract description 32
- 150000007523 nucleic acids Chemical class 0.000 abstract description 32
- 230000003321 amplification Effects 0.000 abstract description 15
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 15
- 230000008569 process Effects 0.000 abstract description 8
- 238000004140 cleaning Methods 0.000 description 19
- 238000005336 cracking Methods 0.000 description 6
- 238000003745 diagnosis Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000012535 impurity Substances 0.000 description 4
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- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 208000016361 genetic disease Diseases 0.000 description 2
- 230000002934 lysing effect Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
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- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
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- 238000003753 real-time PCR Methods 0.000 description 1
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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
- C12Q1/6806—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]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The application relates to a sample extraction and detection device, which comprises a kit, a reaction part and a magnetic attraction part, wherein the kit is provided with a plurality of cavities, different cavities are used for storing different reagents required by sample extraction, at least one cavity is internally provided with the magnetic part, the reaction part is connected with the kit, the reaction part is provided with a reaction cavity, and the reaction cavity is communicated with at least one cavity. The magnetic attraction piece can be inserted into any cavity and is used for attracting or releasing the magnetic piece. The sample extraction and detection device can complete all processes of nucleic acid extraction, amplification, detection and the like in one device, simplifies the whole structure, reduces the probability of sample pollution, and improves the accuracy of sample extraction and detection.
Description
Technical Field
The application relates to the technical field of sample detection, in particular to a sample extraction detection device.
Background
Molecular diagnosis refers to a technique of detecting changes in the structure or expression level of genetic material in a patient using molecular biological methods to make a diagnosis. Molecular diagnosis is a main method for predictive diagnosis, and can be used for diagnosing individual genetic diseases and prenatal diagnosis. Molecular diagnostics refers primarily to the detection of genes encoding various structural proteins, enzymes, antigen antibodies, immunologically active molecules associated with disease.
The main technology of molecular detection is fluorescent quantitative PCR (Polymerase Chain Reaction, PCR) technology, which has high reaction sensitivity, strong specificity and short diagnosis window period, can perform qualitative and quantitative detection, can be widely used for hepatitis, venereal disease, pulmonary infectious diseases, prenatal and postnatal care, genetic disease genes, tumors and the like, fills the detection blank of early immunodetection window period, and provides effective help for early diagnosis, early treatment and safe blood use. The process of detecting a sample by using a PCR technique is generally divided into three steps of sample extraction, amplification and detection. Because of the characteristics of the exponential amplification template of the PCR technology, related sample extraction and detection operations are mostly carried out in an open experimental environment, each step needs to be completed by independent equipment, multiple pieces of equipment are needed in one detection process, samples need to be moved to equipment in a subsequent step after the previous step is completed, the experimental steps are more, the operation is complex, and the sample is easily polluted by the external environment or the sample is polluted by the detection environment in the sample transfer process.
Disclosure of Invention
Based on this, it is necessary to provide a sample extraction and detection device, which can complete all the processes of nucleic acid extraction, amplification, detection and the like in one device, simplify the overall structure, reduce the probability of sample contamination, and improve the accuracy of sample extraction and detection.
A sample extraction detection device, comprising:
the kit is provided with a plurality of cavities, different cavities are used for storing different reagents, and at least one cavity is used for storing a magnetic piece;
the reaction piece is connected with the kit and is provided with a reaction cavity which is communicated with at least one cavity; the method comprises the steps of,
the magnetic attraction piece can be inserted into any cavity and is used for adsorbing or releasing the magnetic piece.
The technical scheme is further described as follows:
in one embodiment, the magnetic attraction piece comprises a sleeve, the sleeve is a hollow pipe body with one end closed and the other end open, the closed end of the magnetic attraction piece can be inserted into any cavity, and the open end of the magnetic attraction piece is used for inserting or withdrawing a magnet so that the sleeve can absorb or release the magnetic piece.
In one embodiment, the sample extraction detection device further comprises a sealing cover, wherein the sealing cover is in sealing connection with the reagent kit and seals each cavity.
In one embodiment, the plurality of cavities comprise a cracking cavity for storing cracking liquid, a magnetic part cavity for storing the magnetic part, a cleaning cavity for storing cleaning liquid, an eluting cavity for storing eluent and a freeze-drying cavity for storing freeze-drying, and the reaction cavity is communicated with the freeze-drying cavity.
In one embodiment, the elution chamber and the lyophilization chamber are arranged in a stacked manner, and a partition is disposed between the elution chamber and the lyophilization chamber, and the magnetic attraction member is capable of penetrating through the partition to enable the elution chamber to be communicated with the lyophilization chamber.
In one embodiment, the plurality of cavities further comprise piston cavities, movable piston pieces are arranged in the piston cavities, the reaction pieces are further provided with liquid inlet channels and liquid outlet channels, the liquid inlet channels and the liquid outlet channels are communicated with the reaction cavities, the liquid inlet channels are communicated with the freeze-drying cavities, the liquid outlet channels are communicated with the piston cavities, and the magnetic attraction pieces can penetrate into the piston cavities and drive the piston pieces to move along the piston cavities.
In one embodiment, a sealing sleeve is arranged in the cavity communicated with the reaction cavity, and the magnetic attraction piece can be in sealing fit with the sealing sleeve to seal the reaction cavity.
In one embodiment, the kit is further provided with a receiving cavity for receiving the magnetic attraction member.
In one embodiment, the kit is provided with a slot, the slot is communicated with at least one of the cavities, the reaction member is inserted into the slot, a clamping member is arranged at the slot, the clamping member is detachably connected with the kit, and the clamping member is matched with the reaction member in a clamping manner so as to limit the reaction member in the slot.
In one embodiment, a connecting hole communicated with at least one cavity is formed in the slot, a sealing ring is arranged in the connecting hole, a connecting pipe communicated with the reaction cavity is convexly arranged on the reaction piece, and the connecting pipe is inserted into the connecting hole and is in sealing fit with the sealing ring.
According to the sample extraction detection device, the plurality of cavities are formed in the kit, so that different reagents required by sample extraction can be respectively preloaded in the different cavities, the magnetic pieces are preloaded in one of the cavities, the magnetic pieces can adsorb nucleic acid of the sample, the magnetic pieces are used for adsorbing the magnetic pieces, then the magnetic pieces are sequentially inserted into the different cavities and released, the magnetic pieces adsorbed with the nucleic acid of the sample can be sequentially transferred into the different cavities, the nucleic acid reacts with the reagents in the different cavities, the extraction of the nucleic acid of the sample is realized, and finally the nucleic acid of the extracted sample is transferred into the reaction tank for reaction by utilizing the cavities communicated with the reaction cavity, so that the amplification and detection of the nucleic acid are finally realized. Therefore, the sample extraction and detection device provided by the embodiment of the application can complete all processes of sample extraction, purification, amplification, detection and the like at one time, reduce different equipment required by sample detection, simplify the overall structure, reduce the probability of sample pollution and improve the accuracy of sample extraction and detection.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application.
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for the description of the embodiments will be briefly described below, and it is apparent that the drawings in the following description are only some embodiments of the present application, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
Moreover, the figures are not drawn to a 1:1 scale, and the relative sizes of various elements are merely exemplary in the figures, and are not necessarily drawn to true scale. In the drawings:
fig. 1 is a schematic structural diagram of a sample extraction detection device according to an embodiment.
Fig. 2 is a top view of the sample volume extraction detection device shown in fig. 1.
FIG. 3 is a cross-sectional view of the sample volume extraction test apparatus shown in FIG. 2 taken along section A-A.
Fig. 4 is a partial enlarged view of a portion a shown in fig. 3.
Fig. 5 is a structural exploded view of the sample body extraction detection apparatus shown in fig. 1.
Reference numerals illustrate:
10. a kit; 11. a lysing chamber; 12. a magnetic member cavity; 13. a first cleaning chamber; 14. a second cleaning chamber; 15. an elution chamber; 151. a partition; 152. sealing sleeve; 16. a lyophilization chamber; 17. a piston chamber; 18. a receiving chamber; 19. a slot; 191. a first connection hole; 192. a second connection hole; 193. a limit part; 20. a reaction member; 21. a reaction chamber; 22. a liquid inlet channel; 23. a liquid discharge channel; 24. a first connection pipe; 25. a second connection pipe; 26. a boss; 30. a magnetic attraction piece; 31. a sleeve; 40. a piston member; 50. and the clamping piece.
Detailed Description
In order that the above objects, features and advantages of the application will be readily understood, a more particular description of the application will be rendered by reference to the appended drawings. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. The present application may be embodied in many other forms than described herein and similarly modified by those skilled in the art without departing from the spirit of the application, whereby the application is not limited to the specific embodiments disclosed below.
In the description of the present application, it should be understood that, if any, these terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc., are used herein with respect to the orientation or positional relationship shown in the drawings, these terms refer to the orientation or positional relationship for convenience of description and simplicity of description only, and do not indicate or imply that the apparatus or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore should not be construed as limiting the application.
Furthermore, the terms "first," "second," and the like, if any, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present application, the terms "plurality" and "a plurality" if any, mean at least two, such as two, three, etc., unless specifically defined otherwise.
In the present application, unless explicitly stated and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly. For example, the two parts can be fixedly connected, detachably connected or integrated; can be mechanically or electrically connected; either directly or indirectly, through intermediaries, or both, may be in communication with each other or in interaction with each other, unless expressly defined otherwise. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art according to the specific circumstances.
In the present application, unless expressly stated or limited otherwise, the meaning of a first feature being "on" or "off" a second feature, and the like, is that the first and second features are either in direct contact or in indirect contact through an intervening medium. Moreover, a first feature being "above," "over" and "on" a second feature may be a first feature being directly above or obliquely above the second feature, or simply indicating that the first feature is level higher than the second feature. The first feature being "under", "below" and "beneath" the second feature may be the first feature being directly under or obliquely below the second feature, or simply indicating that the first feature is less level than the second feature.
It will be understood that if an element is referred to as being "fixed" or "disposed" on another element, it can be directly on the other element or intervening elements may also be present. If 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, if any, are for descriptive purposes only and do not represent a unique embodiment.
An embodiment of the present application provides a sample extraction and detection device for extracting and detecting a sample, where the sample may be a liquid sample or a swab sample. Specifically, referring to fig. 1 to 3, the sample extraction detection device of an embodiment includes a kit 10, a reaction member 20, and a magnetic attraction member 30, where the kit 10 is provided with a plurality of cavities, different cavities are used for storing different reagents required for sample extraction, at least one of the cavities is provided with a magnetic member (not shown), the reaction member 20 is connected with the kit 10, the reaction member 20 is provided with a reaction chamber 21, and the reaction chamber 21 is communicated with at least one of the cavities. The magnetic attraction piece 30 can be inserted into any cavity, and the magnetic attraction piece 30 is used for attracting or releasing the magnetic attraction piece.
According to the sample extraction and detection device, the plurality of cavities are formed in the kit 10, so that different reagents required by sample extraction can be respectively preloaded in the different cavities, the magnetic pieces are preloaded in one of the cavities, the magnetic pieces can adsorb nucleic acid of the sample, the magnetic pieces are adsorbed by the magnetic pieces 30, then the magnetic pieces 30 are sequentially inserted into the different cavities and released, the magnetic pieces adsorbed with the nucleic acid of the sample can be sequentially transferred into the different cavities, the nucleic acid reacts with the reagents in the different cavities, the extraction of the nucleic acid of the sample is realized, the nucleic acid of the extracted sample is transferred into the reaction tank for reaction by the cavities communicated with the reaction cavity 21, and the amplification and detection of the nucleic acid are finally realized. Therefore, the sample extraction and detection device provided by the embodiment of the application can complete all processes of sample extraction, purification, amplification, detection and the like at one time, reduce different equipment required by sample detection, simplify the overall structure, reduce the probability of sample pollution and improve the accuracy of sample extraction and detection.
Referring to fig. 5, alternatively, in an embodiment, the magnetic attraction member includes a sleeve 31, the sleeve 31 is a hollow tube body with one end closed and the other end open, the closed end of the sleeve 31 can be inserted into any cavity, and the open end of the sleeve 31 is used for inserting or withdrawing a magnet (not shown) so that the sleeve 31 attracts or releases the magnetic member. Specifically, when the magnetic member needs to be transferred from a cavity to a target cavity, the magnetic member can be adsorbed to the outer wall of the sleeve 31 by inserting the sleeve 31 into the cavity with the magnetic member and then inserting the magnet from the opening end of the sleeve 31, then the sleeve 31 and the magnet are integrally pulled out of the original cavity and inserted into the target cavity, and finally the magnet is pulled out of the sleeve 31, so that the magnetic member can be released into the target cavity. Preferably, in an embodiment, the magnetic member may be a magnetic bead, and the magnet may be a magnetic rod.
It should be noted that, in other embodiments, the magnetic attraction member 30 may be an electromagnetic member, and the electromagnetic member is electrified or electrified to be electrified or demagnetized, so that the electromagnetic member attracts or releases the magnetic member, and thus the magnetic member can be transferred to different cavities.
Optionally, in an embodiment, the sample extraction detection device further includes a sealing cover (not shown) that is in sealing connection with the kit 10 and seals each cavity, and the magnetic attraction member 30 can be inserted into any cavity through the sealing cover. Preferably, the openings of all the cavities are opened at the same end of the kit 10, and the sealing cover covers the end, so that all the cavities are sealed. By sealing each cavity through the sealing cover, the reagent and the magnetic piece in the cavity are prevented from falling out before the sample extraction detection device is used, for example, in the transportation or storage process, the reagent and the magnetic piece are prevented from being polluted, and the accuracy of subsequent detection is improved. Preferably, the sealing cover may be an aluminum film or a plastic film, further referring to fig. 5, the closed end of the sleeve 31 has a pointed structure so that the sealing cover can be easily pierced through the closed end of the sleeve 31 to open each cavity. It will be appreciated that in other embodiments, the seal caps may also be manually removed to open the chambers.
Specifically, referring to fig. 3 to 5, in the present embodiment, a plurality of chambers include a lysis chamber 11 for storing a lysis solution, a magnetic member chamber 12 for storing a magnetic member, a washing chamber for storing a washing liquid, an elution chamber 15 for storing an elution solution, and a lyophilization chamber 16 for storing lyophilization, and a reaction chamber 21 communicates with the lyophilization chamber 16. The lysis solution is used for fully lysing the sample and releasing nucleic acid so that the nucleic acid of the sample can be adsorbed to the surface of the magnetic piece in a chemical bond combination mode. The cleaning liquid is used for washing away impurities such as cell walls, proteins and the like remained on the surface of the magnetic part, so that the influence of the impurities on experimental results is avoided. The eluent is used to elute the nucleic acids from the magnetic element. And freeze-drying the sample and mixing the sample with the eluent after eluting the nucleic acid to form an amplification reagent, and finally introducing the amplification reagent into a reaction tank to detect the sample.
Optionally, in an embodiment, a plurality of cleaning chambers may be provided, for example, in this embodiment, two cleaning chambers, that is, the first cleaning chamber 13 and the second cleaning chamber 14, respectively, so as to perform more sufficient cleaning on the magnetic member, and better impurity removing effect. It will be appreciated that in other embodiments, one cleaning chamber may be provided.
Further, referring to fig. 3, the eluting chamber 15 is disposed adjacent to the freeze-drying chamber 16, and a partition 151 is disposed between the eluting chamber 15 and the freeze-drying chamber 16, and the magnetic attraction member 30 can pierce the partition 151 to enable the eluting chamber to be communicated with the freeze-drying chamber 16, so that after the eluting solution in the eluting chamber 15 elutes nucleic acid from the magnetic member, the eluting chamber containing nucleic acid can flow into the freeze-drying chamber 16 and be mixed with freeze-drying by piercing the partition 151 between the eluting chamber and the freeze-drying chamber 16 through the magnetic attraction member 30, thereby forming the amplification reagent.
With continued reference to fig. 4 and 5. Further, the plurality of cavities further comprises a piston cavity 17, a movable piston member 40 is arranged in the piston cavity 17, the reaction member 20 is further provided with a liquid inlet channel 22 and a liquid outlet channel 23, the liquid inlet channel 22 and the liquid outlet channel 23 are both communicated with the reaction cavity 21, the liquid inlet channel 22 is communicated with the freeze-drying cavity 16, the liquid outlet channel 23 is communicated with the piston cavity 17, and the closed end of the magnetic attraction member 30 can penetrate into the piston cavity 17 and drive the piston member 40 to move along the piston cavity 17. Specifically, when the magnetic attraction member 30 drives the piston member 40 to move upward, air can be inhaled into the reaction chamber 21, so that the amplification reagent in the lyophilization chamber 16 is sucked into the reaction chamber 21 from the liquid inlet channel 22. When the magnetic attraction member 30 drives the piston member 40 to move downwards, air can be blown into the reaction chamber 21, so that the reagent in the reaction chamber 21 is discharged.
Referring to fig. 3, a sealing sleeve 152 is disposed in a cavity communicating with the reaction chamber 21, and the magnetic attraction member 30 can be in sealing engagement with the sealing sleeve 152 to seal the reaction chamber 21. Specifically, in this embodiment, the sealing sleeve 152 is disposed in the elution chamber 15, and after the amplification reagent in the lyophilization chamber 16 is sucked into the reaction chamber 21 from the liquid inlet channel 22, the magnetic attraction piece 30 is inserted into the elution chamber 15, so that the magnetic attraction piece 30 is in sealing fit with the sealing sleeve to seal the elution chamber 15 and the liquid inlet channel 22, thereby sealing the reaction chamber 21, avoiding the reagent in the reaction chamber 21 from flowing out from the liquid inlet channel 22, and ensuring the accuracy of subsequent detection.
Referring to fig. 3, the kit 10 is further provided with a receiving cavity 18 for receiving the magnetically attractable member 30. By housing the magnetically attractable member 30 within the receiving cavity 18, the likelihood of contamination of the magnetically attractable member 30 prior to use is reduced.
Referring to fig. 4 and 5, the kit 10 is provided with a slot 19, the slot 19 being in communication with at least one of the chambers, in particular, in this embodiment, the slot 19 is in communication with the lyophilization chamber 16 and the piston chamber 17. The reaction member 20 is inserted into the insertion groove 19 so that the reaction chamber 21 in the reaction member 20 can be communicated to the lyophilization chamber 16 and the piston chamber 17. Further, a clamping piece 50 is arranged at the slot 19, the clamping piece 50 is detachably connected with the kit 10, and the clamping piece 50 is in clamping fit with the reaction piece 20 to limit the reaction piece 20 in the slot 19. Specifically, the notch edge of the slot 19 is convexly provided with a limiting portion 193, the periphery of the reaction piece 20 is convexly provided with a boss 26, after the reaction piece 20 is inserted into the slot 19, the boss 26 is positioned in the slot 19, the clamping piece 50 is inserted between the boss 26 and the limiting portion 193, one side of the clamping piece 50 is abutted with the boss 26, and the other side is abutted with the limiting portion 193, so that the reactivity is limited to be separated from the notch of the slot 19.
Further, a connection hole communicating with at least one of the cavities is provided in the slot 19, and a sealing ring (not shown) is provided in the connection hole. The reaction part 20 is provided with a connecting pipe which is convexly provided with a connecting pipe communicated with the reaction cavity 21, and the connecting pipe is inserted into the connecting hole and is matched with the sealing ring in a sealing way. Specifically, referring to fig. 4 and 5, in the present embodiment, two connecting holes, that is, a first connecting hole 191 and a second connecting hole 192, are provided in the slot 19, the first connecting hole 191 is in communication with the freeze-drying chamber 16, and the second connecting hole 192 is in communication with the piston chamber 17. The reaction part 20 is provided with two connecting pipes in a protruding mode, namely a first connecting pipe 24 communicated with the liquid inlet channel 22 and a second connecting pipe 25 communicated with the liquid discharge channel 23, wherein the first connecting pipe 24 is inserted into the first connecting hole 191, so that the freeze-drying cavity 16 is communicated with the reaction cavity 21. The second connection pipe 25 is inserted into the second connection hole 192, thereby allowing the piston chamber 17 to communicate with the reaction chamber 21. Further, sealing rings are respectively arranged in the first connecting hole 191 and the second connecting hole 192, so that the sealing connection between the reaction part 20 and the reagent kit 10 is ensured, and reagent leakage is avoided.
Specifically, the workflow for extracting and detecting the sample by adopting the sample extraction and detection device comprises the following steps:
1. sample addition
Opening a sealing cover of the cracking cavity 11, adding a sample to be detected into the cracking cavity 11, covering the sealing cover of the cracking cavity 11, and placing the sample extraction detection device into an automatic operation instrument according to a preset position.
2. Nucleic acid extraction and PCR detection
1) Puncture sealing cover: the automatically operated instrument pulls the sleeve 31 up from the receiving cavity 18 and then moves over the next cavity and then moves the sleeve 31 down so that the sleeve 31 pierces the seal covers of the corresponding cavity so that the sleeve 31 can extend into the cavity and the above steps are repeated until the seal covers on all cavities are pierced.
2) Sample lysis: the sleeve 31 is moved to above the lysis chamber 11, and then the sleeve 31 is inserted downward into the lysis chamber 11, and the sleeve 31 is moved up and down or rotated in the lysis chamber 11, so that the lysis solution in the lysis chamber 11 is sufficiently mixed with the sample, the sample is sufficiently lysed, and nucleic acid is released.
3) Nucleic acid capture: the sleeve 31 is pulled out of the cracking cavity 11 and moves to the upper part of the magnetic piece cavity 12, then the sleeve 31 is moved downwards to be inserted into the magnetic piece cavity 12, and the sleeve 31 moves up and down or rotates in the magnetic piece cavity 12 so as to realize uniform mixing of the magnetic pieces in the magnetic piece cavity 12; the magnet on the automated instrument is then inserted down into the sleeve 31 to attract the magnetic element to the sleeve 31; the cannula 31 is then lifted up together with the magnet and moved over the lysis chamber 11, and the cannula 31 is then inserted down together with the magnet into the lysis chamber 11; then the magnet is moved upwards to release the magnetic element adsorbed on the sleeve 31, and finally the sleeve 31 is driven to move up and down or rotate in the cavity so as to fully mix the lysis solution containing the nucleic acid with the magnetic element, and the nucleic acid in the sample is adsorbed on the magnetic element.
4) Washing for the first time: the magnet on the automated instrument is again inserted downward into the sleeve 31 to adsorb the magnetic member having nucleic acid adsorbed in the cleavage cavity 11 to the sleeve 31; the sleeve 31 is then moved up out of the lysis chamber 11 together with the magnet and moved over the first washing chamber 13, the sleeve 31 is then inserted down into the first washing chamber 13 together with the magnet, the magnet is then moved up to release the magnetic member adsorbed on the sleeve 31, and the sleeve 31 is then driven to move up and down or rotate in the first washing chamber 13 to mix the washing liquid and the magnetic member in the first washing chamber 13 sufficiently to wash away the cell wall, protein, etc. remaining on the surface of the magnetic member.
5) And (3) washing for the second time: the magnet is inserted down into the sleeve 31 again to attract the magnetic member in the first cleaning chamber 13 to the sleeve 31; the sleeve 31 is then moved up out of the first cleaning chamber 13 together with the magnet and moved over the second cleaning chamber 14, the sleeve 31 is then inserted down into the second cleaning chamber 14 together with the magnet, the magnet is then moved up to release the magnetic member adsorbed on the sleeve 31, and the sleeve 31 is then driven to move up and down or rotate in the second cleaning chamber 14 to mix the cleaning liquid and the magnetic member in the second cleaning chamber 14 sufficiently to further remove impurities from the surface of the magnetic member.
6) Nucleic acid elution: the magnet is again inserted downwardly into the sleeve 31 to attract the magnetic member in the second cleaning chamber 14 to the sleeve 31; the sleeve 31 is then moved up out of the second washing chamber 14 together with the magnet and over the elution chamber 15, the sleeve 31 is then inserted down into the elution chamber 15 together with the magnet, the magnet is then moved up to release the magnetic member attached to the sleeve 31, and the sleeve 31 is then driven to move up and down or rotate within the elution chamber 15 to thoroughly mix the eluate and the magnetic member within the elution chamber 15, eluting nucleic acids from the magnetic member into the eluate.
7) Discarding magnetic beads: the magnet is again inserted down into the cannula 31 to attract the magnetic element in the elution chamber 15 to the cannula 31; the sleeve 31 is then moved up out of the elution chamber 15 together with the magnet and over the second washing chamber 14, the sleeve 31 is then inserted down into the second washing chamber 14 together with the magnet, and the magnet is then moved up to release the magnetic element into the second washing chamber 14. It will be appreciated that in other embodiments the magnetic element may be disposed of in the lysis chamber 11, the magnetic element chamber 12 or the first wash chamber 13, without limitation.
8) Mixing reaction liquid: the sleeve 31 is moved up out of the second wash chamber 14 and over the elution chamber 15, then the sleeve 31 is inserted down into the elution chamber 15, and the sleeve 31 is continued to be moved down until the sleeve 31 pierces the partition 151 between the elution chamber 15 and the lyophilization chamber 16, causing the nucleic acid containing eluate to flow into the lyophilization chamber 16 and mix with the lyophilization, forming an amplification reagent.
9) Nucleic acid detection: the sleeve 31 is moved up out of the elution chamber 15 and over the piston chamber 17, then the sleeve 31 is moved down into the piston chamber 17, and the sleeve 31 is moved down further to press the piston member 40 in the piston chamber 17 down a certain distance, then the sleeve 31 is moved up to move the piston member 40 up, so that the elution solution containing nucleic acid is introduced into the reaction chamber 21 of the reaction member 20 through the first connection hole 191 and the feed channel 22. Then the magnetic attraction piece 30 moves upwards out of the piston cavity 17 and moves above the elution cavity 15, then the sleeve 31 moves downwards into the elution cavity 15, the elution cavity 15 continues to move downwards, the sleeve 31 is matched with the sealing sleeve 152 in the elution cavity 15 to block and seal the liquid inlet channel 22, and then the automatic instrument starts a PCR program to detect nucleic acid of the amplification reagent in the reaction cavity 21.
3. Device discard
And after the PCR is finished, taking the sample extraction and detection device out of the automatic instrument for waste biological treatment.
It should be noted that, in the present embodiment, the movement and the insertion and extraction of the sleeve 31 and the magnet are performed by an automated apparatus, and in other embodiments, the movement and the insertion and extraction of the sleeve 31 and the magnet may be performed manually, which is not limited herein.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the claims. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (10)
1. A sample extraction detection device, comprising:
the kit is provided with a plurality of cavities, different cavities are used for storing different reagents, and at least one cavity is used for storing a magnetic piece;
the reaction piece is connected with the kit and is provided with a reaction cavity which is communicated with at least one cavity; the method comprises the steps of,
the magnetic attraction piece can be inserted into any cavity and is used for adsorbing or releasing the magnetic piece.
2. The sample extraction and detection device according to claim 1, wherein the magnetic attraction member comprises a sleeve, the sleeve is a hollow tube body with one end closed and the other end open, the closed end of the magnetic attraction member can be inserted into any one of the cavities, and the open end of the magnetic attraction member is used for inserting or withdrawing a magnet so that the sleeve adsorbs or releases the magnetic member.
3. The sample extraction testing device of claim 1, further comprising a sealing cap sealingly connected to said kit and closing each of said cavities.
4. The sample extraction detection device of claim 1, wherein the plurality of chambers includes a lysis chamber for storing a lysis solution, a magnetic member chamber for storing the magnetic member, a washing chamber for storing a washing liquid, an elution chamber for storing an eluent, and a lyophilization chamber for storing lyophilization, and the reaction chamber is in communication with the lyophilization chamber.
5. The sample extraction detection device of claim 4, wherein the elution chamber and the lyophilization chamber are stacked, and a partition is disposed between the elution chamber and the lyophilization chamber, and the magnetic attraction member is capable of piercing the partition to communicate the elution chamber with the lyophilization chamber.
6. The sample extraction and detection device according to claim 4, wherein the plurality of cavities further comprises a piston cavity, a movable piston member is arranged in the piston cavity, the reaction member is further provided with a liquid inlet channel and a liquid outlet channel, the liquid inlet channel and the liquid outlet channel are both communicated with the reaction cavity, the liquid inlet channel is communicated with the freeze-drying cavity, the liquid outlet channel is communicated with the piston cavity, and the magnetic attraction member can penetrate into the piston cavity and drive the piston member to move along the piston cavity.
7. The sample extraction and detection device according to claim 1, wherein a sealing sleeve is arranged in the cavity communicated with the reaction cavity, and the magnetic attraction piece can be in sealing fit with the sealing sleeve to seal the reaction cavity.
8. The sample extraction detection device according to claim 1, wherein the kit is further provided with a receiving chamber for receiving the magnetic attraction member.
9. The sample extraction detection device according to claim 1, wherein the kit is provided with a slot, the slot is communicated with at least one of the cavities, the reaction member is inserted into the slot, a clamping member is arranged at the slot, the clamping member is detachably connected with the kit, and the clamping member is in clamping fit with the reaction member to limit the reaction member in the slot.
10. The sample extraction and detection device according to claim 9, wherein a connecting hole communicated with at least one of the cavities is formed in the slot, a sealing ring is arranged in the connecting hole, the reaction piece is provided with a connecting pipe which is convexly arranged and communicated with the reaction cavity, and the connecting pipe is inserted into the connecting hole and is in sealing fit with the sealing ring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310732909.5A CN116640659A (en) | 2023-06-19 | 2023-06-19 | Sample extraction and detection device |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310732909.5A CN116640659A (en) | 2023-06-19 | 2023-06-19 | Sample extraction and detection device |
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| Publication Number | Publication Date |
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| CN116640659A true CN116640659A (en) | 2023-08-25 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202310732909.5A Pending CN116640659A (en) | 2023-06-19 | 2023-06-19 | Sample extraction and detection device |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118879472A (en) * | 2024-07-26 | 2024-11-01 | 湖北寰清检测技术有限公司 | A portable water quality microbial sampling device |
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2023
- 2023-06-19 CN CN202310732909.5A patent/CN116640659A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118879472A (en) * | 2024-07-26 | 2024-11-01 | 湖北寰清检测技术有限公司 | A portable water quality microbial sampling device |
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