CN212316088U - Totally-enclosed multi-index nucleic acid detection device - Google Patents
Totally-enclosed multi-index nucleic acid detection device Download PDFInfo
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- CN212316088U CN212316088U CN202020553383.6U CN202020553383U CN212316088U CN 212316088 U CN212316088 U CN 212316088U CN 202020553383 U CN202020553383 U CN 202020553383U CN 212316088 U CN212316088 U CN 212316088U
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Abstract
The utility model relates to a totally-enclosed multi-index nucleic acid detection device, which comprises a reagent cavity shell, an upper cover, a communicating vessel and a piston cylinder; a reagent cavity is arranged in the reagent cavity shell, an upper cover is arranged on the reagent cavity shell, a sampling hole is formed in the upper cover, a piston cylinder is arranged in the reagent cavity shell, a communicating hole is formed in a base plate of the piston cylinder, a communicating vessel is arranged below the piston cylinder, a communicating groove and a detection hole are formed in the communicating vessel, and an inner cavity of the piston cylinder and a bottom hole of the reagent cavity can be selectively communicated through rotating the communicating hole; the reagent cavity comprises a sample pretreatment cavity, a lysis solution cavity, a magnetic bead cavity, a washing solution cavity and an elution solution cavity, each cavity is a closed cavity and can be communicated with the inner cavity of the piston cylinder through a communicating hole and a communicating groove, and the sample pretreatment cavity corresponds to the sample adding hole; the detection hole is only communicated with the inner cavity of the piston cylinder and the reagent cavity through the communication hole. The utility model discloses a detection when the complete seal of nucleic acid testing process detects, convenient operation and can realize the many indexs of single sample.
Description
Technical Field
The utility model relates to a totally enclosed many indexes nucleic acid detecting device, specifically speaking are collection sample pretreatment and nucleic acid extraction and nucleic acid amplification and detect in an organic whole detection device.
Background
Nucleic acid detection requires the following steps: the method comprises the steps of sample pretreatment, nucleic acid extraction, reagent preparation, amplification and detection, wherein nucleic acid detection is a common detection method at present, but the steps of nucleic acid detection are mainly performed manually or are completed by a semi-automatic instrument in part of the process, the whole process is completed by professional technicians, and meanwhile, professional laboratories are needed, especially samples with high hazard need to be performed in biosafety laboratories of the P2 level, so that the popularization and application of the nucleic acid detection technology are further limited. At present, full-automatic detection instruments exist in the market, but the problems of sample pretreatment, complex operation, high cost, limited detection indexes, incapability of realizing multi-index screening and the like still exist, and the wide-range application is limited.
Disclosure of Invention
The utility model aims at overcoming the not enough of existence among the prior art, provide one kind can realize that the full seal detects, convenient operation and can realize the device that detects when the many indexs of single sample.
According to the technical scheme provided by the utility model, the fully-closed multi-index nucleic acid detection device at least comprises a reagent cavity shell, an upper cover, a communicating vessel, a piston cylinder and a detection hole; the reagent cavity shell is provided with an axial through hole, a reagent cavity is arranged in the reagent cavity shell, a reagent cavity bottom hole is formed in the bottom of the reagent cavity, an upper cover is arranged on the reagent cavity shell, a sample adding hole is formed in the upper cover, a piston cylinder is arranged in the reagent cavity shell, a piston is arranged in the piston cylinder, a communicating hole is formed in a base plate of the piston cylinder, a communicating vessel is arranged below the piston cylinder, a communicating groove and a detection hole are formed in the communicating vessel, one end of the communicating groove is communicated with the inner cavity of the piston cylinder, the other end of the communicating groove is connected with the communicating hole, the communicating hole and the reagent cavity bottom hole are on the same diameter, and the inner cavity of the piston cylinder and the reagent cavity;
the reagent cavity at least comprises a sample pretreatment cavity, a lysis solution cavity, a magnetic bead cavity, a washing solution cavity and an eluent cavity, wherein each cavity is a closed cavity, the bottom of the reagent cavity is provided with a reagent cavity bottom hole and is communicated with the inner cavity of the piston cylinder through a communicating hole and a communicating groove, and the sample pretreatment cavity corresponds to the sample adding hole;
the detection hole is only communicated with the inner cavity of the piston cylinder and the reagent cavity through the communication hole.
Preferably, the reagent chamber further comprises a digestive juice chamber, a combining liquid chamber, a drying chamber, a liquid taking chamber, a dry powder chamber and a magnetic bead removing liquid chamber.
Preferably, a fixing hole for fixing the chamber housing is provided at an inner side or an outer side of the reagent chamber housing.
Preferably, a magnet adsorption hole for magnet adsorption is provided inside or outside the reagent chamber housing.
Preferably, the device further comprises a cavity shell reverse buckle, and the communicating vessel is buckled by the cavity shell reverse buckle.
Preferably, the detection hole is located below the communicating vessel, and the detection hole may be on one device with the communicating vessel.
Preferably, the detection hole is single or multiple, and the detection hole is conical, cylindrical or square.
Preferably, the communicating vessel is freely rotatable.
Preferably, the communicating vessel is integrally connected with the piston cylinder.
The utility model has the advantages of it is following:
the method is suitable for different samples, complex samples do not need pretreatment, and the operation is simpler: the device has the functions of left-right oscillation and high-speed centrifugation, can automatically smash a solid sample, and can liquefy a viscous liquid; the solid-liquid separation can be carried out on the solid-liquid mixed sample by centrifugation.
Full automated inspection, convenient operation: sample pretreatment, nucleic acid extraction, reagent preparation and the like are integrated in one card box, manual sample pretreatment is not needed, and seamless butt joint in the whole process is achieved.
realizes the simultaneous detection of multiple indexes of a single sample: and (3) screening more than 60 indexes at the same time, which is beneficial to the systematic screening of microorganisms.
Closed card box: only need to add the sample, the other operations are all in airtight card box, multiple anti-pollution and prevent leakage reagent, no nucleic acid aerosol pollution risk, no test sample and reagent leakage risk.
Drawings
Fig. 1 is a schematic structural diagram of the present invention.
Figure 2 is a top view of the reagent chamber housing of the present invention.
Fig. 3 is a top view of the upper cover of the present invention.
Fig. 4 is a structure diagram of the connection between the connector and the piston tube base plate of the present invention.
Detailed Description
The utility model discloses a totally-enclosed multi-index nucleic acid detection device, which at least comprises a reagent cavity shell 100, an upper cover 200, a communicating vessel 300, a piston cylinder 400 and a detection hole 302; the reagent chamber shell 100 is provided with an axial through hole, a reagent chamber is arranged in the reagent chamber shell 100, a reagent chamber bottom hole is arranged at the bottom of the reagent chamber, the reagent chamber shell 100 is provided with an upper cover 200, the upper cover 200 is provided with a sample adding hole 201, a piston barrel 400 is arranged in the reagent chamber shell 100, a piston 401 is arranged in the piston barrel 400, a communication hole 402 is arranged on a base plate of the piston barrel 400, a communicating vessel 300 is arranged below the piston barrel 400, the communicating vessel 300 is provided with a communicating groove 301 and a detection hole 302, one end of the communicating groove 301 is communicated with an inner cavity of the piston barrel 400, the other end of the communicating groove is connected with the communication hole 402, the communication hole 402 and the reagent chamber bottom hole are in the same diameter, and the inner cavity of the piston barrel 400 and the reagent chamber;
the reagent cavity at least comprises a sample pretreatment cavity 101, a lysis solution cavity 102, a magnetic bead cavity 105, a washing solution cavity 106 and an eluent cavity 108, each cavity is a closed cavity, a bottom hole of the reagent cavity is formed in the bottom of the reagent cavity and can be communicated with the inner cavity of the piston cylinder 400 through a communication hole 402 and a communication groove 301, and the sample pretreatment cavity 101 corresponds to the sample adding hole 201;
the detection hole 302 is communicated with the inner cavity of the piston cylinder 400 and the reagent cavity only through the communication hole 402.
The reagent chamber further comprises a digestive juice chamber 103, a combining liquid chamber 104, a drying chamber 107, a liquid taking chamber 109, a dry powder chamber 111 and a magnetic bead removing liquid chamber 112.
A fixing hole 113 for fixing the chamber housing is formed at the inner side or the outer side of the reagent chamber housing 100.
A magnet adsorption hole 114 for magnet adsorption is provided inside or outside the reagent chamber housing 100.
The housing undercut 115 is provided and the connector 300 is gripped by the housing undercut 115.
The sensing hole 302 is located below the communicator 300, and the sensing hole 302 may be on one device with the communicator 300.
The detection hole 302 may be a single hole or a plurality of holes, and the detection hole 302 may be conical, cylindrical, or square.
The communicator 300 is freely rotatable.
The communicating vessel 300 and the piston cylinder 400 may be integrally connected.
The utility model discloses in, different reagents can be pre-installed in inclosed reagent intracavity to can realize not having the leakage.
The reagent cavity is connected with the inner cavity of the piston cylinder 400 through the communicating vessel 300, the piston cylinder 400 can be connected with different reagent cavities by rotating the communicating vessel 300, and liquid is transferred back and forth between the reagent cavity and the inner cavity of the piston cylinder 400 through the up-and-down movement of the piston 401.
The fixing of the reagent cavity shell 100 can be realized by means of the module assembly (comprising a fixed reagent cavity shell module, a heating module and a magnet module), the heating of the reagent in the reagent cavity can be realized, and the adsorption and release of the magnetic beads in the cavity can be realized. The transfer of liquid between the reagent chamber and the piston cylinder 400 is achieved by the up and down movement of the rotary communicator 300 and the piston 401.
The structure and the method can be used for crushing and liquefying a sample, cracking cells, extracting nucleic acid, mixing different solutions, amplifying and detecting the nucleic acid. And can also be used for full-automatic detection of chemiluminescence.
The present invention will be further explained by taking sputum liquefaction and nucleic acid extraction as examples.
Opening the upper cover of the sample pretreatment cavity, adding 50-100 microliters of sputum, covering the upper cover (remarks: 200 microliters of liquefied fluid and glass beads with different specifications are pre-filled in the sample pretreatment cavity), and placing the treatment device into the instrument according to the positioning structure.
The start-up procedure was run as follows.
1. Sample pretreatment (sputum liquefaction)
The spindle rotating motor starts to rotate back and forth at the speed of 2m/s-7m/s, the speed is increased by 1m/s every 5 seconds, the maximum speed is increased to 15m/s, the speed is kept for 30s at 15m/s, after the 30s is stopped, the circulation is performed for 2 times (the number of times can be increased or decreased according to the actual situation), and the liquefaction of the sputum sample is realized through the left and right oscillation of the glass beads and the action of the liquefied liquid.
2. Nucleic acid extraction
1) The sample is lysed.
a) The module assembly (including the fixed reagent chamber shell module, the heating module and the magnet module) descends to fix the device chamber shell.
b) The communicating vessel 300 rotates to communicate the bottom hole of the sample pretreatment chamber 101 with the communicating hole 402, the piston rod descends to be connected to the piston 401, the piston 401 ascends, and the supernatant in the sample pretreatment chamber 101 is transferred to the inner cavity of the piston cylinder 400.
c) The communicating vessel 300 is rotated to communicate the bottom hole of the sample pretreatment chamber 101 with the communicating hole 402 by rotating the communicating vessel 300, and the piston 401 is raised to transfer the lysate in the lysate chamber 102 to the inner chamber of the piston cylinder 400.
d) The communicating vessel 300 is rotated to communicate the bottom hole of the digestive juice chamber 103 with the communicating hole 402, the piston 401 is pressed down to transfer the liquid in the piston cylinder 400 to the digestive juice chamber 103, and then the liquid is pumped back to the piston cylinder 400 to reciprocate for 3 times, and finally the liquid is completely pressed into the digestive juice chamber 103.
e) And (3) protease digestion: the module assembly descends to be in contact with the inner wall of the digestive juice cavity 103, the heating device heats the module assembly, the heating device stops heating after heating for 10min at 56 ℃, and thermal cracking and crushing of the sample and digestion of protein are achieved.
2) Nucleic acid capture (nucleic acid bound to magnetic beads).
a) The piston 401 is raised to transfer the sample digest chamber 103 well supernatant to the inner chamber of the piston cylinder 400.
b) The communicating vessel 300 is rotated to communicate the bottom hole of the bonding solution chamber 104 with the communicating hole 402, the piston 401 is pressed down to transfer the liquid to the bonding solution chamber 104, and then the liquid is pumped back to the piston cylinder 400 to reciprocate 3 times. Eventually drawing all of the liquid into the interior chamber of the piston cylinder 400.
c) The communicating vessel 300 is rotated to communicate the magnetic bead cavity 105 with the inner cavity of the piston cylinder 400, the module assembly is lifted to the top of the device cavity shell, the piston 401 is pressed downwards to transfer the liquid to the magnetic bead cavity 105, and then the liquid is pumped back to the piston cylinder 400, the operation is repeated for 3 times, and the operation is static for 30s, so that the magnetic beads and the DNA in the supernatant are fully combined to form a magnetic bead and DNA compound.
d) The module assembly is lowered to the bottom of the device chamber housing, at which time the magnet captures the "magnetic bead + DNA complex" on the inner wall of the piston cylinder 400, resting for 60s, and pressing down the piston 401 to transfer the liquid to the bead chamber 105.
3) First washing
a) The communicating vessel 300 rotates to communicate the first washing liquid chamber 106 (which is close to the magnetic bead chamber 105 and has a volume of 500 microliters) with the device chamber hole, the module assembly ascends to the top of the device chamber shell, the piston 401 is lifted, the washing liquid in the washing liquid chamber 106 is transferred to the inner chamber of the piston cylinder 400, and the washing liquid is transferred between the washing liquid chamber 106 and the piston cylinder 400 back and forth for 3 times, so that impurities are sufficiently washed.
b) The module assembly is lowered to the bottom of the device chamber housing, the plunger 401 is raised and allowed to rest for 15 seconds, at which time the magnet captures the "beads + DNA complexes" on the inner wall of the plunger shaft 400, the plunger 401 is lowered, and the liquid is transferred to the first wash solution chamber 106 (close to the bead chamber 105).
4) Second washing
a) The communicating vessel 300 is rotated to communicate the second washing liquid chamber 106 (with a volume of 500 microliters near the drying chamber 107) with the communicating hole 402, the module assembly is lifted to the top of the device chamber shell, the piston 401 is lifted, the washing liquid in the washing liquid chamber 106 is transferred to the inner chamber of the piston cylinder 400, and the washing liquid is transferred back and forth between the washing liquid chamber 106 and the inner chamber of the piston cylinder 400 for 3 times, so that impurities are sufficiently washed.
b) The module assembly is lowered to the bottom of the device chamber housing, the plunger 401 is raised and allowed to rest for 15 seconds, at which time the magnet captures the "magnetic bead + DNA complex" on the inner wall of the plunger barrel 400, the plunger 401 is lowered, and the liquid is transferred to the wash liquid chamber 106.
5) Magnetic bead drying
The communicating vessel 300 is rotated to communicate the drying chamber 107 with the inner chamber of the piston cylinder 400, and the piston 401 is lifted up and pressed down to be moved back and forth 5 times to sufficiently dry the magnetic beads.
6) Nucleic acid elution
a) Magnetic bead and DNA separation: the communicating vessel 300 is rotated to communicate the bottom hole of the eluent chamber 108 (with a volume of 150 microliters) with the communicating hole 402, the module assembly is lifted to the top of the device chamber housing, the piston 401 is lifted, the eluent in the eluent chamber 108 is transferred to the piston cylinder 400, and the eluent is transferred back and forth between the eluent chamber 108 and the piston cylinder 400 for 3 times, thereby realizing the separation of DNA and magnetic beads.
b) Obtaining of DNA solution: the module descends to the bottom, rises the piston 401, descends 5mm from the bottom after standing for 15 seconds, reciprocates three times, at the moment, the magnet captures the magnetic beads on the inner wall of the piston cylinder 400, the solution is pressed to the eluent cavity 108, and the solution contains high-purity nucleic acid, namely, the nucleic acid in the sample is extracted.
7) Magnetic bead removing liquid
The communicating vessel 300 rotates to communicate the magnetic bead removing liquid cavity with the communicating vessel 300, the piston 401 is lifted upwards, the removing liquid is transferred to the piston cylinder 400, and the removing liquid is transferred between the magnetic bead removing liquid cavity 112 and the piston cylinder 400 for 3 times, so that the magnetic beads are fully mixed. The magnetic beads and the liquid are transferred to the magnetic bead removing liquid cavity 112, and the piston is left at the bottom to remove the magnetic beads.
8) Nucleic acid solution transfer
The transfer connector 300 rotates to connect the eluent chamber 108 and the inner chamber of the plunger 400, and the plunger 401 moves upward to suck the DNA solution into the inner chamber of the plunger 400.
9) Nucleic acid reagent formulation
The communicating vessel 300 is rotated to communicate the piston cylinder 400 with the dry powder chamber 111, and the liquid in the piston cylinder 400 is transferred to the dry powder chamber 111 to dissolve the dry powder. And finally draws the liquid into the interior cavity of the piston cylinder 400.
10) Transfer of nucleic acid reagents to assay wells
The communicating vessel 300 is rotated to communicate the piston cylinder 400 with the detection holes 302, and the reagent in the piston cylinder 400 is transferred to the detection holes 302, and if there are a plurality of detection holes 302, the detection reagent can be sequentially quantitatively added. Or transferring the sample to a reserved reagent hole, and then quantitatively adding the reagent into each detection hole 302 by the way (centrifugal even division, siphon and the like), and mixing the reagent with the reagent (such as a primer, a probe and the like) embedded in the detection hole 302 after the reagent enters the detection hole 302 to realize the preparation of the detection reagent.
11) The communicating vessel 300 is rotated to a zero position where all the communicating holes are sealed to prevent leakage of the reagent and the detecting hole 302 is sealed to prevent contamination of the aerosol.
12) Nucleic acid amplification and detection: the device is transferred to a temperature control and optical detection module, amplification of target gene nucleic acid is realized through the temperature environment provided by an instrument, and real-time detection of nucleic acid can be realized through real-time fluorescence collection of the optical detection module.
Claims (9)
1. A totally-enclosed multi-index nucleic acid detection device is characterized in that: the device at least comprises a reagent cavity shell (100), an upper cover (200), a communicating vessel (300), a piston cylinder (400) and a detection hole (302); the reagent cavity shell (100) is provided with an axial through hole, a reagent cavity is arranged in the reagent cavity shell (100), a reagent cavity bottom hole is arranged at the bottom of the reagent cavity, an upper cover (200) is arranged on the reagent cavity shell (100), a sample adding hole (201) is arranged on the upper cover (200), a piston cylinder (400) is arranged in the reagent cavity shell (100), a piston (401) is arranged in the piston cylinder (400), a communicating hole (402) is arranged on the base plate of the piston cylinder (400), a communicating vessel (300) is arranged below the piston cylinder (400), a communicating groove (301) and a detecting hole (302) are arranged on the communicating vessel (300), one end of the communicating groove (301) is communicated with the inner cavity of the piston cylinder (400), the other end is connected with a communicating hole (402), the communicating hole (402) and the bottom hole of the reagent cavity are on the same diameter, the inner cavity of the piston cylinder (400) can be selectively communicated with the bottom hole of the reagent cavity through the rotary communication hole (402);
the reagent cavity at least comprises a sample pretreatment cavity (101), a lysis solution cavity (102), a magnetic bead cavity (105), a washing solution cavity (106) and an elution solution cavity (108), each cavity is closed, the bottom of the reagent cavity is provided with a bottom hole of the reagent cavity, the reagent cavity can be communicated with the inner cavity of the piston cylinder (400) through a communication hole (402) and a communication groove (301), and the sample pretreatment cavity (101) corresponds to the sample adding hole (201);
the detection hole (302) is only communicated with the inner cavity of the piston cylinder (400) and the reagent cavity through the communication hole (402).
2. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: the reagent cavity further comprises a digestive juice cavity (103), a combining liquid cavity (104), a drying cavity (107), a liquid taking cavity (109), a dry powder cavity (111) and a magnetic bead removing liquid cavity (112).
3. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: and a fixing hole (113) for fixing the cavity shell is arranged on the inner side or the outer side of the reagent cavity shell (100).
4. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: and a magnet adsorption hole (114) for magnet adsorption is arranged on the inner side or the outer side of the reagent cavity shell (100).
5. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: the device also comprises a cavity shell reverse buckle (115), and the communicating vessel (300) is fastened by the cavity shell reverse buckle (115).
6. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: the detection hole (302) is positioned below the communicating vessel (300), and the detection hole (302) and the communicating vessel (300) can be arranged on one device.
7. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1 or 6, wherein: the detection hole (302) is single or multiple, and the detection hole (302) is conical, cylindrical or square.
8. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: the communicator (300) is freely rotatable.
9. The totally enclosed multi-index nucleic acid detecting apparatus according to claim 1, wherein: the communicating vessel (300) is integrally connected with the piston cylinder (400).
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| Application Number | Priority Date | Filing Date | Title |
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| CN202020553383.6U CN212316088U (en) | 2020-04-14 | 2020-04-14 | Totally-enclosed multi-index nucleic acid detection device |
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| CN202020553383.6U CN212316088U (en) | 2020-04-14 | 2020-04-14 | Totally-enclosed multi-index nucleic acid detection device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111394221A (en) * | 2020-04-14 | 2020-07-10 | 无锡科智达科技有限公司 | Totally-enclosed multi-index nucleic acid detection device |
| CN113564044A (en) * | 2021-08-04 | 2021-10-29 | 圣湘生物科技股份有限公司 | Nucleic acid detection device and nucleic acid detection method |
| CN116554998A (en) * | 2023-06-09 | 2023-08-08 | 鲲鹏基因（北京）科技有限责任公司 | Kit for detecting candida |
-
2020
- 2020-04-14 CN CN202020553383.6U patent/CN212316088U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111394221A (en) * | 2020-04-14 | 2020-07-10 | 无锡科智达科技有限公司 | Totally-enclosed multi-index nucleic acid detection device |
| CN113564044A (en) * | 2021-08-04 | 2021-10-29 | 圣湘生物科技股份有限公司 | Nucleic acid detection device and nucleic acid detection method |
| CN113564044B (en) * | 2021-08-04 | 2024-04-30 | 圣湘生物科技股份有限公司 | Nucleic acid detection device and nucleic acid detection method |
| CN116554998A (en) * | 2023-06-09 | 2023-08-08 | 鲲鹏基因（北京）科技有限责任公司 | Kit for detecting candida |
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