CN219079543U - Nucleic acid extraction and detection device for POCT - Google Patents
Nucleic acid extraction and detection device for POCT Download PDFInfo
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- CN219079543U CN219079543U CN202223416760.XU CN202223416760U CN219079543U CN 219079543 U CN219079543 U CN 219079543U CN 202223416760 U CN202223416760 U CN 202223416760U CN 219079543 U CN219079543 U CN 219079543U
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Abstract
The utility model provides a nucleic acid extraction and detection device for POCT, which is characterized by comprising the following components: an extraction member (10), a plurality of reagent containing members (20), a magnetic member (50) and a collection member (60), the extraction member (10) having an extraction chamber and an opening communicating with the extraction chamber; each reagent containing component (20) is communicated with the extraction cavity through a first pipeline, and a first control valve (30) for controlling the trend of liquid and a liquid pump (40) for driving the liquid to flow are arranged on the first pipeline; the collection member (60) is in communication with the extraction chamber via a second conduit, and the collection member (60) is configured to collect a nucleic acid product resulting from the extraction of the nucleic acid. The nucleic acid extraction and detection device provided by the embodiment of the utility model adopts pipeline connection, is a closed type nucleic acid extraction and detection device, and can solve the problem of cross contamination between samples compared with a traditional magnetic rod method full-automatic nucleic acid extraction instrument.
Description
Technical Field
The utility model relates to the field of nucleic acid extraction, in particular to a nucleic acid extraction and detection device for POCT (point-of-care testing).
Background
Nucleic acid detection is one of the very important ways in vitro diagnostic techniques, and has been widely used in a variety of fields such as pathogen detection, early screening of cancer, and genetic disease detection. The premise of nucleic acid detection is that a high-purity nucleic acid sample can be extracted and purified from different biological samples, and then the obtained nucleic acid sample is subjected to related detection by subsequent detection means such as PCR (Polymerase Chain Reaction, chain polymerase reaction), qPCR (quantitative Real-Time PCR, real-Time fluorescent quantitative PCR), RT-qPCR (Reverse Transcription-qPCR, reverse transcription real-Time fluorescent quantitative PCR), NGS (Next-Generation Sequencing, second generation sequencing) and the like. The mode of nucleic acid extraction based on the magnetic bead method is the most widely used nucleic acid extraction scheme at present, however, the conventional full-automatic nucleic acid extractor has the following problems: 1) The extraction process is completed in a deep hole plate, so that cross contamination among nucleic acid samples is easily caused in the cracking process, and false positive results are often caused on samples around positive samples; 2) The flux requirement of the full-automatic nucleic acid extractor is high, and when the pre-loaded magnetic bead method is used for extracting the reagent, the reagent is wasted if the detection flux is smaller. If bottled reagent is adopted, the current filling mode can also cause waste of consumable materials such as deep pore plates, magnetic rod sleeves and the like, and can also cause pollution in the filling process.
Disclosure of Invention
The present utility model aims to solve at least one of the technical problems in the related art to some extent. To this end, the present utility model provides a nucleic acid extraction and detection device for POCT, comprising:
an extraction member having an extraction chamber and an opening communicating with the extraction chamber;
the reagent accommodating parts are communicated with the extracting cavity through first pipelines, and a first control valve for controlling the trend of liquid and a liquid pump for driving the liquid to flow are arranged on the first pipelines;
a magnetic component for adsorbing the magnetic beads within the extraction chamber;
and a collection member in communication with the extraction chamber through a second line, the collection member being for collecting nucleic acid products resulting from nucleic acid extraction.
The nucleic acid extraction and detection device for POCT provided by the embodiment of the utility model adopts pipeline connection, is a closed type nucleic acid extraction and detection device, and can solve the problem of cross contamination between samples compared with a traditional magnetic rod method full-automatic nucleic acid extraction instrument.
Optionally, the opening of the extraction component comprises an effervescent tablet feed inlet and a sample feed inlet.
Optionally, a temperature control rotating component for controlling the temperature in the extraction chamber and stirring the liquid in the extraction chamber is arranged on the extraction chamber, and the magnetic component is arranged on one side of the extraction component.
Optionally, the temperature-controlled rotating component comprises a heating component and a stirring component, the heating component is arranged in the extraction cavity or outside the extraction cavity, and the stirring component is arranged in the extraction cavity.
Optionally, the extracting member has a slope, and the magnetic member is disposed close to the slope.
Optionally, the first pipeline includes collection pipeline and a plurality of branch pipeline, and the one end of a plurality of branch pipelines communicates respectively and holds the part corresponding reagent, and the other end intercommunication first control valve, the one end of collection pipeline communicates first control valve, the other end intercommunication draw the cavity, every all be provided with on the branch pipeline the liquid pump.
Alternatively, the plurality of reagent containing parts include a pure water containing bottle, an ethanol containing bottle, an isopropyl alcohol containing bottle, and a nucleic acid cleaning solution containing bottle.
Optionally, the first control valve is a six-way valve, and the nucleic acid extraction and detection device further comprises a first waste liquid collecting component, and the first waste liquid collecting component is communicated with the six-way valve.
Optionally, a second control valve is arranged on the second pipeline, and the nucleic acid extraction and detection device further comprises a second waste liquid collecting component which is communicated with the second control valve.
Optionally, the nucleic acid extraction and detection device further comprises a PCR module, and the collection member is disposed on the PCR module.
Additional aspects and advantages of the utility model will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the utility model.
Drawings
The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the utility model and together with the description, serve to explain the principles of the utility model.
FIG. 1 is a schematic diagram of a nucleic acid extraction and detection apparatus for POCT according to an embodiment of the present utility model.
Reference numerals:
10-an extraction component; 11-an effervescent tablet feed inlet; 12-a sample adding port; 13-a temperature-controlled rotating member;
20-a reagent containing part;
30-a first control valve;
40-liquid pump;
50-magnetic parts;
60-collecting part;
70-a first waste liquid collection member; 71-a second waste liquid collection member;
an 80-PCR module;
90-a second control valve.
Detailed Description
The technical scheme of the utility model will be further described in detail below with reference to specific embodiments. It is to be understood that the following examples are illustrative only and are not to be construed as limiting the scope of the utility model. All techniques implemented based on the above description of the utility model are intended to be included within the scope of the utility model.
The embodiment provides a nucleic acid extraction and detection device for POCT, which can be used for extracting and purifying a high-purity nucleic acid sample and timely detecting according to the requirement, and the whole nucleic acid extraction and detection device is communicated by adopting a pipeline, so that cross contamination of the sample is avoided.
Referring to FIG. 1, in some embodiments, the nucleic acid extraction, detection apparatus mainly comprises: an extraction part 10, a plurality of reagent containing parts 20, a first control valve 30, a liquid pump 40, a magnetic part 50 and a collection part 60.
Wherein the extraction member 10 has an extraction chamber and an opening communicating with the extraction chamber; the extraction chamber provides space for the sample to decompose and purify. The opening communicating with the extraction chamber may be used for adding samples or other reagents, and the opening may be sealed with a sealing cap to avoid entry of impurities. The extraction member 10 may be a reaction cassette, a reaction box, a reaction bottle, or the like.
Each reagent holding part 20 is communicated with the extraction chamber through a first pipeline, and a first control valve 30 for controlling the trend of liquid and a liquid pump 40 for driving the liquid to flow are arranged on the first pipeline; different reagent holding members 20 may be used to hold different reagents. For example, 4 reagent holding parts 20 are provided in total for holding pure water, ethanol, isopropanol, and nucleic acid washing liquid, respectively, for use in the nucleic acid extraction process, for washing the whole device line, eliminating nucleic acid fragment contamination and rnase contamination remaining in the line. The reagent container 20 may be a reagent bottle having a certain capacity.
The magnetic component 50 is used for adsorbing the magnetic beads in the extraction chamber; the magnetic part 50 can move relative to the extraction part 10 to adsorb the magnetic beads in the extraction chamber, thereby completing the recovery work of the magnetic beads.
The collection unit 60 is in communication with the extraction chamber via a second line, and the collection unit 60 is configured to collect nucleic acid products obtained by nucleic acid extraction, and if amplification reagents are added in advance, subsequent experiments of nucleic acids can be performed in the collection unit 60.
The nucleic acid extraction and detection device of the embodiment adopts pipeline connection, is a closed type nucleic acid extraction and detection device, and can solve the problem of cross contamination between samples compared with a traditional magnetic rod method full-automatic nucleic acid extraction instrument. The device can be used for extracting nucleic acid from various common biological samples such as swab, whole blood, serum, plasma, tissue homogenate and the like.
In some embodiments, the opening of the extraction component 10 includes an effervescent tablet fill port 11 and a sample fill port 12. The effervescent tablet charging port 11 is used for adding effervescent tablets, and the effervescent tablets contain magnetic beads; the sample application port 12 is used for applying a sample to be tested. The effervescent tablet and the sample are respectively added into the extraction cavity at different openings, so that cross contamination of the effervescent tablet and the sample during addition is avoided.
Further, two effervescent tablet charging openings 11 can be provided, so that different types of effervescent tablets can be added respectively, and more effervescent tablet charging openings 11 can be provided as required, namely, the number of the effervescent tablet charging openings 11 is not limited in the scheme protected by the utility model. Likewise, one or more of the sample addition ports 12 may be provided as needed.
In some embodiments, the extraction chamber is provided with a temperature-controlling rotation member 13 for controlling the temperature in the extraction chamber and stirring the liquid in the extraction chamber, namely: the temperature control rotary member 13 has at least two functions of controlling the temperature in the extraction chamber and stirring the liquid in the extraction chamber, respectively.
The magnetic member 50 is provided on the extraction member 10 side. The magnetic member 50 may be a permanent magnet or an electromagnet. The magnetic component 50 can be in sliding fit with the external structure of the extraction component 10, and can be separated from the extraction component 10, and the magnetic component 50 is close to the extraction component 10 when in use, so that the magnetic beads can be adsorbed.
In some embodiments, the temperature-controlled rotary member 13 includes a heating member disposed within or outside the extraction chamber and a stirring member disposed within the extraction chamber. The temperature-controlling rotary member 13 may be a stirring rod having a heating function, which can perform heating work while stirring.
The temperature control rotating part 13 is arranged, so that a plurality of subsequent experimental works such as PCR, qPCR, RT-qPCR and the like of the extracted nucleic acid can be directly realized, and one machine is multipurpose.
In some embodiments, the extraction member 10 has a beveled surface and the magnetic member 50 is disposed adjacent to the beveled surface. The beads in the extraction chamber may move along the inner surface of the ramp with the magnetic member 50 as they are adsorbed.
In some embodiments, the first pipeline includes a collecting pipeline and a plurality of branch pipelines, one ends of the branch pipelines are respectively communicated with the corresponding reagent accommodating components 20, the other ends are communicated with the first control valve 30, one ends of the collecting pipeline are communicated with the first control valve 30, the other ends are communicated with the extracting chamber, and each branch pipeline is provided with a liquid pump 40 respectively used for sucking the reagent of the corresponding reagent accommodating components 20. The first control valve 30 is used to control the flow direction of the reagent.
Specifically, the plurality of reagent holding parts 20 include a pure water holding bottle, an ethanol holding bottle, an isopropyl alcohol holding bottle, and a nucleic acid cleaning liquid holding bottle, which respectively hold pure water, ethanol, isopropyl alcohol, and a nucleic acid cleaning liquid, wherein pure water, ethanol, isopropyl alcohol are used for a nucleic acid extraction process, and the nucleic acid cleaning liquid is used for cleaning the entire device piping, eliminating nucleic acid fragment contamination and rnase contamination thereof remaining in the piping.
Further, the first control valve 30 is a six-way valve, and the nucleic acid extraction and detection apparatus further includes a first waste liquid collecting member 70, and the first waste liquid collecting member 70 communicates with the six-way valve. The waste liquid from the washing in the apparatus may be delivered to the first waste liquid collecting member 70 through the first control valve 30.
In some embodiments, the second control valve 90 is disposed on the second pipeline, and the nucleic acid extraction and detection apparatus further includes a second waste collection member 71, the second waste collection member 71 being in communication with the second control valve 90. The second control valve 90 may be a three-way valve, which communicates with the extraction chamber, the second waste liquid collecting member 71 and the collecting member 60, respectively, and the waste liquid in the extraction chamber may be transferred into the second waste liquid collecting member 71 through the second control valve 90.
In some embodiments, the nucleic acid extraction and detection device further comprises a PCR module 80, and the collection member 60 is disposed on the PCR module 80. The PCR module 80 may comprise temperature control modules, optical fibers, optical signal collection modules, etc. as required in the qPCR process for subsequent PCR detection of the extracted nucleic acid product.
The following describes a procedure for using the nucleic acid isolation and detection device of the above embodiment.
1. Before operation, the nucleic acid scavenger is fed to the whole apparatus through the liquid pump 40 to wash the piping, followed by filling pure water for washing.
2. The sample is added through the sample adding port 12, an instrument operation program is set, and the operation is started.
3. Pure water is pumped into the extraction chamber by the liquid pump 40, and the cracked effervescent tablets enter the extraction chamber through the effervescent tablet feed inlet 11 and start to disintegrate and release after contacting with water. The cracking effervescent tablet is mainly used for promoting the cracking of a sample.
4. By controlling the temperature rotation means 13, the extraction chamber is warmed to a suitable temperature to start the lysis of the sample.
5. Isopropanol (nucleic acid precipitant) is pumped into the extraction chamber by a liquid pump 40, and the nucleic acid and magnetic beads are promoted to be combined by uniformly mixing by the temperature-controlled rotating member 13. In addition to isopropanol, other nucleic acid precipitants may be used.
6. After the completion of the combination, the temperature-controlling rotary member 13 is stopped, the magnetic member 50 is moved to the set position, the magnetic beads are adsorbed, and after the adsorption is completed, the supernatant is discharged to the second waste liquid collecting member 71 through a pipe.
7. The magnetic part 50 is removed, the washing effervescent tablet enters the extraction chamber through the effervescent tablet charging port 11, pure water and ethanol are pumped into the extraction chamber according to a certain proportion through the liquid pump 40, and after the washing effervescent tablet is contacted with water, disintegration release is started. The washing effervescent tablet is mainly used for removing protein or other impurities in the liquid.
8. The mixture is evenly mixed and washed by a temperature-controlled rotating part 13.
9. After the washing is completed, the temperature-controlling rotary member 13 is stopped, the magnetic member 50 is moved to the set position, the magnetic beads are adsorbed, and after the adsorption is completed, the supernatant is discharged to the second waste liquid collecting member 71 through a pipe.
10. The magnetic member 50 is removed and pure water and ethanol are pumped into the extraction chamber in a certain ratio by the liquid pump 40.
11. The mixture is evenly mixed and washed by a temperature-controlled rotating part 13.
12. After the washing is completed, the temperature-controlling rotary member 13 is stopped, the magnetic member 50 is moved to the set position, the magnetic beads are adsorbed, and after the adsorption is completed, the supernatant is discharged to the second waste liquid collecting member 71 through a pipe.
13. And (5) airing the magnetic beads.
14. The magnetic part 50 is removed and pure water is pumped into the extraction chamber.
15. The elution is carried out by mixing at a proper temperature by a temperature-controlled rotary member 13.
16. After the elution is completed, the temperature-controlled rotating part 13 stops operating, the magnetic part 50 moves to a set position to adsorb the magnetic beads, and after the adsorption is completed, the supernatant flows into the collecting part 60 through the pipe to obtain the extracted nucleic acid product.
17. If a subsequent experiment is required, the collection unit 60 may be filled with the corresponding reagent in advance, and the PCR process may be started.
Regarding the above extraction steps, those skilled in the art can appropriately adjust the steps according to actual needs so as to more accurately accomplish the nucleic acid extraction work. Such as the order of the steps, the number of executions of each step, etc.
The nucleic acid extraction and detection device in the present embodiment may be used only for extracting an accounting sample, or may be used for extracting and detecting a nucleic acid sample at the same time, and those skilled in the art may use the device according to actual needs.
In summary, the nucleic acid extraction and detection device for POCT in this embodiment is applicable to effervescent tablet type nucleic acid extraction reagents, is simple and convenient, and can achieve "same sample and detection", and after adding the sample and related reagents, the nucleic acid extraction detection result can be output faster, thus greatly saving detection time and related labor cost. In addition, the whole nucleic acid extraction and detection device adopts pipeline connection, is a closed nucleic acid extraction and detection device, and can solve the problem of cross contamination between samples compared with the traditional magnetic rod method full-automatic nucleic acid extraction instrument. Furthermore, the nucleic acid extraction and detection device can be used for extracting nucleic acids in various common biological samples such as swabs, whole blood, serum, plasma and tissue homogenate, and can be used for directly carrying out various subsequent experimental works such as PCR, qPCR, RT-qPCR (quantitative polymerase chain reaction) on the extracted nucleic acids through a temperature-controlled rotating component at the rear end, so that one machine is multipurpose.
In the description of the present utility model, the terms "first," "second," and the like 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 defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In the description of the present utility model, the meaning of "plurality" means at least two, for example, two, three, etc., unless specifically defined otherwise.
In the present utility model, unless explicitly specified and limited otherwise, the terms "mounted," "connected," "secured," and the like are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally formed; may be mechanically connected, may be electrically connected or may be in communication with each other; 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 utility model can be understood by those of ordinary skill in the art according to the specific circumstances.
In the present utility model, unless expressly stated or limited otherwise, a first feature "up" or "down" a second feature may be the first and second features in direct contact, or the first and second features in indirect contact via 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.
For purposes of this disclosure, the terms "one embodiment," "some embodiments," "example," "a particular example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the utility model. In this specification, schematic representations of the above terms are not necessarily directed to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, the different embodiments or examples described in this specification and the features of the different embodiments or examples may be combined and combined by those skilled in the art without contradiction.
While embodiments of the present utility model have been shown and described, it should be understood that the above embodiments are illustrative and not to be construed as limiting the utility model, and that variations, modifications, alternatives, and variations may be made to the above embodiments by those skilled in the art within the scope and spirit of the utility model, and any such modifications, equivalents, improvements, etc. are intended to be included within the scope of the present utility model.
Claims (10)
1. A nucleic acid extraction and detection device for POCT, comprising:
an extraction member (10), the extraction member (10) having an extraction chamber and an opening communicating with the extraction chamber;
a plurality of reagent containing parts (20), wherein each reagent containing part (20) is communicated with the extraction chamber through a first pipeline, and a first control valve (30) for controlling the trend of liquid and a liquid pump (40) for driving the liquid to flow are arranged on the first pipeline;
-a magnetic member (50), the magnetic member (50) being for attracting magnetic beads within the extraction chamber; and
-a collection member (60), the collection member (60) being in communication with the extraction chamber via a second conduit, the collection member (60) being adapted to collect nucleic acid products resulting from the extraction of nucleic acids.
2. The nucleic acid extraction and detection device for POCT according to claim 1, wherein the opening of the extraction means (10) comprises an effervescent tablet feed inlet (11) and a sample feed inlet (12).
3. The nucleic acid extraction and detection device for POCT according to claim 1, wherein a temperature control rotating member (13) for controlling the temperature in the extraction chamber and stirring the liquid in the extraction chamber is provided on the extraction chamber, and the magnetic member (50) is provided on one side of the extraction member (10).
4. The nucleic acid extraction and detection device for POCT according to claim 3, wherein the temperature-controlled rotation member (13) comprises a heating member and a stirring member, the heating member is disposed in or out of the extraction chamber, and the stirring member is disposed in the extraction chamber.
5. A nucleic acid extraction and detection apparatus for POCT according to claim 3, characterized in that the extraction means (10) has a bevel, the magnetic means (50) being arranged close to the bevel.
6. The nucleic acid extraction and detection device for POCT according to claim 1, wherein the first line comprises a collection line and a plurality of branch lines, one ends of the plurality of branch lines are respectively communicated with the corresponding reagent accommodating parts (20), the other ends are communicated with the first control valve (30), one ends of the collection line are communicated with the first control valve (30), the other ends are communicated with the extraction chamber, and each branch line is provided with the liquid pump (40).
7. The nucleic acid extraction and detection apparatus for POCT according to claim 6, wherein the plurality of reagent holding parts (20) includes a pure water holding bottle, an ethanol holding bottle, an isopropyl alcohol holding bottle, and a nucleic acid washing liquid holding bottle.
8. The nucleic acid extraction and detection device for POCT according to claim 7, characterized in that the first control valve (30) is a six-way valve, the nucleic acid extraction and detection device for POCT further comprising a first waste liquid collection means (70), the first waste liquid collection means (70) being in communication with the six-way valve.
9. The nucleic acid extraction and detection device for POCT according to claim 1, characterized in that a second control valve (90) is provided on the second line, the nucleic acid extraction and detection device for POCT further comprising a second waste liquid collection means (71), the second waste liquid collection means (71) being in communication with the second control valve (90).
10. The nucleic acid extraction and detection device for POCT according to claim 1, further comprising a PCR module (80), wherein the collection means (60) is provided to the PCR module (80).
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116875425A (en) * | 2023-07-24 | 2023-10-13 | 北京万泰生物药业股份有限公司 | Nucleic acid extraction and purification device, reagent combination, nucleic acid extraction and purification system and method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116875425A (en) * | 2023-07-24 | 2023-10-13 | 北京万泰生物药业股份有限公司 | Nucleic acid extraction and purification device, reagent combination, nucleic acid extraction and purification system and method |
| CN116875425B (en) * | 2023-07-24 | 2024-03-12 | 北京万泰生物药业股份有限公司 | Nucleic acid extraction and purification device, reagent combination, nucleic acid extraction and purification system and method |
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