WO2023097922A1 - 一种全封闭全自动核酸提取试剂盒 - Google Patents

一种全封闭全自动核酸提取试剂盒 Download PDF

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Publication number
WO2023097922A1
WO2023097922A1 PCT/CN2022/080032 CN2022080032W WO2023097922A1 WO 2023097922 A1 WO2023097922 A1 WO 2023097922A1 CN 2022080032 W CN2022080032 W CN 2022080032W WO 2023097922 A1 WO2023097922 A1 WO 2023097922A1
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WIPO (PCT)
Prior art keywords
chamber
valve
washing
washing liquid
nucleic acid
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PCT/CN2022/080032
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English (en)
French (fr)
Inventor
张辉
吴学如
李辉
李桂平
Original Assignee
珠海黑马生物科技有限公司
珠海黑马医学仪器有限公司
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Priority claimed from CN202111447749.7A external-priority patent/CN114107035B/zh
Priority claimed from CN202210039444.0A external-priority patent/CN114317255B/zh
Application filed by 珠海黑马生物科技有限公司, 珠海黑马医学仪器有限公司 filed Critical 珠海黑马生物科技有限公司
Publication of WO2023097922A1 publication Critical patent/WO2023097922A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/02—Apparatus for enzymology or microbiology with agitation means; with heat exchange means
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/36—Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
    • C12M1/38—Temperature-responsive control
    • 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

Definitions

  • the technology relates to nucleic acid extraction kits.
  • nucleic acid extraction kit the lysate, washing liquid, and eluent are divided into different functional chambers in the kit.
  • a mechanical device that can move in the three directions of X, Y, and Z is used.
  • the mechanism drives the magnetic bar to move the magnetic beads adsorbing nucleic acid between the various functional chambers.
  • the magnetic bar and the magnetic bar cover also take into account the function of stirring to realize the cracking, adsorption, washing, elution and other steps of the extraction process.
  • the disadvantages of the method are:
  • each functional chamber must be in an open state during the extraction process.
  • the process of magnetic rod adsorption and moving magnetic beads the dripping of the sample liquid, as well as heating and stirring during the extraction process.
  • the generated splash droplets and aerosols can easily cause cross-contamination between parallel samples and laboratory environmental pollution, which seriously interferes with the accuracy of the experiment.
  • the X, Y, and Z three-axis driving device of the magnetic bar and the magnetic bar cover has a complex mechanism, many moving parts, high manufacturing cost, large volume, and many failure points.
  • the waste after nucleic acid extraction stays in the open kit, which is difficult to handle, and if it is not handled properly, it will easily pollute the environment.
  • the purpose of the present invention is to provide a nucleic acid extraction process that does not cause cross-contamination, has a simple structure, is small in size, does not contact the extractor with the sample, does not pollute the extractor, and waste solids and liquids after nucleic acid extraction are sealed in the kit , a fully enclosed fully automatic nucleic acid extraction kit that does not pollute the experimental environment and is easy to handle.
  • the fully-enclosed fully automatic nucleic acid extraction kit of the present invention comprises a box body, and the box body has a plurality of working chambers, namely, a lysing chamber containing a lysate, a suction chamber, a washing liquid chamber containing a washing liquid, and a chamber containing a washing liquid.
  • eluent chamber for eluent eluent
  • the bottom of the suction and washing chamber is connected with the adsorption chamber, and there is nucleic acid adsorption material in the adsorption chamber; the bottom of the lysis chamber, the bottom of the washing liquid chamber, and the bottom of the eluent chamber respectively pass through the lysate pipeline with the lysate valve connected in series, and the wash liquid valve connected in series.
  • the washing solution pipeline and the eluent pipeline connected in series with the eluent valve are connected to the adsorption chamber; the lysate valve, washing solution valve and eluent valve are all non-contact control switch valves;
  • the cracking chamber is provided with a magnetic stirring bar which can rotate under the drive of the magnetic stirring driver;
  • a piston that slides up and down along the inner wall of the suction chamber to seal and contact is arranged in the suction chamber, and the piston is connected with a transmission mechanism that drives the piston to move up and down, and the power input end of the transmission mechanism is arranged outside the box body;
  • the box body has a sample loading cover for covering or opening the upper opening of the lysing chamber, and a sampling cover for covering or opening the upper opening of the eluent chamber.
  • the washing liquid chamber includes a first washing liquid chamber filled with a first washing liquid, a second washing liquid chamber filled with a second washing liquid, and the washing liquid valve includes a first washing liquid valve , the second washing liquid valve, the bottom of the first washing liquid chamber communicates with the adsorption chamber through the first washing liquid pipeline connected in series with the first washing liquid valve, and the bottom of the second washing liquid chamber communicates with the adsorption chamber through the second washing liquid pipeline connected in series with the second washing liquid valve
  • the washing liquid pipeline communicates with the adsorption chamber.
  • the above-mentioned fully enclosed fully automatic nucleic acid extraction kit is provided with a connecting valve on the box body;
  • the connecting valve includes a valve cavity and a connecting valve spool;
  • the air channel of the cracking chamber, the air channel of the suction chamber, the air channel of the washing liquid chamber, and the air channel of the eluent chamber communicate with the valve chamber of the connecting valve;
  • the connecting valve has two working states, that is, the conducting state and the blocking state.
  • the conduction state makes the four air passages connected, and the four air passages are not connected to each other in the cut-off state;
  • the connection valve can be operated from the outside of the kit or controlled by a non-contact control method, so that the connection valve can be separated from the cut-off state. state transitions to the on state.
  • the above-mentioned fully enclosed fully automatic nucleic acid extraction kit has a valve cavity in the upper part of the box body, and the ports communicating with the four air channels and the valve cavity are located on the inner wall of the valve cavity, and the connecting valve core has two different positions in the valve cavity, namely The conduction station and the cutoff station; when the connecting valve spool is at the cutoff station, the surface of the connecting valve spool blocks the four air passage ports, and the four air passages are not connected to each other; when the connecting valve spool is at the conduction station , the surface of the connecting valve spool is not in contact with the ports of the four air passages, and the upper parts of the four working chambers are communicated through four communicating air passages; the connecting valve spool is connected with a driving mechanism extending outside the box body, through which the driving mechanism can be dialed The connecting valve spool in the movable valve chamber moves from the cut-off station to the conduction station.
  • the valve includes a magnetic material spool and a spring.
  • the spring When the spring is in a normal state, the spool is in sealing contact with the inner wall of the pipeline where the spool is located, and the pipeline is in a normally closed and cut-off state; Outside the box, it is used to control a valve.
  • the electromagnet corresponding to the valve position When the electromagnet corresponding to the valve position is energized, it can generate force on the valve core of the valve, so that the valve core moves, and the valve core is no longer in sealing contact with the inner wall of the pipeline.
  • the pipeline is in a conduction state.
  • the transmission mechanism includes a threaded screw that fits with the piston and extends into the suction and washing chamber, and the upper end of the screw that passes through the upper end of the suction and washing chamber is fixed coaxially with the screw as a power input
  • the driving wheel at the end, the structure that prevents the piston from rotating relative to the inner wall of the suction chamber is set between the piston and the inner wall of the suction chamber, so that the piston can only slide up and down relative to the inner wall of the suction chamber.
  • a plurality of working chambers namely, the lysing chamber, the suction chamber, the washing liquid chamber, and the eluent chamber are arranged on the same line, and the lysate valve, the adsorption chamber, the washing liquid valve, the washing chamber
  • the dehydration valves are arranged on the same straight line.
  • one edge of the box body has a chamfer, and the chamfered edge is parallel to the straight line direction in which the plurality of working chambers are arranged.
  • the multiple working chambers arranged sequentially on the same line namely, the lysing chamber, the suction and washing chamber, the washing liquid chamber, and the eluting liquid chamber are called the working chamber row
  • the working chamber row is as follows: Multiple columns arranged side by side, the lysate valves, adsorption chambers, washing liquid valves, and eluent valves arranged in sequence on the same straight line are called valve columns, and the valve columns have multiple columns arranged side by side; each column of working chambers
  • the lower part is a valve row corresponding to the working chamber row; the cracking chamber, suction washing chamber, washing liquid chamber, and eluent chamber in different working chamber rows are arranged in a row, and the lysing liquid valve and adsorption chamber in different valve rows , washing fluid valves, and eluent valves are arranged in a row.
  • the outer wall of the box body has positive and negative contacts of a heating power supply circuit connected to an external power supply;
  • An electric heating device is provided, which can heat the liquid in the cracking chamber and the suction and washing chamber when the electric heating device is powered on, and the electric heating devices in different working chamber columns are all electrically connected to the positive and negative contacts of the heating power supply circuit.
  • a complete electric heating film and a bimetallic temperature controller are arranged in the chamber wall between the rows of lysing chambers arranged in a row and the rows of suction and washing chambers arranged in a row.
  • the membrane is an electric heating device for constant temperature heating of the liquid in each cracking chamber and each suction and washing chamber.
  • the above-mentioned fully enclosed fully automatic nucleic acid extraction kit has 2 columns, 4 columns or 8 columns of working chambers; regardless of whether there are 2 columns, 4 columns or 8 columns of working chambers, the kit has multiple working chambers in one column of working chambers.
  • the widths in the arrangement direction of each working chamber are equal, and the lengths of the kits in the arrangement direction of each working chamber column are equal.
  • the above-mentioned fully enclosed and fully automatic nucleic acid extraction kit has 8 columns of working chambers, and the volume of each working chamber is u; by changing the height of the box body, the size of the volume of each working chamber can be changed.
  • the above-mentioned fully enclosed fully automatic nucleic acid extraction kit also includes a plurality of connecting valves, each connecting valve corresponds to a row of working chambers;
  • the connecting valve includes a valve chamber and a connecting valve spool; each valve chamber is in the upper part of the box body;
  • the upper part of the cracking chamber, the upper part of the suction and washing chamber, the upper part of the washing liquid chamber, and the upper part of the eluent chamber in a row of working chambers respectively pass through the air channel of the cracking chamber, the air channel of the suction and washing chamber, the air channel of the washing liquid chamber, and the air channel of the eluent chamber It communicates with the valve chamber of the communication valve in the working chamber row, and the ports where the four air channels communicate with the valve chamber are located on the inner wall of the valve chamber;
  • the valve core of the communication valve has two different positions in the valve chamber, that is, the conduction work When the connecting valve spool is at the blocking station, the surface of the connecting valve
  • a first electric heating device is arranged in the wall of the lysis chamber, and the first electric heating device can heat the liquid in the lysis chamber when it is energized, and a second electric heating device is arranged in the wall of the suction and washing chamber. The electric heating device, the second electric heating device can heat the liquid in the suction and washing chamber when energized.
  • the above-mentioned fully-enclosed fully automatic nucleic acid extraction kit has a cuboid shape, and one of its edges has a chamfer to prevent the kit from being inserted into the extraction instrument in the wrong direction.
  • Lysis open the sample addition cover, add the sample into the lysis chamber pre-installed with lysate, and close the sample addition cover.
  • the first electric heating device is energized to heat the liquid in the lysis chamber at a constant temperature.
  • the magnetic stirring driver located on the outside of the kit is used to drive the magnetic stirrer in the lysis chamber to rotate. The magnetic stirrer stirs the mixed liquid of the sample and the lysate, and the sample The nucleus or virus in the cell is lysed, releasing the nucleic acid.
  • lysate valve driver VaD located at the outer bottom of the kit, opposite to the lysate valve Va, the valve driver (called lysate valve driver VaD) that controls the lysate valve switch in a non-contact manner drives the lysate valve Va to open, and then,
  • the power device located outside the kit drives the power input end, drives the piston to move upward through the transmission mechanism, and sucks the nucleic acid-dissolved lysis solution from the lysis chamber, through the lysate valve Va, and the adsorption chamber into the suction and washing chamber;
  • the power device drives the piston to move downward through the transmission mechanism, and pumps the lysis solution containing a small amount of nucleic acid from the suction chamber through the adsorption chamber and the lysate valve Va into the lysis chamber; the piston in the suction chamber reciprocates up and down Move several times, drive the liquid to reciprocate between the lysis chamber and the suction chamber through the adsorption chamber with nucleic acid adsorption material to improve the adsorption efficiency of nucleic acid, and finally drive the piston to move down to discharge all the liquid in the suction chamber into the lysis chamber chamber storage, then the lysate valve driver VaD controls the lysate valve Va to close, the lysate pipeline is closed, and the adsorption step is completed.
  • the valve driver located at the outer bottom of the kit and opposite to the washing liquid valve, which controls the opening and closing of the washing liquid valve in a non-contact manner (called the washing liquid valve driver) drives the washing liquid valve to open, and then the power unit passes through
  • the transmission mechanism drives the piston to move upwards, sucking the washing liquid from the washing liquid chamber, through the washing liquid valve and the adsorption chamber into the suction and washing chamber;
  • the washing liquid When the washing liquid flows through the adsorption chamber, it will wash off most of the non-nucleic acid substances adsorbed on the nucleic acid adsorption material.
  • the piston is driven to move downward to discharge the liquid in the suction and washing chamber into the washing liquid chamber for storage, and then the washing liquid valve driver controls the washing liquid valve to close, and the washing liquid pipeline is closed to complete the washing step.
  • valve driver (called eluent valve driver VeD) which is located at the outer bottom of the kit, opposite to the eluent valve Ve, and controls the switch of the eluent valve Ve in a non-contact manner drives the eluent
  • the valve Ve is opened, and then, the power device drives the piston to move upward through the transmission mechanism, and the eluent is sucked into the suction and washing chamber from the eluent chamber through the eluent valve Ve and the adsorption chamber;
  • the eluent When the eluent flows through the adsorption chamber, it will elute the nucleic acid substances adsorbed on the nucleic acid adsorption material. Drive the piston in the suction chamber to reciprocate several times up and down, and the eluent flows through the adsorption chamber repeatedly between the suction chamber and the eluent chamber to elute the nucleic acid adsorbed on the nucleic acid adsorption material to improve Elution efficiency. Finally, the piston is driven to move downward to discharge the liquid in the suction chamber into the eluent chamber for storage, and then the eluent valve driver VeD controls the eluent valve Ve to close, and the eluent pipeline is closed to complete the elution step.
  • step S50 in the extraction process is replaced by steps S51 and S61.
  • the first washing the valve driver located at the outer bottom of the kit, opposite to the first washing liquid valve Vc, and controlling the switch of the first washing liquid valve Vc in a non-contact manner (referred to as the first washing liquid valve driver VcD ) to drive the first washing liquid valve Vc to open, and then, the power device drives the piston to move upward through the transmission mechanism, and the first washing liquid is sucked into the suction washing chamber from the first washing liquid chamber through the first washing liquid valve Vc and the adsorption chamber ;
  • the first washing liquid When the first washing liquid flows through the adsorption chamber, it will wash off part of the non-nucleic acid substances adsorbed on the nucleic acid adsorption material. Drive the piston in the suction and washing chamber to move up and down several times, the first washing liquid flows between the suction and washing chamber and the first washing liquid chamber through the adsorption chamber for many times, and removes most of the non-nucleic acid substances adsorbed on the nucleic acid adsorption material. Wash down to increase washing efficiency.
  • the piston is driven to move down to discharge the liquid in the suction and washing chamber into the first washing liquid chamber for storage, and then the first washing liquid valve driver VcD controls the first washing liquid valve Vc to close, and the first washing liquid pipeline is closed to complete the first washing liquid. wash step.
  • the second washing the valve driver located at the outer bottom of the kit, opposite to the second washing liquid valve Vd, controls the switch of the second washing liquid valve Vd in a non-contact manner (called the second washing liquid valve driver VdD ) drives the second washing liquid valve Vd to open, and then, the power device drives the piston to move upward through the transmission mechanism, and the second washing liquid is sucked into the suction washing chamber from the second washing liquid chamber through the second washing liquid valve Vd and the adsorption chamber ;
  • the second washing liquid When the second washing liquid flows through the adsorption chamber, it will wash off the remaining non-nucleic acid substances adsorbed on the nucleic acid adsorption material.
  • the piston is driven to move down to discharge the liquid in the suction and washing chamber into the second washing liquid chamber for storage, and then the second washing liquid valve driver VdD controls the second washing liquid valve Vd to close, and the second washing liquid pipeline is closed to complete the second washing liquid. wash step.
  • the magnetic stirring driver and the magnetic stirrer driven by the magnetic stirring driver belong to the prior art, and its principle is similar to that of a magnetic stirrer, that is, the same-sex repulsion and opposite-sex attraction principles of the magnetic field are used to drive the magnetic stirrer in a non-contact manner.
  • the magnetic stirring bar placed in the kit runs in a circle, so as to achieve the purpose of stirring the liquid.
  • the magnetic stirring driver, valve driver, power unit, and the control mechanism that controls the state switching of the communication valve are not installed on this kit, they are only used in this kit An external device used to drive the magnetic stirrer, valve, drive wheel, and communication valve in this kit during nucleic acid extraction.
  • magnetic stirring driver, valve driver (including lysate valve driver, washing solution valve driver (including first washing solution valve driver, second washing solution valve driver), eluent valve driver) , the power unit, and the toggle device for moving the connecting valve spool of the toggle connecting valve can be installed on the corresponding position of the extractor, that is to say, when the kit is positioned on the extractor, the magnetic stirring drive can be positioned The position where the magnetic stirrer in the cracking chamber is driven to rotate, the lysate valve driver is located at the position where it can drive the lysate valve to switch, the washing liquid valve driver is located at the position that can drive the washing liquid valve to switch, and the eluent valve driver is at the The position where the eluent valve can be driven to perform switching action, and the power device is located where it can be mechanically connected to the drive wheel.
  • the connecting valve spool can be moved from the blocking position to
  • the sample addition cover, the sampling cover and the box body can be detachably connected, for example, the sample addition cover, the sampling cover, and the box body are mutually independent parts that can be separated from each other.
  • the sample addition cover and the box body can be an integral structure. At this time, in order to be able to open the sample addition cover to add a sample, the sample addition cover can cover the upper opening of the cracking chamber (sealedly connected with the upper opening of the cracking chamber) after adding the sample. It is movably connected with the box body, such as turning over and connecting.
  • sampling cover and the box body can be an integral structure. At this time, in order to open the sampling cover to take a sample, the sampling cover and the box body are movably connected, such as folded.
  • the present invention is provided with an air channel communication control structure including a communication valve and the like.
  • the connecting valve When the connecting valve is in the cut-off state, the air passages of the cracking chamber, the air passage of the suction chamber, the air passage of the washing liquid chamber, the air passage of the eluent chamber, etc. are not connected to each other.
  • the connecting valve When the connecting valve is in the conducting state, these air passages are connected.
  • the connecting valve Before using the kit, if the product leaves the factory, during transportation, and storage, the connecting valve is in an isolated state, so that the pre-installed substances in each working chamber in the kit will not move to other working chambers, and the contents in each working chamber Will not leak or mix with each other. That is, the kit contents do not leak, or contaminate each other.
  • this kit for nucleic acid extraction after opening the sample cap and adding the sample, before performing the subsequent extraction operation, it is necessary to change the connection valve from the cut-off state to the conduction state.
  • the passage, the air passage of the washing liquid chamber, the air passage of the eluent chamber and other air passages are connected, so that when the piston moves up and down, it will not cause resistance to the flow of the liquid due to the different pressures in the working chambers.
  • the communication valve can adopt a non-contact control communication valve, or a communication valve that can be operated from the outside of the reagent box.
  • the kit With the non-contact control communication valve, the kit can be inserted into the extraction instrument, and then the communication valve can be switched to the conduction state in a non-contact control mode.
  • a communication valve that can operate the communication valve from the outside of the kit it is necessary to manually switch the communication valve to the conduction state before the kit is inserted into the extraction instrument, or through the extraction instrument during the insertion of the kit into the extraction instrument. Switch the connecting valve to the conduction state by using the toggle device set on it.
  • the connecting valve spool is connected to a drive mechanism extending outside the box body, through which the connecting valve spool in the valve cavity can be moved from the blocking position to the conducting position. Station.
  • a valve with simple structure, low cost and convenient control includes a magnetic valve core and a spring. Closed flow state.
  • the valve driver corresponding to the valve can be a common and convenient electromagnet.
  • the electromagnet corresponding to the valve installed on the extractor is energized, the spool that absorbs the valve moves against the elastic force of the spring, and the spool is no longer in sealing contact with the inner wall of the pipeline. pass status.
  • the electromagnet is de-energized, the valve core resets under the action of the spring and is in sealing contact with the inner wall of the pipeline where it is located, and the pipeline is in a closed state.
  • the axis of the spool is vertical (the spool moves up and down along the axis of the spool), and the axis of rotation of the magnetic stirrer in the cracking chamber a is horizontal. This prevents drive crosstalk and prevents the magnetic fields of the valve drive and the magnetic stirrer drive from interfering with each other.
  • the power input end of the transmission mechanism is set outside the box body mainly for the convenience of connecting with the power device outside the kit, such as the power device set on the extraction instrument.
  • the transmission mechanism is a screw nut mechanism, and the driving wheel fixed with the screw rod is set on the box body.
  • the outside (such as the top of the box body) is convenient to match and engage with the power device to realize power transmission.
  • Multiple working chambers i.e. cracking chamber, suction chamber, washing liquid chamber, and eluent chamber are arranged on the same line, and the lysate valve, adsorption chamber, washing liquid valve, and eluent valve are arranged on the same line, which is more convenient layout.
  • the straight lines arranged by multiple working chambers may be parallel to or not parallel to the straight lines arranged by the lysate valves, adsorption chambers, washing liquid valves, eluent valves, etc.
  • the positive and negative contacts of the heating power supply circuit are arranged on the outer wall of the box body to facilitate the connection with the power circuit outside the kit, for example, the power circuit provided on the extractor.
  • This kit can use a single-column layout structure with only one column of working chambers to process one sample.
  • this kit adopts an array structure with a multi-column layout, that is, multiple columns of working chambers are arranged side by side.
  • the valve column also has the same number of columns as the working chamber column.
  • the valve actuator columns on the outer bottom of the kit also have the same number of columns as the valve columns.
  • the lysate valve driver, the washing solution valve driver and the eluent valve driver arranged on the same straight line are the valve driver row.
  • each valve row is a valve driver row corresponding to the valve row;
  • the liquid valve driver can drive the corresponding upper lysate valve, washing liquid valve, and eluent valve to switch. That is, each valve has a valve driver that drives the valve switch.
  • valves in one row are in the open state at the same time, and the other valves are in the closed state, so that the liquid will only flow in the corresponding working chamber and related pipelines according to the steps and in the specified pipeline. It can move in the path and working chamber without crosstalk of liquid.
  • the number of magnetic stirring drivers on the outside of the kit is also the same as the number of working chamber columns.
  • Each magnetic stirring driver is located on a column of working chambers (the magnetic stirring driver, cracking chamber, suction washing chamber, washing solution chamber, and eluent chamber are arranged sequentially on the same straight line), and is close to the cracking chamber.
  • Each magnetic stirring driver can drive the magnetic stirring bar in the cracking chamber located on the same straight line to rotate.
  • the valve driver column has 2 columns, 4 columns or 8 columns
  • the magnetic stirring driver has 2, 4 or 8 columns.
  • Nucleic acid extraction is performed on 2, 4 or 8 samples in parallel.
  • the kits have the same width, length, and height. This is to improve the versatility, and the extraction instrument of the same specification can be shared.
  • the height of the box can be increased without changing the length and width of the box.
  • the extraction instrument only needs to There is a height adjuster to adapt to the space height of different capacity reagent boxes, which can be applied to different capacity reagent boxes.
  • this kit does not need to be equipped with an electric heating device.
  • heating is required in the cracking, adsorption and elution processes.
  • a first electric heating device is arranged in the wall of the cracking chamber. Liquid heating.
  • a second electric heating device is arranged in the cavity wall of the suction and washing chamber, and the second electric heating device can heat the liquid in the suction and washing chamber when energized.
  • the advanced temperature control circuit realizes the constant temperature control when the liquid is heated.
  • the cracking chamber and the suction and washing chamber are adjacent, only one electric heating device can be arranged in the chamber wall between the cracking chamber and the suction and washing chamber.
  • the liquid inside is heated at the same time, which can further simplify the structure.
  • the outer wall of the box body has positive and negative contacts of the heating power supply circuit connected to the external power supply, and the electric heating device is electrically connected with the positive and negative contacts of the heating power supply circuit.
  • the box is in the shape of a cuboid, and one of its edges has a chamfer that prevents the kit from being inserted in the reverse direction when it is inserted into the extractor, making it more convenient to use.
  • the kit can be operated manually or fully automatically. It can be combined with the real-time fluorescent PCR module to form an extraction + real-time fluorescent PCR test integrated kit. It can be arranged in a single column or multiple columns in parallel, and a single column layout can process one sample. The multi-column layout can process multiple samples in parallel without interfering with each other.
  • the extraction process is fully enclosed. During the extraction process, the contents of the kit are not in contact with the experimental environment and any parts of the extraction instrument.
  • the nucleic acid adsorption material belongs to the existing materials, and the nucleic acid adsorption materials are concentrated in the adsorption cavity.
  • Nucleic acid adsorption materials such as magnetic beads can also be cemented into an adsorption column and placed in the adsorption chamber, and nano-scale loose magnetic beads can be cemented into an adsorption column to solve the contradiction between the difficulty of controlling discrete magnetic beads and the large adsorption area.
  • the kit is for one-time use, and the box body is made of plastic.
  • This kit can work with the corresponding extraction instrument to complete the automatic and fully enclosed nucleic acid extraction operation.
  • the working principle of the kit of the present invention is different from that of traditional nucleic acid extraction kits.
  • the nucleic acid adsorption material for adsorbing nucleic acid is always retained in the adsorption chamber.
  • the transfer of the liquid between the working chambers is realized, and the liquid passes through the adsorption chamber during the transfer, thereby realizing the adsorption and elution of nucleic acids.
  • the magnetic isolation driving method is adopted between the extractor and the kit. During the extraction process, the material and space inside the kit will not come into contact with any part of the extractor, which avoids sample contamination of the extractor and the experimental environment. The discarded solids and liquids after nucleic acid extraction are sealed in the kit, the waste does not pollute the experimental environment, and it is very convenient for the kit to be disposed of.
  • Fig. 1 is a schematic diagram of a kit in which multiple rows of working chambers are arranged side by side;
  • Fig. 2 is a schematic diagram of a kit of a single row of working chambers (in storage and transportation);
  • Fig. 3 is a schematic diagram of the kit when adding samples
  • Fig. 4 is the schematic diagram of kit when working
  • Fig. 5 is a schematic diagram during kit sampling
  • Figure 6 is a side view of the kit
  • Fig. 7 is a cross-sectional view of the kit
  • Fig. 8 is a schematic diagram of the principle of the kit at work
  • Fig. 9 is a schematic diagram of a piston, a screw rod, etc. of the kit.
  • Fig. 10 is a schematic diagram of the principle of the kit at work
  • Fig. 11 is a schematic diagram of the principle of a constant temperature heater
  • Fig. 12 is a schematic diagram of a state in which five air passages are not connected to each other;
  • Fig. 13 is a schematic diagram of the communication state of five air passages.
  • Fully enclosed fully automatic nucleic acid extraction kits are basically in the shape of a cuboid.
  • kit with multiple rows of working chambers arranged side by side see Figure 1.
  • 8 rows of working chambers are arranged side by side, which are the first working chamber row 1, the second working chamber row 2, and the third working chamber row. 3.
  • the fourth working chamber row 4 the fifth working chamber row 5, the sixth working chamber row 6, the seventh working chamber row 7, and the eighth working chamber row 8, arranged according to numbers from left to right, can be processed simultaneously 8 samples.
  • kits with a single column of chambers see Figures 2-6.
  • working chambers are arranged from left to right in each column of working chambers, which are the lysing chamber a, the suction and washing chamber b, the first washing liquid chamber c, Second washing liquid chamber d, eluent chamber e.
  • the lysis chamber a, the suction chamber b, the first washing liquid chamber c, the second washing liquid chamber d, and the eluent chamber e in different working chamber columns are arranged in a row in the X direction, from left to right, in order Row of lysing chamber, row of suction and washing chamber, row of first washing liquid chamber, row of second washing liquid chamber, row of eluting liquid chamber.
  • the front outer wall of the box body has a heating power supply circuit positive contact 63 connected to an external power supply, and the rear outer wall has a heating power supply circuit negative contact 64.
  • the electric heating film 61 is embedded in the chamber wall between the cracking chamber a and the suction and washing chamber b in the same row of working chambers, and the 8 electric heating films in the kit are connected in parallel to the bimetallic thermostat 62 in series, and then connected to the extractor through the positive contact 63 and the negative contact 64 of the heating power supply circuit, controlled and driven by the extractor, during the extraction related steps, the liquid in the cracking chamber a and the suction chamber b is heated at a constant temperature.
  • the eight electrothermal films 61 can also be a complete electrothermal film that runs through the cavity wall between the row of cracking chambers and the row of suction and washing chambers.
  • the kit mainly includes a box body 10 , a sample adding cover 101 and a sampling cover 102 .
  • the sample addition cover 101 is folded and connected with the box body 10 along the left fold (a kind of movable connection mode), and the lower part of the sample addition cover has a sample injection cover that extends into the upper opening of the cracking chamber and is in sealing contact with the upper opening of the cracking chamber.
  • Cover wings 132 cover wings 132 .
  • the sampling cover 102 is folded and connected with the box body along the right folded part 108 (a kind of flexible connection), and the lower part of the sampling cover has a sampling cover that extends into the upper mouth of the eluent chamber and is in sealing contact with the upper mouth of the eluent chamber. Wings 136.
  • a communication valve 20 is provided on the box body, and the communication valve includes a valve cavity 21 located at the upper part of the box body, and a communication valve spool 22 sliding in the valve cavity.
  • the upper part of the cracking chamber, the upper part of the suction and washing chamber, the upper part of the first washing liquid chamber, the upper part of the second washing liquid chamber, and the upper part of the eluent chamber respectively pass through the air channel 91 of the cracking chamber, the air channel 92 of the suction washing chamber, and the air channel of the first washing liquid chamber.
  • Five air passages, including the air passage 94 of the second washing liquid chamber and the air passage 95 of the eluent chamber communicate with the valve chamber 21 of the communication valve.
  • the ports where the five air passages communicate with the valve cavity are located on the lower inner wall of the valve cavity in contact with the lower surface of the valve core of the communication valve.
  • the valve core 22 of the communication valve has two different positions in the valve cavity 21, that is, the conduction position 23 and Cut-off station 24; when the connecting valve spool 22 is at the blocking station 24, the lower surface of the connecting valve spool blocks 5 air passage ports, the 5 air passages are not connected to each other, and the connecting valve is in an isolated state; the connecting valve spool 22 At the conduction station 23, the lower surface of the spool of the communication valve is not in contact with the ports of the five air passages, the five air passages communicate through the valve cavity, and the communication valve is in the conduction state.
  • each valve cavity The front and rear sides of each valve cavity are provided with a lever groove for sliding of the lever, and the lever 26 is connected with all communication valve spools at the same time, and stretches out of the box body through the lever groove. Toggling the lever 26 can drive all the communication valve spools 22 to move from the cut-off position to the conduction position, so that each communication valve changes from the cut-off state to the conduction state.
  • the driving rod 26 is the driving mechanism capable of moving the spool of the communication valve in the valve cavity.
  • the bottom of the suction chamber communicates with the adsorption chamber 32, and there is an adsorption column 42 formed by glued nucleic acid adsorption materials in the adsorption chamber, and there are gaps between the nucleic acid adsorption materials through which fluid can pass.
  • the bottom of the lysing chamber, the bottom of the first washing liquid chamber, the bottom of the second washing liquid chamber, and the bottom of the eluent chamber respectively pass through the lysing liquid pipeline 41 with the lysing liquid valve 31 connected in series, and the first washing liquid valve 33 connected in series.
  • the washing liquid pipeline 43 , the second washing liquid pipeline 44 connected in series with the second washing liquid valve 34 , and the eluent pipeline 45 connected in series with the eluent valve 35 communicate with the adsorption chamber 32 .
  • the lysate valve 31, the first washing liquid valve 33, the second washing liquid valve 34, and the eluent valve 35 have the same structure, and all include a magnetic material spool 36 and a spring 37.
  • the upper end of the spool 36 is a cone, and the spring is in a normal state.
  • the valve core is in sealing contact with the tapered inner wall of the pipeline where the valve core is located, and the pipeline is in a normally closed and cut-off state.
  • valve driver column On the outside of the kit (for example, on the extractor), a valve driver column is arranged, and the valve driver column is composed of a lysate valve driving electromagnet 51, a first washing liquid valve driving electromagnet 53, and a second washing liquid valve driving electromagnet 54 arranged in sequence. , The eluent valve driving electromagnet 55 is formed.
  • each valve has a corresponding valve driver to control the valve switch, that is to say, the lysate valve 31, the first washing liquid valve 33, the second washing liquid valve 34, the eluent Below the valve 35 there are corresponding lysate valve driving electromagnets 51 , first washing liquid valve driving electromagnets 53 , second washing liquid valve driving electromagnets 54 , and eluent valve driving electromagnets 55 .
  • the electromagnet 51 When the lysate valve driving electromagnet 51 is energized, the electromagnet absorbs the spool 36 in the lysate valve 31 and overcomes the elastic force of the spring 37 and moves downward, the upper end of the spool is no longer in sealing contact with the inner wall of the lysate pipeline, is in the conduction state.
  • the lysate valve driving electromagnet 51 When the lysate valve driving electromagnet 51 is de-energized, the spool 36 moves upward under the elastic force of the spring 37, and the upper end of the spool is in sealing contact with the inner wall of the lysate pipeline, and the lysate pipeline is in a closed state.
  • the magnetic stirrer in the cracking chamber a is rotated and set on the horizontal axis 73.
  • the magnetic stirring bar 74 can be the existing magnetic stirring bars such as plastic stirring blades with built-in magnetic steel.
  • the magnetic stirring driver 70 installed on the extractor outside the reagent box for example, includes a stirring motor 71 and a driving magnet 72 arranged on the output shaft of the stirring motor. After the kit is inserted into the extractor, the stirring motor 71 is coaxial with the horizontal shaft 73, the stirring motor 71 works, the driving magnet 72 rotates, and the magnetic stirrer 74 in the cracking chamber a is driven to rotate through magnetic field coupling.
  • a piston 80 that slides up and down along the inner wall of the suction and washing chamber in sealing contact is arranged in the suction and washing chamber.
  • the screw mandrel 81 is threadedly matched with the piston 80, stretches into the suction and washing chamber, and rotates with the bottom and the upper end of the suction and washing chamber.
  • the screw mandrel 81 upper end that passes through suction washing cavity upper end is fixed with screw mandrel coaxial as the driving wheel 82 of power input end, and driving wheel 82 exposes box body as the power input end that links to each other with external power unit 85.
  • a structure that prevents the piston from rotating relative to the inner wall of the suction and washing chamber is set between the piston and the inner wall of the suction and washing chamber, so that the piston can only slide up and down relative to the inner wall of the suction and washing chamber (such as the inner wall of the suction and washing chamber has a guide groove parallel to the screw rod, the piston There are guide ribs extending into the guide groove, etc.).
  • the power device 85 installed outside the kit, for example, on the extractor, can be connected with the drive wheel 82 to drive the drive wheel 82 to rotate clockwise or counterclockwise around the axis of the screw rod 81, so that the piston 80 moves up and down.
  • the sample addition cover 101 When extracting nucleic acid, the sample addition cover 101 is opened with the left folded part 104 as a pivot, and the sample is added to the lysis chamber a of the kit with a sample addition gun, and then the sample addition cover 101 is tightly closed, and the sample addition cover is surrounded by a protective ring. Wings 133 are in sealing contact with the inner side of the lysis chamber wall.
  • the extractor pops out the kit, and the sampling cover 102 is opened with the right folded part 108 as a pivot, and the nucleic acid extraction product in the kit is extracted with a sampling gun for subsequent nucleic acid testing experiments.
  • Lysis chamber a pre-stored lysate; built-in magnetic stirrer; the upper part has an openable/closed sample cap for adding samples; the lysate valve is arranged at the bottom, and the lysate pipeline arranged in the kit will connect the lysate chamber a 1.
  • the lysate valve is connected in series with the adsorption chamber below the suction and washing chamber b; the upper part of the lysate chamber communicates with the valve chamber of the communication valve on the top of the kit through the air channel of the lysate chamber arranged in the box body.
  • a constant temperature heater composed of an electrothermal film and a bimetallic thermostat for controlling constant temperature is embedded in the partition wall between the cracking chamber a and the suction and washing chamber b. The positive and negative contacts are connected, and the power supply is obtained from the extractor and automatically heated at a constant temperature.
  • Suction and washing chamber b Built-in pipetting drive system: composed of piston, screw rod and driving wheel.
  • the driving wheel is coupled with the power unit of the extractor, and the piston is driven to move up and down along the suction and washing chamber;
  • the bottom is arranged with an adsorption chamber (an adsorption column composed of nucleic acid adsorption materials), and the bottom of the adsorption chamber is directly connected to the lysate pipeline or through the internal pipeline.
  • the first washing liquid pipeline, the second washing liquid pipeline, and the eluent pipeline are connected.
  • the top of the suction and washing chamber b communicates with the valve cavity of the communication valve through the air channel of the suction and washing chamber arranged in the box body.
  • the first washing liquid chamber c the first washing liquid is pre-stored; the bottom layout has a first washing liquid valve, and the first washing liquid chamber c passes through the first washing liquid valve and the first washing liquid pipeline and the adsorption chamber below the suction washing chamber b Connected in series; the top of the first washing liquid chamber c communicates with the valve chamber of the communication valve through the air channel of the first washing liquid chamber arranged in the box body.
  • the second washing liquid chamber d the second washing liquid is pre-stored; the bottom layout has a second washing liquid valve, and the second washing liquid chamber d passes through the second washing liquid valve and the second washing liquid pipeline and the adsorption chamber below the suction washing chamber b Connected in series; the upper side of the second washing liquid chamber d communicates with the valve chamber of the communication valve through the air channel of the second washing liquid chamber arranged in the box body.
  • Eluent chamber e pre-stored eluent; there is a sampling cover on the top, which can be opened/closed, and is used to extract the extraction product after the extraction operation is completed; there is an eluent valve at the bottom, and the eluent chamber e passes through the eluent
  • the valve and the eluent pipeline are connected in series with the adsorption chamber below the suction chamber b.
  • the upper part of the eluent chamber e communicates with the valve chamber of the communication valve through the air channel of the eluent chamber arranged in the box. For storage of extraction product solutions.
  • the appearance of the kit is basically rectangular, with a driving wheel on the top, a sample cap that can be flipped open/closed and a sampling cap, and a +/- electrode contact that is electrically connected to the built-in constant temperature heater on the side, and anti-reverse insertion on one side of the bottom Chamfer.
  • the extraction instrument drives the drive wheel of the kit through mechanical coupling, and drives the piston on the screw rod to reciprocate up and down to drive the liquid inside the kit to move to complete the nucleic acid extraction operation.
  • the bottom layer inside the kit is equipped with multiple valves, which are driven by the valve driver program on the extraction instrument to complete the control of the pipetting flow during the extraction process.
  • the power circuit of the extraction instrument drives the constant temperature heater inside the kit to realize the heating of the liquid during the extraction process through the electrode contacts of the kit.
  • the magnetic stirring bar inside the kit is driven by the magnetic stirring driver in the extractor to complete the liquid stirring work during the extraction process.
  • the operation of the extractor is controlled by the program, which is fully automatic.
  • the program and parameters can be set through the mobile APP, and the extraction operation is completed with one key.
  • the kit can be in the structure of a single column of working chambers only for a single sample, or in the structure of multiple columns of working chambers arranged side by side, so as to realize independent and parallel extraction operations of multiple samples.
  • Each row of working chambers is composed of five independent working chambers: cracking chamber a, suction and washing chamber b, first washing liquid chamber c, second washing liquid chamber d, and eluent chamber e, which is the reaction of the extraction operation process. Chamber. Except for suction chamber b, other chambers are pre-filled with reagents when leaving the factory. Lysis chamber a is pre-filled with lysate, the first washing liquid chamber c is pre-filled with the first washing liquid, the second washing liquid chamber d is pre-filled with the second washing liquid, and the eluent chamber d is pre-filled with eluent.
  • the valve cavity of the connecting valve is located on the top of the reagent box, and there is a connecting valve spool inside.
  • the connecting valve spool is in the partition position in the storage and transportation state, so as to ensure that the reagents in each chamber will not interfere with each other during storage and transportation.
  • the communication valve is in the conduction state, so that the tops of the five chambers of the cracking chamber a, the suction chamber b, the first washing liquid chamber c, the second washing liquid chamber d, and the eluent chamber e are connected with air. Guarantees internal pressure balance during extraction operations.
  • the control part is located at the bottom of the kit, and consists of the lysate valve 31, the first wash liquid valve 33, the second wash liquid valve 31, the first wash liquid valve 33, and the second wash liquid chamber located at the bottom of the lysis chamber a, the first wash liquid chamber c, the second wash liquid chamber d, and the eluent chamber e.
  • An eluent valve 34, an eluent valve 35 and related pipelines are used to control the flow direction of the pipetting liquid during the extraction process.
  • the adsorption chamber is located under the suction and washing chamber b, and contains an adsorption column.
  • sample addition cover is located above the lysis chamber a, which can be opened/closed for nucleic acid extraction Previous sample addition operation.
  • the sampling cover is located above the eluent chamber d, can be opened/closed, and is used for sample removal after nucleic acid extraction.
  • Kit opening Open the kit packaging.
  • the structure such as the stopper on the extractor will use the thrust during insertion to prevent the lever from moving relative to the extractor along with the kit, so that the connecting valve spools of the connecting valves on the upper part of the kit will be changed from the cut-off state
  • the five air passages communicating with the upper parts of the five working chambers are in a connected state to ensure the pressure balance of each chamber during the extraction operation.
  • the driving wheel on the upper part of the suction chamber b is coupled with the power device on the extractor, so that the extractor can drive the piston inside the suction chamber b to move up and down along the suction chamber b.
  • the +/- electrode contacts on the kit are connected to the extractor so that the extractor can supply power to the constant temperature heater in the kit.
  • the valve drivers (solenoid valves) on the extraction instrument are aligned with the corresponding valves in the reagent box, so that the extraction instrument can control the action of each valve according to the extraction procedure.
  • Each magnetic stirring driver on the extractor is aligned with the magnetic stirrer in the cracking chamber a in a row of working chambers, so that the extractor can drive the magnetic stirrer in the kit to stir the liquid.
  • the unilateral chamfer 109 at the lower part of one side of the kit can prevent the kit from being inserted into the extractor in the opposite direction. After the kit is inserted into the extractor, the top is pressed by the pressure plate on the extractor to ensure that the sample addition cap and sampling cap will not pop open during the extraction process.
  • the extractor After the added kit is inserted into the extractor, the extractor will automatically complete the following nucleic acid extraction steps according to the preset program.
  • the extractor supplies power to the constant temperature heater buried in the chamber wall between the lysing chamber a and the suction chamber b through the electrode contacts, and heats the sample and lysate mixture in the lysing chamber a, while the magnetic force in the extractor
  • the stirring driver drives the magnetic stirrer in the lysing chamber a to fully stir the mixed liquid in the lysed chamber a, so as to promote the full reaction between the lysed liquid and the sample, and complete the steps of cell lysing and nucleic acid release.
  • Adsorption The lysate valve located under the lysate valve at the lower part of the lysate chamber a drives the electromagnet to be energized, drives the lysate valve to conduct, and connects the pipeline between the lysate chamber a and the suction chamber b.
  • the power device on the extractor drives the screw rod, which drives the piston to move up, and the lysis mixture in the lysis chamber a is sucked into the suction chamber b through the adsorption column in the adsorption chamber.
  • the nucleic acid in the liquid will be adsorbed by the adsorption column.
  • the extractor drives the piston inside the suction chamber b to reciprocate several times up and down, and drives the liquid to reciprocate through the adsorption column between the cracking chamber a and the suction chamber b to improve the adsorption efficiency of nucleic acids. All the liquid in the chamber b is discharged into the lysis chamber a for storage, and then the lysate valve drives the electromagnet to cut off the power, the lysate valve is closed, the pipeline connecting the lysis chamber a and the suction and washing chamber b is cut off, and the adsorption step is completed.
  • the first washing liquid valve driving electromagnet on the extractor located below the first washing liquid valve 33 at the lower part of the first washing liquid chamber c of the kit is energized to drive the first washing liquid valve conduction, connect the pipeline between the first washing liquid chamber c and the suction and washing chamber b, the power device on the extractor drives the piston in the suction and washing chamber b to move up, and the first washing liquid pre-stored in the first washing liquid chamber c
  • the washing liquid is sucked into the washing chamber b through the adsorption column.
  • the first washing liquid flows through the adsorption column, it will wash off part of the non-nucleic acid substances adsorbed on the adsorption column.
  • the power device will drive the piston in the suction chamber b to reciprocate several times, and drive the first washing liquid to and fro between the suction chamber b and the first washing liquid chamber c to flow through the adsorption column multiple times to improve the washing efficiency.
  • the piston is driven to move down to discharge the liquid in the suction and washing chamber b into the first washing liquid chamber c for storage, the first washing liquid valve drives the electromagnet to cut off the power, releases the spool of the first washing liquid valve 33, and cuts off the connection with the suction and washing chamber b.
  • the pipeline of the first washing liquid chamber c to complete the first washing step.
  • the second washing After the first washing process is completed, the second washing liquid valve drive electromagnet on the extractor located under the valve of the second washing liquid chamber at the lower part of the second washing liquid chamber d of the kit is energized to drive the second washing liquid valve.
  • the washing liquid valve is turned on to communicate with the pipeline between the second washing liquid chamber d and the suction chamber b.
  • the second washing solution is sucked into the washing chamber b through the adsorption column.
  • the second washing liquid flows through the adsorption column, it will wash off the remaining non-nucleic acid substances adsorbed on the adsorption column.
  • the power device will drive the piston in the suction chamber b to reciprocate several times, and drive the second washing liquid to flow through the adsorption column multiple times to improve the washing efficiency. Finally, the piston is driven to move down to discharge the liquid in the suction and washing chamber b into the second washing liquid chamber d for storage.
  • the second washing liquid valve drives the electromagnet to cut off the power, releases the spool in the second washing liquid valve, and cuts off the connection with the suction and washing chamber b.
  • the pipeline of the second washing liquid chamber d to complete the second washing step.
  • the eluent valve drive electromagnet on the extractor located below the eluent valve at the lower part of the eluent chamber d of the kit is energized, and the eluent valve is driven to conduct and connect
  • the pipeline between the eluent chamber d and the suction chamber b, the driving device on the extractor drives the piston in the suction chamber b to move upward, and the eluent pre-stored in the eluent chamber d is pumped through the adsorption column Inhale and wash cavity b.
  • the power device will drive the piston in the suction chamber b to reciprocate several times, and drive the eluent to flow through the adsorption column repeatedly between the suction chamber b and the eluent chamber d, so as to improve the elution efficiency.
  • the piston is driven to move down, and the eluent containing the eluted nucleic acid in the suction chamber b is discharged into the eluent chamber d for storage, and the eluent valve drives the electromagnet to power off, releasing the spool in the eluent valve , cut off the pipeline connecting the suction chamber b and the eluent chamber d, and complete the elution step.
  • the sampling cover can be opened, and the extracted nucleic acid product can be extracted from the eluent chamber d with a sampling gun for subsequent nucleic acid detection operations.
  • each column of working chambers is the same, obviously, the content of the working process described in the present invention is generally for each working chamber in a column of working chambers.

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Abstract

一种核酸提取过程不会发生交叉污染，样品不与提取仪接触，不会污染提取仪，提取核酸后的废弃固体和液体都密封在试剂盒中，不污染实验环境，处理方便的全封闭全自动核酸提取试剂盒。它包括具有加样盖、取样盖的盒体，所述盒体具有裂解腔、吸洗腔、洗涤液腔、洗脱液腔，吸洗腔的底部与吸附腔相通，吸附腔内具有核酸吸附材料；裂解腔、洗涤液腔、洗脱液腔的底部分别通过串联有裂解液阀门的裂解液管路、串联有洗涤液阀门的洗涤液管路、串联有洗脱液阀门的洗脱液管路与吸附腔相通；裂解液阀门、洗涤液阀门、洗脱液阀门均为非接触控制开关的阀门；裂解腔内设置有磁力搅拌子；吸洗腔内设置沿吸洗腔内壁上下滑动密封接触的活塞。

Description

一种全封闭全自动核酸提取试剂盒 技术领域
本技术涉及核酸提取试剂盒。
背景技术
传统的磁珠法核酸提取试剂盒，裂解液，洗涤液，洗脱液，分处于不同的试剂盒内的功能腔室，提取过程中，使用可以在X、Y、Z三个方向移动的机械机构驱动磁棒，将吸附核酸的磁珠，在各个功能腔室之间移动，磁棒及磁棒套还兼顾搅拌的功能，实现提取过程的裂解、吸附、洗涤、洗脱等步骤，这种方法的缺点是：
为了实现磁棒在不同步骤进出不同的功能腔室，各个功能腔室在提取过程中必须处于开放状态，磁棒吸附、移动磁珠过程中，样本液体的滴落，以及提取过程中加热及搅拌产生的飞溅液滴和气溶胶，很容易造成并列样品之间的交叉污染和实验室环境污染，从而严重干扰实验的准确性。
同时，磁棒及磁棒套的X、Y、Z三轴驱动装置，机构复杂，运动部件多，仪器制造成本高，体积大，故障点多。
而且，提取核酸后的废弃物，滞留在开放的试剂盒中，难于处理，处理不好容易污染环境。
发明内容
本发明的目的是提供一种核酸提取过程不会发生交叉污染，结构简单，体积小巧，样品不与提取仪接触，不会污染提取仪，提取核酸后的废弃固体和液体都密封在试剂盒中，不污染实验环境、处理方便的全封闭全自动核酸提取试剂盒。
本发明的全封闭全自动核酸提取试剂盒，包括盒体，所述盒体具有多个工作腔，即装有裂解液的裂解腔、吸洗腔、装有洗涤液的洗涤液腔、装有洗脱液的洗脱液腔，
吸洗腔的底部与吸附腔相通，吸附腔内具有核酸吸附材料；裂解腔底部、洗涤液腔底部、洗脱液腔底部分别通过串联有裂解液阀门的裂解液管路、串联有洗涤液阀门的洗涤液管路、串联有洗脱液阀门的洗脱液管路与吸附腔相通；裂解液阀门、洗涤液阀门、洗脱液阀门均为非接触控制开关的阀门；
裂解腔内设置有能够在磁力搅拌驱动器的带动下转动的磁力搅拌子；
吸洗腔内设置沿吸洗腔内壁上下滑动密封接触的活塞，活塞与带动活塞上下移动的传动机构相连，该传动机构的动力输入端设置在盒体外部；
盒体具有用于盖住或打开裂解腔上口的加样盖，以及用于盖住或打开洗脱液腔上口的取样盖。
上述的全封闭全自动核酸提取试剂盒，洗涤液腔包括装有第一洗涤液的第一洗涤液腔、装有第二洗涤液的第二洗涤液腔，洗涤液阀门包括第一洗涤液阀门、第二洗涤液阀门，第一洗涤液腔底部通过串联有第一洗涤液阀门的第一洗涤液管路与吸附腔相通，第二洗涤液腔底部通过串联有第二洗涤液阀门的第二洗涤液管路与吸附腔相通。
上述的全封闭全自动核酸提取试剂盒，盒体上设置连通阀；连通阀包括阀腔和连通阀阀芯；裂解腔上部、吸洗腔上部、洗涤液腔上部、洗脱液腔上部分别通过裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道与连通阀的阀腔相通；连通阀有两种工作状态，即导通状态和隔断状态，连通阀在导通状态使得该四个空气通道相通，在隔断状态时使得该四个空气通道互不相通；能够从试剂盒外部对连通阀进行操作或者通过非接触控制方式控制连通阀，使得连通阀从隔断状态变换到导通状态。
上述的全封闭全自动核酸提取试剂盒，盒体内上部具有阀腔，四个空气通道与阀腔相通的端口位于阀腔内壁上，连通阀阀芯在阀腔内有两个不同工位，即导通工位和隔断工位；连通阀阀芯在隔断工位时，连通阀阀芯表面封堵四个空气通道端口，四个空气通道互不相通；连通阀阀芯在导通工位时，连通阀阀芯表面与四个空气通道端口不接触，四个工作腔上部通过四个相通的空气通道相通；连通阀阀芯上连接伸出盒体外部的驱动机构，通过该驱动机构能够拨动阀腔内的连通阀阀芯从隔断工位移动到导通工位。
上述的全封闭全自动核酸提取试剂盒，阀门包括磁性材料阀芯、弹簧，弹簧在常态时使得阀芯与阀芯所在管路的内壁密封接触，该管路处于常闭断流状态；在试剂盒外部、用于控制一个阀门、与该阀门位置相对应的电磁铁得电时，能够对该阀门的阀芯产生作用力，使得阀芯移动，阀芯与该管路内壁不再密封接触，管路处于导通状态。
上述的全封闭全自动核酸提取试剂盒，传动机构包括与活塞螺纹配合、伸入到吸洗腔内的丝杆，穿过吸洗腔上端的丝杆上端固定与丝杆同轴的作为动力输入端的驱动轮，活塞与吸洗腔内壁之间设置防止活塞相对 于吸洗腔内壁转动的结构，使得活塞只能相对于吸洗腔内壁上下滑动。
上述的全封闭全自动核酸提取试剂盒，多个工作腔，即裂解腔、吸洗腔、洗涤液腔、洗脱液腔在同一直线上排列，裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门在同一直线上排列。
上述的全封闭全自动核酸提取试剂盒，盒体的一个棱边具有倒角，该具有倒角的棱边与多个工作腔排列的直线方向平行。
上述的全封闭全自动核酸提取试剂盒，在同一直线上按顺序依次排列的多个工作腔即裂解腔、吸洗腔、洗涤液腔、洗脱液腔称为工作腔列，工作腔列有并列排列的多列，在同一直线上按顺序依次排列的裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门称为阀门列，阀门列有并列排列的多列；每列工作腔列的下部是与该工作腔列对应的一阀门列；不同工作腔列中的裂解腔、吸洗腔、洗涤液腔、洗脱液腔分别排成一行，不同阀门列中的裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门分别排成一行。
上述的全封闭全自动核酸提取试剂盒，盒体的外壁上具有与外部电源相连的加热供电电路正负极触点；在一列工作腔列中的裂解腔与吸洗腔之间的腔壁内设置电加热装置，电加热装置在通电时可以对裂解腔和吸洗腔内的液体加热，不同工作腔列内的电加热装置均与加热供电电路正负极触点电连接。
上述的全封闭全自动核酸提取试剂盒，在排成一行的裂解腔行与排成一行的吸洗腔行之间的腔壁内设置一张完整的电热膜和双金属温控器，该电热膜即是对各裂解腔和各吸洗腔内的液体进行恒温加热的电加热装置。
上述的全封闭全自动核酸提取试剂盒，工作腔列有2列、4列或8列；不论工作腔列有2列、4列还是8列，试剂盒在一列工作腔列中多个工作腔的排列方向上宽度相等，试剂盒在各工作腔列排列方向上的长度相等。
上述的全封闭全自动核酸提取试剂盒，有8列工作腔列的试剂盒，各工作腔的容积均为u；有4列工作腔列的试剂盒，各工作腔的容积均为v；有2列工作腔列的试剂盒，各工作腔的容积均为w；w＝2v＝4u；不论工作腔列有2列、4列还是8列，试剂盒高度相等。
上述的全封闭全自动核酸提取试剂盒，试剂盒有8列工作腔列，各工作腔的容积均为u；通过改变盒体的高度，改变各工作腔容积的大小。
上述的全封闭全自动核酸提取试剂盒，它还包括多个连通阀，每个连通阀与一列工作腔列对应；连通阀包括阀腔和连通阀阀芯；各阀腔均在盒体内上部；一列工作腔列中的裂解腔上部、吸洗腔上部、洗涤液腔上部、洗脱液腔上部分别通过裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道与该工作腔列中的连通阀的阀腔相通，该四个空气通道与阀腔相通的端口位于阀腔内壁上；连通阀阀芯在阀腔内有两个不同工位，即导通工位和隔断工位；连通阀阀芯在隔断工位时，该连通阀阀芯表面封堵四个空气通道端口，四个空气通道互不相通；连通阀阀芯在导通工位时，连通阀阀芯表面与四个空气通道端口不接触，四个工作腔上部通过四个相通的空气通道相通；一拨杆同时与各连通阀阀芯连接，并穿过阀腔上所开的拨杆槽伸出盒体；能够从试剂盒外部拨动拨杆，使得各连通阀的连通阀阀芯同时从隔断工位移动到导通工位，连通阀从隔断状态变换到导通状态。
上述的全封闭全自动核酸提取试剂盒，裂解腔的腔壁内设置第一电加热装置，第一电加热装置在通电时可以对裂解腔内液体加热，吸洗腔的腔壁内设置第二电加热装置，第二电加热装置在通电时可以对吸洗腔内液体加热。
上述的全封闭全自动核酸提取试剂盒，盒体成长方体形，其一个棱边具有防止试剂盒插入提取仪时方向插反的倒角。
使用该全封闭全自动核酸提取试剂盒进行核酸提取的步骤：
S10、裂解：打开加样盖，将样品加入到预装有裂解液的裂解腔内，关闭加样盖。第一电加热装置通电对裂解腔内液体进行恒温加热，同时采用位于试剂盒外侧部的磁力搅拌驱动器带动裂解腔内的磁力搅拌子转动，磁力搅拌子对样品和裂解液的混合液体搅拌，样品中的细胞核或病毒裂解，释放出核酸。
S30、吸附：位于试剂盒外底部的、与裂解液阀门Va相对的、用非接触方式控制裂解液阀门开关的阀门驱动器(称之为裂解液阀门驱动器VaD)驱动裂解液阀门Va打开，然后，位于试剂盒外部的动力装置驱动动力输入端，通过传动机构带动活塞向上移动，将溶有核酸的裂解溶液从裂解腔，经裂解液阀门Va、吸附腔吸入到吸洗腔内；
溶有核酸的裂解溶液流经吸附腔的时候，液体中的核酸和部分非核酸物质会被核酸吸附材料吸附。然后， 动力装置再通过传动机构带动活塞向下移动，将溶有少量核酸的裂解溶液从吸洗腔，经吸附腔、裂解液阀门Va泵入到裂解腔内；吸洗腔内的活塞上下往复移动数次，驱动液体在裂解腔、吸洗腔之间往复流经具有核酸吸附材料的吸附腔，以提高核酸的吸附效率，最后驱动活塞向下移动将吸洗腔中的液体全部排入裂解腔存储，然后裂解液阀门驱动器VaD控制裂解液阀门Va关闭，裂解液管路关闭，完成吸附步骤。
S50、洗涤：位于试剂盒外底部的、与洗涤液阀门相对的、用非接触方式控制洗涤液阀门开关的阀门驱动器(称之为洗涤液阀门驱动器)驱动洗涤液阀门打开，然后，动力装置通过传动机构带动活塞向上移动，将洗涤液从洗涤液腔，经洗涤液阀门、吸附腔吸入到吸洗腔内；
洗涤液流经吸附腔时，会将核酸吸附材料上吸附的大部分非核酸物质洗下来。驱动吸洗腔里的活塞上下往复移动数次，洗涤液往返于吸洗腔、洗涤液腔之间多次流经吸附腔，把吸附在核酸吸附材料上的非核酸物质洗下来，以提高洗涤效率。最后驱动活塞向下移动将吸洗腔中的液体排入洗涤液腔存储，然后洗涤液阀门驱动器控制洗涤液阀门关闭，洗涤液管路关闭，完成洗涤步骤。
S70、洗脱：位于试剂盒外底部的、与洗脱液阀门Ve相对的、用非接触方式控制洗脱液阀门Ve开关的阀门驱动器(称之为洗脱液阀门驱动器VeD)驱动洗脱液阀门Ve打开，然后，动力装置通过传动机构带动活塞向上移动，将洗脱液从洗脱液腔，经洗脱液阀门Ve、吸附腔吸入到吸洗腔内；
洗脱液流经吸附腔时，会将核酸吸附材料上吸附的核酸物质洗脱下来。驱动吸洗腔里的活塞上下往复移动数次，洗脱液往返于吸洗腔、洗脱液腔之间多次流经吸附腔，把吸附在核酸吸附材料上的核酸洗脱下来，以提高洗脱效率。最后驱动活塞向下移动将吸洗腔中的液体排入洗脱液腔存储，然后洗脱液阀门驱动器VeD控制洗脱液阀门Ve关闭，洗脱液管路关闭，完成洗脱步骤。
S90、取样：打开取样盖，以移液枪等汲取装置吸取洗脱液腔内的液体，再关闭取样盖。
当然，如果洗涤液腔包括第一洗涤液腔c、第二洗涤液腔d，则前述提取过程中的步骤S50以步骤S51、S61代替。
S51、第一次洗涤：位于试剂盒外底部的、与第一洗涤液阀门Vc相对的、用非接触方式控制第一洗涤液阀门Vc开关的阀门驱动器(称之为第一洗涤液阀门驱动器VcD)驱动第一洗涤液阀门Vc打开，然后，动力装置通过传动机构带动活塞向上移动，将第一洗涤液从第一洗涤液腔，经第一洗涤液阀门Vc、吸附腔吸入到吸洗腔内；
第一洗涤液流经吸附腔时，会将核酸吸附材料上吸附的部分非核酸物质洗下来。驱动吸洗腔里的活塞上下往复移动数次，第一洗涤液往返于吸洗腔、第一洗涤液腔之间多次流经吸附腔，把吸附在核酸吸附材料上的大部分非核酸物质洗下来，以提高洗涤效率。最后驱动活塞向下移动将吸洗腔中的液体排入第一洗涤液腔存储，然后第一洗涤液阀门驱动器VcD控制第一洗涤液阀门Vc关闭，第一洗涤液管路关闭，完成第一次洗涤步骤。
S61、第二次洗涤：位于试剂盒外底部的、与第二洗涤液阀门Vd相对的、用非接触方式控制第二洗涤液阀门Vd开关的阀门驱动器(称之为第二洗涤液阀门驱动器VdD)驱动第二洗涤液阀门Vd打开，然后，动力装置通过传动机构带动活塞向上移动，将第二洗涤液从第二洗涤液腔，经第二洗涤液阀门Vd、吸附腔吸入到吸洗腔内；
第二洗涤液流经吸附腔时，会将核酸吸附材料上吸附的剩余非核酸物质洗下来。驱动吸洗腔里的活塞上下往复移动数次，第二洗涤液往返于吸洗腔、第二洗涤液腔之间多次流经吸附腔，把吸附在核酸吸附材料上的剩余非核酸物质洗下来，以提高洗涤效率。最后驱动活塞向下移动将吸洗腔中的液体排入第二洗涤液腔存储，然后第二洗涤液阀门驱动器VdD控制第二洗涤液阀门Vd关闭，第二洗涤液管路关闭，完成第二次洗涤步骤。
本发明中，磁力搅拌驱动器、磁力搅拌驱动器驱动的磁力搅拌子属于现有技术，其原理类似于磁力搅拌器，即利用磁场的同性相斥、异性相吸的原理，使用磁场以非接触方式带动放置在试剂盒中带磁性的搅拌子进行圆周运转，从而达到搅拌液体的目的。
磁力搅拌驱动器、阀门驱动器、动力装置、控制连通阀进行状态切换的控制机构(或者对连通阀进行操作使其进行状态切换的操作机构)不是安装在本试剂盒上，它们只是在使用本试剂盒提取核酸的过程中，为了驱动本试剂盒中的磁力搅拌子、阀门、驱动轮、连通阀而使用的外部装置。为了使用本试剂盒提取核酸更加方便，磁力搅拌驱动器、阀门驱动器(包括裂解液阀门驱动器、洗涤液阀门驱动器(包括第一洗涤液阀门驱动器、第二洗涤液阀门驱动器)、洗脱液阀门驱动器)、动力装置、拨动连通阀的连通阀阀芯移动的拨动装 置可以安装在提取仪的相应的位置上，也就是说，当把本试剂盒定位在提取仪上时，磁力搅拌驱动器位于能够带动裂解腔内的磁力搅拌子转动的位置，裂解液阀门驱动器位于能够驱动裂解液阀门进行开关动作的位置，洗涤液阀门驱动器位于能够驱动洗涤液阀门进行开关动作的位置，洗脱液阀门驱动器位于能够驱动洗脱液阀门进行开关动作的位置，动力装置位于能够与驱动轮进行机械连接的位置。在试剂盒插入提取仪的过程中，通过提取仪上设置的拨动装置等，可以使连通阀阀芯从隔断工位移动到导通工位，将连通阀从隔断状态切换到导通状态。
加样盖、取样盖与盒体可以是可拆卸连接，例如，加样盖、取样盖、盒体是可以相互分离的相互独立的零部件。
加样盖与盒体可以是一体结构，此时，为了能够打开加样盖加样，在加样后加样盖能够盖住裂解腔上口(与裂解腔上口密封连接)，加样盖与盒体成活动连接，例如翻折连接。
取样盖与盒体可以是一体结构，此时，为了能够打开取样盖取样，取样盖与盒体成活动连接，例如翻折连接。
在使用本试剂盒提取核酸的过程中，为了使得各个工作腔保持空气连通，以保证提取过程中，移液时的各工作腔内部气压平衡，不形成移液阻力，同时也保证工作腔内的液体，不会因为搅拌导致的液体飞溅，污染相邻的工作腔，本发明设置了包括连通阀等在内的空气通道连通控制结构。连通阀在隔断状态时，裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道等空气通道互不相通，连通阀在导通状态时这些空气通道相通。在使用试剂盒之前，如产品出厂、运输、储存过程中，连通阀处于隔断状态，这样试剂盒内各工作腔内预装的物质不会移动到其它的工作腔，各个工作腔内的内容物不会泄漏或互相混合。也就是说，试剂盒内容物不泄漏，或者彼此不污染。在使用该试剂盒进行核酸提取操作时，打开加样盖加样后，进行后续提取操作前，需要把连通阀从隔断状态变换到导通状态，此时，裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道等空气通道相通，这样，活塞上下运动时，不会因为各个工作腔内的压力不同，对液体的流动造成阻力。
为了方便对连通阀进行状态的切换，连通阀可以采用非接触控制的连通阀，或者采用能够从试剂盒外部对连通阀进行操作的连通阀。采用非接触控制的连通阀，本试剂盒可以插入提取仪之后，再以非接触控制方式将连通阀切换到导通状态。采用能够从试剂盒外部对连通阀进行操作的连通阀，需要在试剂盒插入提取仪之前，通过手动方式将连通阀切换到导通状态，或者在试剂盒插入提取仪的过程中，通过提取仪上设置的拨动装置等，将连通阀切换到导通状态。
为了方便从试剂盒外部对连通阀进行操作，连通阀阀芯上连接伸出盒体外部的驱动机构，通过该驱动机构能够拨动阀腔内的连通阀阀芯从隔断工位移动到导通工位。
阀门有多种多样，一种结构简单、成本较低、控制方便的阀门包括磁性材料阀芯、弹簧，弹簧在常态时使得阀芯与阀芯所在管路的内壁密封接触，该管路处于常闭断流状态。此时与阀门对应的阀门驱动器可以是常见的使用方便的电磁铁。例如，安装在提取仪上的与该阀门对应的电磁铁得电，吸附该阀门的阀芯克服弹簧的弹力移动，阀芯与该管路内壁不再密封接触，该阀门所在的管路处于导通状态。当电磁铁断电，阀芯在弹簧的作用下复位又与其所在的管路内壁密封接触，管路处于关闭状态。
阀芯轴线是竖直的(阀芯沿阀芯轴线上下移动)，裂解腔a内的磁力搅拌子转动轴线是水平的。这样可以防止驱动串扰，防止阀门驱动器与磁力搅拌驱动器的磁场相互干扰。
传动机构的动力输入端设置在盒体外部主要是为了方便与试剂盒外部的例如设置在提取仪上的动力装置相连，传动机构为丝杆螺母机构，与丝杆固定的驱动轮设置在盒体外部(如盒体顶部)，方便与动力装置匹配啮合，实现动力传递。
多个工作腔，即裂解腔、吸洗腔、洗涤液腔、洗脱液腔在同一直线上排列，裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门在同一直线上排列，更加方便布置。多个工作腔排列的直线可以与裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门等排列的直线平行，也可以不平行。
盒体的外壁上设置加热供电电路正负极触点，方便与试剂盒外部的电源电路连接，例如设置在提取仪上的电源电路电连接。
本试剂盒可以采用只有一列工作腔列的单列布局结构，处理一个样品。
为了能够同时对多个样品进行核酸提取，本试剂盒采用多列布局的阵列结构，即工作腔列多列并列，此时阀门列也有与工作腔列相同的列数。当然，试剂盒外底部(例如安装在提取仪上)的阀门驱动器列也有与 阀门列相同的列数。排在同一直线上的裂解液阀门驱动器、洗涤液阀门驱动器、洗脱液阀门驱动器为阀门驱动器列。具体在使用本阵列式试剂盒进行核酸提取时，每列阀门列的下部是与该阀门列对应的一阀门驱动器列；每列阀门驱动器列中的裂解液阀门驱动器、洗涤液阀门驱动器、洗脱液阀门驱动器能够驱动其对应上方的裂解液阀门、洗涤液阀门、洗脱液阀门进行开关。也就是说，每个阀门下面都一个驱动该阀门开关的阀门驱动器。
在核酸提取操作过程中，只会有一行中的各阀门同时处于打开状态，其它阀门都处于关闭状态，这样液体只会在相应的工作腔和相关的管路中按照步骤需要，在规定的管路和工作腔中移动，不会出现液体的串扰。
试剂盒外侧部(例如安装在提取仪上)的磁力搅拌驱动器数量也与工作腔列的列数相同。每个磁力搅拌驱动器位于一工作腔列上(磁力搅拌驱动器、裂解腔、吸洗腔、洗涤液腔、洗脱液腔在同一直线上按顺序依次排列)，并靠近裂解腔。每个磁力搅拌驱动器可以带动与其位于同一直线上的裂解腔内的磁力搅拌子转动。
例如工作腔列、阀门列均有2列、4列或8列(相应的，阀门驱动器列均有2列、4列或8列，磁力搅拌驱动器有2、4或8个)，即可同时对2、4或8个样品并行进行核酸提取。不论工作腔列有2列、4列还是8列，试剂盒宽度相等、长度相等、高度相等，这样是为了提高通用性，可以共用同一规格的提取仪。
对于工作腔列、阀门列均有2列、4列或8列的试剂盒，为了改变工作腔的容量，可以增加盒体的高度，而不改变盒体的长度和宽度，这样，提取仪只要有高度调节器适配不同容量试剂盒的空间高度即可适用于不同容量的试剂盒。
有些核酸种类的提取，全程不需要加热，此时，本试剂盒不需要设置电加热装置。但是，大多数核酸的提取过程中，在裂解、吸附和洗脱过程需要加热，此时，裂解腔的腔壁内设置第一电加热装置，第一电加热装置在通电时可以对裂解腔内液体加热。吸洗腔的腔壁内设置第二电加热装置，第二电加热装置在通电时可以对吸洗腔内液体加热。采用内嵌在试剂盒内的第一电加热装置(如电加热膜)、第二电加热装置(如电加热膜)实现提取过程的液体加热，当然还可以在电加热装置上附上属于现有技术的温度控制电路，实现液体加热时的恒温控制。当然，如果裂解腔和吸洗腔相邻时，可以在裂解腔与吸洗腔之间的腔壁内仅仅设置在一个电加热装置，该电加热装置在通电时可以对裂解腔和吸洗腔内的液体同时加热，这样可以进一步简化结构。盒体的外壁上具有与外部电源相连的加热供电电路正负极触点，电加热装置均与加热供电电路正负极触点电连接。在使用本试剂盒的过程中，具体是否加热，在哪个环节加热，由外部电源控制，例如，由位于提取仪上的外部电源控制。
盒体成长方体形，其一个棱边具有防止试剂盒插入提取仪时方向插反的倒角，使用更加便利。
本试剂盒可手动操作，可全自动操作。可以与实时荧光PCR模组结合，构成提取+实时荧光PCR测试一体化试剂盒。可单列/多列并列布局，单列布局处理一个样品。多列布局可以并列同步处理多个样品，样品之间互不干涉。提取过程全封闭，提取过程中，试剂盒内容物不与实验环境和提取仪的任何部件接触。核酸吸附材料属于现有材料，将核酸吸附材料集中置于吸附腔内。也可以将核酸吸附材料例如磁珠胶结为吸附柱置于吸附腔内，将纳米级松散的磁珠胶结成吸附柱，解决离散的磁珠难控制和大吸附面积的矛盾。
试剂盒为一次性使用，盒体采用塑料材质。
本试剂盒可以与相应的提取仪配合工作，完成核酸全自动、全封闭提取操作。
本发明的试剂盒，工作原理与传统的核酸提取试剂盒不同，整个提取过程中，吸附核酸的核酸吸附材料，始终被滞留在吸附腔内。通过活塞的上下移动，实现液体在工作腔之间的转移，液体在转移时通过吸附腔，从而实现对核酸的吸附、洗脱。
由于省略了传统的转移磁珠的、需要不断进出各个工作腔的磁棒，整个提取过程，试剂盒各个样品之间，试剂盒与实验环境之间处于封闭隔离状态，不会发生交叉污染和实验室污染。
提取仪与试剂盒之间采用磁力隔离驱动方式，提取过程中，试剂盒内部的物质及空间，不会与提取仪的任何部件有接触，避免了样品污染提取仪及实验环境。提取核酸后的废弃固体和液体，都密封在试剂盒中，废弃物不污染实验环境，对试剂盒处理也十分方便。
附图说明
图1是多列工作腔列并列的试剂盒示意图；
图2是单列工作腔列的试剂盒示意图(储运时)；
图3是试剂盒在加样时示意图；
图4是试剂盒在工作时的示意图；
图5是试剂盒取样时示意图；
图6是试剂盒侧视图；
图7是试剂盒剖视图；
图8是试剂盒在工作时原理示意图；
图9是试剂盒的活塞、丝杆等示意图；
图10是试剂盒在工作时原理示意图；
图11是恒温加热器原理示意图；
图12是5个空气通道互不相通状态的示意图；
图13是5个空气通道连通状态的示意图。
图中，裂解腔a，吸洗腔b，第一洗涤液腔c，第二洗涤液腔d，洗脱液腔e，第一工作腔列1，第二工作腔列2，第三工作腔列3，第四工作腔列4，第五工作腔列5，第六工作腔列6，第七工作腔列7，第八工作腔列8，盒体10，加样盖101，取样盖102，左翻折处104，右翻折处108，倒角109，加样盖护翼132，取样盖护翼136，连通阀20，阀腔21，连通阀阀芯22，导通工位23，隔断工位24，拨杆26；裂解液阀门31，吸附腔32，第一洗涤液阀门33，第二洗涤液阀门34，洗脱液阀门35，阀芯36，弹簧37，裂解液管路41，吸附柱42，第一洗涤液管路43，第二洗涤液管路44，洗脱液管路45，裂解液阀门驱动电磁铁51，第一洗涤液阀门驱动电磁铁53，第二洗涤液阀门驱动电磁铁54，洗脱液阀门驱动电磁铁55，电热膜61，双金属温控器62，加热供电电路正极触点63，加热供电电路负极触点64，磁力搅拌驱动器70，搅拌电机71，原动磁铁72，水平轴73，磁力搅拌子74，活塞80，丝杆81，驱动轮82，动力装置85，裂解腔空气通道91，吸洗腔空气通道92，第一洗涤液腔空气通道93，第二洗涤液腔空气通道94，洗脱液腔空气通道95。
具体实施方式
全封闭全自动核酸提取试剂盒，不论是多列工作腔列并列的试剂盒，还是单列工作腔列的试剂盒，基本都成长方体形。对于多列工作腔列并列的试剂盒参见图1，在X方向，并列8列工作腔列(样品组)，分别是第一工作腔列1、第二工作腔列2、第三工作腔列3、第四工作腔列4、第五工作腔列5、第六工作腔列6、第七工作腔列7、第八工作腔列8，从左到右，按编号排列，可以同步处理8个样品。对于单列工作腔列的试剂盒，参见图2-6。
参见图7，在试剂盒的Y方向(宽度方向)，每列工作腔列中从左到右，排列5个工作腔，依次为裂解腔a、吸洗腔b、第一洗涤液腔c、第二洗涤液腔d、洗脱液腔e。不同工作腔列中的裂解腔a、吸洗腔b、第一洗涤液腔c、第二洗涤液腔d、洗脱液腔e，分别在X方向排成一行，从左到右，依次是裂解腔行、吸洗腔行、第一洗涤液腔行、第二洗涤液腔行、洗脱液腔行。
参见图1、6，盒体的前侧外壁上具有与外部电源相连的加热供电电路正极触点63，后侧外壁上具有加热供电电路负极触点64。
参见图9-11，同一列工作腔列内的裂解腔a与吸洗腔b之间的腔壁内预埋电热膜61，该试剂盒内的8个电热膜并联后与双金属温控器62串联，再通过加热供电电路正极触点63及负极触点64与提取仪相连，由提取仪控制驱动，在提取相关步骤，对裂解腔a与吸洗腔b内液体进行恒温加热。当然，8个电热膜61也可以是一个完整的在裂解腔行与吸洗腔行之间的腔壁内贯穿的电热膜。
试剂盒主要包括盒体10、加样盖101、取样盖102。
加样盖、取样盖的结构基本相同。参见图7，加样盖101沿左翻折处与盒体10成翻折连接(一种活动连接方式)，加样盖下部具有伸入裂解腔上口与裂解腔上口密封接触的加样盖护翼132。同样，取样盖102沿右翻折处108与盒体成翻折连接(一种活动连接方式)，取样盖下部具有伸入洗脱液腔上口与洗脱液腔上口密封接触的取样盖护翼136。
参见图7、12、13，盒体上设置连通阀20，连通阀包括位于盒体内上部的阀腔21、在阀腔内滑动的连通阀阀芯22。裂解腔上部、吸洗腔上部、第一洗涤液腔上部、第二洗涤液腔上部、洗脱液腔上部分别通过裂解腔空气通道91、吸洗腔空气通道92、第一洗涤液腔空气通道93、第二洗涤液腔空气通道94、洗脱液腔空气通道95等5个空气通道与连通阀的阀腔21相通。
5个空气通道与阀腔相通的端口位于与连通阀阀芯下表面接触的阀腔下内壁上，连通阀阀芯22在阀腔21内有两个不同工位，即导通工位23和隔断工位24；连通阀阀芯22在隔断工位24时，连通阀阀芯下表面封堵5个空气通道端口，5个空气通道互不相通，连通阀处于隔断状态；连通阀阀芯22在导通工位23时，连 通阀阀芯下表面与5个空气通道端口不接触，5个空气通道通过阀腔相通，连通阀处于导通状态。每个阀腔的前后侧面上均开有供拨杆滑动的拨杆槽，拨杆26同时与所有的连通阀阀芯连接，并穿过拨杆槽伸出盒体。拨动拨杆26能够带动所有的连通阀阀芯22从隔断工位移动到导通工位，使得各连通阀从从隔断状态变换到导通状态。拨杆26即是能够拨动阀腔内的连通阀阀芯移动的驱动机构。
参见图7、8、10、12，吸洗腔的底部与吸附腔32相通，吸附腔内具有胶接一起的核酸吸附材料形成的吸附柱42，核酸吸附材料之间具有流体可以穿过的空隙；裂解腔底部、第一洗涤液腔底部、第二洗涤液腔底部、洗脱液腔底部分别通过串联有裂解液阀门31的裂解液管路41、串联有第一洗涤液阀门33的第一洗涤液管路43、串联有第二洗涤液阀门34的第二洗涤液管路44、串联有洗脱液阀门35的洗脱液管路45与吸附腔32相通。
裂解液阀门31、第一洗涤液阀门33、第二洗涤液阀门34、洗脱液阀门35结构相同，均包括磁性材料阀芯36、弹簧37，阀芯36上端是锥面，弹簧在常态时使得阀芯与阀芯所在管路的锥形内壁密封接触，该管路处于常闭断流状态。
在试剂盒外部(例如提取仪上)设置有阀门驱动器列，阀门驱动器列由依次排列的裂解液阀门驱动电磁铁51、第一洗涤液阀门驱动电磁铁53、第二洗涤液阀门驱动电磁铁54、洗脱液阀门驱动电磁铁55组成。
当试剂盒插入提取仪内后，每一个阀门下面有控制该阀门开关的对应的阀门驱动器，也就是说，裂解液阀门31、第一洗涤液阀门33、第二洗涤液阀门34、洗脱液阀门35的下方分别有相对应的裂解液阀门驱动电磁铁51、第一洗涤液阀门驱动电磁铁53、第二洗涤液阀门驱动电磁铁54、洗脱液阀门驱动电磁铁55。
裂解液阀门驱动电磁铁51得电时，电磁铁吸附裂解液阀门31中的阀芯36克服弹簧37的弹力向下移动，阀芯上端与裂解液管路内壁不再密封接触，裂解液管路处于导通状态。当裂解液阀门驱动电磁铁51断电时，阀芯36在弹簧37的弹力向上移动，阀芯上端与裂解液管路内壁又密封接触，裂解液管路处于关闭状态。
参见图8、10，裂解腔a内磁力搅拌子转动设置水平轴73上。磁力搅拌子74可以是内置磁钢的塑料搅拌叶片等现有的磁力搅拌子。在试剂盒外部的例如安装在提取仪上的磁力搅拌驱动器70包括搅拌电机71、设置在搅拌电机输出轴上的原动磁铁72。当试剂盒插入提取仪后，搅拌电机71与水平轴73同轴，搅拌电机71工作，原动磁铁72转动，通过磁场耦合，带动裂解腔a内磁力搅拌子74转动。
参见图9、10，吸洗腔内设置沿吸洗腔内壁上下滑动密封接触的活塞80。丝杆81与活塞80螺纹配合，伸入到吸洗腔内，与吸洗腔底部和上端转动配合。穿过吸洗腔上端的丝杆81上端固定与丝杆同轴的作为动力输入端的驱动轮82，驱动轮82作为与外部的动力装置85相连的动力输入端露出盒体。活塞与吸洗腔内壁之间设置防止活塞相对于吸洗腔内壁转动的结构，使得活塞只能相对于吸洗腔内壁上下滑动(如吸洗腔内壁上具有平行于丝杆的导向槽，活塞上具有伸入导向槽内的导向棱等等)。在试剂盒外部的例如安装在提取仪上的动力装置85，能够与驱动轮82连接，带动驱动轮82绕丝杆81轴线顺时针或逆时针转动，使得活塞80上下移动。
提取核酸时，将加样盖101以左翻折处104为枢轴翻开，用加样枪将样品加入试剂盒的裂解腔a，然后再盖紧加样盖101，加样盖环周护翼133与裂解腔腔壁内侧密封接触。
将加样后的试剂盒放入提取仪，启动提取程序，进行全自动核酸提取。
提取完成后，提取仪弹出试剂盒，以右翻折处108为枢轴翻开取样盖102，用加样枪抽取试剂盒里的核酸提取产物，用于后继的核酸测试实验。
下面描述一下各工作腔的用途。
裂解腔a：预存裂解液；内置磁力搅拌子；上部有可开启/关闭的加样盖，用于加注样品；下方布局裂解液阀门，布局在试剂盒内的裂解液管路将裂解腔a、裂解液阀门与吸洗腔b下方的吸附腔串联起来；裂解腔上方通过在盒体内布设的裂解腔空气通道与试剂盒顶部的连通阀的阀腔连通。裂解腔a与吸洗腔b的间隔腔壁中埋藏电热膜及控制恒温的双金属温控器串联构成的恒温加热器，恒温加热器通过埋藏在试剂盒内的线路与试剂盒前后侧下方的正负极触点连接，从提取仪上获取供电并自动恒温加热。
吸洗腔b：内置移液驱动系统：由活塞、丝杆和驱动轮构成。驱动轮与提取仪的动力装置耦合，驱动活塞沿吸洗腔上下移动；底部布局有吸附腔(内容核酸吸附材料组成的吸附柱)，吸附腔底部直接或经过内部管路分别与裂解液管路、第一洗涤液管路、第二洗涤液管路、洗脱液管路连通。吸洗腔b上方通过在盒体内布设的吸洗腔空气通道与连通阀的阀腔连通。
第一洗涤液腔c：预存第一洗涤液；底部布局有第一洗涤液阀门，第一洗涤液腔c通过第一洗涤液阀门 及第一洗涤液管路与吸洗腔b下方的吸附腔串联连通；第一洗涤液腔c上方通过在盒体内布设的第一洗涤液腔空气通道与连通阀的阀腔连通。
第二洗涤液腔d：预存第二洗涤液；底部布局有第二洗涤液阀门，第二洗涤液腔d通过第二洗涤液阀门及第二洗涤液管路与吸洗腔b下方的吸附腔串联连通；第二洗涤液腔d上方通过在盒体内布设的第二洗涤液腔空气通道与连通阀的阀腔连通。
洗脱液腔e：预存洗脱液；顶部有取样盖，可翻开/关闭，用于在提取操作完成后抽取提取产物；底部布局有洗脱液阀门，洗脱液腔e通过洗脱液阀门及洗脱液管路与吸洗腔b下方的吸附腔串联连通。洗脱液腔e上方通过在盒体内布设的洗脱液腔空气通道与连通阀的阀腔连通。用于存储提取产物溶液。
试剂盒外观基本为矩形体，顶部有驱动轮、可翻盖打开/关闭的加样盖和取样盖，侧面有与内置恒温加热器电连接的+/-电极触点，底部单侧面有防反插倒角。提取仪经过机械耦合驱动试剂盒的驱动轮，带动丝杆上的活塞上下往复移动驱动试剂盒内部的液体移动，完成核酸提取操作。试剂盒内部的底层，布局有多个阀门，由提取仪上的阀门驱动器程控驱动，完成提取过程中移液流向的控制。提取仪的电源电路经过试剂盒的电极触点，驱动试剂盒内部的恒温加热器实现提取过程中的液体加热需要。试剂盒内部的磁力搅拌子，由提取仪内的磁力搅拌驱动器驱动，完成提取过程中的液体搅拌工作。提取仪运行由程序控制，全自动完成，程序和参数，可以通过手机APP设置，提取操作一键完成。
试剂盒可以是只针对单个样品的单列工作腔列的结构，也可以是多列工作腔列并列的结构，以实现多样品独立并行提取操作。
每列工作腔列，由裂解腔a、吸洗腔b、第一洗涤液腔c、第二洗涤液腔d、洗脱液腔e五个独立工作腔腔室构成，是提取操作过程的反应腔室。除吸洗腔b之外，其他腔室出厂时预装试剂。裂解腔a预装裂解液，第一洗涤液腔c预装第一洗涤液，第二洗涤液腔d预装第二洗涤液，洗脱液腔d预装洗脱液。
连通阀的阀腔，位于试剂盒顶部，内部有连通阀阀芯，连通阀阀芯在储运状态处于隔断工位，保证各个腔室的内容试剂在储运过程中不会互相串扰。在试剂盒工作状态连通阀处于导通状态，使裂解腔a、吸洗腔b、第一洗涤液腔c、第二洗涤液腔d、洗脱液腔e五个腔室的顶部空气连通，保证提取操作过程中内部压力平衡。
控制部分位于试剂盒的底部，由位于裂解腔a、第一洗涤液腔c、第二洗涤液腔d、洗脱液腔e下部的裂解液阀门31、第一洗涤液阀门33、第二洗涤液阀门34、洗脱液阀门35以及相关的管路构成，用于控制提取过程中移液的流向。吸附腔位于吸洗腔b下方，内容吸附柱，其他腔室通过阀门、管路与吸附腔及吸洗腔b串联；加样盖位于裂解腔a的上方，可开启/关闭，用于提取核酸之前的样品加入操作。取样盖位于洗脱液腔d的上方，可开启/关闭，用于提取核酸之后的样品取出操作。
核酸提取工作流程：
试剂盒开封：打开试剂盒包装。
翻开加样盖，将待提取核酸的样品，用移液枪加入裂解腔a，盖好加样盖。
将加样后、盖好盖的试剂盒插入提取仪。插入操作时，提取仪上的挡块等结构，会借助插入时的推力，阻挡拨杆随试剂盒一起相对于提取仪移动，从而将试剂盒上部各连通阀的连通阀阀芯从隔断状态变换到导通状态，这时，每列工作腔列中，与5个工作腔上部相通的5个空气通道处于连通状态，保证提取操作过程中各腔室的压力平衡。吸洗腔b上部的驱动轮与提取仪上的动力装置耦合，使得提取仪可以驱动吸洗腔b内部的活塞沿吸洗腔b上下移动。试剂盒上+/-电极触点与提取仪接通，使提取仪可以给试剂盒中的恒温加热器供电。提取仪上的各阀门驱动器(电磁阀)对准试剂盒中对应的阀门，使得提取仪可以依据提取程序控制各个阀门动作。提取仪上的每个磁力搅拌驱动器对准一列工作腔列中的裂解腔a内的磁力搅拌子，使得提取仪可以驱动试剂盒中的磁力搅拌子搅拌液体。试剂盒一侧下部的单边倒角109，可以防止试剂盒插入提取仪时方向插反。试剂盒插入提取仪后，顶部被提取仪上的压板等机构压住，保证提取过程中，加样盖和取样盖不会弹开。
加样后的试剂盒插入提取仪后，提取仪会依据预设的程序，全自动完成如下核酸提取操作步骤。
裂解：提取仪通过电极触点给裂解腔a与吸洗腔b之间的腔壁内埋藏的恒温加热器供电，给裂解腔a内的样品和裂解液混合液加热，同时提取仪中的磁力搅拌驱动器驱动裂解腔a中的磁力搅拌子充分搅拌裂解腔a中的混合液，促使裂解液与样品充分反应，完成细胞裂解、释放核酸的步骤。
吸附：位于裂解腔a下部的裂解液阀门下方的、提取仪上的裂解液阀门驱动电磁铁通电，驱动裂解液阀门导通，连通裂解腔a与吸洗腔b之间的管路，此时提取仪上的动力装置驱动丝杆，带动活塞上移，将裂解 腔a中的裂解混合液经过吸附腔内的吸附柱吸入吸洗腔b，混合液流经吸附柱的时候，液体中的核酸会被吸附柱吸附。提取仪驱动吸洗腔b内部的活塞上下往复移动数次，驱动液体在裂解腔a、吸洗腔b之间往复流经吸附柱，以提高核酸的吸附效率，最后驱动活塞下移将吸洗腔b中的液体全部排入裂解腔a存储，然后裂解液阀门驱动电磁铁断电，截止裂解液阀门，截断连通裂解腔a、吸洗腔b的管路，完成吸附步骤。
第一次洗涤：完成吸附流程后，位于试剂盒第一洗涤液腔c下部的第一洗涤液阀门33下方的、提取仪上的第一洗涤液阀门驱动电磁铁通电，驱动第一洗涤液阀门导通，连通第一洗涤液腔c和吸洗腔b之间的管路，提取仪上的动力装置驱动吸洗腔b中的活塞上移，将第一洗涤液腔c中预存的第一洗涤液，经过吸附柱吸入吸洗腔b。第一洗涤液流经吸附柱时，会将吸附柱上吸附的部分非核酸物质洗下来。动力装置会驱动吸洗腔b里的活塞往复数次，驱动第一洗涤液往返于吸洗腔b、第一洗涤液腔c之间多次流经吸附柱，以提高洗涤效率。最后驱动活塞下移将吸洗腔b中的液体排入第一洗涤液腔c存储，第一洗涤液阀门驱动电磁铁断电，释放第一洗涤液阀门33的阀芯，截断连通吸洗腔b、第一洗涤液腔c的管路，完成第一次洗涤步骤。
第二次洗涤：完成第一次洗涤流程后，位于试剂盒第二洗涤液腔d下部的第二洗涤液腔阀门下方的、提取仪上的第二洗涤液阀门驱动电磁铁通电，驱动第二洗涤液阀门导通，连通第二洗涤液腔d和吸洗腔b之间的管路，提取仪上的动力装置驱动吸洗腔b中的活塞上移，将第二洗涤液腔d中预存的第二洗涤液，经过吸附柱吸入吸洗腔b。第二洗涤液流经吸附柱时，会将吸附柱上吸附的剩余非核酸物质洗下来。动力装置会驱动吸洗腔b里的活塞往复数次，驱动第二洗涤液多次流经吸附柱，以提高洗涤效率。最后驱动活塞下移将吸洗腔b中的液体排入第二洗涤液腔d存储，第二洗涤液阀门驱动电磁铁断电，释放第二洗涤液阀门中的阀芯，截断连通吸洗腔b、第二洗涤液腔d的管路，完成第二次洗涤步骤。
洗脱：完成第二次洗涤流程后，位于试剂盒洗脱液腔d下部的洗脱液阀门下方的、提取仪上的洗脱液阀门驱动电磁铁通电，驱动洗脱液阀门导通，连通洗脱液腔d和吸洗腔b之间的管路，提取仪上的驱动装置驱动吸洗腔b中的活塞上移，将洗脱液腔d中预存的洗脱液，经过吸附柱抽入吸洗腔b。洗脱液流经吸附柱时，会将吸附柱上吸附的核酸物质洗脱下来。动力装置会驱动吸洗腔b里的活塞往复数次，驱动洗脱液往返吸洗腔b、洗脱液腔d之间多次流经吸附柱，以提高洗脱效率。最后驱动活塞下移将吸洗腔b中溶有洗脱下来的核酸的洗脱液排入洗脱液腔d存储，洗脱液阀门驱动电磁铁断电，释放洗脱液阀门中的阀芯，截断连通吸洗腔b、洗脱液腔d的管路，完成洗脱步骤。
弹出试剂盒：提取流程完成之后，提取仪会将试剂盒弹出。
取样：提取仪完成提取流程弹出试剂盒后，可以打开取样盖，用加样枪从洗脱液腔d抽取提取的核酸产物，用于后继的核酸检测操作。
废弃：取样之后，关闭取样盖，提取过程的废弃物全部封闭在试剂盒中，废弃的试剂盒不会出现泄漏污染环境的危险。
每列工作腔列的工作过程相同，显然，本发明中描述的工作过程的内容一般是对一列工作腔列中的各工作腔而言的。

Claims (17)

  1. 一种全封闭全自动核酸提取试剂盒，包括盒体，所述盒体具有多个工作腔，即装有裂解液的裂解腔、吸洗腔、装有洗涤液的洗涤液腔、装有洗脱液的洗脱液腔，其特征是：
    吸洗腔的底部与吸附腔相通，吸附腔内具有核酸吸附材料；裂解腔底部、洗涤液腔底部、洗脱液腔底部分别通过串联有裂解液阀门的裂解液管路、串联有洗涤液阀门的洗涤液管路、串联有洗脱液阀门的洗脱液管路与吸附腔相通；裂解液阀门、洗涤液阀门、洗脱液阀门均为非接触控制开关的阀门；
    裂解腔内设置有能够在磁力搅拌驱动器的带动下转动的磁力搅拌子；
    吸洗腔内设置沿吸洗腔内壁上下滑动密封接触的活塞，活塞与带动活塞上下移动的传动机构相连，该传动机构的动力输入端设置在盒体外部；
    盒体具有用于盖住或打开裂解腔上口的加样盖，以及用于盖住或打开洗脱液腔上口的取样盖。
  2. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：洗涤液腔包括装有第一洗涤液的第一洗涤液腔、装有第二洗涤液的第二洗涤液腔，洗涤液阀门包括第一洗涤液阀门、第二洗涤液阀门，第一洗涤液腔底部通过串联有第一洗涤液阀门的第一洗涤液管路与吸附腔相通，第二洗涤液腔底部通过串联有第二洗涤液阀门的第二洗涤液管路与吸附腔相通。
  3. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：盒体上设置连通阀；连通阀包括阀腔和连通阀阀芯；裂解腔上部、吸洗腔上部、洗涤液腔上部、洗脱液腔上部分别通过裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道与连通阀的阀腔相通；连通阀有两种工作状态，即导通状态和隔断状态，连通阀在导通状态使得该四个空气通道相通，在隔断状态时使得该四个空气通道互不相通；能够从试剂盒外部对连通阀进行操作或者通过非接触控制方式控制连通阀，使得连通阀从隔断状态变换到导通状态。
  4. 如权利要求3所述的全封闭全自动核酸提取试剂盒，其特征是：盒体内上部具有阀腔，四个空气通道与阀腔相通的端口位于阀腔内壁上，连通阀阀芯在阀腔内有两个不同工位，即导通工位和隔断工位；连通阀阀芯在隔断工位时，连通阀阀芯表面封堵四个空气通道端口，四个空气通道互不相通；连通阀阀芯在导通工位时，连通阀阀芯表面与四个空气通道端口不接触，四个工作腔上部通过四个相通的空气通道相通；连通阀阀芯上连接伸出盒体外部的驱动机构，通过该驱动机构能够拨动阀腔内的连通阀阀芯从隔断工位移动到导通工位。
  5. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：阀门包括磁性材料阀芯、弹簧，弹簧在常态时使得阀芯与阀芯所在管路的内壁密封接触，该管路处于常闭断流状态；在试剂盒外部、用于控制一个阀门、与该阀门位置相对应的电磁铁得电时，能够对该阀门的阀芯产生作用力，使得阀芯移动，阀芯与该管路内壁不再密封接触，管路处于导通状态。
  6. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：传动机构包括与活塞螺纹配合、伸入到吸洗腔内的丝杆，穿过吸洗腔上端的丝杆上端固定与丝杆同轴的作为动力输入端的驱动轮，活塞与吸洗腔内壁之间设置防止活塞相对于吸洗腔内壁转动的结构，使得活塞只能相对于吸洗腔内壁上下滑动。
  7. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：多个工作腔，即裂解腔、吸洗腔、洗涤液腔、洗脱液腔在同一直线上排列，裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门在同一直线上排列。
  8. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：盒体的一个棱边具有倒角，该具有倒角的棱边与多个工作腔排列的直线方向平行。
  9. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：在同一直线上按顺序依次排列的多个工作腔即裂解腔、吸洗腔、洗涤液腔、洗脱液腔称为工作腔列，工作腔列有并列排列的多列，在同一直线上按顺序依次排列的裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门称为阀门列，阀门列有并列排列的多列；每列工作腔列的下部是与该工作腔列对应的一阀门列；不同工作腔列中的裂解腔、吸洗腔、洗涤液腔、洗脱液腔分别排成一行，不同阀门列中的裂解液阀门、吸附腔、洗涤液阀门、洗脱液阀门分别排成一行。
  10. 如权利要求9所述的全封闭全自动核酸提取试剂盒，其特征是：盒体的外壁上具有与外部电源相连的加热供电电路正负极触点；在一列工作腔列中的裂解腔与吸洗腔之间的腔壁内设置电加热装置，电加热装置在通电时可以对裂解腔和吸洗腔内的液体加热，不同工作腔列内的电加热装置均与加热供电电路正负极触点电连接。
  11. 如权利要求10所述的全封闭全自动核酸提取试剂盒，其特征是：在排成一行的裂解腔行与排成一行 的吸洗腔行之间的腔壁内设置一张完整的电热膜和双金属温控器，该电热膜即是对各裂解腔和各吸洗腔内的液体进行恒温加热的电加热装置。
  12. 如权利要求9所述的全封闭全自动核酸提取试剂盒，其特征是：工作腔列有2列、4列或8列；不论工作腔列有2列、4列还是8列，试剂盒在一列工作腔列中多个工作腔的排列方向上宽度相等，试剂盒在各工作腔列排列方向上的长度相等。
  13. 如权利要求12所述的全封闭全自动核酸提取试剂盒，其特征是：有8列工作腔列的试剂盒，各工作腔的容积均为u；有4列工作腔列的试剂盒，各工作腔的容积均为v；有2列工作腔列的试剂盒，各工作腔的容积均为w；w＝2v＝4u；不论工作腔列有2列、4列还是8列，试剂盒高度相等。
  14. 如权利要求9所述的全封闭全自动核酸提取试剂盒，其特征是：试剂盒有8列工作腔列，各工作腔的容积均为u；通过改变盒体的高度，改变各工作腔容积的大小。
  15. 如权利要求9所述的全封闭全自动核酸提取试剂盒，其特征是：它还包括多个连通阀，每个连通阀与一列工作腔列对应；连通阀包括阀腔和连通阀阀芯；各阀腔均在盒体内上部；一列工作腔列中的裂解腔上部、吸洗腔上部、洗涤液腔上部、洗脱液腔上部分别通过裂解腔空气通道、吸洗腔空气通道、洗涤液腔空气通道、洗脱液腔空气通道与该工作腔列中的连通阀的阀腔相通，该四个空气通道与阀腔相通的端口位于阀腔内壁上；连通阀阀芯在阀腔内有两个不同工位，即导通工位和隔断工位；连通阀阀芯在隔断工位时，该连通阀阀芯表面封堵四个空气通道端口，四个空气通道互不相通；连通阀阀芯在导通工位时，连通阀阀芯表面与四个空气通道端口不接触，四个工作腔上部通过四个相通的空气通道相通；一拨杆同时与各连通阀阀芯连接，并穿过阀腔上所开的拨杆槽伸出盒体；能够从试剂盒外部拨动拨杆，使得各连通阀的连通阀阀芯同时从隔断工位移动到导通工位，连通阀从隔断状态变换到导通状态。
  16. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：裂解腔的腔壁内设置第一电加热装置，第一电加热装置在通电时可以对裂解腔内液体加热，吸洗腔的腔壁内设置第二电加热装置，第二电加热装置在通电时可以对吸洗腔内液体加热。
  17. 如权利要求1所述的全封闭全自动核酸提取试剂盒，其特征是：盒体成长方体形，其一个棱边具有防止试剂盒插入提取仪时方向插反的倒角。
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