CN114317255B - Totally-enclosed full-automatic nucleic acid extraction kit - Google Patents

Abstract

The invention provides a totally-enclosed full-automatic nucleic acid extraction kit which does not generate cross contamination in the nucleic acid extraction process, does not contact and pollute the extractor when a sample is not contacted with the extractor, seals waste solid and liquid after nucleic acid extraction in the kit, does not pollute the experimental environment and is convenient to treat. The kit comprises a kit body with a sample adding cover and a sampling cover, wherein the kit body is provided with a cracking cavity, a suction washing cavity, a washing liquid cavity and an eluent cavity, the bottom of the suction washing cavity is communicated with an adsorption cavity, and a nucleic acid adsorption material is arranged in the adsorption cavity; the bottoms of the cracking cavity, the washing liquid cavity and the elution liquid cavity are respectively communicated with the adsorption cavity through a cracking liquid pipeline in series connection with a cracking liquid valve, a washing liquid pipeline in series connection with a washing liquid valve and an elution liquid pipeline in series connection with an elution liquid valve; the cracking liquid valve, the washing liquid valve and the eluent valve are all valves of non-contact control switches; a magnetic stirrer is arranged in the cracking cavity; the suction washing cavity is internally provided with a piston which slides up and down along the inner wall of the suction washing cavity in a sealing way.

Description

Totally-enclosed full-automatic nucleic acid extraction kit

Technical Field

The present technology relates to nucleic acid extraction kits.

Background

Traditional magnetic bead method nucleic acid extraction kit, the lysate, the washing liquid, the eluant, the branch is in the functional cavity in different reagent boxes, and the extraction in-process uses the mechanical mechanism drive bar magnet that can remove in X, Y, the three direction of Z, will adsorb the magnetic bead of nucleic acid, removes between each functional cavity, and bar magnet cover still compromise the function of stirring, realize steps such as the schizolysis of extraction process, absorption, washing, elution, and the shortcoming of this kind of method is:

in order to realize that the magnetic rod enters and exits different functional chambers in different steps, each functional chamber must be in an open state in the extraction process, and cross contamination between parallel samples and environmental pollution of a laboratory are easily caused by dropping of sample liquid, splashing liquid drops and aerosol generated by heating and stirring in the extraction process in the processes of magnetic rod adsorption and magnetic bead movement, so that the accuracy of the experiment is seriously disturbed.

Meanwhile, the X, Y and Z three-axis driving devices of the magnetic rods and the magnetic rod sleeves have complex mechanisms, more moving parts, high manufacturing cost of instruments, large volume and more fault points.

Furthermore, the waste after nucleic acid extraction is left in an open kit, which is difficult to handle and is not easy to handle, thus causing environmental pollution.

Disclosure of Invention

The invention aims to provide a totally-enclosed full-automatic nucleic acid extraction kit which has the advantages of no cross contamination in the nucleic acid extraction process, simple structure, small volume, no contact between a sample and an extraction instrument, no pollution to the experimental environment because waste solid and liquid after nucleic acid extraction are sealed in the kit, and convenient treatment.

The invention discloses a totally-enclosed full-automatic nucleic acid extraction kit, which comprises a kit body, wherein the kit body is provided with a plurality of working cavities, namely a cracking cavity filled with cracking liquid, an absorption and washing cavity, a washing liquid cavity filled with washing liquid, an elution liquid cavity filled with elution liquid,

the bottom of the suction washing cavity is communicated with the adsorption cavity, and a nucleic acid adsorption material is arranged in the adsorption cavity; the bottom of the cracking cavity, the bottom of the washing liquid cavity and the bottom of the elution liquid cavity are respectively communicated with the adsorption cavity through a cracking liquid pipeline in series connection with a cracking liquid valve, a washing liquid pipeline in series connection with a washing liquid valve and an elution liquid pipeline in series connection with an elution liquid valve; the cracking liquid valve, the washing liquid valve and the eluent valve are all valves of non-contact control switches;

a magnetic stirrer capable of rotating under the drive of a magnetic stirring driver is arranged in the cracking cavity;

a piston which slides up and down along the inner wall of the suction washing cavity to be in sealing contact with the suction washing cavity is arranged in the suction washing cavity, the piston is connected with a transmission mechanism which 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 is provided with a sample adding cover for covering or opening the upper opening of the lysis cavity and a sampling cover for covering or opening the upper opening of the eluent cavity.

The totally-enclosed full-automatic nucleic acid extraction kit comprises a washing liquid cavity, a first washing liquid cavity and a second washing liquid cavity, wherein the first washing liquid cavity is filled with a first washing liquid, the second washing liquid cavity is filled with a second washing liquid, the washing liquid valve comprises a first washing liquid valve and a second washing liquid valve, the bottom of the first washing liquid cavity is communicated with an adsorption cavity through a first washing liquid pipeline in series connection with the first washing liquid valve, and the bottom of the second washing liquid cavity is communicated with the adsorption cavity through a second washing liquid pipeline in series connection with the second washing liquid valve.

In the totally-enclosed full-automatic nucleic acid extraction kit, the box body is provided with the communicating valve; the communicating valve comprises a valve cavity and a communicating valve core; the upper part of the cracking cavity, the upper part of the suction washing cavity, the upper part of the washing liquid cavity and the upper part of the elution liquid cavity are respectively communicated with the valve cavity of the communicating valve through a cracking cavity air passage, a suction washing cavity air passage, a washing liquid cavity air passage and an elution liquid cavity air passage; the communicating valve has two working states, namely a conducting state and a blocking state, the communicating valve enables the four air channels to be communicated in the conducting state, and the four air channels are not communicated with each other in the blocking state; the communication valve can be operated from the outside of the reagent cartridge or controlled in a non-contact control manner so that the communication valve is switched from a blocked state to a conducting state.

The totally-enclosed full-automatic nucleic acid extraction kit is characterized in that a valve cavity is formed in the upper part in the box body, ports of the four air channels communicated with the valve cavity are positioned on the inner wall of the valve cavity, and a valve core of the communicating valve is provided with two different stations, namely a communicating station and a separating station, in the valve cavity; when the communicating valve core is at the partition station, the surface of the communicating valve core plugs four air channel ports, and the four air channels are not communicated with each other; when the communicating valve core is in a conducting station, the surface of the communicating valve core is not contacted with the ports of the four air channels, and the upper parts of the four working cavities are communicated through the four air channels; the communicating valve core is connected with a driving mechanism extending out of the box body, and the communicating valve core in the valve cavity can be stirred to move from the partition station to the conduction station through the driving mechanism.

The valve comprises a magnetic material valve core and a spring, the spring makes the valve core contact with the inner wall of the pipeline where the valve core is positioned in a sealing way in a normal state, and the pipeline is in a normally closed cutoff state; when the electromagnet corresponding to the position of the valve, which is used for controlling the valve outside the kit, is electrified, acting force can be generated on the valve core of the valve, so that the valve core moves, the valve core is not in sealing contact with the inner wall of the pipeline, and the pipeline is in a conducting state.

The transmission mechanism comprises a screw rod which is in threaded fit with the piston and extends into the suction and washing cavity, a driving wheel which is coaxial with the screw rod and serves as a power input end penetrates through the upper end of the screw rod at the upper end of the suction and washing cavity, and a structure which is used for preventing the piston from rotating relative to the inner wall of the suction and washing cavity is arranged between the piston and the inner wall of the suction and washing cavity, so that the piston can only slide up and down relative to the inner wall of the suction and washing cavity.

The totally-enclosed full-automatic nucleic acid extraction kit comprises a plurality of working cavities, namely a cracking cavity, an adsorption cavity, a washing liquid cavity and an eluent cavity which are arranged on the same straight line, and a cracking liquid valve, an adsorption cavity, a washing liquid valve and an eluent valve are arranged on the same straight line.

In the totally-enclosed full-automatic nucleic acid extraction kit, one edge of the kit body is provided with the chamfer, and the edge is parallel to the linear direction in which the plurality of working cavities are arranged.

The totally-enclosed full-automatic nucleic acid extraction kit comprises a plurality of working chambers, namely a cracking chamber, an adsorption chamber, a washing liquid chamber and an elution liquid chamber which are sequentially arranged on the same straight line in sequence, wherein the working chambers are a working chamber row, a plurality of columns are arranged in parallel, a cracking liquid valve, an adsorption chamber, a washing liquid valve and an eluent valve which are sequentially arranged on the same straight line in sequence are a valve row, and the valve row is a plurality of columns which are arranged in parallel; the lower part of each row of working cavity rows is a valve row corresponding to the working cavity row; the cracking cavity, the absorbing and washing cavity, the washing liquid cavity and the eluent cavity in different working cavity rows are respectively arranged in a line, and the cracking liquid valve, the absorbing cavity, the washing liquid valve and the eluent valve in different valve rows are respectively arranged in a line.

In the totally-enclosed full-automatic nucleic acid extraction kit, the outer wall of the box body is provided with a positive electrode contact and a negative electrode contact of a heating power supply circuit connected with an external power supply; an electric heating device is arranged in the cavity wall between the cracking cavity and the suction washing cavity in one working cavity row, the electric heating device can heat liquid in the cracking cavity and the suction washing cavity when being electrified, and the electric heating devices in different working cavity rows are electrically connected with the anode contact and the cathode contact of the heating power supply circuit.

The totally-enclosed full-automatic nucleic acid extraction kit is characterized in that a complete electrothermal film and a bimetallic temperature controller are arranged in the cavity wall between the cracking cavity rows which are arranged in a line and the absorbing and washing cavity rows which are arranged in a line, and the electrothermal film is an electric heating device for heating the liquid in each cracking cavity and each absorbing and washing cavity at constant temperature.

The totally-enclosed full-automatic nucleic acid extraction kit has 2, 4 or 8 working cavity columns; no matter the working cavity rows have 2 rows, 4 rows or 8 rows, the reagent box has the same width in the arrangement direction of the working cavities in one working cavity row, and has the same length in the arrangement direction of the working cavity rows.

The totally-enclosed full-automatic nucleic acid extraction kit comprises 8 rows of kits, and the volume of each working cavity is u; the kit is provided with 4 working cavity rows, and the volume of each working cavity is v; the kit comprises 2 working cavity rows, wherein the volume of each working cavity is w; w =2v =4u; the reagent box height is equal no matter the working chamber is arranged in 2 rows, 4 rows or 8 rows.

The totally-enclosed full-automatic nucleic acid extraction kit is provided with 8 working cavity rows, and the volume of each working cavity is u; the size of each working cavity volume is changed by changing the height of the box body.

The totally-enclosed full-automatic nucleic acid extraction kit also comprises a plurality of communicating valves, and each communicating valve corresponds to one row of working cavity; the communicating valve comprises a valve cavity and a communicating valve core; each valve cavity is arranged at the upper part in the box body; the upper parts of the cracking cavities, the upper parts of the washing liquid cavities and the upper parts of the eluent cavities in the working cavity row are respectively communicated with the valve cavities of the communicating valves in the working cavity row through cracking cavity air channels, washing liquid cavity air channels and eluent cavity air channels, and ports of the four air channels communicated with the valve cavities are positioned on the inner walls of the valve cavities; the valve core of the communicating valve is provided with two different stations, namely a communicating station and a separating station, in the valve cavity; when the valve core of the communicating valve is at a blocking station, the surface of the valve core of the communicating valve blocks four air channel ports, and the four air channels are not communicated with each other; when the communicating valve core is in a conducting station, the surface of the communicating valve core is not contacted with the ports of the four air channels, and the upper parts of the four working cavities are communicated through the four air channels; a deflector rod is simultaneously connected with the valve cores of the communicating valves and passes through a deflector rod groove formed on the valve cavity to extend out of the box body; the shifting lever can be shifted from the outside of the kit, so that the valve cores of the communicating valves move to the communicating station from the separating station simultaneously, and the communicating valves are switched to the communicating state from the separating state.

Foretell totally closed full-automatic nucleic acid extraction kit sets up first electric heater unit in the chamber wall in schizolysis chamber, and first electric heater unit can be to the heating of schizolysis intracavity liquid when the circular telegram, inhales to wash the intracavity set up second electric heater unit in the chamber wall in chamber, and second electric heater unit can be to inhaling when circular telegram and wash intracavity liquid heating.

The totally-enclosed full-automatic nucleic acid extraction kit has a cuboid box body, and one edge of the box body is provided with a chamfer for preventing the kit from being inserted reversely when the kit is inserted into the extraction instrument.

The method comprises the following steps of using the totally-enclosed full-automatic nucleic acid extraction kit to extract nucleic acid:

s10, cracking: and opening the sample adding cover, adding the sample into the cracking cavity pre-filled with the lysate, and closing the sample adding cover. First electric heater unit circular telegram carries out constant temperature heating to the intracavity liquid that splits, adopts the magnetic stirring driver that is located the kit outside portion to drive the magnetic stirrers rotation of the intracavity that splits simultaneously, and the magnetic stirrers stirs the mixed liquid of sample and lysate, and cell nucleus or virus schizolysis in the sample release nucleic acid.

S30, adsorption: a valve driver (called as a lysate valve driver VaD) which is positioned at the outer bottom of the kit, is opposite to the lysate valve Va and controls the opening and closing of the lysate valve in a non-contact mode to drive the lysate valve Va to be opened, then a power device positioned outside the kit drives a power input end, a piston is driven to move upwards through a transmission mechanism, and a lysate solution dissolved with nucleic acid is sucked into a suction washing cavity from a lysis cavity through the lysate valve Va and an adsorption cavity;

when the lysis solution containing the nucleic acid is passed through the adsorption chamber, the nucleic acid in the solution is adsorbed by the nucleic acid adsorbing material. The power device drives the piston to move downwards through the transmission mechanism, and the lysis solution dissolved with the nucleic acid is pumped into the lysis cavity from the adsorption cavity through the lysis solution valve Va; and finally, the piston is driven to move downwards to discharge all the liquid in the absorption and washing cavity into the cracking cavity for storage, then a lysate valve driver VaD controls a lysate valve Va to be closed, a lysate pipeline is closed, and the adsorption step is completed.

S50, washing: a valve driver (called as a washing liquid valve driver) which is positioned at the outer bottom of the kit, is opposite to the washing liquid valve and controls the washing liquid valve to be opened and closed in a non-contact mode drives the washing liquid valve to be opened, then, a power device drives a piston to move upwards through a transmission mechanism so as to suck the washing liquid into a suction washing cavity from a washing liquid cavity through the washing liquid valve and an adsorption cavity;

when the washing solution flows through the adsorption chamber, the non-nucleic acid substances adsorbed on the nucleic acid adsorbing material are washed off. The piston in the suction washing cavity is driven to reciprocate up and down for a plurality of times, and the washing liquid flows through the adsorption cavity for a plurality of times between the suction washing cavity and the washing liquid cavity to improve the washing efficiency. And finally, driving the piston to move downwards to discharge the liquid in the suction washing cavity into the washing liquid cavity for storage, and controlling the washing liquid valve to be closed by the washing liquid valve driver, so that the washing step is completed.

S70, elution: a valve driver (called as an eluent valve driver VeD) which is positioned at the outer bottom of the kit, is opposite to the eluent valve Ve and controls the opening and closing of the eluent valve Ve in a non-contact mode drives the eluent valve Ve to open, then, a power device drives a piston to move upwards through a transmission mechanism, and eluent is absorbed into an absorption and elution cavity from an eluent cavity through the eluent valve Ve and an adsorption cavity;

when the eluent flows through the adsorption cavity, the nucleic acid substances adsorbed on the nucleic acid adsorption material are eluted. The piston in the suction washing cavity is driven to reciprocate up and down for a plurality of times, and the eluent flows through the adsorption cavity for a plurality of times between the suction washing cavity and the eluent cavity to improve the elution efficiency. And finally, driving the piston to move downwards to discharge the liquid in the absorption and washing cavity into an eluent cavity for storage, and controlling the closing of an eluent valve Ve and the closing of an eluent pipeline by an eluent valve driver VeD to finish the elution step.

S90, sampling: the sampling cover is opened, liquid in the eluent cavity is absorbed by a drawing device such as a pipette gun and the like, and then the sampling cover is closed.

Of course, if the washing liquid chamber includes the first washing liquid chamber c and the second washing liquid chamber d, the step S50 in the aforementioned extraction process is replaced with the steps S51 and S61.

S51, first washing: a valve driver (called as a first washing liquid valve driver VcD) which is positioned at the outer bottom of the kit, is opposite to the first washing liquid valve Vc, controls the switch of the first washing liquid valve Vc in a non-contact mode, drives the first washing liquid valve Vc to be opened, then, a power device drives a piston to move upwards through a transmission mechanism, and first washing liquid is sucked into a suction washing cavity from a first washing liquid cavity through the first washing liquid valve Vc and an adsorption cavity;

when the first washing solution flows through the adsorption chamber, the non-nucleic acid substances adsorbed on the nucleic acid adsorbing material are washed off. The piston in the suction washing cavity is driven to reciprocate up and down for a plurality of times, and the first washing liquid flows through the adsorption cavity for a plurality of times between the suction washing cavity and the washing liquid cavity to improve the washing efficiency. And finally, driving the piston to move downwards to discharge the liquid in the suction washing cavity into the first washing liquid cavity for storage, and then controlling the first washing liquid valve Vc to be closed by the first washing liquid valve driver VcD, closing the first washing liquid pipeline, and finishing the first washing step.

S61, washing for the second time: a valve driver (called as a second washing liquid valve driver Vdd) which is positioned at the outer bottom of the kit, is opposite to the second washing liquid valve Vd and controls the opening and closing of the second washing liquid valve Vd in a non-contact mode drives the second washing liquid valve Vd to be opened, then, a power device drives a piston to move upwards through a transmission mechanism, and second washing liquid is sucked into a suction washing cavity from a second washing liquid cavity through the second washing liquid valve Vd and an adsorption cavity;

when the second washing solution flows through the adsorption chamber, the non-nucleic acid substances adsorbed on the nucleic acid adsorbing material are washed off. The piston in the suction washing cavity is driven to reciprocate up and down for a plurality of times, and the second washing liquid flows through the adsorption cavity for a plurality of times between the suction washing cavity and the washing liquid cavity to improve the washing efficiency. And finally, driving the piston to move downwards to discharge the liquid in the suction washing cavity into a second washing liquid cavity for storage, and then controlling a second washing liquid valve Vd to be closed by a second washing liquid valve driver Vdd and closing a second washing liquid pipeline to finish a second washing step.

The magnetic stirring driver and the magnetic stirrer driven by the magnetic stirring driver belong to the prior art, the principle of the magnetic stirring driver is similar to that of the magnetic stirring driver, namely the magnetic stirring driver pushes the magnetic stirrer placed in the kit to perform circumferential operation in a non-contact way by utilizing the principle that like poles repel and opposite poles attract of a magnetic field, so that the aim of stirring liquid is fulfilled.

The magnetic stirrer driver, the valve driver, the power unit, and the control mechanism for controlling the communication valve to switch the state (or the operation mechanism for operating the communication valve to switch the state) are not attached to the present kit, but they are external devices used for driving the magnetic stirrer, the valve, the driving wheel, and the communication valve in the present kit only in the process of extracting nucleic acid using the present kit. In order to use this kit to extract nucleic acid more convenient, the magnetic stirring driver, the valve driver (including schizolysis liquid valve driver, washing liquid valve driver (including first washing liquid valve driver, second washing liquid valve driver), eluent valve driver), power device, the striking gear of the intercommunication valve case removal of striking intercommunication valve can be installed on the corresponding position of drawing the appearance, that is to say, when this kit location is on drawing the appearance, the magnetic stirring driver is located the magnetic stirring son pivoted position that can drive the schizolysis intracavity, the schizolysis liquid valve driver is located the position that can drive the schizolysis liquid valve and carry out the switch action, washing liquid valve driver is located the position that can drive the washing liquid valve and carry out the switch action, eluent valve driver is located the position that can drive the eluent valve and carry out the switch action, power device is located the position that can carry out mechanical connection with the drive wheel. In the process that the kit is inserted into the extraction instrument, the valve core of the communicating valve can be moved to the communicating station from the partition station through a shifting device and the like arranged on the extraction instrument, and the communicating valve is switched to the communicating state from the partition state.

The sample adding cover, the sample taking cover and the box body can be detachably connected, for example, the sample adding cover, the sample taking cover and the box body are mutually independent parts which can be mutually separated.

The application of sample lid can be a body structure with the box body, and at this moment, in order to open the application of sample of application of sample lid, application of sample lid can cover schizolysis chamber upper shed (with schizolysis chamber upper shed sealing connection) after the application of sample, and application of sample lid becomes swing joint with the main part lid, turns over the connection for example.

The sample lid can be a body structure with the box body, and at this moment, in order to open the sample of sample lid, the sample lid becomes swing joint, for example turns over to be connected with the main part lid.

In the process of extracting nucleic acid by using the kit, in order to ensure that each working chamber is kept in air communication so as to ensure that the internal air pressure of each working chamber is balanced during pipetting, pipetting resistance is not formed, and liquid in the working chambers is ensured to avoid splashing of the liquid caused by stirring to pollute the adjacent working chambers, the invention is provided with an air channel communication control structure comprising a communication valve and the like. When the communicating valve is in a cut-off state, air channels such as a cracking cavity air channel, a suction washing cavity air channel, a washing liquid cavity air channel, an eluent cavity air channel and the like are not communicated with each other, and when the communicating valve is in a conduction state, the air channels are communicated with each other. Before the reagent kit is used, such as in the processes of product delivery, transportation and storage, the communication valve is in a blocking state, so that substances pre-filled in each working cavity in the reagent kit cannot move to other working cavities, and the contents in each working cavity cannot be leaked or mixed with each other. That is, the kit contents do not leak out, or contaminate one another. When the kit is used for nucleic acid extraction, after the sample adding cover is opened for sample adding, before subsequent extraction operation, the communicating valve needs to be switched to a conducting state from a partition state, and at the moment, air channels such as a cracking cavity air channel, a suction and washing cavity air channel, a washing liquid cavity air channel, an elution liquid cavity air channel and the like are communicated with each other, so that resistance to the flow of liquid cannot be caused due to different pressures in each working cavity when the piston moves up and down.

In order to facilitate switching of the state of the communication valve, the communication valve may be a communication valve that is controlled in a non-contact manner, or a communication valve that can be operated from the outside of the reagent cartridge. By adopting the communicating valve controlled by non-contact, the kit can be inserted into the extractor and then switched to a conducting state by a non-contact control mode. The communication valve that can operate the communication valve from the kit outside is adopted, needs before the kit inserts the extraction appearance, switches the communication valve to the conducting state through manual mode, perhaps inserts the in-process of extraction appearance at the kit, through the toggle device that sets up on the extraction appearance etc. with the communication valve switch to the conducting state.

In order to operate the communicating valve from the outside of the kit conveniently, the communicating valve core is connected with a driving mechanism extending out of the box body, and the communicating valve core in the valve cavity can be stirred to move from the partition station to the conduction station through the driving mechanism.

The valve has various types, and the valve with simple structure, lower cost and convenient control comprises a magnetic material valve core and a spring, wherein the spring enables the valve core to be in sealing contact with the inner wall of a pipeline where the valve core is positioned in a normal state, and the pipeline is in a normally closed cutoff state. The valve drive associated with the valve can then be a conventional electromagnet which is easy to use. For example, an electromagnet corresponding to the valve and installed on the extraction instrument is electrified, a valve core for adsorbing the valve moves against the elastic force of a spring, the valve core is not in sealing contact with the inner wall of the pipeline any more, and the pipeline where the valve is located is in a conducting state. When the electromagnet is powered off, the valve core is reset under the action of the spring and is in sealing contact with the inner wall of the pipeline where the valve core is located, and the pipeline is in a closed state.

The axis of the valve core is vertical (the valve core moves up and down along the axis of the valve core), and the rotation axis of the magnetic stirrer in the cracking cavity a is horizontal. Therefore, drive crosstalk can be prevented, and the mutual interference of the magnetic fields of the valve driver and the magnetic stirring driver can be prevented.

The power input end of the transmission mechanism is arranged outside the box body, so that the transmission mechanism is connected with an external power device, for example, the power device is arranged on the extraction instrument, the transmission mechanism is a screw rod and nut mechanism, and a driving wheel fixed with a screw rod is arranged outside the box body (such as the top of the box body), so that the transmission mechanism is conveniently matched and meshed with the power device, and power transmission is realized.

The plurality of working cavities, namely the cracking cavity, the adsorption cavity, the washing liquid cavity and the eluent cavity are arranged on the same straight line, and the cracking liquid valve, the adsorption cavity, the washing liquid valve and the eluent valve are arranged on the same straight line, so that the arrangement is more convenient. The straight line of the plurality of working cavities can be parallel to the straight line of the cracking liquid valve, the adsorption cavity, the washing liquid valve and the like, and also can be nonparallel.

Set up the positive negative pole contact of heating power supply circuit on the outer wall of box body, conveniently be connected with the outside power supply circuit of kit, for example the power supply circuit electricity that sets up on drawing the appearance is connected.

The kit can adopt a single-row layout structure with only one working chamber row to process one sample.

In order to extract nucleic acid from multiple samples simultaneously, the kit adopts an array structure with multi-column layout, namely, multiple columns of working cavity columns are arranged in parallel, and the valve columns also have the same column number as the working cavity columns. Of course, the column of valve actuators at the outer bottom of the cartridge (e.g., mounted on the extractor) also has the same number of columns as the valve columns. The valve drivers of the lysis solution valve driver, the washing solution valve driver and the eluent valve driver which are arranged on the same straight line are valve driver columns. Specifically, when the array type kit is used for extracting nucleic acid, the lower part of each valve row is a valve driver row corresponding to the valve row; the lysate valve driver, the washing liquid valve driver and the eluent valve driver in each row of valve driver row can drive the lysate valve, the washing liquid valve and the eluent valve above the corresponding row to be switched. That is, under each valve is a valve actuator that actuates the valve to open and close.

In the nucleic acid extraction operation process, each valve in one row is in an open state at the same time, and other valves are in closed states, so that liquid can only move in the corresponding working chamber and the related pipeline according to the step requirements in the specified pipeline and the working chamber, and the crosstalk of the liquid can not occur.

The number of magnetic stirring actuators on the outside of the cartridge (e.g., mounted on the extractor) is also the same as the number of columns in the array of working chambers. Each magnetic stirring driver is positioned on a working cavity array (the magnetic stirring driver, the cracking cavity, the suction washing cavity, the washing liquid cavity and the elution liquid cavity are sequentially arranged on the same straight line) and is close to the cracking cavity. Each magnetic stirring driver can drive the magnetic stirrers in the cracking cavity which are positioned on the same straight line with the magnetic stirring drivers to rotate.

For example, the working chamber column and the valve column have 2 columns, 4 columns or 8 columns (correspondingly, the valve driver column has 2 columns, 4 columns or 8 columns, and the magnetic stirring driver has 2, 4 or 8), so that the nucleic acid extraction can be simultaneously carried out on 2, 4 or 8 samples in parallel. No matter the working chambers are arranged in 2 columns, 4 columns or 8 columns, the reagent boxes are equal in width, length and height, so that the extraction instruments with the same specification can be shared for improving the universality.

For reagent boxes with 2 columns, 4 columns or 8 columns of working cavity columns and valve columns, in order to change the capacity of the working cavity, the height of the box body can be increased without changing the length and the width of the box body, so that the extraction instrument can be suitable for reagent boxes with different capacities as long as a height regulator is matched with the space heights of the reagent boxes with different capacities.

Some nucleic acid species are extracted without heating in the whole process, and at the moment, the kit does not need to be provided with an electric heating device. However, in the extraction process of most nucleic acids, heating is needed in the processes of cracking, adsorption and elution, at the moment, a first electric heating device is arranged in the cavity wall of the cracking cavity, the first electric heating device can heat liquid in the cracking cavity when being electrified, and a second electric heating device is arranged in the cavity wall of the adsorption cavity, and the second electric heating device can heat liquid in the adsorption cavity when being electrified. The first electric heating device (such as an electric heating film) and the second electric heating device (such as an electric heating film) which are embedded in the reagent box are adopted to realize the liquid heating in the extraction process, and a temperature control circuit belonging to the prior art can be attached to the electric heating devices to realize the constant temperature control in the liquid heating process. Of course, if the cracking chamber and the suction washing chamber are adjacent, only one electric heating device can be arranged in the chamber wall between the cracking chamber and the suction washing chamber, and the electric heating device can heat the liquid in the cracking chamber and the suction washing chamber simultaneously when being electrified, so that the structure can be further simplified. The outer wall of the box body is provided with a heating power supply circuit anode contact and a heating power supply circuit cathode contact which are connected with an external power supply, and the electric heating device is electrically connected with the heating power supply circuit anode contact and the heating power supply circuit cathode contact. In the use of the kit, whether or not it is heated, and at that point, is controlled by an external power source, e.g., an external power source located on the extractor.

The box body is cuboid, and one edge of the box body is provided with a chamfer which prevents the kit from being inserted reversely when being inserted into the extraction instrument, so that the use is more convenient.

The kit can be operated manually and can be operated fully automatically. Can be combined with a real-time fluorescent PCR module to form an extraction and real-time fluorescent PCR test integrated kit. Can be arranged in a single-row/multi-row parallel way, and a sample can be processed by the single-row arrangement

The multi-column layout can process a plurality of samples in parallel and synchronously without mutual interference among the samples. The extraction process is totally closed, and the contents in the kit do not contact with the experimental environment and any part of the extractor in the extraction process. The nucleic acid adsorption material belongs to the existing material, and is intensively placed in the adsorption cavity. The nucleic acid adsorption material such as magnetic beads can also be bonded into an adsorption column which is arranged in the adsorption cavity, and the nano-scale loose magnetic beads are bonded into the adsorption column, so that the contradiction between the difficulty in controlling the discrete magnetic beads and the large adsorption area is solved.

The kit is disposable, and the kit body is made of plastic materials.

The kit can be matched with a corresponding extractor to work, and complete the full-automatic and full-closed extraction operation of nucleic acid.

The kit of the invention has a working principle different from that of the traditional nucleic acid extraction kit, and the nucleic acid adsorbing material for adsorbing nucleic acid is always retained in the adsorbing cavity in the whole extraction process. The liquid is transferred between the working chambers by the up-and-down movement of the piston, and the liquid passes through the adsorption chamber during the transfer, so that the adsorption and elution of the nucleic acid are realized.

Because the traditional magnetic bar for transferring magnetic beads and needing to continuously enter and exit each working cavity is omitted, the whole extraction process is carried out, and the reagent kit and the experimental environment are in a closed isolation state among all samples of the reagent kit, so that cross contamination and laboratory pollution can not occur.

The liquid stirring is realized by a built-in magnetic stirrer.

The extraction instrument and the kit are driven in a magnetic isolation mode, and substances and space in the kit cannot contact with any part of the extraction instrument in the extraction process, so that the sample is prevented from polluting the extraction instrument and the experimental environment.

The waste solid and liquid after nucleic acid extraction are sealed in the kit, so that the waste does not pollute the experimental environment and is convenient to treat.

Drawings

FIG. 1 is a schematic diagram of a kit with multiple rows of working chambers side-by-side;

FIG. 2 is a schematic view of a single row of working chamber rows (during storage and transport);

FIG. 3 is a schematic view of the kit as it is loaded;

FIG. 4 is a schematic view of the cartridge in operation;

FIG. 5 is a schematic view of a cartridge when it is sampled;

FIG. 6 is a side view of the kit;

FIG. 7 is a sectional view of the kit;

FIG. 8 is a schematic view of the cartridge in operation;

FIG. 9 is a schematic view of a piston, screw, etc. of the cartridge;

FIG. 10 is a schematic view of the cartridge in operation;

FIG. 11 is a schematic view of a thermostatic heater;

FIG. 12 is a schematic view showing a state where 5 air passages are not communicated with each other;

fig. 13 is a schematic view showing a state where 5 air passages communicate.

In the figure, a lysis chamber a, a suction washing chamber b, a first washing liquid chamber c, a second washing liquid chamber d, an elution liquid chamber e,

a first working cavity row 1, a second working cavity row 2, a third working cavity row 3, a fourth working cavity row 4, a fifth working cavity row 5, a sixth working cavity row 6, a seventh working cavity row 7, an eighth working cavity row 8,

a box body 10, a sample adding cover 101, a sampling cover 102, a front folding part 104, a back folding part 108, a chamfer 109, a sample adding cover wing 132, a sampling cover wing 136,

the device comprises a communication valve 20, a valve cavity 21, a communication valve core 22, a conducting station 23, a blocking station 24 and a deflector rod 26;

a lysis solution valve 31, an adsorption cavity 32, a first washing solution valve 33, a second washing solution valve 34, an eluent valve 35, a valve core 36, a spring 37,

an adsorption column 42, a lysate line 41, a first washing liquid line 43, a second washing liquid line 44, an eluent line 45,

a lysate valve driving electromagnet 51, a first wash solution valve driving electromagnet 53, a second wash solution valve driving electromagnet 54, an eluent valve driving electromagnet 55,

an electric heating film 61, a bimetal temperature controller 62, a heating power supply circuit anode contact 63, a heating power supply circuit cathode contact 64,

a magnetic stirring driver 70, a stirring motor 71, a motive magnet 72, a horizontal shaft 73, a magnetic stirrer 74,

a piston 80, a screw 81, a driving wheel 82, a power device 85,

a lysis chamber air channel 91, an aspiration and wash chamber air channel 92, a first wash solution chamber air channel 93, a second wash solution chamber air channel 94, and an eluent chamber air channel 95.

Detailed Description

The totally-enclosed full-automatic nucleic acid extraction kit is basically cuboid, regardless of whether the kit is provided with a plurality of parallel working cavities or a single working cavity. For the reagent kit with multiple parallel working chambers, referring to fig. 1, in the X direction, 8 working chamber columns (sample groups) are parallel, namely a first working chamber column 1, a second working chamber column 2, a third working chamber column 3, a fourth working chamber column 4, a fifth working chamber column 5, a sixth working chamber column 6, a seventh working chamber column 7 and an eighth working chamber column 8, and from left to right, the 8 samples can be synchronously processed by numbering. See fig. 2-6 for a single working chamber row kit.

Referring to fig. 7, in the Y direction (width direction) of the kit, 5 working chambers are arranged in each working chamber row from left to right, and are a lysis chamber a, an imbibing chamber b, a first washing solution chamber c, a second washing solution chamber d, and an eluent chamber e in sequence. The cracking cavity a, the suction washing cavity b, the first washing liquid cavity c, the second washing liquid cavity d and the elution liquid cavity e in different working cavity rows are respectively arranged in a line in the X direction, and from left to right, the cracking cavity line, the suction washing cavity line, the first washing liquid cavity line, the second washing liquid cavity line and the elution liquid cavity line are sequentially arranged.

Referring to fig. 1 and 6, the front outer wall of the box body is provided with a heating and power supply circuit positive contact 63 connected with an external power supply, and the rear outer wall is provided with a heating and power supply circuit negative contact 64.

Referring to fig. 9-11, an electric heating film 61 is pre-embedded in the cavity wall between the cracking cavity a and the absorbing and washing cavity b in the same working cavity row, 8 electric heating films in the reagent box are connected in parallel and then connected in series with a bimetallic temperature controller 62, and then connected with an extraction instrument through a heating power supply circuit anode contact 63 and a cathode contact 64, and driven by the control of the extraction instrument, and liquid in the cracking cavity a and the absorbing and washing cavity b is heated at constant temperature in relevant extraction steps. Of course, the 8 electrothermal films 61 may also be a complete electrothermal film penetrating through the cavity wall between the cracking cavity row and the suction and washing cavity row.

The kit mainly comprises a box body 10, a sample adding cover 101 and a sampling cover 102.

The structures of the sample adding cover and the sample sampling cover are basically the same. Referring to fig. 7, the sample application cover 101 is connected with the case 10 along the front folding part in a folding manner (a movable connection manner), and the lower part of the sample application cover is provided with a sample application cover wing 132 extending into the upper opening of the lysis chamber and contacting with the upper opening of the lysis chamber in a sealing manner. Similarly, the sampling cap 102 is connected to the case body along the rear folded part 108 in a folding manner (a movable connection manner), and the lower part of the sampling cap has a sampling cap wing 136 extending into the upper opening of the eluent chamber and contacting with the upper opening of the eluent chamber in a sealing manner.

Referring to fig. 7, 12 and 13, the box body is provided with a communicating valve 20, and the communicating valve comprises a valve cavity 21 positioned at the upper part in the box body and a communicating valve core 22 sliding in the valve cavity. The upper part of the cracking cavity, the upper part of the suction washing cavity, the upper part of the first washing liquid cavity, the upper part of the second washing liquid cavity and the upper part of the eluent cavity are respectively communicated with the valve cavity 21 of the communicating valve through 5 air passages, such as an air passage 91 of the cracking cavity, an air passage 92 of the suction washing cavity, an air passage 93 of the first washing liquid cavity, an air passage 94 of the second washing liquid cavity, an air passage 95 of the eluent cavity and the like.

The ports of the 5 air passages communicated with the valve cavity are positioned on the lower inner wall of the valve cavity contacted with the lower surface of the valve core of the communicating valve, and the valve core 22 of the communicating valve is provided with two different stations in the valve cavity 21, namely a communicating station 23 and a separating station 24; when the communicating valve core 22 is at the blocking station 24, the lower surface of the communicating valve core blocks 5 air channel ports, the 5 air channels are not communicated with each other, and the communicating valve is in a blocking state; when the communicating valve spool 22 is in the communicating position 23, the lower surface of the communicating valve spool is not in contact with the 5 air channel ports, the 5 air channels are communicated through the valve cavity, and the communicating valve is in a communicating state. The front side and the rear side of each valve cavity are provided with a deflector rod groove for a deflector rod to slide, and the deflector rod 26 is connected with all the valve cores of the communicating valves at the same time and penetrates through the deflector rod grooves to extend out of the box body. The poking rod 26 can drive all the communicating valve cores 22 to move from the partition station to the conduction station, so that each communicating valve is changed from the partition state to the conduction state. The dial 26 is a driving mechanism capable of moving the communicating valve element in the dial valve chamber.

Referring to FIGS. 7, 8, 10 and 12, the bottom of the adsorption and washing chamber is communicated with the adsorption chamber 32, and the adsorption chamber is provided with an adsorption column 42 formed by nucleic acid adsorbing materials which are glued together, and a gap through which fluid can pass is arranged between the nucleic acid adsorbing materials; the bottom of the cracking cavity, the bottom of the first washing liquid cavity, the bottom of the second washing liquid cavity and the bottom of the elution liquid cavity are respectively communicated with the adsorption cavity 32 through a cracking liquid pipeline 41 connected with a cracking liquid valve 31 in series, a first washing liquid pipeline 43 connected with a first washing liquid valve 33 in series, a second washing liquid pipeline 44 connected with a second washing liquid valve 34 in series and an elution liquid pipeline 45 connected with an elution liquid valve 35 in series.

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 respectively comprise a magnetic material valve core 36 and a spring 37, the upper end of the valve core 36 is a conical surface, the spring enables the valve core to be in sealing contact with the conical inner wall of a pipeline where the valve core is located in a normal state, and the pipeline is in a normally closed cut-off state.

A valve driver column is arranged outside the kit (for example, on an extraction instrument), and consists of a lysate valve driving electromagnet 51, a first washing solution valve driving electromagnet 53, a second washing solution valve driving electromagnet 54 and an eluent valve driving electromagnet 55 which are sequentially arranged.

After the reagent kit is inserted into the extraction instrument, a corresponding valve driver for controlling the valve switch is arranged below each valve, that is, a lysate valve driving electromagnet 51, a first washing liquid valve driving electromagnet 53, a second washing liquid valve driving electromagnet 54 and an eluent valve driving electromagnet 55 are respectively arranged below the lysate valve 31, the first washing liquid valve 33, the second washing liquid valve 34 and the eluent valve 35.

When the lysate valve drives the electromagnet 51 to be electrified, the electromagnet adsorbs the valve core 36 in the lysate valve 31 to move downwards against the elastic force of the spring 37, the upper end of the valve core is not in sealing contact with the inner wall of the lysate pipeline any more, and the lysate pipeline is in a conducting state. When the cracking liquid valve drives the electromagnet 51 to be powered off, the valve core 36 moves upwards under the elastic force of the spring 37, the upper end of the valve core is in sealing contact with the inner wall of the cracking liquid pipeline, and the cracking liquid pipeline is in a closed state.

Referring to fig. 8 and 10, the magnetic stirrer inside the lysis chamber a is rotatably arranged on a horizontal shaft 73. The magnetic stirrer can be a plastic stirring blade with built-in magnetic steel, and the like. The magnetic stirring driver outside the cartridge, for example, mounted on the extractor, includes a stirring motor 71, a motive magnet 72 provided on the output shaft of the stirring motor. When the reagent box is inserted into the extraction instrument, the stirring motor 71 is coaxial with the horizontal shaft 73, the stirring motor 71 works, the motive magnet 72 rotates, and the magnetic stirrer in the cracking cavity a is driven to rotate through magnetic field coupling.

Referring to fig. 9 and 10, a piston 80 is disposed in the suction washing chamber to slide up and down along the inner wall of the suction washing chamber in a sealing manner. The screw rod 81 is in threaded fit with the piston 80, extends into the suction washing cavity, and is in running fit with the bottom and the upper end of the suction washing cavity. The upper end of a screw rod 81 which penetrates through the upper end of the suction washing cavity is fixed with a driving wheel 82 which is coaxial with the screw rod and is used as a power input end, and the driving wheel 82 is used as a power input end connected with an external power device 85 and is exposed out of the box body. The structure for preventing the piston from rotating relative to the inner wall of the suction washing cavity is arranged between the piston and the inner wall of the suction washing cavity, so that the piston can only slide up and down relative to the inner wall of the suction washing cavity (for example, a guide groove parallel to the screw rod is arranged on the inner wall of the suction washing cavity, and a guide edge extending into the guide groove is arranged on the piston). A power means 85, for example mounted on the extractor, external to the cartridge, can be connected to the drive wheel 82 to rotate the drive wheel 82 clockwise or anticlockwise about the axis of the screw 81, so that the piston 80 moves up and down.

When extracting nucleic acid, the sample-adding cover 101 is turned over with the front turning part 104 as a pivot, a sample is added into the cracking cavity a of the kit by a sample-adding gun, then the sample-adding cover 101 is tightly covered, and the circumferential protective wing 133 of the sample-adding cover is in sealing contact with the inner side of the wall of the cracking cavity.

And (3) putting the kit after sample addition into an extractor, starting an extraction program, and performing full-automatic nucleic acid extraction.

After extraction, the extraction instrument ejects the kit, the sampling cover 102 is opened by taking the turnover part 108 as a pivot, and the nucleic acid extraction product in the kit is extracted by using a sample adding gun for subsequent nucleic acid test experiments.

The purpose of each working chamber is described below.

Lysis chamber a:

pre-storing a lysate;

a magnetic stirrer is arranged in the device;

the upper part is provided with an openable/closable sample adding cover for adding a sample;

a cracking liquid valve is arranged below the reagent box, and a cracking liquid pipeline arranged in the reagent box connects the cracking cavity a, the cracking liquid valve and the adsorption cavity below the adsorption cavity b in series;

the upper part of the cracking cavity is communicated with the valve cavity of a communicating valve at the top of the kit through a cracking cavity air passage distributed in the box body.

The constant temperature heater is formed by embedding an electrothermal film and a bimetal temperature controller for controlling constant temperature in series in the wall of a separation cavity of the cracking cavity a and the absorbing and washing cavity b, the constant temperature heater is connected with positive and negative contacts below the front side and the rear side of the kit through a circuit embedded in the kit, and power is supplied from the extraction instrument and is heated at constant temperature automatically.

Suction and washing cavity b:

built-in pipetting drive system: consists of a piston, a screw rod and a driving wheel. The driving wheel is coupled with a power device of the extraction instrument and drives the piston to move up and down along the suction washing cavity;

the bottom is distributed with an adsorption cavity (containing an adsorption column composed of nucleic acid adsorption materials) which is respectively communicated with a lysate pipeline, a first washing liquid pipeline, a second washing liquid pipeline and an eluent pipeline through an internal pipeline.

The upper part of the suction and washing cavity b is communicated with the valve cavity of the communicating valve through a suction and washing cavity air passage arranged in the box body.

First washing liquid chamber c:

pre-storing a first washing solution;

a first washing liquid valve is distributed at the bottom, and a first washing liquid cavity c is communicated with the adsorption cavity below the adsorption cavity b in series through the first washing liquid valve and a first washing liquid pipeline;

the upper part of the first washing liquid cavity c is communicated with the valve cavity of the communicating valve through a first washing liquid cavity air passage distributed in the box body.

Second washing liquid chamber d:

pre-storing a second washing solution;

a second washing liquid valve is distributed at the bottom, and a second washing liquid cavity d is communicated with the adsorption cavity below the adsorption cavity b in series through the second washing liquid valve and a second washing liquid pipeline;

the upper part of the second washing liquid cavity d is communicated with the valve cavity of the communicating valve through a second washing liquid cavity air passage distributed in the box body.

Eluent chamber e:

pre-storing an eluent;

a sampling cover is arranged at the top and can be turned over/closed, and is used for extracting an extraction product after the extraction operation is finished;

the bottom is distributed with an eluent valve, and an eluent cavity e is communicated with the adsorption cavity below the suction and washing cavity b in series through the eluent valve and an eluent pipeline.

The upper part of the eluent cavity e is communicated with the valve cavity of the communicating valve through an eluent cavity air passage distributed in the box body.

For storing the extract product solution.

The appearance of the kit is basically a rectangular cube, the top of the kit is provided with a driving wheel, a sample adding cover and a sampling cover, the sample adding cover can be opened/closed by a turnover cover, the side of the kit is provided with +/-electrode contacts electrically connected with a built-in constant temperature heater, and the single side of the bottom of the kit is provided with an anti-reverse insertion chamfer.

The extractor drives a driving wheel of the kit through mechanical coupling to drive a piston on a screw rod to reciprocate up and down to drive liquid in the kit to move, so that the nucleic acid extraction operation is completed.

The bottom layer in the kit is provided with a plurality of valves in a layout manner, and the valves are driven by a valve driver on the extraction instrument in a program control manner to complete the control of the flow direction of the liquid-transfering in the extraction process.

A power circuit of the extraction instrument drives a constant temperature heater in the kit to meet the requirement of heating liquid in the extraction process through an electrode contact of the kit.

The magnetic stirrer in the kit is driven by a magnetic stirring driver in the extraction instrument to complete the liquid stirring work in the extraction process.

The operation of the extraction instrument is controlled by a program and is fully automatically completed, and the program and parameters can be set through a mobile phone APP, so that the extraction operation can be completed by one key.

The kit can be a structure of a single-row working cavity row only aiming at a single sample, and can also be a structure of a plurality of rows of working cavities in parallel, so that the independent and parallel extraction operation of a plurality of samples is realized.

Each row of working cavity is composed of five independent cavities, namely a cracking cavity a, a suction washing cavity b, a first washing liquid cavity c, a second washing liquid cavity d and an elution liquid cavity e, and is a reaction cavity in the extraction operation process. Except the adsorption and washing cavity b, other cavities are pre-filled with reagents when being delivered. And a lysis chamber a filled with lysis solution. And a first washing liquid chamber c pre-filled with a first washing liquid. And a second washing liquid chamber (d) filled with a second washing liquid. And an eluent chamber d chamber is pre-filled with eluent.

The valve cavity of the communicating valve is positioned at the top of the kit, the communicating valve core is arranged in the communicating valve, and the communicating valve core is positioned at a partition station in a storage and transportation state, so that the content reagents of each cavity can not be mutually interfered in the storage and transportation process. In kit operating condition, the intercommunication valve is in the conducting state, makes the top air intercommunication of five cavities of schizolysis chamber a, inhale and wash chamber b, first washing liquid chamber c, second washing liquid chamber d, eluant chamber e, guarantees to draw the operation in-process, and internal pressure is balanced.

The control part is positioned at the bottom of the kit and consists of a lysis solution valve 31, a first washing solution valve 33, a second washing solution valve 34, an eluent valve 35 and related pipelines which are positioned at the lower parts of the lysis chamber a, the first washing solution chamber c, the second washing solution chamber d and the eluent chamber e, and is used for controlling the flow direction of pipetting in the extraction process.

The adsorption cavity is positioned below the adsorption and washing cavity b, the adsorption column is arranged in the adsorption cavity, and other cavities are connected with the adsorption cavity and the adsorption and washing cavity b in series through a valve and a pipeline; and a sample adding cover which is positioned above the lysis cavity a, can be opened/closed and is used for sample adding operation before extracting nucleic acid. And a sampling cover which is positioned above the d chamber of the eluent chamber and can be opened/closed, and is used for sample taking operation after extracting nucleic acid.

Nucleic acid extraction workflow:

unsealing the kit: opening a kit package

And (3) turning over the sample adding cover, adding the sample of nucleic acid to be extracted into the cracking cavity a by using a pipette, and covering the sample adding cover.

And inserting the reagent kit which is subjected to sample addition and covered by the cover into an extraction instrument.

During the operation of inserting, draw structures such as dog on the appearance, can stop the driving lever and remove for drawing the appearance along with the kit with the help of the thrust when inserting to dial the intercommunication valve case of each communicating valve in kit upper portion from the state of cutting off to the on-state, at this moment, in every working chamber row, be in the on-state with the communicating 5 air passage in 5 working chamber upper portions, guarantee to draw the pressure balance of each cavity in the operation process.

The driving wheel on the upper part of the suction washing cavity b is coupled with the power device on the extraction instrument, so that the extraction instrument can drive the piston in the suction washing cavity b to move up and down along the suction washing cavity b.

The +/-electrode contact on the reagent box is communicated with the extractor, so that the extractor can supply power to the constant temperature heater in the reagent box.

And each valve driver (electromagnetic valve) on the extractor aligns to the corresponding valve in the kit, so that the extractor can control the action of each valve according to the extraction program.

The magnetic stirring driver on the extractor is aligned with the magnetic stirrer in the lysis chamber a, so that the extractor can drive the magnetic stirrer in the kit to stir the liquid.

Unilateral chamfer 109 of kit one side lower part can prevent that the direction from inserting when the kit inserts and draws the appearance and inserting instead.

After the kit is inserted into the extraction instrument, the top of the kit is pressed by a pressing plate of the extraction instrument, and the sampling cover cannot be bounced off in the extraction process.

After the reagent kit after the sample adding is inserted into the extractor, the extractor can fully automatically complete the following nucleic acid extraction operation steps according to a preset program:

cracking: the extraction instrument supplies power to a constant temperature heater buried in the cavity wall between the cracking cavity a and the suction washing cavity b through an electrode contact, the mixed liquid of the sample and the lysate in the cracking cavity a is heated, and meanwhile, a magnetic stirring driver in the extraction instrument drives a magnetic stirrer in the cracking cavity a to fully stir the mixed liquid in the cracking cavity a, so that the lysate and the sample are fully reacted, cell cracking is completed, and nucleic acid is released.

Adsorption: in extracting the appearance, the lysate valve drive electro-magnet below the lysate valve that is located schizolysis chamber a lower part is circular telegram, and the drive lysate valve switches on, and intercommunication schizolysis chamber a and inhale the pipeline between the chamber b, power device drive on the extraction appearance this moment inhale the lead screw that washes chamber b, drive the piston and shift up, inhale and wash chamber b with the schizolysis mixed liquid in the schizolysis chamber a through the adsorption column in the adsorption cavity, when mixed liquid flows through the adsorption column, nucleic acid in the liquid can be adsorbed by the adsorption column. The extractor drives the piston in the suction washing cavity b to reciprocate up and down for several times, the driving liquid flows through the adsorption column between the cracking cavity a and the suction washing cavity b in a reciprocating manner so as to improve the adsorption efficiency of nucleic acid, finally the driving piston discharges all the liquid in the suction washing cavity b into the cracking cavity a for storage, then the cracking liquid valve drives the electromagnet to be powered off, the cracking liquid valve is cut off, the pipeline communicating the cracking cavity a and the suction washing cavity b is cut off, and the adsorption step is completed.

Washing for the first time: after the adsorption process is completed, in the extraction instrument, the electromagnet is driven to be electrified by the first washing liquid valve below the first washing liquid valve 33 positioned at the lower part of the first washing liquid cavity c of the kit, the first washing liquid valve is driven to be conducted, the first washing liquid cavity c is communicated with a pipeline between the first washing liquid cavity c and the adsorption cavity b, the piston in the adsorption cavity b is driven to move upwards by the power device on the extraction instrument, the first washing liquid prestored in the first washing liquid cavity c is sucked into the adsorption cavity b through the adsorption column, and when the first washing liquid flows through the adsorption column, the non-nucleic acid substances adsorbed on the adsorption column can be washed down. The power device can drive the piston in the suction washing cavity b to reciprocate for a plurality of times, and drive the first washing liquid to reciprocate between the suction washing cavity b and the first washing liquid cavity c and flow through the adsorption column for a plurality of times so as to improve the washing efficiency. And finally, driving the piston to move downwards to discharge the liquid in the suction washing cavity b into the first washing liquid cavity c for storage, driving the electromagnet to be powered off by the first washing liquid valve, releasing the valve core of the first washing liquid valve 33, and cutting off a pipeline for communicating the suction washing cavity b with the first washing liquid cavity c to finish the first washing step.

And (3) second washing: after the first washing process is finished, in the extraction instrument, a second washing liquid valve below a second washing liquid cavity valve at the lower part of a second washing liquid cavity d of the kit is driven to be electrified to drive the second washing liquid valve to be conducted, the second washing liquid cavity d is communicated with a pipeline between the second washing liquid cavity d and the adsorption washing cavity b, a power device on the extraction instrument drives a piston in the adsorption washing cavity b, second washing liquid prestored in the second washing liquid cavity d is sucked into the adsorption washing cavity b through an adsorption column, and when the second washing liquid flows through the adsorption column, non-nucleic acid substances adsorbed on the adsorption column can be washed down. The power device can drive the piston in the suction washing cavity b to reciprocate for a plurality of times, and drive the second washing liquid to flow through the adsorption column for a plurality of times so as to improve the washing efficiency. And finally, driving the piston to discharge the liquid in the suction washing cavity b into a second washing liquid cavity d for storage, powering off the electromagnet driven by the second washing liquid valve, releasing the valve core in the second washing liquid valve, and cutting off a pipeline for communicating the suction washing cavity b with the second washing liquid cavity d to finish the second washing step.

And (3) elution: after the second washing process is finished, in the extraction instrument, an eluent valve driving electromagnet below an eluent valve positioned at the lower part of an eluent cavity d of the kit is electrified to drive the eluent valve to be conducted, a pipeline between the eluent cavity d and the adsorption and washing cavity b is communicated, a driving device on the extraction instrument drives a piston in the adsorption and washing cavity b, the eluent prestored in the eluent cavity d is pumped into the adsorption and washing cavity b through an adsorption column, and when the eluent flows through the adsorption column, the nucleic acid substances adsorbed on the adsorption column are eluted. The power device can drive the piston in the suction cavity b to reciprocate for a plurality of times, and drive the eluent to flow through the adsorption column for a plurality of times between the suction cavity b and the eluent cavity d, so as to improve the elution efficiency. Finally, the piston is driven to discharge the eluent dissolved with the eluted nucleic acid in the absorbing and washing cavity b into the eluent cavity d for storage, the electromagnet is driven by the eluent valve to be powered off, the valve core in the eluent valve is released, a pipeline communicating the absorbing and washing cavity b with the eluent cavity d is cut off, and the elution step is finished

Popping up a kit: after the extraction process is completed, the extractor will pop up the kit.

Sampling: after the extraction instrument is taken out and the reagent box is popped up after the extraction process is completed, the sampling cover can be opened, and the extracted nucleic acid product is extracted from the eluent cavity d by the sample adding gun and is used for subsequent nucleic acid detection operation.

Abandonment: after the sample, close the sample lid, will draw the whole closures of the discarded object of process in the kit, the danger of revealing the polluted environment can not appear in the discarded kit.

The working process is the same for each working chamber row, and it is clear that the content of the working process described in the present invention is generally for each working chamber in one working chamber row.

Claims (15)

1. The utility model provides a totally closed full-automatic nucleic acid extraction kit, includes the box body, the box body has a plurality of working chamber, is equipped with the schizolysis chamber of lysate promptly, inhales and washes the chamber, is equipped with the washing liquid chamber of washing liquid, is equipped with the eluant chamber of eluant, characterized by:

the bottom of the absorbing and washing cavity is communicated with the absorbing cavity, and a nucleic acid absorbing material is arranged in the absorbing cavity; the bottom of the cracking cavity, the bottom of the washing liquid cavity and the bottom of the elution liquid cavity are respectively communicated with the adsorption cavity through a cracking liquid pipeline in series connection with a cracking liquid valve, a washing liquid pipeline in series connection with a washing liquid valve and an elution liquid pipeline in series connection with an elution liquid valve; the cracking liquid valve, the washing liquid valve and the eluent valve are all valves of non-contact control switches;

a magnetic stirring bar which can rotate under the drive of a magnetic stirring driver is arranged in the cracking cavity;

a piston which slides up and down along the inner wall of the suction washing cavity and is in sealing contact with the suction washing cavity is arranged in the suction washing cavity, the piston is connected with a transmission mechanism which 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 is provided with a sample adding cover for covering or opening the upper opening of the lysis cavity and a sampling cover for covering or opening the upper opening of the eluent cavity;

the box body is provided with a communicating valve; the communicating valve comprises a valve cavity and a communicating valve core which are positioned at the upper part in the box body; the upper part of the cracking cavity, the upper part of the suction washing cavity, the upper part of the washing liquid cavity and the upper part of the elution liquid cavity are respectively communicated with the valve cavity of the communicating valve through a cracking cavity air passage, a suction washing cavity air passage, a washing liquid cavity air passage and an elution liquid cavity air passage; the ports of the four air channels communicated with the valve cavity are positioned on the inner wall of the valve cavity, and the communicating valve core is provided with two different stations, namely a communicating station and a separating station, in the valve cavity; when the valve core of the communicating valve is at the partition station, the surface of the valve core of the communicating valve blocks four air channel ports, the four air channels are not communicated with each other, and the communicating valve is in a partition state; when the valve core of the communicating valve is in a conducting state, the surface of the valve core of the communicating valve is not contacted with the ports of the four air channels, the upper parts of the four working cavities are communicated through the four air channels, and the communicating valve is in a conducting state; the communicating valve core is connected with a driving mechanism extending out of the box body, and the communicating valve core in the valve cavity can be stirred to move from the partition station to the conduction station through the driving mechanism, so that the communicating valve is changed from the partition state to the conduction state.

2. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, characterized in that: the washing liquid cavity comprises a first washing liquid cavity filled with first washing liquid and a second washing liquid cavity filled with second washing liquid, the washing liquid valve comprises a first washing liquid valve and a second washing liquid valve, the bottom of the first washing liquid cavity is communicated with the adsorption cavity through a first washing liquid pipeline connected with the first washing liquid valve in series, and the bottom of the second washing liquid cavity is communicated with the adsorption cavity through a second washing liquid pipeline connected with the second washing liquid valve in series.

3. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, which is characterized in that: the valve comprises a magnetic material valve core and a spring, wherein the spring enables the valve core to be in sealing contact with the inner wall of a pipeline where the valve core is located in a normal state, and the pipeline is in a normally closed cut-off state; when the electromagnet which is arranged outside the kit and is used for controlling one valve and corresponds to the position of the valve is electrified, acting force can be generated on the valve core of the valve, so that the valve core moves, the valve core is not in sealing contact with the inner wall of the pipeline, and the pipeline is in a conducting state.

4. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, which is characterized in that: the transmission mechanism comprises a screw rod which is in threaded fit with the piston and extends into the suction and washing cavity, a driving wheel which is coaxial with the screw rod and serves as a power input end is fixed at the upper end of the screw rod which penetrates through the upper end of the suction and washing cavity, and a structure which is prevented from rotating relative to the inner wall of the suction and washing cavity is arranged between the piston and the inner wall of the suction and washing cavity, so that the piston can only slide up and down relative to the inner wall of the suction and washing cavity.

5. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, which is characterized in that: the plurality of working cavities, namely the cracking cavity, the adsorption cavity, the washing liquid cavity and the eluent cavity, are arranged on the same straight line, and the cracking liquid valve, the adsorption cavity, the washing liquid valve and the eluent valve are arranged on the same straight line.

6. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, characterized in that: one edge of the box body is provided with a chamfer, and the edge is parallel to the linear direction of the arrangement of the working cavities.

7. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, characterized in that: the multiple working cavities, namely the cracking cavity, the adsorption cavity, the washing liquid cavity and the elution liquid cavity, which are sequentially arranged on the same straight line are called working cavity columns, the working cavity columns are provided with multiple columns which are arranged in parallel, the cracking liquid valve, the adsorption cavity, the washing liquid valve and the elution liquid valve which are sequentially arranged on the same straight line are called valve columns, and the valve columns are provided with multiple columns which are arranged in parallel; the lower part of each working cavity row is a valve row corresponding to the working cavity row; the cracking cavity, the absorbing and washing cavity, the washing liquid cavity and the eluent cavity in different working cavity rows are respectively arranged in a line, and the cracking liquid valve, the absorbing cavity, the washing liquid valve and the eluent valve in different valve rows are respectively arranged in a line.

8. The totally enclosed full-automatic nucleic acid extraction kit according to claim 7, characterized in that: the outer wall of the box body is provided with a positive contact and a negative contact of a heating power supply circuit connected with an external power supply; an electric heating device is arranged in the cavity wall between the cracking cavity and the suction washing cavity in one working cavity row, the electric heating device can heat liquid in the cracking cavity and the suction washing cavity when being electrified, and the electric heating devices in different working cavity rows are electrically connected with the anode contact and the cathode contact of the heating power supply circuit.

9. The totally enclosed full-automatic nucleic acid extraction kit according to claim 8, characterized in that: the wall between the splitting cavity row and the absorbing and washing cavity row is provided with a complete electric heating film and a bimetal temperature controller, and the electric heating film is an electric heating device for heating the liquid in the splitting cavities and the absorbing and washing cavities at constant temperature.

10. The totally enclosed full-automatic nucleic acid extraction kit according to claim 7, characterized in that: the working cavity rows are 2 rows, 4 rows or 8 rows; no matter the working cavity rows have 2 rows, 4 rows or 8 rows, the reagent boxes have equal width in the arrangement direction of a plurality of working cavities in one working cavity row, and the reagent boxes have equal length in the arrangement direction of each working cavity row.

11. The totally enclosed full-automatic nucleic acid extraction kit according to claim 10, characterized in that: the kit is provided with 8 working cavity rows, and the volume of each working cavity is u; the kit is provided with 4 working cavity rows, and the volume of each working cavity is v; the kit comprises 2 working cavity rows, wherein the volume of each working cavity is w; w =2v =4u; the reagent box height is equal no matter the working chamber is arranged in 2 rows, 4 rows or 8 rows.

12. The totally enclosed full-automatic nucleic acid extraction kit according to claim 7, characterized in that: the kit is provided with 8 working cavity rows, and the volume of each working cavity is u; the size of the volume of each working cavity is changed by changing the height of the box body.

13. The totally enclosed full-automatic nucleic acid extraction kit according to claim 7, characterized in that: the device also comprises a plurality of communicating valves, and each communicating valve corresponds to one working cavity row; the upper parts of the cracking cavities, the upper parts of the washing liquid cavities and the upper parts of the eluent cavities in the working cavity row are respectively communicated with the valve cavities of the communicating valves in the working cavity row through an air channel of the cracking cavity, an air channel of the washing liquid cavity and an air channel of the eluent cavity; a deflector rod is simultaneously connected with the valve cores of the communicating valves and passes through a deflector rod groove formed on the valve cavity to extend out of the box body; the shifting lever can be shifted from the outside of the kit, so that the valve cores of the communicating valves move to the conducting station from the partition station simultaneously, and the communicating valves are changed to the conducting state from the partition state.

14. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, which is characterized in that: set up first electric heater unit in the chamber wall in schizolysis chamber, first electric heater unit can be to the heating of schizolysis intracavity liquid when the circular telegram, inhales and washes the intracavity and set up second electric heater unit in the chamber wall in chamber, and second electric heater unit can be to the heating of inhaling the intracavity liquid when circular telegram.

15. The totally enclosed full-automatic nucleic acid extraction kit according to claim 1, characterized in that: the box body is cuboid, and one edge of the box body is provided with a chamfer which prevents the kit from being inserted reversely when the kit is inserted into the extraction instrument.

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