CN113308366A - Full-automatic nucleic acid extraction device and method - Google Patents
Full-automatic nucleic acid extraction device and method Download PDFInfo
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- CN113308366A CN113308366A CN202110731487.0A CN202110731487A CN113308366A CN 113308366 A CN113308366 A CN 113308366A CN 202110731487 A CN202110731487 A CN 202110731487A CN 113308366 A CN113308366 A CN 113308366A
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Abstract
A full-automatic nucleic acid extraction device comprises a suction head carrier, a sample carrier, a reagent carrier, a PCR refrigeration module and a multipurpose carrier, wherein an extraction module is arranged at the rear side of the PCR refrigeration module, the reagent carrier is a bulk extraction reagent carrier, the multipurpose carrier can be used as a deep-hole plate carrier or a pre-packaged extraction reagent carrier, a clamping mechanism and a sampling mechanism can carry out automatic clamping and sampling, the full-automatic continuous extraction work of nucleic acid is completed at the extraction module, and the extraction device also comprises a carrier in-place detection system. The full-automatic nucleic acid extraction device provided by the invention realizes continuous and automatic extraction in the whole process, can realize continuous and automatic extraction of multiple samples and continuous and automatic extraction of a small number of samples, and can effectively avoid interference between the sampling needle and the sample cup in the sampling process due to the fact that the carrier is not pushed in place.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a full-automatic nucleic acid extraction device and an extraction method thereof.
Background
The nucleic acid extractor is an instrument which uses a matched nucleic acid extraction reagent to complete the extraction work of sample nucleic acid. The method is widely applied to various fields of disease control center, clinical disease diagnosis, blood transfusion safety, forensic medicine identification, environmental microorganism detection, food safety detection, animal husbandry, molecular biology research and the like. At present, a magnetic bead method is widely adopted for extracting nucleic acid, namely, the magnetic bead is used as a carrier, the principle that the magnetic bead adsorbs nucleic acid under high salt and low pH value and is separated from the nucleic acid under low salt and high pH value is utilized, and then the whole extraction and purification process of the nucleic acid is realized by moving the magnetic bead or transferring liquid. The existing nucleic acid extraction technology has no continuity, manual intervention is needed in the process, errors can exist in extraction results due to manual pollution while the accuracy is influenced, and most of the existing nucleic acid extraction instruments are used for extracting a small amount of samples, if a large amount of samples to be extracted exist, frequent operation is needed, the extraction efficiency is influenced, and the error rate can be increased.
Disclosure of Invention
In order to solve the defects of the prior art, the invention provides a full-automatic nucleic acid extraction device and an extraction method, which can realize continuous automatic extraction in the whole process, can realize continuous automatic extraction of multiple samples, and can also realize continuous automatic extraction of a small amount of samples.
In order to achieve the purpose, the invention provides the following technical scheme:
a full-automatic nucleic acid extraction device comprises a bottom frame and a supporting vertical beam formed by extending upwards along four corners of the bottom frame, wherein the supporting vertical beam is fixedly connected with a supporting base parallel to the bottom frame and two X-direction guide rails parallel to each other, the X-direction guide rails are positioned above the supporting base, a first Y-direction guide rail and a second Y-direction guide rail parallel to each other are connected between the two X-direction guide rails along the vertical direction, the first Y-direction guide rail is connected with a clamping mechanism, and the second Y-direction guide rail is connected with a sampling mechanism;
the first Y-direction guide rail and the second Y-direction guide rail respectively drive the clamping mechanism and the sampling mechanism to slide along the X-direction guide rail, the clamping mechanism and the sampling mechanism slide along the Y-direction guide rail, and the clamping mechanism and the sampling mechanism slide in the Z direction perpendicular to the Y-direction guide rail;
the upper part of the supporting base is provided with a sucker carrier, a sample carrier, a reagent carrier, a PCR refrigeration module and a multipurpose carrier from the left side to the right side in sequence, the rear side of the PCR refrigeration module is provided with an extraction module, the reagent carrier is a bulk extraction reagent carrier, and the multipurpose carrier can be used as a deep hole plate carrier or a pre-packaged extraction reagent carrier.
Preferably, the full-automatic nucleic acid extraction device further comprises a carrier in-place detection device, the carrier in-place detection device comprises a first magnetic steel sheet fixing position and a positioning column fixing position which are arranged at the rear end of the sample carrier, the supporting base corresponds to a positioning baffle plate fixedly connected with the sample carrier, and the positioning baffle plate corresponds to the first magnetic steel sheet fixing position and the positioning column fixing position and is fixedly connected with a second magnetic steel sheet fixing position and a first Hall sensor fixing position respectively.
Preferably, full-automatic nucleic acid extraction element still includes sample scanning device, sample scanning device is including setting up support the base up end sweep ink recorder, connect support the base under the induction plate of terminal surface and set up and be in the fixed position of third magnetic steel piece of terminal surface under the sample carrier, set up on the induction plate with the fixed position of pilot lamp and second hall sensor that sample carrier quantity corresponds, the fixed position of pilot lamp, second hall sensor and the position of the fixed position of third magnetic steel piece is the straight line.
Preferably, support vertical beam fixed connection X to driving motor, first Y is to guide rail and second Y to guide rail integrated into one piece, X passes through the hold-in range to driving motor and connects first Y is to guide rail and second Y to the guide rail.
Preferably, first Y is to driving motor to the first Y of first Y to guide rail fixed connection, first Y passes through the hold-in range to driving motor and connects fixture, second Y is to guide rail fixed connection second Y to driving motor, second Y passes through the hold-in range to driving motor and connects sampling mechanism.
Preferably, the clamping mechanism comprises a clamping Z-direction driving motor and a clamping assembly, the clamping Z-direction driving motor drives the clamping assembly to slide in the Z direction through a screw rod fixedly connected with the clamping mechanism, the clamping assembly comprises a screw rod, a sliding plate, a clamping motor fixedly installed at the side end of the sliding plate and a support fixedly installed at the lower end part of the sliding plate, a clamping piece is sleeved outside the screw rod, two ends of the screw rod penetrate through the support, and the clamping motor is connected with the screw rod through a synchronous belt.
Preferably, the sampling mechanism comprises a plurality of sampling Z-direction driving motors, a plurality of sampling assemblies and spacing adjusting assemblies, the number of the sampling Z-direction driving motors corresponds to that of the sampling Z-direction driving motors, and the sampling Z-direction driving motors are fixedly connected with the spacing adjusting assemblies and are connected with the sampling assemblies through synchronous belts;
the sampling assembly comprises a connecting plate, the connecting plate is connected with a sampling driving motor through a screw rod, and the sampling driving motor is sequentially connected with a sampling pump and a sampling needle;
the distance adjusting assembly comprises a distance adjusting motor and a cam, the adjusting motor is connected with the cam through a synchronous belt, the sampling assembly is connected with the cam through a sliding plate of a clamping column fixedly connected with the lower end of the sampling assembly, and the clamping column is clamped in a groove of the cam.
Preferably, the extraction module comprises an extraction base and a support frame vertically fixed on the extraction base, the upper end face of the extraction base is respectively connected with an extraction support plate in a sliding manner and is arranged in a liquid receiving cover above the extraction support plate, the front side end of the support frame is respectively connected with a magnetic rod carrier and a magnetic rod sleeve carrier in a sliding manner, the lower end of the magnetic rod sleeve carrier is fixedly provided with a spring steel ball combination used for clamping a magnetic rod sleeve, and the extraction support plate is used for placing the deep-hole plate carrier or pre-packaging extraction reagent. The heating assembly is connected to the lower portion of the extracting carrier plate and comprises a polyimide heating sheet, a control panel, a temperature control switch fixing position and a temperature sensor fixing position, wherein the polyimide heating sheet and the control panel are sequentially connected to the lower portion of the extracting carrier plate, and the temperature control switch fixing position and the temperature sensor fixing position are arranged on the control panel.
Preferably, an extraction carrier driving motor and a liquid receiving cover driving motor are fixedly connected below the extraction base, the extraction carrier driving motor is connected with the extraction carrier through a synchronous belt, the liquid receiving cover driving motor is connected with the liquid receiving cover through a synchronous belt, the rear side end of the support frame is fixedly connected with a magnetic rod carrier driving motor and a magnetic rod sleeve carrier driving motor, the magnetic rod carrier driving motor is connected with the magnetic rod carrier through a synchronous belt, and the magnetic rod sleeve carrier driving motor is connected with the magnetic rod sleeve carrier through a synchronous belt;
preferably, the fully automatic nucleic acid isolation apparatus further comprises a needle washing carrier and a needle releasing carrier disposed between the reagent carrier and the multipurpose carrier.
The invention also provides a full-automatic nucleic acid extraction method, which adopts the extraction device and comprises the following steps:
the method comprises the following steps: preparing, clamping the deep hole plate carrier onto an extraction carrier plate of an extraction module by a clamping mechanism, and automatically loading a magnetic rod sleeve on a magnetic rod sleeve carrier;
step two: cracking, clamping a pre-packaged lysate carrier positioned on a multipurpose carrier onto an extraction carrier plate of an extraction module by a clamping mechanism, extracting a sample from a sample carrier and adding the sample into lysate of the extraction module after a sucker is loaded by a sucker carrier by a sampling mechanism, and uniformly mixing the lysate and the sample by utilizing the up-and-down movement of a magnetic rod sleeve; the clamping mechanism clamps the lysate carrier mixed with the sample to a multipurpose carrier for standby;
step three: moving magnetism, clamping a pre-packaged magnetic bead liquid carrier positioned on the multipurpose carrier onto an extraction carrier plate of an extraction module by a clamping mechanism, stirring the magnetic bead liquid up and down by matching a magnetic rod and a magnetic rod sleeve, and adsorbing magnetic beads in the magnetic bead liquid to the outer surface of the magnetic rod sleeve; the clamping mechanism clamps the lysate carrier mixed with the sample onto the extraction carrier plate of the extraction module again, the magnetic rod sleeve carrier and the magnetic rod carrier which are adsorbed with the magnetic beads simultaneously descend, move up and down in the lysate and stir, and adsorb the nucleic acid in the lysate onto the magnetic beads;
step four: rinsing, wherein a pre-packaged rinsing liquid carrier positioned on the multipurpose carrier is clamped onto an extraction carrier of the extraction module by a clamping mechanism, a magnetic rod sleeve carrier and a magnetic rod carrier, which are adsorbed with nucleic acid and magnetic beads, simultaneously descend, move up and down in rinsing liquid for rinsing, and then simultaneously ascend;
step five: eluting, wherein a pre-packaged eluent carrier positioned on the multipurpose carrier is clamped onto an extraction carrier of the extraction module by a clamping mechanism, a magnetic rod sleeve carrier and a magnetic rod carrier, which are adsorbed with nucleic acid and magnetic beads, simultaneously descend and move up and down in the eluent, and the nucleic acid is separated from the magnetic beads to the eluent;
step six: extracting, wherein a sampling mechanism loads a suction head from a suction head carrier to sample the eluent mixed with the nucleic acid into a PCR tube in a PCR refrigeration module for waiting for amplification and analysis and interpretation;
step seven: and (4) sterilizing and taking off the needle in the needle washing carrier and the needle taking-off carrier in sequence by the sampling mechanism.
The invention achieves the following beneficial effects:
1) the full-automatic nucleic acid extraction device can realize the continuous and automatic extraction of nucleic acid in the whole process;
2) the multi-purpose carrier can be used for placing a pre-packaged extraction reagent, so that continuous and automatic extraction of nucleic acid of multiple samples can be realized, rapid extraction is realized, and repeated absorption of bulk reagents is avoided;
3) the arranged bulk extraction reagent carrier can realize continuous automatic extraction of nucleic acid of a small amount of samples, and waste caused by using a pre-packaged extraction reagent is avoided;
4) the carrier in-place detection device can effectively avoid the interference between the sampling needle and the sample cup in the sampling process due to the fact that the carrier is not pushed in place;
5) the sample scanning device can realize automatic sample scanning in the process of pushing the sample carrier, so that manual intervention is reduced;
6) the heating component of the extraction carrier plate can realize heating in the nucleic acid extraction process and has the function of preventing overheating;
7) the magnet bar sleeve carrier can automatically clamp the magnet bar sleeve, and avoids pollution caused by manual installation of the magnet bar sleeve.
Drawings
FIG. 1 is a schematic diagram showing the overall structure of a fully automatic nucleic acid isolation apparatus according to the present invention;
FIG. 2 is a schematic view of the overall structure of the upper end face of the support base according to the present invention;
FIG. 3 is a schematic view of the overall construction of the clamping mechanism and the sampling mechanism of the present invention;
FIG. 4 is a schematic view of the overall construction of the clamping mechanism of the present invention;
FIG. 5 is a schematic view of the overall construction of the sampling mechanism of the present invention;
FIG. 6 is a bottom view of the sampling mechanism of the present invention;
FIG. 7 is a schematic diagram of the overall structure of the extraction module of the present invention;
FIG. 8 is a rear view of the extraction module of the present invention;
FIG. 9 is a schematic view of the overall construction of the present invention for a sleeve carrier;
fig. 10 is a bottom view of an extraction carrier of the present invention;
FIG. 11 is a rear view of the sample carrier of the present invention;
FIG. 12 is a bottom view of the sample carrier of the present invention;
FIG. 13 is a rear view of the positioning baffle of the present invention;
FIG. 14 is a schematic view of an induction plate of the present invention;
in the figure, 1 chassis; 2 supporting the vertical beam; 3 supporting the base; 4X-direction guide rails; 5 a first Y-direction guide rail; 6 second Y-direction guide rail; 7, a clamping mechanism; 8, a sampling mechanism; 9 suction head carrier; 10 sample carriers; 11 a reagent carrier; 12PCR refrigeration module; 13 a multipurpose carrier; 14 an extraction module; a 15X-direction driving motor; 16 a first Y-direction drive motor; 17 a second Y-direction drive motor; 18 clamping a Z-direction driving motor; 19 clamping the screw rod; 20, a screw rod; 21 a sliding plate; 22 a clamping motor; 23, a support; 24 a clamp; 25 sampling a Z-direction driving motor; 26 a connecting plate; 27 sampling screw; 28 sampling a driving motor; 29 a sampling pump; 30 sampling needles; 31 spacing adjustment motor; 32 cams; 33, clamping the column; 34 extracting the base; 35 a support frame; 36 extracting the carrier plate; 37, a liquid receiving cover; 38 a magnetic rod carrier; 39 a magnet bar sleeve carrier; 40, combining spring steel balls; 41 extracting a carrier plate driving motor; 42 a liquid receiving cover driving motor; 43 magnetic bar carrier driving motor; 44 magnetic bar sleeve carrier driving motor; 45, washing a needle carrier; 46 a needle release carrier; 47 normal temperature carrier; 48 a cover plate carrier; 49 reagent refrigeration carrier; 50 positioning a baffle plate; 51, fixing a first magnetic steel sheet; 52, positioning the positioning column for fixing; 53, fixing the second magnetic steel sheet; 54 first hall sensor fixed position; 55 a polyimide heating sheet; 56 a control panel; 57 the temperature control switch is fixed; 58 temperature sensor fixing position; 59 code scanning machine; 60 a sensing plate; 61 an indicator light; 62, fixing the second Hall sensor; and 63, fixing the third magnetic steel sheet.
Detailed Description
Other advantages and features of the present invention will become readily apparent to those skilled in the art from the following detailed description, wherein it is to be understood that the invention is not limited to the specific embodiments disclosed, but is to be construed as limited only by the appended claims. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
To facilitate an understanding of the present invention by those skilled in the art, specific embodiments thereof are described below with reference to the accompanying drawings.
Referring to fig. 1 to 3, a fully automatic nucleic acid extracting apparatus is disclosed, comprising a chassis 1 and a vertical support beam 2 extending upwards along four corners of the chassis, wherein a support base 3 parallel to the chassis 1 and two parallel X-direction rails 4 are fixedly connected to the vertical support beam 2, the X-direction rails 4 are positioned above the support base 3, a first Y-direction rail 5 and a second Y-direction rail 6 parallel to each other are connected between the two X-direction rails 4 along a vertical direction, the first Y-direction rail 5 is connected with a clamping mechanism 7, and the second Y-direction rail 6 is connected with a sampling mechanism 8; the first Y-direction guide rail 5 and the second Y-direction guide rail 6 respectively drive the clamping mechanism 7 and the sampling mechanism 8 to slide along the X-direction guide rail 4, the clamping mechanism 7 and the sampling mechanism 8 respectively slide along the first Y-direction guide rail 5 and the second Y-direction guide rail 6, and the clamping mechanism 7 and the sampling mechanism 8 perform Z-direction sliding perpendicular to the first Y-direction guide rail 5;
the upper part of the supporting base 3 is provided with a suction head carrier 9, a sample carrier 10, a reagent carrier 11, a PCR refrigeration module 12 and a multipurpose carrier 13 in sequence from the left side to the right side, the rear side of the PCR refrigeration module 12 is provided with an extraction module 14, the reagent carrier 11 is a bulk extraction reagent carrier, and the multipurpose carrier 13 can be used as a deep hole plate carrier or a pre-packaged extraction reagent carrier.
On one hand, the arranged multipurpose carrier 13 can be used for placing prepackaged extraction reagents, can realize continuous automatic extraction of 96 multi-sample nucleic acids, realizes quick extraction, avoids repeatedly absorbing bulk reagents, and can greatly improve the extraction efficiency; on the other hand, the arranged bulk extraction reagent carrier 11 can realize continuous automatic extraction of nucleic acid of a small amount of samples, and waste caused by using a pre-packaged extraction reagent is avoided.
Further, the full-automatic nucleic acid extracting device further comprises a carrier in-place detection device, referring to fig. 11 to 14, the carrier in-place detection device comprises a first magnetic steel sheet fixing position 51 and a positioning column fixing position 52 which are arranged at the rear end of the sample carrier 10, the supporting base 3 is fixedly connected with a positioning baffle 50 corresponding to the sample carrier 10, and a second magnetic steel sheet fixing position 53 and a first hall sensor fixing position 54 are respectively arranged at the positions of the positioning baffle 50 corresponding to the first magnetic steel sheet fixing position 51 and the positioning column fixing position 52. If the sample carrier 10 is pushed in place, the first magnetic steel sheet and the second magnetic steel sheet attract each other, the positioning column positioned on the positioning column fixing position 52 is contacted with the first hall sensor positioned on the first hall sensor fixing position 54, the first hall sensor sends out a signal, the signal is transmitted to the indicator light main control board, and the indicator light main control board controls the indicator light to operate. The carrier in-place detection system can effectively avoid the interference between the sampling needle 30 and the sample cup in the sampling process due to the fact that the carrier is not pushed in place. In the same manner, the tip carrier 9, reagent carrier 11 and multipurpose carrier 13 can be arranged to detect the presence.
The full-automatic nucleic acid extraction device further comprises a sample scanning device, the sample scanning device comprises a code scanning machine 59 arranged on the upper end face of the support base 3, an induction plate 60 connected to the lower end face of the support base 3 and a third magnetic steel sheet fixing position 63 arranged on the lower end face of the sample carrier 10, the induction plate is provided with indicating lamps 61 and second Hall sensor fixing positions 62 corresponding to the number of the sample carriers, the positions of the indicating lamps 61, the second Hall sensor fixing positions 62 and the third magnetic steel sheet fixing positions 63 are on the same straight line, the sample carrier 10 is pushed in, the second Hall sensor senses the third magnetic steel sheet on the lower end face of the sample carrier 10, the second Hall sensor sends out a signal and transmits the signal to a code scanning main control board, and the code scanning machine 59 is controlled by the code scanning main control board to operate. The sample scanning device can realize automatic sample scanning in the process of pushing the sample carrier, and manual intervention is reduced.
Further, support vertical beam 2 fixed connection X to driving motor 15, first Y to guide 5 and second Y to guide 6 integrated into one piece, X to driving motor 15 through the hold-in range connection first Y to guide 5 and second Y to guide 6 to drive first Y to guide 5 and second Y to guide 6 and slide along X to guide 4.
As shown in fig. 3, the first Y-direction rail 5 is fixedly connected to a first Y-direction driving motor 16, the first Y-direction driving motor 16 is connected to the clamping mechanism 7 through a timing belt and drives the clamping mechanism 7 to slide along the first Y-direction rail 5, the second Y-direction rail 6 is fixedly connected to a second Y-direction driving motor 17, and the second Y-direction driving motor 17 is connected to the sampling mechanism 8 through a timing belt and drives the sampling mechanism 8 to slide along the second Y-direction rail 6.
As shown in fig. 4, the clamping mechanism 7 includes a clamping Z-direction driving motor 18 and a clamping assembly, the clamping Z-direction driving motor 18 drives the clamping assembly to perform Z-direction sliding through a clamping screw 19 fixedly connected to the clamping mechanism, the clamping assembly includes a screw rod 20, a sliding plate 21, a clamping motor 22 fixedly mounted at a side end of the sliding plate 21, and a support 23 fixedly mounted at a lower end of the sliding plate 21, a clamping member 24 is sleeved outside the screw rod 20, two ends of the screw rod 20 penetrate through the support 23, the clamping motor 22 is connected to the screw rod 20 through a synchronous belt, and the clamping motor 22 drives the screw rod 20 to move through the synchronous belt, so as to drive the clamping member 24 sleeved outside.
Further, referring to fig. 5 and 6, the sampling mechanism 8 includes a plurality of sampling Z-direction driving motors 25, a plurality of sampling assemblies corresponding to the number of the sampling Z-direction driving motors 25, and a spacing adjustment assembly, wherein the sampling Z-direction driving motors 25 are fixedly connected to the spacing adjustment assembly and connected to the sampling assemblies through synchronous belts;
specifically, the sampling assembly comprises a connecting plate 26, the connecting plate 26 is connected with a sampling driving motor 28 through a sampling screw 27, and the sampling driving motor 28 is sequentially connected with a sampling pump 29 and a sampling needle 30;
specifically, interval adjustment assembly includes interval adjustment motor 31, cam 32, and interval adjustment motor 31 passes through the hold-in range and connects cam 32, and sampling component passes through the sliding plate and the cam 32 of lower extreme fixedly connected with joint post 33 and is connected, and joint post 33 joint is in the recess of cam 32, and interval adjustment motor 31 moves through hold-in range drive cam 32 to make the interval between the sampling component change through the motion of joint post 33 in the recess of cam 32.
Further, referring to fig. 7 to 10, the extraction module 14 includes an extraction base 34 and a support frame 35 vertically fixed on the upper end of the extraction base 34, the upper end surface of the extraction base 34 is respectively connected with an extraction carrier 36 and a liquid receiving cover 37 disposed above the extraction carrier 36 in a sliding manner, the front end of the support frame 35 is respectively connected with a magnetic rod carrier 38 and a magnetic rod sleeve carrier 39 in a sliding manner, the lower end of the magnetic rod sleeve carrier 39 is fixed with a spring steel ball assembly 40 for engaging with a magnetic rod sleeve, and the extraction carrier 36 is used for placing the deep-well plate carrier or pre-packaging extraction reagent. The magnetic rod sleeve carrier 39 can automatically clamp the magnetic rod sleeve, and pollution caused by manual installation of the magnetic rod sleeve is avoided.
Further, a heating assembly is connected below the extraction carrier plate 36, and the heating assembly comprises a polyimide heating sheet 55, a control board 56, a temperature control switch fixing position 57 and a temperature sensor fixing position 58, which are sequentially connected below the extraction carrier plate 36, and the temperature control switch fixing position 57 and the temperature sensor fixing position 58 are arranged on the control board 56. The polyimide heating sheet 55 is used for heating in the nucleic acid extraction process, and the temperature control switch fixing position 57 and the temperature sensor fixing position 58 are used for fixing the temperature control switch and the temperature sensor respectively, and are used for preventing overheating in a matching manner.
Specifically, draw base 34 below fixed connection and draw support plate driving motor 41 and connect liquid lid driving motor 42, draw support plate driving motor 41 and connect through the hold-in range and draw support plate 36, connect liquid lid driving motor 42 to connect through the hold-in range connection connect liquid lid 37, support frame 35 back side end fixed connection bar magnet carrier driving motor 43 and bar cover carrier driving motor 44, bar magnet carrier driving motor 43 passes through the hold-in range connection bar magnet carrier 38, bar cover carrier driving motor 44 passes through the hold-in range connection bar cover carrier 39. An extraction carrier driving motor 41 and a liquid receiving cover driving motor 42 respectively drive the extraction carrier 36 and the liquid receiving cover 37 to move in the Y direction, a magnetic rod carrier driving motor 43 and a magnetic rod sleeve carrier driving motor 44 respectively drive the magnetic rod carrier 38 and the magnetic rod sleeve carrier 39 to move in the Z direction, and magnetic rods on the magnetic rod carrier 38 can be inserted into magnetic rod sleeves; the liquid receiving cover 37 is used for preventing the liquid on the outer surface of the magnetic rod sleeve from dripping to cause pollution.
Further, the full-automatic nucleic acid extracting apparatus further includes a needle washing carrier 45, a needle releasing carrier 46, a normal temperature carrier 47, a cover carrier 48, and a reagent cooling carrier 49 provided between the reagent carrier 11 and the multipurpose carrier 13.
The invention also provides a full-automatic nucleic acid extraction method, which adopts the extraction device and comprises the following steps:
1) preparing: the clamping mechanism 7 clamps the deep hole plate carrier (provided with the magnetic rod sleeve) positioned on the multipurpose carrier 13 onto the extraction carrier 36 of the extraction module 14, the extraction carrier 36 moves to the lower part of the magnetic rod sleeve carrier 39, and after the magnetic rod sleeve carrier 39 automatically loads the magnetic rod sleeve, the clamping mechanism 7 clamps the deep hole plate carrier onto the multipurpose carrier 13;
2) cracking:
a, the clamping mechanism 7 clamps the pre-packaged lysate carrier positioned on the multipurpose carrier 13 onto the extraction carrier 36 of the extraction module 14;
b, after the suction head carrier 9 is loaded with the suction head, the sampling mechanism 8 extracts a sample from the sample carrier 10 and adds the sample into the lysate of the extraction module 14, the extraction carrier 36 moves to the lower part of the magnetic rod sleeve carrier 39, the magnetic rod sleeve carrier 39 moves downwards, the lysate and the sample are uniformly mixed by utilizing the up-and-down movement of the magnetic rod sleeve, and the magnetic rod sleeve carrier 39 moves upwards;
c, the clamping mechanism 7 clamps the lysate carrier mixed with the sample to a multi-purpose carrier 13 for standby;
3) moving magnetism:
a, a clamping mechanism 7 clamps a pre-packaged magnetic bead liquid carrier positioned on a multipurpose carrier 13 onto an extraction carrier 36 of an extraction module 14, the extraction carrier 36 moves to the position below a magnetic rod sleeve carrier 39, the magnetic rod sleeve carrier 39 moves downwards, a magnetic rod sleeve moves upwards and downwards to mix magnetic bead liquid, then the magnetic rod carrier 38 moves downwards, a magnetic rod is inserted into the magnetic rod sleeve, the magnetic rod and the magnetic rod sleeve are matched to move upwards and downwards to stir the magnetic bead liquid, magnetic beads in the magnetic bead liquid are adsorbed to the outer surface of the magnetic rod sleeve, and the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 move upwards at the same time;
b, the clamping mechanism 7 clamps the magnetic bead liquid carrier which has absorbed the magnetic beads to the multipurpose carrier 13;
c, the clamping mechanism 7 clamps the lysate carrier mixed with the sample again onto the extraction carrier 36 of the extraction module 14, the extraction carrier 36 moves to the lower part of the magnetic rod loading rack 39, the magnetic rod loading rack 39 and the magnetic rod carrier 38 adsorbing the magnetic beads simultaneously move downwards and stir up and down in the lysate, the nucleic acid in the lysate is adsorbed onto the magnetic beads, and the magnetic rod loading rack 39 and the magnetic rod carrier 38 simultaneously move upwards;
d the holding mechanism 7 holds the lysate carrier from which the nucleic acid has been sucked to the multi-purpose carrier 13;
4) rinsing:
a, the clamping mechanism 7 clamps a pre-packaged rinsing liquid carrier positioned on the multipurpose carrier 13 onto an extraction carrier 36 of the extraction module 14, the extraction carrier 36 moves to the lower part of a magnetic rod sleeve carrier 39, the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbing nucleic acid and magnetic beads simultaneously move downwards and perform up-and-down rinsing in the rinsing liquid, and then the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 simultaneously move upwards;
b the clamping mechanism 7 clamps the rinsing liquid carrier to the multipurpose carrier 13;
5) and (3) elution:
a, the clamping mechanism 7 clamps the pre-packaged eluent carrier positioned on the multipurpose carrier 13 onto the extraction carrier 36 of the extraction module 14, the extraction carrier 36 moves to the lower part of the magnetic rod sleeve carrier 39, the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbing nucleic acid and magnetic beads simultaneously move downwards and move upwards and downwards in the eluent, the nucleic acid is separated from the magnetic beads into the eluent, and the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbing only the magnetic beads simultaneously move upwards; discarding the magnetic beads;
6) extraction:
a, a sampling mechanism 8 loads a suction head from a suction head carrier 9 to sample the eluent mixed with nucleic acid into a PCR tube in a PCR refrigeration module 12 for waiting amplification and analysis interpretation;
7) and (3) disinfection:
a the sampling mechanism 8 completes the disinfection and the needle removal in the needle washing carrier 45 and the needle removal carrier 46 in sequence.
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed. The above-described embodiments of the present invention do not limit the scope of the present invention. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.
Claims (10)
1. The utility model provides a full-automatic nucleic acid extraction element, includes chassis (1) and support vertical beam (2) that upwards extends formation along the chassis four corners, its characterized in that:
the supporting vertical beam (2) is fixedly connected with a supporting base (3) parallel to the underframe (1) and two X-direction guide rails (4) parallel to each other, the X-direction guide rails (4) are positioned above the supporting base (3), a first Y-direction guide rail (5) and a second Y-direction guide rail (6) parallel to each other are connected between the two X-direction guide rails (4) along the vertical direction, the first Y-direction guide rail (5) is connected with a clamping mechanism (7), and the second Y-direction guide rail (6) is connected with a sampling mechanism (8);
the first Y-direction guide rail (5) and the second Y-direction guide rail (6) respectively drive the clamping mechanism (7) and the sampling mechanism (8) to slide along the X-direction guide rail (4), the clamping mechanism (7) and the sampling mechanism (8) respectively slide along the first Y-direction guide rail (5) and the second Y-direction guide rail (6), and the clamping mechanism (7) and the sampling mechanism (8) slide in a Z direction perpendicular to the first Y-direction guide rail (5);
the upper end face of the supporting base (3) is sequentially provided with a sucker carrier (9), a sample carrier (10), a reagent carrier (11), a PCR refrigeration module (12) and a multipurpose carrier (13) from the left side to the right side, the rear side of the PCR refrigeration module (12) is provided with an extraction module (14), the reagent carrier (11) is a bulk extraction reagent carrier, and the multipurpose carrier (13) can be used as a deep-hole plate carrier or a pre-packaged extraction reagent carrier.
2. The full-automatic nucleic acid extraction device according to claim 1, further comprising a carrier in-place detection device, wherein the carrier in-place detection device comprises a first magnetic steel sheet fixing position (51) and a positioning column fixing position (52) which are arranged at the rear end of the sample carrier (10), the supporting base (3) is fixedly connected with a positioning baffle plate (50) corresponding to the sample carrier (10), and the positioning baffle plate (50) is respectively provided with a second magnetic steel sheet fixing position (53) and a first Hall sensor fixing position (54) corresponding to the first magnetic steel sheet fixing position (51) and the positioning column fixing position (52).
3. The automatic nucleic acid extracting device according to claim 1, wherein the automatic nucleic acid extracting device further comprises a sample scanning device, the sample scanning device comprises a code scanner (59) disposed on the upper end surface of the supporting base (3), a sensing plate (60) connected to the lower end surface of the supporting base (3), and a third magnetic steel sheet fixing position (63) disposed on the lower end surface of the sample carrier (10), the sensing plate is provided with indicating lamps (61) and second hall sensor fixing positions (62) corresponding to the number of the sample carriers, and the positions of the indicating lamps (61), the second hall sensor fixing positions (62), and the third magnetic steel sheet fixing position (63) are linear.
4. The automatic nucleic acid isolation apparatus according to claim 1, wherein the support vertical beam (2) is fixedly connected to an X-direction drive motor (15), the first Y-direction guide rail (5) and the second Y-direction guide rail (6) are integrally formed, and the X-direction drive motor (15) is connected to the first Y-direction guide rail (5) and the second Y-direction guide rail (6) through a timing belt; first Y is to guide rail (5) first Y of fixed connection to driving motor (16), first Y is to driving motor (16) through the hold-in range connection fixture (7), second Y is to guide rail (6) fixed connection second Y to driving motor (17), second Y is to driving motor (17) through the hold-in range connection sampling mechanism (8).
5. The automatic nucleic acid extracting apparatus according to claim 1, wherein the clamping mechanism (7) comprises a clamping Z-direction driving motor (18) and a clamping assembly, the clamping Z-direction driving motor (18) drives the clamping assembly to perform Z-direction sliding through a clamping screw (19) fixedly connected, the clamping assembly comprises a sliding plate (21), a screw rod (20), a clamping motor (22) fixedly installed at a side end of the sliding plate (21), and a support (23) fixedly installed at a lower end of the sliding plate (21), a clamping member (24) is sleeved outside the screw rod (20), two ends of the screw rod (20) penetrate through the support (23), and the clamping motor (22) is connected with the screw rod (20) through a synchronous belt.
6. The automatic nucleic acid extracting device according to claim 1, wherein the sampling mechanism (8) comprises a plurality of sampling Z-direction driving motors (25), a plurality of sampling assemblies corresponding to the number of the sampling Z-direction driving motors, and a spacing adjusting assembly, wherein the sampling Z-direction driving motors (25) are fixedly connected with the spacing adjusting assembly and connected with the sampling assemblies through synchronous belts;
the sampling assembly comprises a connecting plate (26), the connecting plate (26) is connected with a sampling driving motor (28) through a sampling screw rod (27), and the sampling driving motor (28) is sequentially connected with a sampling pump (29) and a sampling needle (30);
the interval adjusting assembly comprises an interval adjusting motor (31) and a cam (32), the interval adjusting motor (31) is connected with the cam (32) through a synchronous belt, the sampling assembly is connected with the cam (32) through a sliding plate of a clamping column (33) fixedly connected with the lower end, and the clamping column (33) is clamped in a groove of the cam (32).
7. The automatic nucleic acid extracting device according to claim 1, wherein the extracting module (14) comprises an extracting base (34) and a support frame (35) vertically fixed on the upper end of the extracting base (34), the upper end surface of the extracting base (34) is respectively connected with an extracting carrier plate (36) and a liquid receiving cover (37) arranged above the extracting carrier plate (36) in a sliding manner, the front end of the support frame (35) is respectively connected with a magnetic rod carrier (38) and a magnetic rod sleeve carrier (39) which correspond to each other in a sliding manner, the lower end of the magnetic rod sleeve carrier (39) is fixed with a spring steel ball assembly (40) for clamping a magnetic rod sleeve, and the extracting carrier plate (36) is used for placing the deep hole plate carrier or pre-packaging extracting reagent.
8. The automatic nucleic acid extracting apparatus according to claim 7, wherein a heating assembly is connected below the extracting carrier plate (36), and the heating assembly comprises a polyimide heating plate (55), a control plate (56), and a temperature control switch fixing position (57) and a temperature sensor fixing position (58) which are arranged on the control plate (56), wherein the polyimide heating plate (55) and the control plate (56) are sequentially connected below the extracting carrier plate (36).
9. The apparatus for automatically extracting nucleic acid according to claim 7, wherein an extraction carrier driving motor (41) and a liquid receiving lid driving motor (42) are fixedly connected below the extraction base (34), the extraction carrier driving motor (41) is connected to the extraction carrier (36) through a timing belt, the liquid receiving lid driving motor (42) is connected to the liquid receiving lid (37) through a timing belt, a magnetic rod carrier driving motor (43) and a magnetic rod sleeve carrier driving motor (44) are fixedly connected to the rear side end of the support frame (35), the magnetic rod carrier driving motor (43) is connected to the magnetic rod carrier (38) through a timing belt, and the magnetic rod sleeve carrier driving motor (44) is connected to the magnetic rod sleeve carrier (39) through a timing belt.
10. A fully automatic nucleic acid extraction method using the fully automatic nucleic acid extraction apparatus according to any one of claims 1 to 9, comprising the steps of:
the method comprises the following steps: preparing, the clamping mechanism 7 clamps the deep hole plate carrier onto the extraction carrier plate 36 of the extraction module 14, and the magnetic rod sleeve carrier 39 automatically loads the magnetic rod sleeve;
step two: cracking, wherein a pre-packaged lysate carrier positioned on the multipurpose carrier 13 is clamped onto an extraction carrier plate 36 of an extraction module 14 by a clamping mechanism 7, samples are extracted from a sample carrier 10 and added into lysate of the extraction module 14 after suction heads are loaded on a suction head carrier 9 by a sampling mechanism 8, and the lysate and the samples are uniformly mixed by utilizing the up-and-down movement of a magnetic rod sleeve; the clamping mechanism 7 clamps the lysate carrier mixed with the sample to a multi-purpose carrier 13 for standby;
step three: moving magnetism, clamping the pre-packaged magnetic bead liquid carrier positioned on the multipurpose carrier 13 onto an extraction carrier plate 36 of an extraction module 14 by a clamping mechanism 7, and stirring the magnetic bead liquid up and down by matching a magnetic rod and a magnetic rod sleeve, wherein magnetic beads in the magnetic bead liquid are adsorbed to the outer surface of the magnetic rod sleeve; the clamping mechanism 7 clamps the lysate carrier mixed with the sample again onto the extraction carrier 36 of the extraction module 14, the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbed with the magnetic beads simultaneously descend, move up and down in the lysate and stir, so that the nucleic acid in the lysate is adsorbed onto the magnetic beads;
step four: rinsing, the clamping mechanism 7 clamps the pre-packaged rinsing liquid carrier positioned on the multipurpose carrier 13 onto the extraction carrier 36 of the extraction module 14, the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbing nucleic acids and magnetic beads simultaneously descend, move up and down in the rinsing liquid for rinsing, and then simultaneously ascend the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38;
step five: eluting, wherein the clamping mechanism 7 clamps the pre-packaged eluent carrier positioned on the multipurpose carrier 13 onto the extraction carrier 36 of the extraction module 14, the magnetic rod sleeve carrier 39 and the magnetic rod carrier 38 adsorbing nucleic acid and magnetic beads simultaneously descend and move up and down in the eluent, and the nucleic acid is separated from the magnetic beads to the eluent;
step six: extracting, loading the suction head from the suction head carrier 9 by the sampling mechanism 8, sampling the eluent mixed with the nucleic acid into a PCR tube in the PCR refrigeration module 12 for waiting for amplification and analysis interpretation;
step seven: and (4) sterilizing and needle releasing are finished in the needle washing carrier 45 and the needle releasing carrier 46 by the sampling mechanism 8 in sequence.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110731487.0A CN113308366A (en) | 2021-06-30 | 2021-06-30 | Full-automatic nucleic acid extraction device and method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113943633A (en) * | 2021-10-25 | 2022-01-18 | 苏州缔因安生物科技有限公司 | Nucleic acid extraction device based on ultrasonic waves and use method thereof |
| CN116731856A (en) * | 2023-04-24 | 2023-09-12 | 扬州市职业大学(扬州开放大学) | High-flux automatic sorting device and sorting method thereof |
-
2021
- 2021-06-30 CN CN202110731487.0A patent/CN113308366A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113943633A (en) * | 2021-10-25 | 2022-01-18 | 苏州缔因安生物科技有限公司 | Nucleic acid extraction device based on ultrasonic waves and use method thereof |
| CN113943633B (en) * | 2021-10-25 | 2023-11-14 | 苏州缔因安生物科技有限公司 | Ultrasonic-based nucleic acid extraction device and application method thereof |
| CN116731856A (en) * | 2023-04-24 | 2023-09-12 | 扬州市职业大学(扬州开放大学) | High-flux automatic sorting device and sorting method thereof |
| CN116731856B (en) * | 2023-04-24 | 2023-12-01 | 扬州市职业大学(扬州开放大学) | High-flux automatic sorting device and sorting method thereof |
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