WO2023018193A1 - Nucleic acid extraction apparatus in which reagents are dispensed in bulk - Google Patents

Abstract

The present invention relates to an apparatus for separating and concentrating pathogens and extracting nucleic acid materials by automatically dispensing reagents and, more particularly, to a nucleic acid extraction apparatus in which reagents are dispensed in an integral bulk, the apparatus comprising: a reagent inlet for inserting a large-capacity bulk bottle; a reagent reservoir for receiving and storing reagents supplied from the reagent inlet; and a dispensing syringe for suctioning the reagents in the reagent reservoir and dispensing the reagents into a cartridge, wherein the apparatus can receive reagents from the large-capacity bulk bottle to process several reagents at once, and can batch process separation, concentration, and purification of pathogens and a nucleic acid extraction process, thereby not requiring a separate pathogen concentration apparatus, nucleic acid purification apparatus, and nucleic acid extraction apparatus that are commonly used.

Description

Nucleic acid extraction device dispensing reagents in bulk

The present invention relates to an apparatus for automatically dispensing reagents to isolate and concentrate pathogens and to extract nucleic acid materials, and more particularly, to a reagent inlet device for inserting a large-capacity bulk bottle; a reagent reservoir for receiving and storing reagents supplied from the reagent inlet; A dispensing syringe for inhaling the reagents in the reagent reservoir and dispensing them to the cartridge, and receiving reagents from a large-capacity bulk bottle to process several reagents at once, separating, concentrating and purifying pathogens and nucleic acids The present invention relates to a nucleic acid extraction device capable of dispensing reagents in an integral bulk that does not require a commonly used pathogen enrichment device, nucleic acid purification device, and nucleic acid extraction device because the extraction process can be batch-processed.

In general, devices for extracting or purifying nucleic acids or biological substances from various biological samples such as human tissues and blood are widely used in various fields such as medicine and biotechnology, and recently Polymerase Chain Reaction (PCR) As the molecular diagnostic method for detecting mutations in genes by amplifying them through amplification has become commonplace, devices for separating or purifying nucleic acids or various biological substances efficiently and with high purity are being developed one after another.

In addition, this device can be applied not only to humans but also to samples collected from animals, plants, insects, seaweeds, microorganisms, bacteria, fungi, viruses, and the like.

In order to extract or purify nucleic acids or various biological substances, various steps such as lysis, binding, washing, and elutting are required. In addition, there is a risk of infection when used, and reliability is deteriorated due to the inconsistent quantity and quality of isolated or purified nucleic acids or various biological substances, and thus, it has also been developed as an automated device. Among these devices, as an advantageous method for application to automated devices, a method using magnetic particles, which are magnetic materials having a property of adsorbing nucleic acids or various biological substances, is widely spread in the field of molecular diagnosis.

As a tool for carrying out this process, a conventional kit for extracting or purifying nucleic acids or biological substances includes a magnet mounting rod having a magnet installed at the bottom, a mounting rod cover into which the magnet mounting rod is inserted into an open top, and a biological sample. It is composed of a cartridge having a plurality of wells necessary for extraction or purification, such as wells for dispensing magnetic materials, wells for dispensing washing liquid, and wells for dispensing eluent after dispensing, and in automated devices, magnet mounting rods and mounting rod covers The lifting means and the moving means of the multi-well are additionally installed. In order to simultaneously extract or purify nucleic acids or various biological substances from a plurality of biological samples, a cartridge having a plurality of wells required for extraction or purification in a plurality of rows, a mounting rod cover arranged in the number of rows, and a magnet mounting rod may be used.

In a kit for extracting nucleic acids or biological substances using a polymerase chain reaction, reagents are dispensed into each well, and a sealing operation is performed to seal the reagents at an open top of the cartridge.

However, in the case of a cartridge equipped with multi-wells consisting of a plurality of rows and columns, there are many openings formed at the upper end, so that the sealing paper for sealing the reagent lacks space for bonding to the upper end of the cartridge, making it difficult to seal each well.

Due to the limitation of the space where the sealing paper is attached to the top of the cartridge, a high degree of technical skill is required, and if the sealing process fails, the reagents dispensed inside are transferred to other wells or leaked to the outside of the cartridge, making it unusable.

In addition, ethanol, among reagents used to extract nucleic acids or biological substances, has a low boiling point, and thus has a disadvantage in that evaporation from a liquid state to a gaseous state occurs easily.

Therefore, ethanol is prepared by dispensing an appropriate amount into the wells of the kit, and if the kit is not completely sealed during transport, ethanol is vaporized and evaporated or mixed into wells that perform other functions, adversely affecting the quality and performance of the kit. works as

In addition, since the kit prepared by dispensing reagents appropriately into each well requires dispensing reagents of different types and capacities according to the nucleic acid and biological material to be extracted, a corresponding kit must be separately prepared according to the type of biological sample to be extracted. should be provided This acts as a factor in increasing the price of the kit, which inevitably incurs a lot of cost, which increases inefficiency.

Therefore, in order to extract a specific substance from a biological sample in a constant amount and with improved quality, various types of reagents are automatically dispensed into each well, and reagents are dispensed into each well, making it difficult to seal and providing a kit that can be used only in a single sample. Development of a nucleic acid extraction device that provides necessary reagents in the form of a bulk bottle that can be easily sealed and dispenses a set amount of reagents according to each biological sample is emerging.

[Prior art literature]

[Patent Literature]

(Patent Document 1) KR 10-2110647 (B1) 2020.05.07.

(Patent Document 2) JP 2006-223175 (A) 2006.08.31.

In order to solve the above problems, an object of the present invention is to automatically dispense a reagent for extracting nucleic acid from a biological sample into a cartridge and to automatically inhale the reagent from a reagent reservoir by manually moving the dispensing syringe up and down. It is an object of the present invention to provide a nucleic acid extraction device that dispenses reagents in bulk, reducing errors occurring during operation and maintaining a constant quantity and quality of extracted nucleic acids.

Another object of the present invention is to provide a nucleic acid extraction device for dispensing reagents in bulk, which is easy to seal and can reduce the sealing cost required for nucleic acid extraction, since reagents required in the nucleic acid extraction process can be moved in a bulk bottle. is to provide

Another object of the present invention is to provide a nucleic acid extraction device capable of dispensing reagents in bulk, which can be used for various samples by dispensing the type and amount of reagents to a cartridge at a set value according to a biological sample.

Another object of the present invention is to provide a nucleic acid extraction device for dispensing reagents in bulk, in which the amount of reagent sucked into the tip is determined according to the height at which the piston of the dispensing syringe rises, so that the dispensing capacity of the reagent solution can be conveniently adjusted.

Another object of the present invention is to form a partition wall protruding upward in the central part of the reagent storage chamber, which is a space supplied and stored from a bulk bottle, so that a small amount of reagent forms a high water level, thereby confirming whether the reagent has flowed in by a target amount. It is to provide a nucleic acid extracting device in which reagents are dispensed in bulk capable of inhaling reagents on both sides around the septum.

Another object of the present invention is to provide a plurality of tips that are detachable from the injection tip of the dispensing syringe, and to separate and mount the tips according to the type of reagent to be sucked, so that different types of reagents do not mix with each other, thereby improving the quality of nucleic acid extraction It is to provide a nucleic acid extraction device in which reagents are dispensed in bulk capable of maintaining

In addition, another object of the present invention is a bulk that can automate the attachment and detachment of the tip by separating the tip from the injection tip while the piston of the dispensing syringe further descends and discharges the remaining amount after dispensing the reagent inside the tip. To provide a nucleic acid extraction device that is dispensed with reagents.

Another object of the present invention is to automatically lift and move an extractor that separates and extracts nucleic acids from a sample at a location opposite to the dispensing syringe, so that reagent dispensing and nucleic acid extraction processes are performed simultaneously, greatly reducing the time required for nucleic acid extraction. It is to provide a nucleic acid extraction device in which reagents are dispensed in a reduced bulk.

The present invention for achieving the above object is a reagent inlet 100 in which a bulk bottle 20 containing reagents is inserted therein, and reagents inside the bulk bottle 20 are discharged through a reagent hole 110 formed at the lower side. ; a transfer line 200 having one end communicating with the reagent hole 110 so that the reagent flows along the hollow interior and controlling the flow of the reagent by a valve 210 provided on one side; a reagent reservoir 300 in which a plurality of reagent storage chambers 310 having different sizes are formed to receive and store reagents from the transfer line 200; The piston 431 is combined with the cylinder 441 to inhale and discharge the reagent stored in the reagent reservoir 300 while moving up and down, and the injection tip 442 is disposed at the end of the cylinder 441 in a plurality of rows and columns. A dispensing syringe 400; A tip 710 that is disposed on the side of the reagent reservoir 300, is attached to the lower end of the injection tip 442 and moves, and the injection tip 442 descends and is inserted into the reagent reservoir 300 to suck the reagent. ) is placed in a plurality of rows and columns (700); A cartridge 600 in which the reagent contained in the tip 710 is discharged while the piston 431 descends and contained therein, and nucleic acid extraction is performed; and a storage cartridge 800 storing the nucleic acid extracted from the cartridge 600.

In addition, the number of columns of the tips 710 placed on the tip plate 700 of the present invention is the same as the number of columns of the injection tips 442 of the dispensing syringe 400, and the number of rows of the tips 710 is greater than the number of rows of the scanning tips 442.

In addition, the reagent storage chamber 310 of the present invention has the same height as the neighboring reagent storage chamber 310, but has different lengths.

In addition, a partition wall 311 protruding upward is formed in the central portion of the reagent storage chamber 310 of the present invention, and the partition wall 311 is smaller than the length of the reagent storage chamber 310 so that the reagent passes through the partition wall 311. The tip 710, which surrounds and forms a water level of the same height and is attached to the injection tip 442, consists of two rows, and the distance between the rows at the end of the tip 710 is greater than the width of the partition wall 311. It is larger and smaller than the width of the reagent storage chamber 310 and descends on both sides with the partition wall 311 interposed therebetween to suck in the reagent.

In addition, a water level sensor 120 is provided in the reagent inlet 100 or the reagent reservoir 300 of the present invention to detect whether the reagent level reaches a target level.

In addition, the reagents stored in any one of the reagent storage chambers 310 of the present invention are simultaneously sucked into the tip 710 attached to the injection tip 442 composed of two rows, and the reagents are drawn into the cylinder 441. The piston 431 enters and is dispensed into the cartridge 600 at the same time.

In addition, the dispensing syringe 400 of the present invention is a detachable plate 450 for separating the tip 710 mounted on the injection tip 442 while descending; And a push bar 432 that moves up and down together with the piston 431 and presses the detachable plate, and while the push bar 432 spaced apart from the detachable plate 450 descends, the detachable plate 450 The tip 710 is separated while the detachable plate 450 descends downward while being in contact with.

In addition, the present invention further includes a piston plate 430 for fixing the piston 431 and a cylinder plate 440 for fixing the cylinder 441, and the detachable plate 450 is the cylinder plate 440 When the push bar 432 descends, the push bar 432 penetrates the cylinder plate 440 and presses the detachable plate 450 to lower it, and the detachable plate 450 Silver descends from the outside of the injection tip 442 and separates the tip 710 coupled with the injection tip 442 .

The nucleic acid extracting apparatus for dispensing reagents in bulk of the present invention automatically dispenses reagents for extracting nucleic acids from biological samples into cartridges and inhales the reagents from the reagent reservoir by automatically performing the process of lifting and lowering the dispensing syringe manually. It has the advantage of minimizing errors occurring during manipulation and maintaining a constant quantity and quality of extracted nucleic acids.

In addition, the present invention has the advantage of being able to move the reagents dispensed in the nucleic acid extraction process in a state contained in a bulk bottle, so that sealing is easy and the sealing cost required for nucleic acid extraction can be reduced.

In addition, the present invention has the advantage of high versatility because it can be used for various samples by dispensing the type and amount of reagents to the cartridge at a set value according to the biological sample.

In addition, the present invention has the advantage of conveniently adjusting the dispensing capacity of the reagent because the amount of the reagent sucked into the tip is determined according to the height at which the piston of the dispensing syringe rises.

In addition, the present invention forms a partition wall protruding upward in the central part of the reagent storage chamber, which is a space supplied and stored from a bulk bottle, so that a small amount of reagent forms a high water level, so that it can be confirmed whether the reagent has flowed in by a target amount and the partition wall It has the advantage of being able to aspirate reagents from both sides of the center.

In addition, the present invention is provided with a plurality of tips that are detachable from the injection tip of the dispensing syringe, and the tips are separately installed according to the type of reagent to be sucked, thereby preventing different types of reagents from mixing with each other, thereby maintaining the quality of nucleic acid extraction. There are advantages to being

In addition, the present invention has the advantage of automating the attachment and detachment of the tip by separating the tip from the injection tip while the piston of the dispensing syringe further descends and discharges the remaining amount after dispensing the reagent inside the tip.

In addition, the present invention is provided with an extractor that separates and extracts nucleic acids from a sample at a location opposite to the dispensing syringe to automatically lift and move, so that the reagent dispensing and nucleic acid extraction processes can be performed simultaneously, significantly reducing the time required for nucleic acid extraction. There are advantages to being

1 is an overall perspective view of a nucleic acid extracting device in which reagents are dispensed in bulk according to the present invention.

Figure 2 is a perspective view showing a reagent take-in device in the nucleic acid extraction device in which reagents are dispensed in bulk according to the present invention.

Figure 3 is a perspective view showing a reagent reservoir in the nucleic acid extraction device in which reagents are dispensed in bulk according to the present invention.

Figure 4 shows a reagent reservoir in the nucleic acid extraction device in which reagents are dispensed in bulk of the present invention, (a) is a plan view, (b) is a front view.

Figure 5 is a perspective view showing a dispensing syringe in the nucleic acid extraction device in which reagents are dispensed in bulk according to the present invention.

Figure 6 shows the operating state of the dispensing syringe in the nucleic acid extraction device in which reagents are dispensed in bulk according to the present invention, (a) is a rear view before descending, (b) is a state in which the lift plate is lowered by the first motor rear view of

Figure 7 is a partially enlarged view to show the operating state of the dispensing syringe in the nucleic acid extraction device in which reagents are dispensed in bulk of the present invention, (a) is a front view of a state in which the lift plate is lowered by the first motor, (b) ) is a front view of a state in which the piston plate is lowered by the second motor.

Figure 8 is a partially enlarged view to show the operating state of the dispensing syringe in the nucleic acid extraction device in which reagents are dispensed in bulk of the present invention, (a) is a front view of a state in which the tip is mounted on the injection tip, (b) is a front view A front view of the detachable plate separating the tip.

9 is a perspective view of a state in which a cartridge, a storage cartridge, and a tip plate are provided on a support plate in the nucleic acid extraction device in which reagents are dispensed in bulk according to the present invention.

Figure 10 is a perspective view showing an extractor in the nucleic acid extracting device in which reagents are dispensed in bulk according to the present invention.

Hereinafter, with reference to the accompanying drawings, a preferred embodiment of the present invention will be described so that those skilled in the art can easily implement it.

As shown in FIGS. 1 to 10, the nucleic acid extracting apparatus for dispensing reagents in bulk according to the present invention is an apparatus for extracting or purifying nucleic acids or biological substances from a biological sample using a polymerase chain reaction,

a reagent inlet 100 into which a bulk bottle 20 containing reagents is inserted, and reagents inside the bulk bottle 20 are discharged through a reagent hole 110 formed at a lower side;

a transfer line 200 having one end communicating with the reagent hole 110 so that the reagent flows along the hollow interior and controlling the flow of the reagent by a valve 210 provided on one side;

a reagent reservoir 300 in which a plurality of reagent storage chambers 310 having different sizes are formed to receive and store reagents from the transfer line 200;

The piston 431 is combined with the cylinder 441 to inhale and discharge the reagent stored in the reagent reservoir 300 while moving up and down, and the injection tip 442 is disposed at the end of the cylinder 441 in a plurality of rows and columns. A dispensing syringe 400;

A tip 710 that is disposed on the side of the reagent reservoir 300, is attached to the lower end of the injection tip 442 and moves, and the injection tip 442 descends and is inserted into the reagent reservoir 300 to suck the reagent. ) is placed in a plurality of rows and columns (700);

A cartridge 600 in which the reagent contained in the tip 710 is discharged while the piston 431 descends and contained therein, and nucleic acid extraction is performed; and

A storage cartridge 800 for storing the nucleic acid extracted from the cartridge 600; includes.

A nucleic acid extraction device for dispensing reagents in bulk according to the present invention is a device for extracting nucleic acid materials by manipulating and purifying a biological sample using a polymerase chain reaction. In order to extract a desired nucleic acid material, a biological sample undergoes various steps such as lysis, binding, washing, and eluting, and various reagents are introduced to perform the process.

The reagent is supplied in the form of a large-capacity bulk bottle 20, and the bulk bottle 20 is mounted on the reagent inlet 100 of the present invention and transferred to the reagent reservoir 300 through the transfer line 200. move The reagent is aspirated by the dispensing syringe 400 by a set amount and dispensed into the cartridge 600, and then the nucleic acid extraction process is performed in the extractor 500. Finally, the separated nucleic acid is moved to the storage cartridge 800 by the extractor 500 .

Since the entire process from the dispensing of reagents to the transfer of extracted nucleic acids is operated automatically and human intervention is minimized, nucleic acid materials can be extracted with more accurate quantity and quality.

In addition, since the reagent is supplied in the form of a large-capacity bulk bottle 20, there is no difficulty in sealing the reagent to prevent vaporization, and at the same time, the quality of nucleic acid extraction due to the vaporization of the reagent can be maintained constant without deterioration.

In addition, since reagents are dispensed according to values set according to the types and amounts of reagents required for nucleic acid extraction according to biological samples, nucleic acid materials can be extracted from various biological samples.

As shown in FIG. 2, the nucleic acid extracting apparatus for dispensing reagents in bulk according to the present invention is provided with a reagent inlet 100 on one side, which is inserted so that the inlet of the bulk bottle 20 containing a large amount of reagent faces downward. A plurality of bulk bottles 20 may be mounted in the reagent inlet 100, and five bulk bottles 20 are mounted according to an embodiment of the present invention, but is not limited thereto.

A reagent hole 110 through which the reagent inside the bulk bottle 20 passes is formed at the bottom of the reagent inlet 100 . The reagent hole 110 is formed in the center of each reagent inlet 100 . The lower part of each compartment in which the bulk bottle 20 is mounted is inclined downward toward the reagent hole 110 .

A water level sensor 120 for detecting whether the level of the reagent inside the bulk bottle 20 has reached a target level is further formed in the reagent inlet 100 . The water level sensor 120 is composed of a pair of a transmitter and a receiver.

When the reagent inside the bulk bottle 20 is reduced and the water level is formed lower than the height at which the water level sensor 120 is located, or when the reagent is exhausted, even if one of the water level sensors 120 transmits an electrical signal, there is no medium, so that the other Since it is not received by one water level sensor 120, it is possible to detect that the level of the reagent is not formed above a certain level.

The reagent inside the bulk bottle 20 flows through the reagent hole 110 and is supplied to the reagent reservoir 300 through the transfer line 200 . The transfer line 200 communicates with the reagent hole 110 and controls the flow of the reagent through the valve 210 provided on one side. Through the opening/closing operation of the valve 210, the reagent contained in the large-capacity bulk bottle 20 flows into the reagent reservoir 300 in an appropriate amount.

One end of the transfer line 200 is in communication with the reagent hole 110, so that the reagent flows into the bulk bottle 20. In addition, the other end of the transfer line 200 is connected to one side of the reagent reservoir 300 so that the reagent is supplied to the reagent reservoir 300 .

3 and 4, the reagent reservoir 300 forms a plurality of reagent storage chambers 310. The reagent storage chamber 310 may be formed in different sizes to store reagents in different amounts according to reagents.

The reagent storage chamber 310 is formed long in the longitudinal direction, and by forming different lengths, the amount of reagents that can be stored is different. Here, the reagent storage chamber 310 is formed to have the same height as all neighboring reagent storage chambers 310 .

The number of reagent storage chambers 310 is less than the number of transfer lines 200 by one. Therefore, according to an embodiment of the present invention, 5 reagent inlets 100 into which bulk bottles 20 are inserted are provided and 5 transfer lines 200 are also provided, and in the case of reagents with similar functions, even a little reagent mixing Since there is no deterioration in the performance of nucleic acid extraction, the number of reagent storage chambers 310 is less than the number of transfer lines 200 by one less than four.

Each reagent storage chamber 310 is connected to the transfer line 200 so that a reagent is introduced therein, and two transfer lines 200 are connected to any one reagent storage chamber 310 . Accordingly, different types of reagents are introduced into any one reagent storage chamber 310 .

The reagent flowing into the reagent storage chamber 310 to which the two transfer lines 200 are connected is a washing reagent for cleaning the cartridge 600, and the remaining amount of any one of the washing reagents is partially present inside the reagent storage chamber 310. Therefore, even if slightly mixed with another washing reagent, it does not cause an effect on its performance, so that it can be jointly stored in one reagent storage chamber 310.

A pair of water level sensors 120 are provided inside each reagent storage chamber 310 . The water level sensor performs a function of detecting whether the water level of the reagent in the reagent storage chamber 310 has reached a target height value.

The water level sensor 120 has a different height according to the size of the reagent storage chamber 310 provided therein. In FIG. 4, the height of the water level sensor 120 is shown as distances (a, b, c, d) from the upper surface. a is the height of the water level sensor 120 provided in the reagent storage chamber 310 having the smallest storage capacity, the water level sensor 120 is disposed at the highest position, and d is the reagent storage chamber having the largest storage capacity At the height of the water level sensor 120 provided in 310, the water level sensor 120 is disposed at the lowest position. In the reagent storage chamber 310 having a large storage capacity, there is an amount of reagent that can be sucked even if the water level is set to a lower level, so the water level sensor 120 checks the remaining amount of reagent at a lower position. Since the water level sensor 120 has the same technical concept as the water level sensor 120 provided in the reagent injector 100, a detailed description thereof will be omitted.

The reagent storage chamber 310 is formed with a barrier rib 311 protruding upward from a bottom surface. The barrier rib 311 is formed to be long along the length direction of the reagent storage chamber 310 but smaller than the length of the reagent storage chamber 310 . In addition, the barrier rib 311 does not come into contact with the inner side surface of the reagent storage chamber 310 and is formed independently in the central portion.

Reagents stored in the reagent storage chamber 310 have the same water level in the chamber while surrounding the partition wall 311 . Since the partition wall 311 does not divide the storage space, reagents located on one side of the partition wall 311 can move to the other side.

The barrier rib 311 is for forming a high level of the reagent stored in the reagent storage chamber 310, and since the amount of reagent required for nucleic acid extraction is small, it is contained in the storage space to check the presence or absence of the reagent. In order to check the water level for the reagent, a partition wall 311 is provided at the center so that the water level can be sufficiently high even for a small amount of reagent.

In addition, since the level of the reagent is sufficiently high inside the reagent storage chamber 310, the tip 710 inserted into the injection tip 442 of the dispensing syringe 400, which will be described later, contains the reagent stored inside the reagent storage chamber 310. It can be sufficiently immersed in the piston 431, and when sucking due to the rise of the piston 431, only the liquid can be sucked without the inflow of air.

When the dispensing syringe 400 located above the reagent reservoir 300 descends and sucks the reagent inside the reagent storage chamber 310, it descends while the tip 710 is attached to the injection tip 442. 710 are located on both sides of the partition wall 311 of the reagent storage chamber 310. Since the tips 710 are mounted in two rows, eight in each row, in the dispensing syringe 400, the tips 710 enter the inside of the reagent storage chamber 310, and the eight tips located in one row on one side of the partition wall 311 ( 710) enters, and eight tips 710 located in two rows enter the other side of the partition wall 311.

At this time, the distance between the ends of the tip 710 is greater than the width of the partition wall 311 and smaller than the width of the reagent storage chamber 310, so that the tip 710 is placed with the partition wall 311 therebetween. A total of 16 tips 710 enter both sides of the partition wall 311 and simultaneously suck the reagents stored in the reagent storage chamber 310 . Accordingly, reagents stored in one of the reagent storage chambers 310 are sucked while being divided into 16 tips 710 at the same time.

5 to 8 show a dispensing syringe in the nucleic acid extracting device in which reagents are dispensed in bulk according to the present invention.

The reagent stored in the reagent reservoir 300 is sucked by the dispensing syringe 400 located at the top. The dispensing syringe 400 inhales the reagent and dispenses it to each well 610 of the cartridge 600 .

As shown in FIGS. 5 to 8, the dispensing syringe 400 is provided to be movable in a vertical or horizontal direction, and sucks or dispenses reagents by lifting a piston 431, which moves in a horizontal direction. a dividing frame 410; an elevation plate 420 that moves up and down; a piston plate 430 fixed to a lower portion of a plurality of pistons 431 for sucking and dispensing reagents while moving up and down; Connected to the lift plate 420 by a connecting table L, a cylinder 441 in which the piston 431 moves inside is fixed on the upper part, and a needle 442 communicating with the cylinder 441 is provided on the lower part. It includes a cylinder plate 440 to be.

One side of the dividing frame 410 is coupled to move along the horizontal bar 10 disposed across the horizontal direction. In addition, the dividing frame 410 is coupled to the first belt B1 meshed with the motor M, and moves in the disposition direction of the horizontal bar 10 by the motor M.

Accordingly, the dispensing syringe 400 inhales the reagent from the reagent reservoir 300 and then moves to the top of the cartridge 600 to dispense the reagent into each well 610 .

A first motor M1 for lifting the elevator plate 420 up and down is provided on the upper side of the dispensing frame 410 . The first motor M1 rotates the first rotation shaft A1 formed long downward. The first rotating shaft A1 has a screw formed on its outer periphery and engages with the first connector C1 formed on the lifting plate 420 to move the lifting plate 420 up and down.

The first connector (C1) and connectors 2, 3, and 4, which will be described later, are configured to move up and down by receiving rotational motion from a rotating shaft, and convert power from rotational motion into linear motion.

The first connector (C1) has a first rotational shaft (A1) passing through the inside, and since the screw on the outer periphery of the first connector (C1) is engaged with the inner periphery of the first connector (C1), the first rotational shaft ( As A1) rotates by the first motor M1, the first connector C1 moves up and down.

Since the first connector C1 is coupled to the lift plate 420, the lift plate 420 moves up and down as the first connector C1 moves up and down.

The lifting plate 420 moves up and down and is located on the top of the cartridge 600, the top of the tip plate 700, or the top of the cartridge 600 to perform operations necessary for dispensing reagents.

The lifting plate 420, which moves up and down by the first motor M1 provided in the dispensing frame 410, is disposed inside the dispensing frame 410 and is coupled to the cylinder plate 440 by the connecting rod L. has been The cylinder plate 440 is disposed below the lift plate 420 at a predetermined interval, and the upper end of the connecting table L is coupled to the lift plate 420 and the lower end is coupled to the cylinder plate 440 . Therefore, the cylinder plate 440 can also move up and down according to the up and down movement of the elevating plate 420 .

A second motor M2 for moving the piston plate 430 up and down is provided at the center of the lift plate 420 . The second motor M2 is meshed with and connected to idle gears G provided on both sides of the lifting plate 420 through a timing belt B to rotate the idle gear G. A second rotational shaft A2 is formed that extends downward from the idle gears G on both sides, and a screw is formed on the outer periphery of the second rotational shaft A2.

The second rotation shaft A2 penetrates and engages the inside of the second connector C2 formed on the piston plate 430, and as the second rotation shaft A2 rotates, the second connector C2 transmits rotational motion. Take it and move yourself up and down in a straight line.

The piston plate 430 has a plurality of columns and rows of pistons 431 formed downwardly on the lower surface, and the piston 431 is coupled with the cylinder 441 to move up and down inside. When the piston 431 rises, the dispensing syringe 400 sucks in the reagent, and when the piston 431 descends, it can discharge the sucked reagent.

In addition, as shown in FIGS. 7 and 8 , a push bar 432 pushing a detachable plate 450 to be described later is formed on the piston plate 430 and extends downward.

The cylinder plate 440 disposed downwardly from the piston plate 430 by a predetermined distance is coupled to the lift plate 420 through a connecting table L. When the lift plate 420 moves up and down by the first motor M1, the cylinder plate 440 also moves up and down.

In the upper part of the cylinder plate 440, the cylinder 441 in which the piston 431 moves up and down is provided in the same row and row as the piston 431, and is disposed at the end of the cylinder 441 in the lower part. The injection tips 442 communicating with the cylinder 441 are arranged in the same plurality of rows and columns as the cylinder 441 . The lower end of the injection tip 442 is formed such that the tip 710 containing the reagent to be sucked is detachable.

A detachable plate 450 is provided on the lower surface of the cylinder plate 440 to further form a hole through which the injection tip 442 passes. The detachable plate 450 pushes and separates the tip 710 mounted on the injection tip 442 . As the piston plate 430 descends, the push bar 432 presses and pushes the detachable plate 450, and while the detachable plate 450 descends by the push bar 432, the tip mounted on the injection tip 442 Disconnect (710).

At this time, the push bar 432 is always spaced apart from the detachable plate 450 at a certain distance, and when the piston plate 430 descends by a certain distance, it comes into contact and further descends at the contacted descending height. When the detachable plate 450 is pushed down by the push bar 432, the tip 710 mounted on the injection tip 442 is pushed and separated.

After the detachable plate 450 descends to separate the tip 710 from the injection tip 442, the piston plate 430 equipped with the push bar 432 rises and the pressing force to press the detachable plate 450 increases. When removed, a restoring pin 451 for returning the detachable plate 450 to an original position is further provided. The restoring pin 451 is formed in an upper direction of the detachable plate 450 . Since the spring is located between the head of the restoring pin 451 and the cylinder plate 440, when the detachable plate 450 descends, the spring is compressed to store restoring force, and when the push bar 432 is pressurized, it is removed. The restoring force is released to return the detachable plate 450 to the lower surface of the piston plate 430.

In the dispensing syringe 400, the dispensing frame 410 is connected to the horizontal bar 10 and is responsible for horizontal movement, and the lifting plate 420 is the first motor M1 provided in the dispensing frame 410. It is responsible for moving up and down by When the lifting plate 420 moves up and down, the cylinder plate 440 and the piston plate 430 move up and down together.

The piston plate 430 moves up and down by the second motor M2, and at this time, the cylinder plate 440 is fixed without moving up and down by the second motor M2.

When the piston plate 430 is at a high position, the push bar 432 does not come into contact with the detachable plate 450, and when the piston plate 430 descends to a certain height, the push bar 432 moves to the cylinder plate 440. While penetrating, it comes into contact with the detachable plate 450. When further descending, the detachable plate 450 descends due to the pressure of the push bar 432 and separates and drops the tip 710 mounted on the injection tip 442.

Looking at the operation of the dispensing syringe 400 according to an embodiment of the present invention, the reagent contained in the bulk bottle 20 flows into the reagent reservoir 300 through the transfer line 200, and the reagent reservoir 300 The reagent stored in the reagent storage chamber 310 of ) is sucked from the dispensing syringe 400.

First, the dispensing syringe 400 is located on the top of the tip plate 700 having a plurality of tips 710, and while the lift plate 420, which is lifted by the first motor M1, moves down, the tip ( 710) is installed.

The dispensing syringe 400 moves to the top of the reagent reservoir 300 again, and the tip 710 inserted into the injection tip 442 is lowered into the reagent storage chamber 310 containing the reagent to be sucked, and the tip 710 is removed. dipped in reagent

As the second motor M2 rotates, the piston plate 430 rises, and the reagent is sucked into the tip 710. At this time, the rising height of the piston 431 is raised only to a height according to a predetermined value so that only the amount required for dispensing can be sucked. Since the capacity required for dispensing is different for each reagent, the rising height of the piston 431 is set differently for each reagent. Accordingly, the rising height of the piston 431 may be set differently by inputting the amount of reagents required according to the types of biological samples and reagents to the control unit (not shown).

In addition, it is preferable that the reagent is stored only inside the tip 710 without passing through the injection tip 442 and flowing into the cylinder 441 .

Then, the dispensing syringe 400 moves from the reagent reservoir 300 to the top of the cartridge 600 to be dispensed. An operation of dispensing the reagent contained in the tip 710 to each well 610 of the cartridge 600 by lowering and pressurizing the piston plate 430 is performed.

As shown in FIG. 9, a cartridge 600 is provided on the lower surface of the nucleic acid extracting apparatus for dispensing reagents in bulk according to the present invention, in which a process of extracting nucleic acids from a biological sample is performed. Each cartridge 600 is comprised of a number of wells 610. According to one embodiment of the present invention, a plurality of cartridges 600 composed of 96 wells 610 are provided.

In the cartridge 600, the reagent contained in the tip 710 is discharged while the piston 431 descends, and is contained therein.

In addition, the plurality of tips 710 arranged inside the tip plate 700 disposed on the side of the reagent reservoir 300 are arranged in a plurality of columns and rows. The tips 710 have the same number of rows as the scan tips 442 and more rows than the scan tips 442 .

The number of rows of the tips 710 is more than four times the number of reagent storage chambers in the number of rows of the injection tips 442 . According to one embodiment of the present invention, since the number of rows of injection tips 442 is two and four reagent storage chambers are provided, the number of rows of tips is eight.

The tip mounted on the injection tip 442 descends as the injection tip 442 descends and is inserted into the reagent reservoir 300 to aspirate the reagent.

Therefore, the injection tip 442 is equipped with a part of the tip 710 when aspirating and dispensing any one reagent, and when changing the reagent to be aspirated and dispensed, the previously mounted tip 710 is separated and other Mount some tips 710 located in rows to aspirate and dispense reagents.

Therefore, by classifying the tip 710 used according to the type of reagent, mixing of the residual amount of reagent remaining inside the tip 710 with other reagents can be prevented.

Reagent dispensing and nucleic acid extraction are performed in the cartridge 600, and a storage cartridge 800 for storing the extracted nucleic acid material is provided. The storage cartridge 800 is further provided with a cover 810 so as to safely store the extracted nucleic acid material to prevent foreign matter from entering.

The cartridge 600, the storage cartridge 800, and the tip plate 700 are coupled to the upper surface of the support plate 900, and the support plate 900 is moved in the horizontal direction by the motor M shown in FIG. formed to be able to

Therefore, the cartridge 600, the storage cartridge 800, and the tip plate 700 may be positioned directly below the dispensing syringe 400 and the extractor 500 through horizontal movement. The support plate 900 may be provided while being fixed without moving, and in this case, the dispensing syringe 400 and the extractor 500 are configured to be movable in a horizontal direction. It is sufficient if the dispensing syringe 400, the extractor 500, and the supporting plate 900 have a structure capable of relative movement to each other so that the nucleic acid extraction process can be performed.

As shown in FIG. 10, the other side of the dispensing syringe 400 is inserted into and withdrawn from the well 610 of the cartridge 600 in which the reagent is dispensed, and the extractor 500 for extracting nucleic acids can be lifted up and down. formed into a structure

The extractor 500 is coupled to the horizontal bar 10, the extraction frame 510 moving in the horizontal direction; A mounting rod plate 520 in which a plurality of magnet mounting rods 521 are formed downward; and a cover plate 530 having a plurality of mounting rod covers 531 into which the magnet mounting rods 521 are inserted and withdrawn.

The extraction frame 510 is coupled to the second belt (B2) on one side, and the second belt (B2) is moved by the motor (M). Therefore, the extraction frame 510 can move in the direction in which the horizontal bar 10 is elongated and reciprocate the cartridge 600 or each well 610, or the nucleic acid extracted from the cartridge 600 to the storage cartridge 800. can be moved.

On the upper side of the extraction frame 510, a third motor (M3) and a fourth motor (M4) for lifting the mounting rod plate 520 and the cover plate 530 up and down are disposed, and each motor is rotated around the central axis. A third rotation axis A3 and a fourth rotation axis A4 are formed.

The third rotary shaft A3 rotated by the third motor M3 has a screw formed on the outer periphery and is engaged through the third connector C3 provided on the mounting plate 520. When the third rotary shaft A3 is rotated by the third motor M3, the third connector C3 receives the rotational motion and converts it into linear motion to lift the mounting plate 520 up and down.

As the mounting rod plate 520 moves up and down, the magnet mounting rod 521 also moves up and down. The magnet mounting bar 521 is inserted into and pulled out of the mounting bar cover 531 .

Similarly, the fourth rotary shaft A4 rotated by the fourth motor M4 is coupled to the fourth connector C4 provided on the cover plate 530, and is rotated by the fourth motor M4. The cover plate 530 moves up and down. Since the lifting mechanism has the same technical concept as that described in the dispensing syringe 400 above, a detailed description thereof will be omitted.

As the cover plate 530 moves up and down, the mounting rod cover 531 is also inserted into and pulled out of the well 610 of the cartridge 600 . At this time, it moves independently of the magnet attaching bar 521, but moves mutually in conjunction with each other to perform a nucleic acid extraction operation. Since the nucleic acid extraction operation using the magnet mounting bar 521 and the mounting bar cover 531 borrows a conventional technique, the description of the process of extracting nucleic acid while moving each will be omitted.

A magnet is formed at the lower end of the magnet mounting rod 521 to collect nucleic acid material by magnetic force and reciprocates inside the mounting rod cover 810 . The upper end of the mounting rod cover 810 is open, and magnetized nucleic acid material can be attached to the outer surface by the magnetic force of the magnet mounting rod 521. In the extractor 500, the magnet mounting bar 521 and the mounting bar cover 531 may borrow conventional technical ideas and configurations.

In the present invention, the dispensing syringe 400 and the extractor 500 are provided on both sides separately and operate independently. After the reagent is dispensed into the well 610 of the cartridge 600 in the dispensing syringe 400, the reagent for the next process may be dispensed into another well 610, and in the extractor 500, the reagent is dispensed well ( 610) may perform a nucleic acid extraction process. Accordingly, reagent dispensing and nucleic acid extraction are performed independently and simultaneously, so that the time required can be greatly reduced.

A nucleic acid extraction process in the nucleic acid extracting apparatus in which reagents are dispensed in bulk according to the present invention will be described as an example.

To extract nucleic acids from biological samples, different types of bulk bottles 20 are mounted on the reagent take-in device 100. The reagent inside the bulk bottle 20 flows into the reagent hole 110 and flows into the reagent reservoir 300 through the transfer line 200 .

The reagent reservoir 300 receives reagents from the transfer line 200 connected to one side and stores them in the reagent storage chamber 310 . Due to the partition wall 311 formed in the central part of the reagent storage chamber 310, the reagent is mainly stored on both sides of the partition wall 311 and forms a high water level even in a small amount.

The dispensing syringe 400 moves to the top of the tip plate 700 along the horizontal bar 10, and the tip 710 is mounted on the injection tip 442 while the lifting plate 420 is lowered by the first motor M1. do. After that, it rises again and moves to the top of the reagent reservoir 300 .

The dispensing syringe 400 to which the tip 710 is attached to the injection tip 442 descends to a height at which the tip 710 is submerged in the reagent inside the reagent storage chamber 310 . As the piston plate 430, which is raised and lowered by the second motor M2, rises, the piston 431 rises inside the cylinder 441, and the reagent inside the reagent storage chamber 310 is sucked into the tip 710. . At this time, the height at which the piston 431 rises is increased by a predetermined height according to the amount of reagent required, and an appropriate amount of reagent is sucked.

In addition, the tips 710 are arranged in two rows and submerged into one of the reagent storage chambers 310 while the reagents are divided into a plurality of tips 710 and sucked simultaneously.

Thereafter, the dispensing syringe 400 rises and moves to the top of the cartridge 600, and then descends to the well 610 to be dispensed, and the reagent stored in the tip 710 is transferred to the well 610 while the piston plate 430 descends. ) is busy At this time, the reagent inside the tip 710 is simultaneously dispensed by the pressure of the piston 431 . At this time, the suction and dispensing operations are performed while the piston 431 moves up and down inside the cylinder 441 .

After the dispensing is finished, the tip 710 is exchanged for inhalation of different types of reagents. The tip 710 mounted on the injection tip 442 is separated by pushing the detachable plate 450 . Thereafter, a new tip 710 suitable for the reagent to be sucked is mounted on the injection tip 442 and moved to the reagent reservoir 300 to suck another type of reagent.

Then, the operation of dispensing to the well 610 of the cartridge 600 described above is repeated.

When the reagent is dispensed into the well 610 containing the biological sample, the mounting rod plate 520 and the cover plate 530 in the extractor 500 move up and down while the magnet mounting rod 521 and the mounting rod cover 810 move toward the cartridge 600. A series of operations for extracting nucleic acids (borrowed from prior art) are performed, and the extracted nucleic acids are attached to the outer surface of the attaching rod cover 531 and transferred to the storage cartridge 800.

In the nucleic acid extracting apparatus for dispensing reagents in bulk according to the present invention, reagents are not provided in the form of a cartridge 600 in which reagents are dispensed into each well 610 for nucleic acid extraction work, but are placed in a bulk bottle 20 and stored in a biological sample. Since the capacity is automatically dispensed according to the method, sealing is simple, nucleic acid extraction is possible quickly from various samples, and versatility is high.

In addition, partition walls 311 are formed in each reagent storage chamber 310 of the reagent reservoir 300, so even a small amount of reagent can be formed high, so that the reagent can be easily sucked from the dispensing syringe 400.

In addition, while the reagent is dispensed from the dispensing syringe 400 to the well 610 of the cartridge 600, the extractor 500 simultaneously performs a process for extracting nucleic acids from each well 610 to which the reagent is dispensed. It has the advantage of being able to extract nucleic acids quickly by reducing the time required for work.

[Description of code]

M: motor M1: 1st motor M2: 2nd motor M3: 3rd motor M4: 4th motor G: idle gear

A1: 1st rotational axis A2: 2nd rotational axis A3: 3rd rotational axis A4: 4th rotational axis

B: Timing belt B1: 1st belt B2: 2nd belt

C1: 1st connector C2: 2nd connector C3: 3rd connector C4: 4th connector

10: horizontal bar 20: bulk bottle

100: reagent inlet 110: reagent hole 120: water level sensor

200: transfer line 210: valve

300: reagent reservoir 310: reagent storage chamber 311: bulkhead

400: dispensing syringe 410: dispensing frame 420: lift plate 430: piston plate 431: piston

432: Push bar 440: Cylinder plate 441: Cylinder 442: Injection tip 450: Detachable plate 451: Restoration pin

500: Extractor 510: Extraction frame 520: Mounting plate 521: Magnet mounting rod 530: Cover plate

531: mounting rod cover

600: cartridge 610: well 700: tip plate 710: tip 800: storage cartridge 810: cover

900: support plate

Claims (8)

1. a reagent inlet 100 into which a bulk bottle 20 containing reagents is inserted, and reagents inside the bulk bottle 20 are discharged through a reagent hole 110 formed at a lower side;

   a transfer line 200 having one end communicating with the reagent hole 110 so that the reagent flows along the hollow interior and controlling the flow of the reagent by a valve 210 provided on one side;

   a reagent reservoir 300 in which a plurality of reagent storage chambers 310 having different sizes are formed to receive and store reagents from the transfer line 200;

   The piston 431 is combined with the cylinder 441 to inhale and discharge the reagent stored in the reagent reservoir 300 while moving up and down, and the injection tip 442 is disposed at the end of the cylinder 441 in a plurality of rows and columns. The dispensing syringe 400;

   A tip 710 that is disposed on the side of the reagent reservoir 300, is attached to the lower end of the injection tip 442 and moves, and the injection tip 442 descends and is inserted into the reagent reservoir 300 to suck the reagent. ) is placed in a plurality of rows and columns (700);

   A cartridge 600 in which the reagent contained in the tip 710 is discharged while the piston 431 descends and contained therein, and nucleic acid extraction is performed; and

   A storage cartridge 800 for storing the nucleic acid extracted from the cartridge 600;

   A nucleic acid extraction device that dispenses reagents in bulk.
2. According to claim 1,

   The number of rows of tips 710 placed on the tip plate 700 is equal to the number of rows of injection tips 442 of the dispensing syringe 400,

   The number of rows of the tips 710 is greater than the number of rows of the injection tips 442.

   A nucleic acid extraction device that dispenses reagents in bulk.
3. According to claim 1,

   The reagent storage chamber 310 has the same height as the adjacent reagent storage chamber 310, but its length is different from each other.

   A nucleic acid extraction device that dispenses reagents in bulk.
4. According to claim 1,

   A partition wall 311 protruding upward is formed at the center of the reagent storage chamber 310, and the partition wall 311 is smaller than the length of the reagent storage chamber 310 so that the reagents surround the partition wall 311 and have the same height. form the level of

   The tip 710 attached to the injection tip 442 is composed of two rows, and the distance between the rows at the end of the tip 710 is greater than the width of the partition wall 311 and the width of the reagent storage chamber 310. It is smaller than the partition wall 311 and descends to both sides to inhale the reagent.

   A nucleic acid extraction device that dispenses reagents in bulk.
5. According to claim 1,

   In the reagent inlet 100 or the reagent reservoir 300,

   A water level sensor 120 is provided to detect whether the level of the reagent has reached a target height value.

   A nucleic acid extraction device that dispenses reagents in bulk.
6. According to claim 1,

   The reagent stored in one of the reagent storage chambers 310 is simultaneously sucked into the tip 710 attached to the injection tip 442 formed of two rows, and the piston 431 moves toward the inside of the cylinder 441. enters and is simultaneously dispensed into the cartridge 600

   A nucleic acid extraction device that dispenses reagents in bulk.
7. According to claim 1,

   The dispensing syringe 400 is

   A detachable plate 450 separating the tip 710 mounted on the injection tip 442 while descending; And a push bar 432 that moves up and down together with the piston 431 and presses the detachable plate,

   While the push bar 432 spaced apart from the detachable plate 450 descends and contacts the detachable plate 450, the detachable plate 450 descends downward to separate the tip 710.

   A nucleic acid extraction device that dispenses reagents in bulk.
8. According to claim 7,

   Further comprising a piston plate 430 for fixing the piston 431 and a cylinder plate 440 for fixing the cylinder 441,

   The detachable plate 450 is disposed under the cylinder plate 440,

   When the push bar 432 descends, the push bar 432 penetrates the cylinder plate 440 and presses the detachable plate 450 to lower it, and the detachable plate 450 moves the injection tip 442 Descending from the outside of and separating the tip 710 coupled with the injection tip 442

   A nucleic acid extraction device that dispenses reagents in bulk.

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