Kit integrated with waste liquid bag, liquid path system and gene sequencer

Abstract

The utility model provides a kit integrated with a waste liquid bag, a liquid path system and a gene sequencer, and relates to the technical field of gene sequencer, the kit integrated with the waste liquid bag comprises a kit body, a reagent bag and a waste liquid bag, the reagent bag and the waste liquid bag are arranged in the kit body, and the reagent bag and the waste liquid bag are arranged in the kit body. The internal space of the kit body is arranged to at least contain the reagent bag in a full-load state, the reagent bag and the waste liquid bag respectively comprise a flexible bag and a reagent hole, and the reagent holes are formed in the kit body. According to the technical scheme, the technical problems that in the prior art, reagents in a kit are inconvenient to suck, and the space occupied by the kit and a waste liquid box is large are solved.

Description

TECHNICAL FIELD

[0001] The utility model relates to the field of gene sequencing technology, especially to the reagent box of integrated waste liquid bag, liquid path system and gene sequencer. BACKGROUND

[0002] The reagent box of gene sequencer usually has multiple reagent hole positions, and is a fixed structure, usually a needle or a pipette pierces the reagent hole sealing device from top to bottom to extend into the reagent box to suck reagent, which needs a certain stroke, and the negative pressure of the needle sucking reagent is provided by the pump or other power device at the rear end. The instrument also needs to be equipped with a corresponding waste liquid box, and the volume of the waste liquid box is not less than the volume of the reagent box. The reagent box is usually disposable, and the waste liquid box can be reused, which is very dirty and difficult to clean after long-term use.

[0003] The stroke of the needle piercing the reagent hole sealing device will significantly increase the volume of the instrument.

[0004] The reagent box and the waste liquid box are both fixed volume structures, and the reagent box and the waste liquid box occupy a large volume of the instrument. If the volume of the reagent box and the waste liquid box is approximately equal to the volume, the volume of the waste liquid box should be not less than the volume of the reagent box. Ideally, assuming that the volume of the reagent box and the waste liquid box is V0, the total space of the reagent box and the waste liquid box dedicated instrument is a fixed value 2*V0.

[0005] The current sequencer usually uses negative pressure sampling, and excessive negative pressure is easy to cause bubbles to be precipitated in the liquid path system, which limits the liquid suction speed and has a great influence on the whole sequencing time. SUMMARY

[0006] The utility model provides the reagent box of integrated waste liquid bag, liquid path system and gene sequencer to solve the technical problem of inconvenient reagent suction of the reagent box in the prior art, and large space occupied by the reagent box and the waste liquid box.

[0007] On the one hand, the utility model provides the reagent box of integrated waste liquid bag, including box body, reagent bag and waste liquid bag, the reagent bag and waste liquid bag set up in the box body, the inside space of the box body is arranged as at least can contain the reagent bag of full load state, the reagent bag and waste liquid bag all include flexible bag and reagent hole, the reagent hole sets up on the box body.

[0008] Optionally, a movable partition is arranged between the reagent bag and the waste liquid bag.

[0009] Optionally, an elastic device is arranged on one side of the partition to enable the partition to obtain a force towards the side of the reagent bag.

[0010] Optionally, the reagent hole is clamped on the box body.

[0011] Optionally, the reagent hole is slidingly arranged on the box body.

[0012] Optionally, the reagent bags are arranged in multiple, the multiple reagent bags are arranged side by side, and the waste liquid bag is arranged side by side with the reagent bags or is arranged on the side of the reagent bags in the vertical direction of the reagent bags.

[0013] Optionally, the reagent hole opening of the reagent bag is sealed by a sealing part.

[0014] Optionally, a quick release joint is arranged at the reagent hole opening.

[0015] In another aspect, the utility model provides a liquid path system, including the reagent box of integrated waste liquid bag as any above described, the liquid inlet end of the liquid path system is connected the reagent hole of the reagent bag, and the waste liquid discharge end of the liquid path system is connected the reagent hole of the waste liquid bag.

[0016] In still another aspect, the utility model provides a gene sequencer, including the liquid path system as described above.

[0017] In order to make the features and advantages of the present application more obvious and easy to understand, some embodiments are described below, and the accompanying drawings are described in detail as follows. BRIEF DESCRIPTION OF DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the utility model or the prior art, the following will briefly introduce the drawings needed to be used in the embodiment or prior art description, and obviously, the drawings in the following description are only some embodiments of the utility model, and for those skilled in the art, other drawings can also be obtained according to these drawings without creative labor.

[0019] Figure 1 It is a structural schematic view of the gene sequencer;

[0020] Figure 2 It is a front view of the reagent box of integrated waste liquid bag according to an embodiment of the utility model;

[0021] Figure 3 It is a right view of the reagent box of integrated waste liquid bag according to an embodiment of the utility model;

[0022] Figure 4 It is a top view of the reagent box of integrated waste liquid bag according to an embodiment of the utility model;

[0023] Figure 5 It is a structural schematic view of the reagent box of integrated waste liquid bag according to an embodiment of the utility model in the state that the reagent bag is empty and the waste liquid bag is full;

[0024] Figure 6A structure schematic view of a reagent hole part of a reagent box integrated with a waste liquid bag according to an embodiment of the present utility model;

[0025] Figure 7 A structure schematic view when a reagent hole of a reagent box integrated with a waste liquid bag according to an embodiment of the present utility model is connected with a liquid path;

[0026] Figure 8 A top view of a structure of a reagent box integrated with a waste liquid bag according to another embodiment of the present utility model;

[0027] Figure 9 A side view of a reagent box integrated with a waste liquid bag according to still another embodiment of the present utility model.

[0028] In the figure: 1, box body;2, reagent bag;2-1, liquid outlet hole;2-2, sealing part 3, waste liquid bag;3-1, liquid inlet hole;4, partition;5, spring;10, chip;20, chip platform;30, reagent storage container;40, flow guide system;50, optical detection system;60, computer system. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical scheme and advantages of the present utility model more clear, the technical scheme of the present utility model will be described in detail below. Obviously, the described embodiments are only a part of the embodiments of the present utility model, not all the embodiments. Based on the embodiments in the present utility model, all other embodiments obtained by those skilled in the art without creative labor belong to the scope of protection of the present utility model.

[0030] It should be noted that when an element is referred to as being "fixed to" or "disposed on" another element, it can be directly on the other element or a middle element can be present at the same time. When an element is referred to as being "connected to" another element, it can be directly connected to the other element or a middle element can be present at the same time.

[0031] Figure 1 A structure schematic view of a gene sequencer. As shown in Figure 1 The gene sequencer provided by the present disclosure includes: a chip 10, a chip platform 20, a reagent storage container 30, a flow guide system 40, an optical detection system 50 and a computer system 60.

[0032] Among them, the chip 10 is attached with one or more sequencing objects;Here, the sequencing object can be a DNA / RNA fragment, which includes a certain length of base sequence, for example, 150bp length;The sequencing object can also be a DNA / RNA molecule.

[0033] The chip platform 20 is configured to fix and support the chip 10;

[0034] a reagent storage container 30 configured to store one or more reagents; here, the reagent can exemplarily include a polymerase chain reaction (PCR) fluorescent reagent.

[0035] a flow guiding system 40 configured to controllably deliver the one or more reagents from the reagent storage container 30 into the chip 10 so as to be in contact with the sequencing object and to occur a chemical reaction, so that the sequencing object is labeled with a fluorescent label;

[0036] an optical detection system 50 configured to excite the fluorescent label on the sequencing object and to detect a fluorescent signal generated by the excited fluorescent label;

[0037] a computer system 60 configured to acquire a fluorescent image from the optical detection system 50 and to identify a gene sequence of the sequencing object according to the fluorescent image.

[0038] Regarding the reactions related to the liquid system in the gene sequencer, the following are exemplarily amplification reactions and sequencing reactions.

[0039] Amplification reaction

[0040] Taking the amplification of a DNA library by bridge PCR (Polymerase chain reaction) as an example, after the library is constructed (usually, the library construction is completed outside the sequencer, for example, the library is obtained by experimental operation in the laboratory), the library can be inoculated on the sequencing chip and amplified on the sequencing chip. Among them, the adapters at both ends of the library are complementary to the first amplification primer on the sequencing chip, so the library can be inoculated on the sequencing chip by complementary hybridization.

[0041] After the library is inoculated onto the sequencing chip, an amplification reaction can be performed using the library as a template strand. For example, the amplification reaction can be performed by first adding dNPs and polymerase to the sequencing chip. The polymerase will synthesize a new DNA strand from the first amplification primer, which is completely complementary to the template strand, and the new DNA strand will be covalently attached to the sequencing chip. Next, NaOH solution is added to the sequencing chip to wash away the template strand, and the complementary strand remains covalently attached to the sequencing chip. A neutral solution is then added to neutralize the NaOH solution, and the entire environment in the sequencing chip becomes neutral. The other end of the complementary strand then hybridizes with the second amplification primer on the sequencing chip. dNPs and polymerase are added, and the polymerase synthesizes a new DNA strand from the second amplification primer, which is completely complementary to the complementary strand and identical to the template strand. NaOH solution is then added to separate the two strands, and two strands that are covalently attached to the sequencing chip and complementary to each other are obtained. This process is repeated, and the number of DNA strands increases exponentially.

[0042] After amplification, both DNA strands identical to the template strand and the complementary strand are retained on the sequencing chip. A specific reagent is then added to the sequencing chip to cleave the DNA strand synthesized from one of the amplification primers, for example, the DNA strand identical to the complementary strand. The DNA strand identical to the template strand is retained. NaOH solution is then added to the sequencing chip to separate the two strands, and the cleaved DNA strand is washed away with the NaOH solution. Finally, only DNA single strands are retained on the sequencing chip, and the number of DNA single strands is exponentially increased from the beginning of the amplification. A DNA cluster is formed, and all DNA single strands in the DNA cluster are identical. A neutral solution is then added to sequence all DNA single strands in the DNA cluster in a neutral solution environment.

[0043] It should be noted that the above amplification reaction can be completed outside the sequencer, such as amplifying the library through experimental operation in the laboratory, or can be completed inside the sequencer. When the amplification reaction is completed inside the sequencer, the library and various reaction reagents (for example, dNP, polymerase, NaOH alkali solution, neutral solution, etc.) participating in the amplification reaction can be added to the sequencing chip through the liquid channel system of the sequencer. In addition, the above amplification reaction is only exemplary, and the present application is not limited to the bridge PCR amplification method, and other amplification methods such as loop-mediated isothermal amplification (LAMP, loop-mediated isothermal amplification), nucleic acid sequence-based amplification (NASBA, Nuclear acid sequence-based amplification), rolling circle amplification (RCA, Rolling Circle Amplification), multiplex probe amplification (MPA, Multiplex Probe Amplification) and the like can also be used for amplification.

[0044] sequencing reaction

[0045] For example, using the principle of sequencing by synthesis, when sequencing, 4 dNTPs with fluorescent groups are added to the sequencing chip through the liquid channel system, each dNTP can only be synthesized with one of the four bases ATCG, and the 3' end of the dNTP has been blocked by a blocking group (the blocking group includes but is not limited to azido group). Then, through the liquid channel system, polymerase is continuously added to the sequencing chip. Through the action of the polymerase, one of the four dNTPs will be synthesized with the complementary base of the single strand being sequenced. Since the 3' end of the dNTP is blocked by a blocking group, only one dNTP can be elongated on the single strand being sequenced each time. After synthesis, a specific chemical reagent is added to the sequencing chip through the liquid channel system, thereby washing away the excess dNTP and polymerase. Next, the fluorescent group of the dNTP synthesized on the single strand can be excited by the optical detection system, so that the fluorescent group emits a fluorescent signal. Since the fluorescent group of each dNTP on a cluster in the single strand emits the same fluorescent signal, the fluorescent signal is amplified, and therefore the optical detection system can collect the fluorescent signal and generate a fluorescent image.

[0046] The fluorescent image is processed and analyzed by a computer system to determine which dNTP is synthesized on the single strand being sequenced, and then according to the complementary principle, it can be deduced which base on the single strand being sequenced is synthesized with the dNTP. Thus, one sequencing cycle is completed.

[0047] Down the road through the liquid path system to the sequencing chip to add specific chemical reagents, will be cut off the blocking group and fluorescent group, and then the 3' end of the hydroxyl group of dNTP is exposed.

[0048] Next into the next sequencing cycle, and repeat the above process.

[0049] It can be understood that a sequencing cycle can detect a base, and after multiple sequencing cycles, multiple bases in the single strand to be sequenced can be detected. Specifically, the number of sequencing cycles can be determined according to the set sequencing read length, for example, 150, 300 sequencing cycles.

[0050] Of course, it should be noted that the above sequencing reaction is only exemplary, and the present application is not limited to using the principle of sequencing by synthesis, but also other sequencing principles.

[0051] In Figure 1 In the structure of the gene sequencer shown in the figure, the reagents in the reagent storage container 30 are transported to the relevant parts of the gene sequencer through the liquid path system in the flow guide system 40, and the waste liquid is returned to the actual storage container 30 through the liquid path system. The reagents in the actual storage container 30 are stored in the reagent box, and the waste liquid is stored in the waste liquid box. In the prior art, the reagent in the reagent box needs to be extracted by a long needle to suck liquid, and related structures need to be set to balance the negative pressure generated by the reagent flowing out of the reagent box. The structure is complex, the reagent is not easy to suck, and the reagent box and the waste liquid box occupy a large volume of the instrument.

[0052] To solve the above technical problems, the present application provides a reagent box integrated with a waste liquid bag.

[0053] Figures 2 to 4 As shown in the figure, the structure of the reagent box integrated with the waste liquid bag of the embodiment of the utility model is shown.

[0054] The utility model provides a reagent box integrated with a waste liquid bag, including box body 1, reagent bag 2 and waste liquid bag 3, reagent bag 2 and waste liquid bag 3 are arranged in box body 1, the inside space of box body 1 is set as at least can accommodate the reagent bag 2 of full load state, reagent bag 2 and waste liquid bag 3 all include flexible pouch and reagent hole, reagent hole is set up on box body 1.

[0055] Specifically, the reagent and waste liquid storage device are both bags made of flexible material, the reagent bag 2 and the waste liquid bag 3 are accommodated by the reagent box box body 1, the box body 1 is made of ordinary solid material, and the reagent bag 2 and the waste liquid bag 3 can be arranged in an upper-lower or left-right structure, such as Figure 8 、 Figure 9Arrangement. The inside space of the box 1 can accommodate the full reagent bag 2, so that the reagent bag 2 can be full of reagent in the initial state, and the waste liquid bag 3 is empty. During use, the reagent is extracted while the waste liquid is injected into the waste liquid bag 3, which not only meets the storage and supply of reagent, but also solves the storage of waste liquid reflux, and the space for storing reagent and waste liquid is overlapped, thereby saving the space occupied by the waste liquid box and the reagent box.

[0056] The reagent bag 2 of flexible material can be deformed to balance the pressure inside and outside the bag during liquid absorption in the liquid path system, without being pierced to contact the reagent with the atmosphere. The waste liquid bag 3 can also be discharged without being pierced. Therefore, the reagent box in the present application does not need to be pierced by a needle during use, and does not need to be designed with corresponding mechanism movement stroke. Of course, the hole positions on the reagent box can also be designed in the form of needing to be pierced by a needle, and the needle can be sealed after being pierced, and can be pierced upward, downward, leftward, rightward or obliquely. The length of the needle does not need to be too long, and the needle can pass through the reagent hole.

[0057] Optionally, the space occupied by the reagent bag 2 and the waste liquid bag 3 in the box 1 is complementary.

[0058] Specifically, in the initial state, the reagent bag 2 is full, and the waste liquid bag 3 is empty. The liquid path system continuously extracts the reagent in the reagent bag 2, part of the volume of the reagent fills the liquid path system, and the rest is gradually discharged into the waste liquid bag 3. The above steps are equivalent to the reagent in the reagent bag 2 being gradually transferred to the waste liquid bag 3, and the total volume of the reagent bag 2 and the waste liquid bag 3 does not change substantially during this process. The total volume of the reagent bag 2 and the waste liquid bag 3 is approximately equal to the volume of the reagent box. Assuming that the initial volume of the reagent bag 2 is V0, the volume of the waste liquid bag 3 is 0, and the volume of the reagent box is V0. During the operation of the reagent box, the volume of the reagent bag 2 changes from V0 to 0, and the volume of the waste liquid bag 3 changes from 0 to V0, that is, Figure 2 to the state shown in Figure 5 .

[0059] Optionally, a movable partition 4 is arranged between the reagent bag 2 and the waste liquid bag 3.

[0060] Specifically, reagent bag 2 and waste liquid bag 3 can be in direct contact or can be designed with a partition 4. The partition 4 divides the reagent kit into at least two parts, one part containing reagent bag 2 and the other part containing waste liquid bag 3. In the design where the partition 4 is pressurized within the reagent kit, when the liquid path system uses negative pressure to aspirate liquid from reagent bag 2, the positive pressure applied by the partition 4 to reagent bag 2 can offset part of the negative pressure, reducing the volume and number of reagent bubbles precipitated within the liquid path system. It also helps to further improve the aspiration speed at the design level without excessively increasing the negative pressure within the liquid path system.

[0061] Optionally, an elastic device is provided on one side of the partition 4 so that the partition 4 receives a force toward the reagent bag 2.

[0062] Specifically, the partition 4 on the side of the waste liquid bag 3 can be designed with a spring 5 or other device that can apply a certain pressure to the partition 4, so that the reagent bag 2 can withstand a certain squeezing force during use.

[0063] Optionally, the reagent orifice is snapped onto the housing 1.

[0064] Specifically, such as Figure 6 As shown, a slot is provided on the reagent well, and the reagent well is engaged with the reagent kit through the slot.

[0065] Optionally, the reagent orifice is slidably disposed on the housing 1.

[0066] Specifically, because the volume of reagent bag 2 and waste liquid bag 3 changes during use, the position of the reagent hole connected to reagent bag 2 and waste liquid bag 3 may need to be moved. Therefore, the reagent hole is slidably mounted on the box body 1. As for the power for the sliding of the reagent hole on the box body 1, it can be driven by the volume change of reagent bag 2 and waste liquid bag 3 themselves, or a separate driving component can be provided to actively drive the sliding of the reagent hole according to the volume change of reagent bag 2 and waste liquid bag 3 during operation.

[0067] Optionally, multiple reagent bags 2 are provided, arranged side by side, and the waste liquid bag 3 is arranged side by side with the reagent bags 2, such as... Figure 9 As shown; or the waste liquid bag 3 is arranged on the side of the reagent bag 2 along the vertical direction of the arrangement of the reagent bags 2, such as... Figure 3 As shown.

[0068] Specifically, waste liquid bag 3 requires only one flexible sac to contain waste liquid, while reagent bag 2 requires one or more (usually multiple) flexible sacs to contain different reagents. Multiple reagent flexible sacs can be arranged horizontally or as required by design.

[0069] Optionally, the reagent hole opening of the reagent bag 2 is sealed by the sealing part 2-2.

[0070] Specifically, as shown in Figure 6 、 Figure 7 For the design of the kit and the liquid path system in a split type, the reagent holes of the kit can be designed at any position required on the kit, each hole position is designed to be sealed and docked with the liquid path system, the reagent holes of the kit are sealed by a sealing part 2-2 made of flexible material to prevent leakage of reagents during transportation of the kit. The liquid path system is provided with a short needle, the short needle can pierce the sealing part 2-2 made of flexible material, the sealing part 2-2 made of flexible material wraps the short needle to form a seal, and the short needle has a flange-like structure to extrude and strengthen the seal with the sealing material. After the kit is connected with the liquid path system, the connection is maintained. The waste liquid bag 3 and the reagent bag 2 are discarded together after use, and there is no need to consider the cleaning or pouring of the waste liquid box.

[0071] For the design of the kit and the liquid path system in an integrated type, the kit and the liquid path system are an integral whole, and there is no need to consider the docking and separation of the kit and the liquid path system, and the kit and the liquid path system can be discarded as a whole after use.

[0072] In some embodiments, the reagent holes of the reagent bag 2 and the waste liquid bag 3 are sealed by a sealing part 2-2 at the opening of the reagent holes. In this way, the sealing of the waste liquid bag and the leakage of the waste liquid in the waste liquid bag can be ensured.

[0073] Optionally, a quick release connector is arranged at the opening of the reagent hole.

[0074] By arranging the quick release connector at the opening of the reagent hole, the kit 1 can be conveniently connected or disconnected with the liquid path system of the sequencer. The above-mentioned quick release connector can be a mature quick release connector in the prior art, such as a buckle structure.

[0075] In another aspect, the utility model provides a liquid path system, including the reagent kit of integrated waste liquid bag 3 as any above described, the liquid inlet end of the liquid path system is connected with the reagent hole of the reagent bag 2, and the waste liquid discharge end of the liquid path system is connected with the reagent hole of the waste liquid bag 3.

[0076] In still another aspect, the utility model provides a gene sequencer, including the liquid path system as described above.

[0077] Based on the above technical scheme, the following beneficial effects are obtained:

[0078] (1) The total volume of the kit and the waste liquid box is reduced, the stroke required for the needle to pierce the reagent hole is reduced, and finally the volume of the instrument is reduced.

[0079] (2) The inner partition plate 4 of the kit with the initial pressure design can provide a certain positive pressure for the liquid inlet, improve the liquid inlet efficiency, shorten the sequencing time and reduce the bubble release.

[0080] (3), the waste liquid box is disposable, convenient and clean.

[0081] In the description of the utility model, it is explained that, the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "internal", "external" and the like indicate the orientation or position relationship based on the orientation or position relationship shown in the drawings, and are only for the convenience of describing the utility model and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as limiting the utility model. In addition, the terms "first", "second", "third" are only for the purpose of description, and cannot be understood as indicating or implying relative importance.

[0082] In the description of the utility model, it is explained that, unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connection" should be broadly understood, for example, it can be fixed connection, or detachable connection, or integrally connected; can be mechanical connection, or electrical connection; can be directly connected, or indirectly connected through an intermediate medium, can be the communication between two elements. For ordinary skilled in the art, the specific meaning of the above terms in the utility model can be understood according to the specific circumstances.

[0083] The above is only a specific embodiment of the utility model, but the protection scope of the utility model is not limited to this, any skilled in the art can easily think of changes or replacements within the technical range disclosed by the utility model, which should be covered in the protection scope of the utility model. Therefore, the protection scope of the utility model should be subject to the protection scope of the claims.

Claims

1. A reagent kit with an integrated waste liquid bag, characterized in that, The device includes a box body, reagent bags, and waste liquid bags. The reagent bags and waste liquid bags are disposed within the box body. The internal space of the box body is configured to accommodate the reagent bags at least when fully loaded. Each reagent bag and waste liquid bag includes a flexible pouch and a reagent hole, which is located on the box body.

2. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, A movable partition is provided between the reagent bag and the waste liquid bag.

3. The reagent kit with an integrated waste liquid bag according to claim 2, characterized in that, An elastic device is provided on one side of the partition so that the partition receives a force toward the reagent bag.

4. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, The reagent port is snapped onto the box body.

5. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, The reagent orifice is slidably disposed on the box body.

6. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, The reagent bags are configured in multiple ways, and the multiple reagent bags are arranged side by side. The waste liquid bag is arranged side by side with the reagent bags or is arranged on the side of the reagent bags along the vertical direction of the arrangement of the reagent bags.

7. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, The reagent bag's reagent hole opening is sealed by a sealing part.

8. The reagent kit with an integrated waste liquid bag according to claim 1, characterized in that, A quick-release connector is provided at the opening of the reagent well.

9. A fluid circuit system, characterized in that, The reagent kit includes an integrated waste liquid bag as described in any one of claims 1-8, wherein the inlet end of the liquid path system is connected to the reagent port of the reagent bag, and the waste liquid outlet end of the liquid path system is connected to the reagent port of the waste liquid bag.

10. A gene sequencer, characterized in that, Includes the fluid system as described in claim 9.

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