CN114682310A

CN114682310A - Liquid path system, sequencing system and method

Info

Publication number: CN114682310A
Application number: CN202011617771.7A
Authority: CN
Inventors: 吴平; 万雪峰; 姜泽飞
Original assignee: Genemind Biosciences Co Ltd
Current assignee: Genemind Biosciences Co Ltd
Priority date: 2020-12-31
Filing date: 2020-12-31
Publication date: 2022-07-01
Anticipated expiration: 2040-12-31
Also published as: CN114682310B

Abstract

The application discloses a liquid path system, a sequencing system and a method, wherein the liquid path system comprises: a plurality of flow paths fluidly connected to the flow cell to support a target analyte when the flow cell is installed in the fluidic system; a flow path selector valve connected to the flow path, the flow path selector valve including a manifold provided with a common port, a plurality of first ports and a plurality of second ports, and a first spool provided with a communication groove, the first spool being rotatable or slidable relative to the manifold, the first ports being communicated with the second ports through the communication groove when the flow path selector valve is in the first valve position, the first ports being communicated with the common port through the communication groove when the flow path selector valve is in the second valve position; a pump fluidly connected to the flow cell when the flow cell is installed in the fluid path system; and a control circuit operatively coupled to the flow path selection valve, the control circuit for controlling the flow path selection valve to select a specified flow path.

Description

Liquid path system, sequencing system and method

Technical Field

The present application relates to the field of mechanical and fluidic technology, and more particularly to a fluid path system, a sequencing system and a method.

Background

In the related art, nucleic acid detection of a nucleic acid sample disposed on a solid substrate, for example, nucleic acid sequencing (sequencing) based on chip detection, generally includes sample processing before loading and loading, where the sample processing before loading is generally referred to as processing a nucleic acid sample to be detected so as to meet the loading requirement, for example, attaching the nucleic acid sample to the surface of the solid substrate, and the loading refers to entering the processed nucleic acid sample to be detected into a sequencer for sequencing.

Current commercial sequencing platforms typically include two or more separate or functionally linked devices to accomplish the above-described sample processing and sequencing. For sequencing platforms comprising two mating devices, typically one of the devices used to process the nucleic acid sample to be tested comprises attaching it to a flow cell, such as a chip, the device often being referred to as a hybridization instrument, a sample introduction instrument, a library construction instrument, a sample processing instrument, or a sample preparation instrument; the other device detects the chip connected with the nucleic acid sample to be detected and output by the last device, thereby realizing the determination of the nucleic acid sequence, and the device is the most main hardware of the sequencing platform, namely a sequencing instrument generally.

Current commercial sequencing platforms, these stand-alone and functionally linked devices can be sold or purchased together or separately/in part; purchasing multiple associated devices is generally expensive, and purchasing one or a portion of the devices, such as purchasing only one sequencer, typically requires more user operations or involvement in manual operations or selection of a commercial sample processing service.

With the development of sequencing technology, automation and other related technologies and the expectation of the market on simple operation and the like, a sequencer integrating at least a part of pre-computer sample processing and sequencing functions is a development direction (hereinafter, the sequencer with the integrated function is referred to as an integrated sequencer or an integrated sequencing platform for short), and particularly, compared with the sequencing platform before integration, the integrated sequencer is not weakened in performance, equivalent or smaller in volume and simpler and more convenient to operate.

Few currently mature integrated sequencer commodities with integrated functions are limited by at least the following factors: sample processing and sequencing before loading generally comprises a plurality of steps and a plurality of reactions, and relates to switching of a plurality of reagents, a plurality of serial or parallel liquid inlet and outlet sequence control and microfluidic control.

Therefore, how to construct a set of system integrating at least a part of the pre-processing sample processing and sequencing functions, and the system or the integrated sequencer including the system has performance equivalent to that before integration or has a certain quality level and cost within a certain range, is a concern.

Disclosure of Invention

The inventor of the present application finds that, in the process of designing and testing an integrated circuit for pre-machine sample processing and on-machine sequencing, the integrated circuit capable of accurately controlling a trace amount of liquid, particularly the integrated circuit capable of independently or alternately controlling reactions on a plurality of reactors (flow cells/flow cells) and/or controlling liquid environments of reactions at different stages in different regions of one reactor, involves a considerable number of valve bodies, so that the number of associated pipelines is large and relatively complicated to interleave, the integrated circuit constructed relatively occupies a large space, and when the integrated circuit is matched with other functional modules or systems to construct a complete machine (integrated sequencer), it is difficult to achieve good matching at a predicted volume.

Accordingly, embodiments of the present application provide a fluid path system, a sequencing system, and a method to address, at least to some extent, at least some of the problems discussed above.

The fluid path system of the embodiment of the present application includes: a plurality of flow paths fluidly connected with the flow cell to support a target analyte when the flow cell is installed in the fluidic circuit system; a flow path selector valve connected to the flow path, the flow path selector valve including a manifold provided with a common port, a plurality of first ports, and a plurality of second ports, and a first spool provided with a communication groove, the first spool being rotatable or slidable with respect to the manifold to switch the flow path selector valve between a first valve position, through which the first ports communicate with the second ports, and a second valve position, through which the first ports communicate with the common port when the flow path selector valve is in the first valve position, and the second valve position, through which the first ports communicate with the common port; a pump fluidly connected to the flow cell when the flow cell is installed in the fluid path system and flowing liquid through the flow path selected by the flow path selection valve during an analysis operation; and a control circuit operatively coupled to the flow path selection valve, the control circuit for controlling the flow path selection valve to select a specified flow path.

The sequencing system of the embodiments of the present application includes the fluid path system of any of the embodiments described above.

The liquid path system of the embodiment can realize independent or parallel three-way control of the plurality of liquid paths by utilizing the flow path selection valve, and can avoid simultaneously arranging a plurality of three-way valves in the liquid path system, thereby reducing the cost of the liquid path system, reducing the volume, reducing the reagent consumption, improving the reliability, and being convenient for maintenance, repair and control. The fluid path system described above is particularly suitable for systems comprising multiple flow paths and requiring high precision in fluid control and delivery, such as sequencing systems (nucleic acid sequencing systems). The sequencing system comprising the liquid path system can realize independent control of different reaction processes of different parts/areas of the flow cell, and has low preparation cost.

A control system of an embodiment of the present application implements a method of sequencing, the system comprising a plurality of flow paths, a flow cell connected to the plurality of flow paths, a flow path selection valve and pump, a first reservoir, and a second reservoir,

the flow path selection valve comprises a manifold and a first valve core, the manifold is provided with a common port, a plurality of first ports and a plurality of second ports, the first valve core is provided with a communication groove, the first valve core can rotate or slide relative to the manifold so that the communication groove can selectively communicate the common port with the first ports or the first ports with the second ports, so as to select different flow paths, the first storage device stores a sample solution to be detected, the first storage device is connected with the second ports, the second storage device stores reaction reagents, the second storage device is connected with the common port, the first storage device carries a first reaction liquid, the first reaction liquid contains nucleic acid molecules, the second storage device carries a second reaction liquid, and the second reaction liquid contains components required for nucleic acid sequencing, the pump for flowing liquid through a flow path selected by the flow path selection valve, the method comprising:

switching the flow path selector valve to a first valve position to cause the communication groove to communicate the first port and the second port to communicate the first reservoir and the flow cell; controlling the pump to operate to cause the first reaction solution to enter the flow cell to perform a first reaction including attaching at least a portion of the nucleic acid molecule to the flow cell with the flow path selection valve in the first valve position; switching the flow path selector valve to a second valve position to cause the communication groove to communicate the first port and the common port to communicate a second reservoir and a flow cell; controlling the pump to operate to cause the second reaction solution to enter the flow cell to perform a second reaction including causing a nucleic acid molecule in the flow cell after the first reaction is performed to interact with the second reaction solution to undergo a polymerization reaction and detecting a signal from the reaction with the flow cell in a state where the flow path selection valve is in the second valve position to effect sequencing of the nucleic acid molecule.

The method realizes the proceeding of various reactions by designing and controlling a liquid path system, comprising configuring a flow path selection valve and controlling the flow path selection valve to be at a first valve position or a second valve position to utilize a designated flow channel to communicate a solution storage and a flow cell; the method integrates and simplifies a liquid path system for multiple reactions by configuring a specific flow path selection valve, so that the multiple reactions can be realized by simply operating the flow path selection valve or the liquid path system comprising the flow path selection valve, and the method is particularly suitable for an automatic biomolecule detection system related to the multiple reactions, such as a nucleic acid sequencing system.

The method can perform a plurality of reactions by connecting the flow path selection valve to a specified flow channel at a specified valve position, and can realize nucleic acid Sequencing related to the solid phase carrier, including connecting or fixing nucleic acid molecules to the surface of the solid phase carrier and Sequencing the nucleic acid molecules on the surface of the solid phase carrier by Sequencing By Synthesis (SBS) or Sequencing By Ligation (SBL). The hardware configuration required for automatically realizing the method is simple and easy to realize, and is beneficial to batch preparation and industrialization.

Additional aspects and advantages of embodiments of the present application will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of embodiments of the present application.

Drawings

The above and/or additional aspects and advantages of the present application will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:

FIG. 1 is an exploded schematic view of a flow path selector valve according to an embodiment of the present application;

fig. 2 is a schematic structural view of a fluid path system according to an embodiment of the present application;

FIG. 3 is a schematic perspective view of a flow path selector valve of an embodiment of the present application;

FIG. 4 is a schematic perspective view of a manifold according to an embodiment of the present application;

FIG. 5 is a side schematic view of a manifold of an embodiment of the present application;

FIG. 6 is a schematic cross-sectional view of the manifold of FIG. 5 taken along the line A-A;

FIG. 7 is a schematic plan view of a first valve spool of an embodiment of the present application;

FIG. 8 is a schematic plan view of a second valve spool of an embodiment of the present application;

FIG. 9 is a schematic illustration of a first spool engaged with a second spool in a first position according to an embodiment of the present application;

FIG. 10 is a schematic illustration of the first valve spool engaged with the second valve spool and in a second position in accordance with an embodiment of the present application;

FIG. 11 is a schematic perspective view of a slider according to an embodiment of the present application;

FIG. 12 is a schematic structural view of a housing according to an embodiment of the present application;

FIG. 13 is a schematic structural view of a flow cell assembly according to an embodiment of the present application;

fig. 14 is a schematic structural view of a fluid path system according to an embodiment of the present application;

FIG. 15 is a schematic block diagram of a system according to an embodiment of the present application;

FIG. 16 is a schematic flow chart of a method of an embodiment of the present application;

FIG. 17 is a block schematic diagram of a fluid path system according to an embodiment of the present application;

FIG. 18 is a schematic flow chart of a method of an embodiment of the present application;

fig. 19 is a schematic flow diagram of a method of an embodiment of the present application.

Description of the main element symbols:

the flow path selection valve 10, the fluid passage system 12, the pump 14, the common port 16, the first port 18, the second port 20, the communication groove 21, the first communication groove 22, the manifold 24, the first spool 26, the second communication groove 23, the second spool 28, the first passage 30, the second passage 32, the third passage 34, the housing chamber 36, the first connection port 38, the second connection port 40, the third connection port 42, the fastener 55, the driving member 56, the slider 57, the housing 58, the guide groove 59, the connection portion 60, the guide rail portion 61, the flow cell assembly 101, the plurality of flow paths 102, the flow cell 62, the control circuit 63, the reservoir 64, the first reservoir 66, the second reservoir 68, the first flow cell 70, the second flow cell 72, the first flow path selection valve 74, the second flow path selection valve 76, the first reaction region 78, the second reaction region 80, the reagent selection valve 84, the fluid inlet 86, the fluid inlet, A liquid outlet 88, a liquid collector 89 and a sequencing system 90.

Detailed Description

Embodiments of the present application will be further described below with reference to the accompanying drawings. The same or similar reference numbers in the drawings identify the same or similar elements or elements having the same or similar functionality throughout.

In addition, the embodiments of the present application described below in conjunction with the accompanying drawings are exemplary and are only for the purpose of explaining the embodiments of the present application, and are not to be construed as limiting the present application.

In the present application, "first", "second", "third", "fourth" and "fifth" are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance, order or implicit indication of the number of technical features indicated. Thus, features defined as "first," "second," etc. may explicitly or implicitly include one or more of the described features. In the description of the present application, "a plurality" means two or more unless otherwise limited.

In this application, unless expressly stated or limited otherwise, "mounted" and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; may be mechanically connected, may be electrically connected or may be in communication with each other; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art as appropriate.

In this application, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through intervening media. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.

In the present application, the terms "nucleic acid sequencing", "sequencing" and "sequencing" are equivalent and include DNA sequencing and/or RNA sequencing, including long fragment sequencing and/or short fragment sequencing; the so-called "sequencing reaction" is the same as the sequencing reaction. Generally, in nucleic acid sequence determination, a base/nucleotide on a template nucleic acid can be identified/determined by one round of sequencing reaction, the base being selected from at least one of A, T, C, G and U. In sequencing-by-synthesis (SBS) and/or sequencing-by-ligation (SBL), one round of sequencing reactions includes extension reactions (base extension), signal detection (e.g. photo/image capture) and group excision (excision of detectable and/or blocking groups). The so-called "nucleotide analogs", i.e., substrates, also known as reversible terminators (reversible terminators), are analogs of A, T, C, G and/or U nucleotides that are capable of pairing with a particular type of base following the base complementarity principle, while being capable of blocking or inhibiting the binding of the nucleotide/substrate to the next nucleotide position of the template strand. One round of sequencing reaction includes one or several repeated reactions consisting of extension reaction (base extension), signal detection and radical excision; for example, four nucleotide analogs carry the same detectable label, e.g., the same fluorescent molecule, and a round of sequencing reactions involves four repetitive reactions, performed sequentially, corresponding to the four nucleotides; for another example, four nucleotide analogs carry two detectable labels separated by a detectable signal region, two detectable labels are the same, one round of sequencing reaction can comprise two repeated reactions, namely, two nucleotides with different detectable labels carry out extension reaction, signal detection and radical excision in the same reaction system, and the two repeated reactions realize the base type identification of one position of the template after one round of sequencing reaction; as another example, four nucleotide analogs each carry four detectable labels with a signal detectable region, or three of the four nucleotide analogs each carry three detectable labels with a signal detectable region, and another nucleotide analog does not carry a detectable label, and a round of sequencing reactions involves subjecting the four nucleotide analogs to a single repetitive reaction in the same reaction system.

Referring to fig. 1-2, the present disclosure provides a flow path selection valve 10, where the flow path selection valve 10 may be applied to a fluid path system 12, or the fluid path system 12 includes the flow path selection valve 10, to implement merging, splitting, flow path switching, and/or flow rate control.

Referring to fig. 1 and 3, the flow path selection valve 10 of the present embodiment is configured to switch between a first valve position and a second valve position, and the flow path selection valve 10 has a common port 16, a plurality of first ports 18, a plurality of second ports 20, and a plurality of communication grooves 21. The communication groove 21 selectively communicates the common port 16 and the first port 18 or the first port 18 and the second port 20.

With the flow path selector valve 10 in the first valve position, the first port 18 and the second port 20 communicate through the communication groove 21. With the flow path selector valve 10 in the second valve position, the first port 18 communicates with the common port 16 through the communication groove 21.

In the flow path selector valve 10 and the fluid passage system 12 of the embodiment of the present application, in the case where the flow path selector valve 10 is in the first valve position, each first port 18 communicates with the corresponding second port 20 through the corresponding communication groove 21, and in the case where the flow path selector valve 10 is in the second valve position, each first port 18 communicates with the common port 16 through the corresponding communication groove 21, so that the flow path selector valve 10 has two or more passages; the flow path selector valve 10 is applicable to any fluid path system requiring the use of three-way valves, particularly multiple three-way valves, for example those involving the need to change the direction of one or more flow paths and/or dispense multiple liquids/solutions. The flow path selection valve is applied to the liquid path system 12 comprising a plurality of liquid paths/flow paths, independent or parallel three-way control over the plurality of liquid paths can be achieved, and a plurality of three-way valves can be prevented from being arranged in the liquid path system 12 at the same time, so that the cost of the liquid path system 12 is reduced, the size is reduced, the reagent consumption is reduced, the reliability is improved, and the maintenance, the maintenance and the control are convenient.

Specifically, the number of the common ports 16 is one, and the common ports 16 may serve as liquid inlets or outlets of the flow path selection valve 10. Alternatively, liquid may enter or exit the flow routing valve 10 from the common port 16, thereby achieving either a split or a parallel flow. Here, the common port 16 serves as a liquid inlet of the flow path selection valve 10, and liquid can enter the flow path selection valve 10 from the common port 16. The shape of the common port 16 may be a regular shape such as a circle, a polygon, or an irregular shape. In the present embodiment, the common port 16 is circular in shape to facilitate forming, manufacturing, and/or connecting the common port 16 to a common conduit.

The plurality of second ports 20 correspond one-to-one to the plurality of first ports 18. The plurality of communication grooves 21 correspond one-to-one to the plurality of first ports 18. The number of first ports 18 may be 2, 3, 4, 5, 6, 7, 8, or more. In the embodiment shown in fig. 1, the number of first ports 18 is 8 to accommodate elements or devices containing 8 outlets or inlets upstream or downstream of the flow path selector valve 10.

As shown in fig. 2, liquid can enter the flow path selection valve 10 from the common port 16, flow out from the 8 first ports 18, and then enter a flow cell (flow cell)62 containing 8 channels or 8 reaction regions, so that the parallel control of the inlet of the 8 channels or 8 reaction regions of the flow cell 62 can be realized. The shape of the first port 18 may be a regular shape such as a circle, a polygon, or an irregular shape. In the present embodiment, the first port 18 has a circular shape. The first port 18 may serve as a liquid outlet or an inlet of the flow path selector valve 10.

Similarly, the number of the second ports 20 is not limited in the present application, and the number of the second ports 20 may be 2, 3, 4, 5, 6, 7, 8 or more. In the embodiment shown in fig. 3, the number of the second ports 20 is 8 to adapt an element or device including 8 outlets or inlets upstream or downstream of the flow path selection valve 10, such as the flow cell 62 including 8 channels or 8 reaction regions downstream of the flow path selection valve 10 in fig. 2, to achieve parallel control of the 8 channels or 8 reaction regions of the flow cell 62. The number of second ports 20 is equal to the number of first ports 18. The shape of the second port 20 may be a regular shape such as a circle, a polygon, or an irregular shape. In the present embodiment, the second port 20 has a circular shape. The second port 20 may serve as a liquid inlet or an outlet of the flow path selector valve 10. For ease of manufacture, the second port 20 may be the same size as the first port 18.

The number of the communication grooves 21 is the same as the number of the first ports 18 and/or the second ports 20. In the present embodiment, the number of the communication grooves 21 is also 8. The communication groove 21 is not limited in shape as long as it communicates the first port 18 and the second port 20 when the flow path selector valve 10 is in the first valve position, and may have a curved or rectangular shape, for example.

In summary, in the first valve position, each of the first ports 18 communicates with a corresponding one of the second ports 20, and at this time, liquid enters the flow path selector valve 10 from the second port 20 and flows out from the corresponding first port 18. That is, the flow path selection valve 10 also has a function of simultaneously supplying liquid to a plurality of paths and simultaneously discharging liquid from the plurality of paths.

In the second valve position, the common port 16 communicates with each of the first ports 18 through a corresponding one of the communication grooves 21, and at this time, the liquid can enter the flow path selector valve 10 from the common port 16, pass through the communication groove 21, and then flow out from the first port 18. Alternatively, in the second valve position, fluid enters the flow path selector valve 10 from a common port 16 and exits a plurality of first ports 18. That is, the flow path selector valve 10 has a function of one-way liquid inlet and multi-way liquid outlet.

In the present embodiment, with the flow path selector valve 10 in the first valve position, both the first port 18 and the second port 20 are blocked from the common port 16; with the flow path selector valve 10 in the second valve position, the common port 16 and the first port 18 are both blocked from the second port 20 to form separate first or second flow passages. The first flow channel and the second flow channel are multiple, so that parallel control of liquid inlet and outlet of a plurality of reaction areas is facilitated, and independent control of liquid inlet and outlet of different reaction processes on the reaction areas is facilitated.

In some embodiments, the flow path selector valve 10 may be in a third valve position, in which case the common port 16, the plurality of first ports 18, and the plurality of second ports 20 are isolated from one another. As such, the flow path selector valve 10 may also perform the function of controlling the fluid shutoff with the flow path selector valve 10 in the third valve position.

In this embodiment, the flow path selector valve 10 has three modes, that is, a mode in which the first port 18 and the second port 20 communicate when the flow path selector valve 10 is in the first valve position; a mode in which the common port 16 and the first port 18 communicate when the flow path selector valve 10 is in the second valve position; a mode in which the common port 16, the first port 18, and the second port 20 are blocked from each other when the flow path selector valve 10 is in the third valve position.

It should be noted that reference herein to a plurality of ports being "isolated" means that there is no communication between the plurality of ports, or that fluid cannot enter a given port or ports and exit another given port or ports.

Referring to fig. 1 and 3, in some embodiments, the flow path selector valve 10 includes a manifold 24 and a first spool 26. Referring to fig. 4-6, the manifold 24 has a common port 16, a plurality of first ports 18, and a plurality of second ports 20. Referring to fig. 7, the first valve core 26 is provided with a communication groove 21. The first spool 26 may rotate or slide relative to the manifold 24 to switch the flow path selector valve 10 between the first and second valve positions.

As such, during rotation or sliding of the first spool 26, the communication groove 21 may communicate the common port 16 and the first port 18, or may communicate the first port 18 and the second port 20, so that liquid may enter the flow path selector valve 10 from the manifold 24 and exit the flow path selector valve 10 from the manifold 24.

Specifically, the manifold 24 is a flow splitting or converging module, and the manifold 24 may split the liquid entering from the common port 16 and then flow out from the plurality of first ports 18, or may guide the liquid entering from the second port 20 and then flow out from the corresponding first ports 18. It will be appreciated that the interior of the manifold 24 has flow passages communicating with each port.

In the present example, the first spool 26 is in the shape of a square block, and the first spool 26 is slidable with respect to the manifold 24. The communication groove 21 is linear, and the communication groove 21 may have a bent shape, a curved shape, or the like, and the specific shape of the communication groove 21 is not limited herein.

The number of the communication grooves 21 is schematically the same as the number of the first ports 18, and in the present embodiment, the number of the communication grooves 21 is 8. Of course, in other embodiments, the number of the communication grooves 21 may be other numbers such as 2, 3, 4, 5, and 6.

Referring to fig. 1, 3, and 8, in some embodiments, the flow path selector valve 10 includes a second spool 28 disposed on the manifold 24, the first spool 26 is disposed on the second spool 28, the second spool 28 is provided with a first passage 30, a second passage 32, and a third passage 34, the first passage 30 is in communication with the first port 18, the second passage 32 is in communication with the second port 20, the third passage 34 is in communication with the common port 16,

the communication groove 21 communicates the first passage 30 and the second passage 32 to enable the first port 18 and the second port 20 to communicate with each other with the flow path selector valve 10 in the first valve position, and the communication groove 21 communicates the first passage 30 and the third passage 34 to enable the first port 18 to communicate with the common port 16 with the flow path selector valve 10 in the second valve position.

In this manner, the first spool 26 selectively communicates the first port 18 with the common port 16, or communicates the first port 18 with the second port 20, via the second spool 28, which facilitates preparation of the flow stream through the selector valve.

Specifically, the second spool 28 is in the shape of a block, and the volumes of the first and

second spools

26 and 28 are each smaller than the volume of the manifold 24. Since the manifold 24 and the first valve element 26 are manufactured separately, the accuracy of the two may not be such that the manifold 24 and the first valve element 26 meet the mating requirements and a liquid leak occurs. The first and

second spools

26, 28 are relatively regular in shape and are easy to manufacture. Therefore, the present embodiment connects the manifold 24 and the first spool 26 through the second spool 28, which may make the flow path selecting valve 10 easier to manufacture and more stable.

Of course, in other embodiments, the second spool 28 may be omitted. At this point, the first spool 26 is in mating connection with the manifold 24.

In the embodiment of the present application, the first channel 30, the second channel 32, and the third channel 34 are independent channels, and each channel is separately disposed. The first passage 30, the second passage 32, and the third passage 34 are all linear, and all penetrate the second spool 28 in the thickness direction of the second spool 28.

Of course, in other embodiments, at least one of the first channel 30, the second channel 32, and the third channel 34 may have a bent shape, a curved shape, and the like, and the specific shapes of the first channel 30, the second channel 32, and the third channel 34 are not limited herein.

In the present example, the number of first passages 30 is the same as the number of first ports 18, and is 8. The number of the second passages 32 is the same as the number of the second ports 20, and each is 8. The number of the third passages 34 is 8. Of course, in other embodiments, the number of first channels 30, second channels 32, and third channels 34 may be other numbers such as 2, 3, 4, 5, 6, etc.

As in the example of fig. 8, the first passages 30, the second passages 32, and the third passages 34 are arranged in a matrix, 8 first passages 30, 8 second passages 32, and 8 third passages 34 are all spaced apart along the length of the second spool 28, 8 first passages 30 are arranged along a first line, 8 second passages 32 are arranged along a second line, and 8 third passages 34 are arranged along the third passages 34. The first passages 30 and the third passages 34 are aligned one by one in the width direction of the second spool 28, and the first passages 30 and the second passages 32 are arranged offset from each other.

Of course, in other embodiments, the first channel 30, the second channel 32, and the third channel 34 may be arranged in other manners, and the arrangement manner of the first channel 30, the second channel 32, and the third channel 34 is not limited herein.

In the embodiment of the present application, the second spool 28 is fixedly provided with respect to the manifold 24. The first spool 26 slides on a surface of the second spool 28 relative to the second spool 28 so that the first spool 26 can slide between the first position and the second position, so that the communication groove 21 can selectively communicate the first passage 30 and the second passage 32, or the first passage 30 and the third passage 34.

Referring to fig. 3 and 4, in some embodiments, manifold 24 defines a cavity 36, and second spool 28 is at least partially received within cavity 36. In this manner, the manifold 24 and the second spool 28 cooperate more compactly, and the volume of the flow path selector valve 10 can be reduced, which contributes to miniaturization of the flow path selector valve 10.

Specifically, the shape of the cavity 36 is adapted to the shape of the second valve element 28, and in the embodiment of the present application, the cavity 36 is also rectangular. As shown in fig. 3, the second valve spool 28 is partially received in the cavity 36, or a portion of the second valve spool 28 protrudes from the cavity 36.

Referring to fig. 4, in some embodiments, the bottom surface of the cavity 36 is provided with a first connection port 38, a second connection port 40 and a third connection port 42. The first connection port 38 communicates the first port 18 and the first passage 30. The second connection port 40 communicates the second port 20 with the second passage 32. The third connection port 42 communicates the common port 16 and the third passage 34.

In this way, the second valve element 28 is communicated with the first port 18, the second port 20 and the common port 16 through the connection port arranged at the bottom surface of the cavity 36, so that the contact area between the second valve element 28 and the cavity 36 can be increased, and liquid leakage can be prevented.

In order to enable the first connection port 38 to be in butt joint with the first channel 30, the second connection port 40 to be in butt joint with the second channel 32, and the third connection port 42 to be in butt joint with the third channel 34, the arrangement of the first connection port 38, the second connection port 40, and the third connection port 42 is the same as the arrangement of the first channel 30, the second channel 32, and the third channel 34, and the arrangement of the first connection port 38, the second connection port 40, and the third connection port 42 is not described herein again.

Referring to fig. 4 and 5, in some embodiments, at least two of the first port 18, the second port 20, and the common port 16 are located on different sides of the manifold 24. For example, the first port 18 and the second port 20 are located on the same side of the manifold 24, and the common port 16 is located on the other side of the manifold 24. As another example, first port 18 and common port 16 are located on the same side of manifold 24, and second port 20 is located on the other side of manifold 24. As another example, first port 18, second port 20, and common port 16 are each located on a different side of manifold 24.

In the orientation illustration of FIG. 5, the first port 18 is located on the left side of the manifold 24, the second port 20 is located on the bottom side of the manifold 24, and the common port 16 is located on the right side of the manifold 24. In this manner, the ports are disposed on different sides of the manifold 24, which makes full use of the space of the manifold 24, so that the manifold 24 is more compact and the connection of the manifold 24 to other components is facilitated.

Referring to fig. 7, in some embodiments, the communication groove 21 includes a first communication groove 22 and a second communication groove 23 that are spaced apart from each other. With the flow path selector valve 10 in the first valve position, the first port 18 communicates with the second port 20 through the first connecting channel 22, as shown in fig. 9. With the flow path selector valve 10 in the second valve position, the first port 18 and the common port 16 communicate through the second communication groove 23, as shown in fig. 10. Alternatively, the communication groove 21 includes two discontinuous portions, which facilitates control of the communication of the first port 18 with the second port 20 and the communication of the first port 18 with the common port 16 through the communication groove 21.

The arrows in fig. 9 and 10 represent the flow direction of the liquid. As shown in fig. 9, after the liquid enters the flow path selector valve 10 from the second port 20, the liquid flows out from the first port 18 after passing through the second passage 32, the first connecting groove 22, and the first passage 30 in this order.

In fig. 10, after the liquid enters the flow path selector valve 10 from the common port 16, the liquid flows out from the first port 18 after passing through the third passage 34, the second communication groove 34, and the first passage 30 in this order.

As in the example of fig. 7, in order to accommodate the arrangement of the first passage 30, the second passage 32, and the third passage 34, the first communication groove 22 extends linearly obliquely with respect to the width direction of the first spool 26, and the second communication groove 23 extends linearly in the width direction of the first spool 26.

It is understood that fig. 7 is only one example of the first and

second communication grooves

22 and 23. In other embodiments, the shapes of the first and

second communication grooves

22 and 23 may be an arc shape, a bent shape, or other shapes.

In addition, in other embodiments, one of the first communicating groove 22 and the second communicating groove 23 may be omitted, or the communicating groove 21 is a groove having a continuous structure. At this time, in one example, the first channel 30 may be disposed between the second channel 32 and the third channel 34, and the first channel 30, the second channel 32, and the third channel 34 are arranged along the same straight line. The communication groove 21 has a linear shape. When the first spool 26 is located at the first position, one end of the communication groove 21 communicates with the first passage 30 and the other end communicates with the second passage 32 to communicate the first port 18 with the second port 20; when the second spool 28 is located at the second position, one end of the communication groove 21 communicates with the third passage 34 and the other end communicates with the first passage 30, so that the common port 16 communicates with the first port 18.

Referring again to fig. 1 and 3, in some embodiments, the flow path selector valve 10 includes a drive member 56, and the drive member 56 is configured to drive the first valve element 26 to rotate or slide. As such, the drive member 56 may move the first valve spool 26 such that the first valve spool 26 may be positioned in different positions to enable different functions of the flow path selector valve 10.

In particular, the drive component 56 may provide electromagnetic drive. For example, the driving part 56 may be a motor having a screw structure when the first spool 26 slides with respect to the manifold 24, and the driving part 56 may be a motor having a rotation shaft when the first spool 26 rotates with respect to the manifold 24.

Referring to fig. 1, 3 and 11, in some embodiments, the flow path selector valve 10 includes a slider 57 connected to the first valve element 26, the slider 57 is connected to a driving member 56, and the driving member 56 drives the first valve element 26 to slide through the slider 57. In this way, the driving member 56 is facilitated to drive the first valve element 26 to slide.

Specifically, the slider 57 is detachably connected to the first spool 26. For example, the slider 57 is connected to the first valve element 26 via the positioning pin 54, and when the slider 57 receives the driving force of the driving member 56, the slider 57 can drive the first valve element 26 to slide via the positioning pin. Of course, the slider 57 and the first valve core 26 may be connected by a snap-fit structure or the like, and the application does not limit the specific connection manner of the slider 57 and the first valve core 26.

When the driving part 56 is a screw motor, the slider 57 may be sleeved on a screw of the screw motor. During the rotation of the screw, the slider 57 can move relative to the screw, thereby pushing the first valve core 26 to move.

Referring to fig. 1 and 3, in some embodiments, the flow path selector valve 10 includes a housing 58 removably coupled to the manifold 24, and the first valve element 26 and the slider 57 are received in the housing 58. In this way, the housing 58 can protect the first valve element 26 and the slider 57 and provide pressing force to the first valve element 26 and the second valve element 28, thereby avoiding the undesirable phenomenon of leakage of the first valve element 26 and the second valve element 28.

Specifically, the housing 58 may be coupled to the manifold 24 via fasteners 55. In one example, the flow routing valve 10 is assembled by first installing the second valve spool 28 into the manifold 24, then installing the first valve spool 26 on the second valve spool 28, then trapping the first valve spool 26 and the slider 57, then covering the housing 58 on the manifold 24 so that both the slider 57 and the first valve spool 26 are located in the housing 58, and finally passing the fasteners 55 through the manifold 24 and tightening the housing 58 so that the housing 58 is secured to the manifold 24.

Referring to fig. 11 and 12, in some embodiments, the housing 58 is provided with a guide groove 59, and the slider 57 includes a connecting portion 60 and a guide portion 61 connected to the connecting portion 60. The connecting portion 60 is connected to the first spool 26. The guide rail portion 61 cooperates with the guide groove 59 to guide the sliding block 57 to slide the first valve core 26. In this manner, the guide track portion 61 and the guide groove 59 cooperate to enable the slider 57 to slide along a predetermined track, thereby driving the first valve core 26 to slide between the first position and the second position.

Specifically, the connection portion 60 may be connected to the first spool 26 by the dowel pin 54. The profile of the connection portion 60 may be adapted to the shape of the first spool 26 to facilitate connection with the first spool 26. The rail portion 61 protrudes from the surface of the connecting portion 60. The cross-sectional area of the guide rail portion 61 is smaller than that of the connecting portion 60.

Referring to fig. 13, a flow cell assembly 101 according to an embodiment of the present application includes a flow path selection valve 10 according to any one of the above embodiments and a flow cell 62, and the flow cell 62 is connected to the flow path selection valve 10. The flow cell 62 includes a plurality of passages 621 arranged in parallel, one end of the passages 621 communicating with the first port 18. In this manner, the flow routing valve 10 and the flow cell 62 may be formed as an integral module for ease of installation.

The flow cell 62 referred to herein provides a biochemical reaction site, and the flow cell 62 may be specifically a chip, and the flow cell 62 is removably connected to the flow path selection valve 10. Specifically, the flow cell 62 may be provided with a plug formed with a channel 621, which may be inserted into the first port 18, thereby enabling the channel 621 of the flow cell 62 to be directly connected with the first port 18. Of course, in other embodiments, the flow cell 62 and the flow path selector valve 10 may be in communication via tubing.

In some embodiments, the flow cell assembly 101 includes two flow path selection valves 10 and one flow cell 62, one end of the channel 621 communicates with the first port 18 of one of the flow path selection valves 10, and the other end of the channel 621 communicates with the first port 18 of the other flow path selection valve 10.

In some embodiments, one or the other end of the channel 621 is ductless connected to the first port 18. In this way, the flow cell 62 is directly connected to the flow path selector valve 10, and the connected piping can be omitted, reducing the risk of liquid leakage.

Referring again to fig. 2, the fluid path system 12 of one embodiment of the present application includes the flow path selector valve 10 of any of the above embodiments, the pump 14, the plurality of flow paths 102, and the control circuit 63. A plurality of flow paths 102 are fluidly connected to flow cell 62 to support the target analyte when flow cell 62 is installed in fluid path system 12.

Pump 14 is fluidly connected to flow cell 62 when flow cell 62 is installed in fluid system 12 and flows liquid through flow path 102 selected by flow path selection valve 10 during analysis operations.

The control circuit 63 is operatively coupled to the flow path selection valve 10, the control circuit 63 having one or more processors and memory storing computer-executable instructions that, when executed by the processors, control the processors to command the flow path selection valve 10 to select a specified flow path 102.

In the fluid path system 12 of the embodiment of the present application, the flow path selector valve 10 can independently or parallelly control the plurality of flow paths 102 in a three-way manner, and it can be avoided that the plurality of three-way valves are simultaneously arranged in the fluid path system 12, so that the cost of the fluid path system 12 is reduced, the volume is reduced, the reagent consumption is reduced, the reliability is improved, and the maintenance, the maintenance and the control are convenient. The fluid path system 12 described above is particularly suited for use in systems requiring high precision in fluid control and delivery, such as sequencing systems.

Specifically, the pump 14 may power the fluid path system 12 such that fluid may flow. The number of pumps 14 may comprise a plurality, such as two, where one pump 14 may be in communication with the common port 16 and the other pump 14 is in communication with the second port 20, either in split or in parallel. A pump in communication with the common port 16 may provide power to the liquid from the common port 16 to the first port 18, and a pump in communication with the second port 20 may provide power to the liquid from the second port 20 to the first port 18.

In one embodiment, the number of the first ports 18 is 8, and the pump 14 includes 8 pumps equal to the number of the first ports 18, such as a negative pressure eight-way pump, which is located downstream of the fluid path system 12, specifically downstream of the flow cell 62, and is capable of independently providing a negative pressure to the fluid after passing through the flow path selection valve 10, so that the dynamic magnitude of the fluid in each of the flow paths 102 can be independently controlled, which facilitates fine control of the flow rate and/or flow velocity of the fluid in each of the flow paths.

Referring to fig. 2, in the present embodiment, the fluid path system 12 further includes a storage 64, the storage 64 can store a plurality of solutions, including reaction solutions, buffers, washing solutions and/or purified water, etc., including reagents for different reactions or different steps of a reaction, and the fluid path system 12 including the pump 14 can enable one or more solutions to flow toward the flow cell 62 sequentially or simultaneously.

In certain embodiments, the reservoir 64 includes a first reservoir 66 and a second reservoir 68, the first reservoir 66 carrying the biological sample solution and the second reservoir 68 carrying the reaction solution. With the flow routing valve 10 in the first valve position, the flow routing valve 10 communicates the first reservoir 66 and the flow cell 62, and the pump 14 induces the flow of the biological sample solution toward the flow cell 62. With the flow path selector valve 10 in the second valve position, the flow path selector valve 10 communicates the second reservoir 68 and the flow cell 62, and the pump 14 induces the reaction liquid to flow through the second flow passage toward the flow cell 62.

That is, the flow path selection valve 10 may allow the reaction solution and the biological sample solution to enter the flow cell 62 independently and time-divisionally to sequentially perform corresponding reactions, such as a sample loading reaction (immobilization and/or hybridization of nucleic acids) and a sequencing reaction.

Illustratively, the first reaction is, for example, an immobilization and/or hybridization reaction, i.e., a nucleic acid to be tested is immobilized or attached to a channel or reaction area of the flow cell 62, and the biological sample solution is a solution containing the nucleic acid to be tested, which may enter the flow cell 62 through the second port 20 and the first port 18 in order to perform the first reaction. In one example, the flow cell 62 is a sandwich-like structure having three layers, or a structure with an upper layer (close to the objective lens) and a lower layer, wherein the upper layer is a transparent glass layer, the middle layer or the lower layer is a transparent glass layer or an opaque substrate layer, the middle layer or the lower layer is provided with a plurality of channels arranged in an array, the channels can accommodate liquid to provide physical space for reaction, each channel is provided with an independent liquid inlet and an independent liquid outlet, the number of the channels is equal to that of the first ports 18, a plurality of biological sample solutions can simultaneously and independently enter one channel of the flow cell 62 through one second port 20 and one first port 18, thus, the flow path selection valve 10 or the fluid path system 12 including the flow path selection valve 10 can perform loading and detection analysis of a plurality of biological samples without combining other means such as labeling different biological samples with labels. Specifically, the flow cell 62 is, for example, a solid phase substrate having a functional group on the surface and/or a solid phase substrate having a probe attached to the surface, the solid phase substrate having a functional group on the surface and/or the solid phase substrate having a probe attached to the surface are also generally referred to as a chip or a microsphere, for example, the channel of the upper surface (upper glass layer of the lower surface) and/or the channel of the lower surface (middle or lower structure of the upper surface) has a functional group or connected with a probe (oligonucleotide), the functional group can be connected with the nucleic acid to be detected, and/or the probe at least a portion can be complementary with the nucleic acid to be detected and paired, to immobilize or attach the nucleic acid to be detected to the surface of the solid phase substrate, for performing a subsequent detection analysis, such as a sequencing reaction, on the nucleic acid to be detected immobilized or attached to the surface of the solid phase substrate.

The second reaction is, for example, a sequencing reaction, that is, a nucleic acid sequencing reaction, more specifically, a sequencing-by-synthesis reaction using a reversible terminator based on chip detection, and accordingly, the reaction solution includes one or more reagents including a substrate (reversible terminator), a polymerase catalyst, a cleavage reagent (group excision reagent), an imaging reagent, and a washing reagent, and the reagents/reaction solutions may sequentially or simultaneously enter the flow cell 62 through the common port 16 and the first port 18 to perform the second reaction, and specifically, the reagents may sequentially or simultaneously flow to the flow cell 62 after passing through the flow path selection valve 10 to perform a plurality of reaction steps in the flow cell 62 to achieve the sequencing reaction; the flow cell 62 is, for example, a solid phase substrate to which nucleic acid to be detected is attached on a surface, and the solid phase substrate to which nucleic acid to be detected is attached is, for example, a chip or a microsphere.

The above-described first reaction and second reaction can be performed in the flow cell 62 by controlling one flow path selection valve 10 to switch the flow paths, so that a nucleic acid sequencing system (integrated system) including the first reaction and the second reaction can be made to have a simpler structure. Thus, the first reaction and the second reaction do not need to be carried out in different systems/devices/apparatuses, the operation of a user is simpler, and the cost for constructing the integrated nucleic acid sequence measuring system is far lower than the sum of the cost for constructing a nucleic acid sequence measuring system (sample loading device) for separately realizing the first reaction and the cost for separately realizing the second reaction.

The first reaction of the present embodiment includes a reaction of attaching a biomolecule to the flow cell 62, including, for example, an immobilization, hybridization, or sampling reaction. The biomolecules include DNA and/or RNA and the like, including ribonucleotides, deoxyribonucleotides and analogs thereof, including A, T, C, G and U and analogs thereof. Wherein C represents cytosine or a cytosine analogue, G represents guanine or a guanine analogue, A represents adenine or an adenine analogue, T represents thymine or a thymine analogue, and U represents uracil or a uracil analogue.

The second reaction includes a reaction for detecting a biomolecule, for example, a nucleic acid, attached to the flow cell 62, and the second reaction may be a sequencing reaction, so-called sequencing in general, including determining the primary structure or sequence of DNA or RNA, etc., including determining the order of nucleotides/bases of a given nucleic acid fragment. The second reaction may include one or more sub-reactions. In one example, sequencing the DNA, the second reaction is sequencing, sequencing by synthesis or sequencing by ligation, in particular, sequencing by synthesis, for example, based on chip detection, using a modified nucleotide with a detectable label, such as a dNTP or a dNTP analogue with a detectable label, the sequencing comprising a plurality of sub-reactions, including a base extension reaction, signal acquisition and detection group excision, to effect determination of the base type at a position on the nucleic acid sequence to be detected; performing the plurality of sub-reactions once may be referred to as performing one repeat reaction or one round of reaction, and sequencing comprises performing a plurality of repeat reactions or multiple rounds of reactions to determine the nucleotide/base order of at least one sequence of the nucleic acid molecule (template). The modified nucleotide is said to carry a fluorescent molecule which in a particular context is capable of being excited to fluoresce for detection by an optical system, and when bound to the test nucleic acid, the modified nucleotide label prevents base/nucleotide binding to the next position in the test nucleic acid, for example a dNTP having a chemically cleavable moiety at the 3' hydroxyl terminus or a dNTP having a molecular conformation which prevents the binding of the next nucleotide to the test nucleic acid, the dNTP or dNTP analogue being four deoxyribonucleotides comprising bases A, T/U, C and G respectively.

For sequencing-by-synthesis (SBS) or sequencing-by-ligation (SBL) based on chip detection, the base extension reaction involves binding of a nucleotide (including the modified nucleotides) to the nucleic acid molecule to be detected on the basis of the base complementation principle on a flow cell 62 in which the nucleic acid molecule to be detected is immobilized, under the action of polymerase or ligase, and collecting the corresponding reaction signal. The modified nucleotide may be a nucleotide with a detectable label that allows the modified nucleotide to be detected under certain circumstances, for example, a nucleotide with a fluorescent molecular label that fluoresces when excited by a laser of a particular wavelength; typically, for SBS, the engineered nucleotide also has the function of inhibiting the binding of another nucleotide to the next position of the same nucleic acid molecule, e.g. with a blocking group that prevents the binding of other nucleotides to the next position of the template, so that each extension reaction is a single base extension reaction to enable the acquisition of the corresponding signal from a single secondary base extension, the blocking group being e.g. an engineered azide (-N) attached at the 3' position of the sugar residue of the nucleotide₃)。

Detection analysis of biomolecules, generally, biomolecules are first connected to the flow cell 62, and then the biomolecules connected to the flow cell 62 are detected; specifically, in any of the above embodiments, the first reaction is performed before the second reaction, i.e., the sequencing reaction is performed after the nucleic acid to be detected is attached to the flow cell 62. Thus, by using the fluid path system 12, it is possible to perform a plurality of types of reactions including sampling and sample detection in one nucleic acid sequence measurement system.

In some embodiments, the first reaction is a sampling reaction, the second reaction is a sequencing reaction, the first reaction is performed before the second reaction, and the nucleic acid molecule to be detected is contained in a micro-biological sample solution, for example, on a microliter scale, for example, 20 microliters; before the first reaction, the liquid path system 12 is cleaned by using a cleaning solution, for example, a solution that does not affect the subsequent reaction, and the liquid path system 12 is filled with the cleaning solution; before the first reaction is started, a section of air is firstly injected to separate a biological sample solution flowing in subsequently and a cleaning solution in a liquid path system so as to prevent a trace biological sample from being diffused and/or diluted to influence the connection of a nucleic acid molecule to be detected to the flow cell 62 and the subsequent detection of the nucleic acid molecule to be detected, and the separation of the biological sample solution flowing in subsequently and the cleaning solution in the liquid path system is also favorable for observing the sample injection condition, and is favorable for observing whether a chip comprising a plurality of channels of the flow cell 62 is normal or not, whether the liquid path system 12 is normal or not and the like.

The substrate can be any solid support useful for immobilizing nucleic acid sequences, such as nylon membranes, glass sheets, plastics, silicon wafers, magnetic beads, and the like. Probes can be randomly distributed on the surface of the substrate, can be a section of DNA and/or RNA sequence and the like, and can also be called as a primer, a capture chain or a fixed chain. The first reaction may fixedly attach the biomolecule to the probe, for example based on the base-complementary principle, to attach the biomolecule to the flow cell 62.

Acquiring signals comprises acquiring signals emitted by the engineered nucleotides bound to the nucleic acid molecules, for example, by laser irradiation of a specific region in the flow cell 62 after the base extension reaction using an optical imaging assembly/system, the fluorescent molecular labels in the specific region being excited to emit fluorescence, and then photographing/image acquiring the region to record the biochemical reaction signals as image information. The sequencing in turn comprises converting the image information obtained from the multiple rounds/repeats of the reaction into sequence information, i.e.determining the base type based on the image information, so-called base-calling.

Group excision involves removal of the detectable label and/or blocking group bound to the engineered nucleotide of the nucleic acid molecule after the base extension reaction to enable the binding of other nucleotides (including engineered nucleotides) to the next position of the nucleic acid molecule for the next repeat reaction or round of reaction.

A wash reagent may also be introduced to remove residual unreacted materials, materials that interfere with the reaction or signal acquisition in flow cell 62 or in fluid path system 12 after the previous round or previous sub-reaction or previous step is completed and before the next round or subsequent sub-reaction or next step is begun.

In certain embodiments, the flow path selector valve 10 is disposed upstream of the flow cell 62. Thus, the flow path selection valve 10 can control the solution to enter the flow cell 62, and the liquid path system 12 can control the entering and exiting of various solutions to realize various reactions only by using one power assembly (e.g., a pump) or only providing power in one direction, which is beneficial to further reducing the volume of the liquid path system 12 and improving the integration degree thereof, and is beneficial to industrialization.

In certain embodiments, pump 14 is positioned downstream of flow cell 62 to provide negative pressure. In this manner, pump 14 may create a negative pressure in flow cell 62 and flow routing valve 10, thereby allowing solution to enter flow cell 62. In addition, the negative pressure created by pump 14 can remove air from flow cell 62, thereby preventing air from affecting the normal reaction of flow cell 62. The fluid path system 12 including the pump 14 located downstream of the flow cell 62 can provide a uniform power direction for the inlet and outlet of a plurality of types of reactions, and is particularly suitable for the fluid path system 12 including pressure-sensitive elements/components, for example, the flow cell 62 is a chip including thin glass and a multi-layer sheet structure bonded by glue, the chip may further include independent reaction regions/channels, and the change of the power/pressure direction is easy to deform the chip or generate liquid leakage or cross-channel phenomenon.

Referring to fig. 14, in some embodiments, flow cell 62 includes a first flow cell 70 and a second flow cell 72, and flow routing valve 10 includes a first flow routing valve 74 and a second flow routing valve 76, with first flow routing valve 74 and second flow routing valve 76 communicating with first flow cell 70 and second flow cell 72, respectively. In this way, the first flow path selection valve 74 and the second flow path selection valve 76 can independently control the reactions of the first flow cell 70 and the second flow cell 72, so that the first flow cell 70 and the second flow cell 72 can be staggered to perform different reactions or different steps/sub-reactions of the same reaction, thereby improving the reaction efficiency. In addition, the fluid path system 12 including the flow routing valve 10, in combination with the plurality of flow cells 62 or the flow cells 62 having a plurality of independent reaction regions, facilitates improved detection throughput and/or enables detection of multiple samples at a time.

Specifically, the first flow cell 70 and the second flow cell 72 may be of a separate structure or may be of an integral structure. In the example shown in fig. 14, first flow cell 70 and second flow cell 72 are of unitary construction. First flow cell 70 and/or second flow cell 72 may include one or more reaction zones. Wherein each reaction zone can effect a reaction, the reaction zones can be intersecting, continuous or separate zones.

As in the example of fig. 14, the first flow cell 70 includes eight first reaction zones 78. The eight first reaction areas 78 correspond one-to-one to the eight first ports 18 of the first flow path selector valve 74. Eight first reaction zones 78 are separately provided. Similarly, the second flow cell 72 includes eight second reaction zones 80. Eight second reaction regions 80 are in one-to-one correspondence with the eight second ports of the second flow path selector valve 76. Eight second reaction regions 80 are separately provided.

It should be noted that the first flow path selector valve 74 and the second flow path selector valve 76 may be operated simultaneously, individually or in a staggered manner, so that the reactions or steps in the first flow cell 70 and the second flow cell 72 may be performed in a time-sharing staggered manner or simultaneously, which is beneficial to improving the efficiency of the reactions and saving the consumption of detection reagents and/or time.

Referring to FIG. 15, in some embodiments, the fluid path system 12 includes a reagent selection valve 84 and the flow cell 62, the reagent selection valve 84 selecting a reagent from a plurality of reagents according to an analytical protocol. The flow path selection valve 10 is fluidly connected between the reagent selection valve 84 and the flow cell 62, the flow path selection valve 10 for selecting a flow path of the flow cell 62 from a plurality of flow paths through the flow cell 62 according to an analysis protocol and directing the selected reagent through the flow cell 62. The pump 14 flows the selected reagent through the selected flow path according to the analytical protocol.

Specifically, the reagent selection valve 84 is provided with a plurality of liquid inlets 86 and a liquid outlet 88, the liquid outlet 88 is selectively communicated with one of the liquid inlets 86, and the liquid inlet 86 is communicated with the reagent selection valve 82. As such, the multiple loading ports 86 of the reagent selection valve 84 allow the flow cell to be loaded with different liquids to achieve multiple rounds/repeats of reactions.

As shown in fig. 15, in certain embodiments, fluid path system 12 includes a liquid trap 89, and liquid trap 89 collects liquid flowing from flow cell 62. For example, the liquid trap 89 collects the liquid after the first reaction and the second reaction are performed.

In summary, in one embodiment of the present application, the fluid circuit system 12 includes a flow path selector valve 10, a pump 14, a plurality of flow paths 102, and a control circuit 63. The flow path selector valve 10 includes a manifold 24 and a first spool 26. The manifold 24 is provided with a common port 16, a plurality of first ports 18 and a plurality of second ports 20. The first valve element 26 is provided with a communication groove 21. The first spool 26 may rotate or slide relative to the manifold 24 to switch the flow path selector valve 10 between the first and second valve positions. With the flow path selector valve 10 in the first valve position, the first port 18 and the second port 20 communicate through the communication groove 21. With the flow path selector valve 10 in the second valve position, the first port 18 communicates with the common port 16 through the communication groove 21. A plurality of flow paths 102 are fluidly connected to flow cell 62 to support the target analyte when flow cell 62 is installed in fluid path system 12. Pump 14 is fluidly connected to flow cell 62 when flow cell 62 is installed in fluid system 12 and flows liquid through flow path 102 selected by flow path selection valve 10 during analysis operations. The control circuit 63 is operatively coupled to the flow path selection valve 10, the control circuit 63 having one or more processors and memory storing computer-executable instructions that, when executed by the processors, control the processors to command the flow path selection valve 10 to select a specified flow path 102.

Referring to fig. 15, the present application further provides a sequencing system 90, for example, the sequencing system 90 is a nucleic acid determination system, and the sequencing system 90 includes the fluid path system 12 of any of the above embodiments.

In one example, the control circuit 63 is configured to control the flow path selector valve 10 to rotate to a first valve position to communicate the first reservoir 66 and the flow cell 62, the first reservoir 66 carrying a first reaction liquid, the first reaction liquid comprising nucleic acid molecules; and configured to pass a first reaction liquid into the flow cell 62 to perform a first reaction with the flow path selection valve 10 in the first valve position, the first reaction including attaching at least a portion of the nucleic acid molecule to the flow cell 62; and a second valve position configured to rotate the flow path selector valve 10 to communicate the second reservoir 68 and the flow cell 62, the second reservoir 68 carrying a second reaction solution containing a component necessary for performing nucleic acid sequencing; and a second reaction liquid is caused to enter the flow cell 62 through the second flow channel to perform a second reaction including causing the nucleic acid molecule in the flow cell 62 after performing the first reaction to interact with the second reaction liquid to undergo a polymerization reaction and detecting a signal from the reaction to effect sequencing of the nucleic acid molecule, with the flow path selection valve 10 in the second valve position. The sequencing system 90 is a machine such as a sequencer or a sequencing platform.

In one example, the first reservoir 66 carries a biological sample solution containing nucleic acid molecules, the second reservoir 68 carries a reaction solution containing components required to perform a polymerization reaction, the control circuit 63 is configured to control the flow path selection valve 10 to allow the biological sample solution in the first reservoir 66 to enter the flow cell 62 to perform a first reaction including at least a portion of the nucleic acid molecules being connected to the flow cell 62, and the control circuit 63 is configured to control the flow path selection valve 10 to allow the reaction solution in the second reservoir 68 to enter the flow cell 62 to perform a second reaction including interaction of the nucleic acid molecules in the flow cell 62 with the reaction solution to perform the polymerization reaction after performing the first reaction.

The sequencing system 90 operates in accordance with commands that implement a prescribed protocol for testing, validation, analysis (e.g., including sequencing), and the like. The prescribed protocol will be established in advance and includes a series of events or operations for activities such as aspirating reagents, aspirating air, aspirating other fluids, ejecting such reagents, air and fluids, etc. The protocol will allow for coordination of such fluidic operations with other operations of the instrument, such as reactions occurring in flow cell 62, imaging of flow cell 62 and its site, and the like.

The present application also provides a system comprising the sequencing system 90 of any of the above embodiments.

The present application also provides a method for controlling a system to perform sequencing, wherein the system can be the above fluid path system 12, for example, the system comprises a plurality of flow paths 102, a flow cell 62 connected to the plurality of flow paths 102, a flow path selection valve 10, a pump 14, a first storage 66 and a second storage 68. The flow path selector valve 10 includes a manifold 24 and a first spool 26. The manifold 24 is provided with a common port 16, a plurality of first ports 18 and a plurality of second ports 20. The first valve element 26 is provided with a communication groove 21. The first spool 26 is rotatable or slidable relative to the manifold 24 to selectively communicate the communication groove 21 with the common port 16 and the first port 18 or the first port 18 and the second port 20 to select different flow paths. The first memory 66 is connected to the second port 18. The second memory 68 is connected to the common port 16. The first reservoir 66 carries a first reaction liquid containing nucleic acid molecules. The second reservoir 68 carries a second reaction solution containing components necessary for nucleic acid sequencing. The pump 14 is used to flow liquid through the flow path selected by the flow path selection valve 10.

Referring to fig. 16, the method includes: s110, switching the flow path selector valve 10 to the first valve position so that the communication groove 21 communicates the first port 18 and the second port 20 to communicate the first reservoir 66 and the flow cell 62; s120, with the flow path selection valve 10 in the first valve position, controlling the pump 14 to work to make the first reaction liquid enter the flow cell 62 to perform a first reaction, wherein the first reaction comprises connecting at least a part of the nucleic acid molecules to the flow cell 62; s130, switching the flow path selector valve 10 to the second valve position to cause the communication groove 21 to communicate the first port 18 and the common port 16 to communicate the second reservoir 68 and the flow cell 62; s140, with the flow path selection valve 10 in the second valve position, controlling the pump 14 to operate to cause the second reaction liquid to enter the flow cell 62 to perform a second reaction, the second reaction including causing the nucleic acid molecule in the flow cell 62 after the first reaction to interact with the second reaction liquid to undergo a polymerization reaction and detecting a signal from the reaction, to thereby effect sequencing of the nucleic acid molecule.

The method realizes the first reaction and the second reaction by enabling one flow path selection valve 10 to be in different valve positions to realize the switching of different flow paths/reagents, and is particularly suitable for the operation control of a system or equipment with high integration.

Current generation high throughput sequencing platforms or single molecule sequencing platforms generally require processing a sample to be tested before on-line sequencing, such as adapting to a designated sequencing platform, processing the sample to be tested to convert it into a library adapted to the sequencing platform, and loading the library into a designated area, such as a flow cell 62, so as to place the flow cell 62 containing the sample to be tested into a sequencer for automated sequencing. Currently commercially available sequencing platforms, the processing of the sample to be tested before being loaded on the machine is generally separated from the on-machine sequencing, for example, by manually performing sample processing/library preparation in a reagent tube, or by performing the processing and loading of the sample to be tested on a sample processing device. The method can realize sample processing and sequencing before the computer is operated, realize the switching and the in-and-out control of different flow channels of a plurality of reagents by enabling the flow path selection valve 10 to be in different valve positions, and is particularly suitable for an integrated sequencing platform integrating the sample processing and sequencing functions before the computer is operated.

Referring to fig. 17, in some embodiments, the system further includes a third memory 104, the third memory 104 is connected to the common port 16, and the third memory 104 carries a third reaction solution containing a component required for amplification. The method further comprises performing the following after performing step S120 and before performing S140 (after performing the first reaction and before performing the second reaction): pump 14 is controlled to operate to cause the third reaction solution in third storage 104 to enter flow cell 62 to perform a third reaction, which includes allowing the nucleic acid molecule in flow cell 62 after performing the first reaction to interact with the third reaction solution to achieve amplification of the nucleic acid molecule.

The term "amplification" refers to cloning a nucleic acid molecule, for example, by replicating thousands or even millions of copies of the nucleic acid molecule into a cluster (cluster) by polymerase chain reaction, any one of the thousands or millions of copies/cluster being identical in sequence to the original nucleic acid molecule, such that the signal from the nucleic acid molecule can be amplified by increasing the number of molecules, facilitating detection of the nucleic acid molecule; in particular, in subsequent sequencing, the signal emitted by the tens of thousands of millions of molecules (clusters) is equivalent to the signal from the single nucleic acid molecule, greatly enhancing the signal of the molecule, facilitating detection.

Second generation high throughput sequencing platforms currently on the market, such as ILLUMINA sequencing platform, Ion Torrent sequencing platform and chinese genom sequencing platform, require amplification of the signal from the molecule to be detected by amplification, such as bridge PCR or rolling circle amplification, prior to sequencing, to obtain a stronger signal that is easily recognized and detected (or not susceptible to interference).

Referring to fig. 17, in some embodiments, the system further includes a fourth reservoir 106, wherein the fourth reservoir 106 is connected to the common port 16, and the fourth reservoir carries a washing solution. The method further comprises, prior to performing step S120 (prior to performing the first reaction): with the flow path selector valve 10 in the first valve position, the pump 14 is controlled to operate to cause the wash solution in the fourth reservoir 106 to enter the flow cell 62. Thus, the washing solution can rinse the flow cell 62, thereby preventing the first reaction solution from being contaminated by the last residual substance and/or reducing unnecessary consumption of the first reaction solution (e.g., filling the conduit of the fluid path system 12) such as a trace amount of the biological sample solution.

In certain embodiments, the method further comprises performing (prior to the first reaction) the following steps prior to performing step S120: with the flow path selector valve 10 in the first valve position, the pump 14 is controlled to operate to vent air to the flow cell 62. Thus, before the first reaction is initiated, a section of air is injected to separate the first reaction solution flowing in subsequently from the cleaning solution in the liquid path system, so as to prevent the micro biological sample from being diffused and/or diluted, thereby preventing the nucleic acid molecules to be detected from being connected to the flow cell 62 and the subsequent detection of the nucleic acid molecules to be detected from being influenced.

In certain embodiments, the method further comprises performing the following steps before performing step S120 and/or before performing step S140 (before performing the first reaction and/or before performing the second reaction): pump 14 is controlled to operate to move the wash solution in fourth reservoir 106 into flow cell 62. Thus, the washing solution can clean the flow cell 62, and the subsequent first reaction solution and/or second reaction solution can be prevented from being affected by the previous reaction or the residue of the previous step.

In certain embodiments, flow cell 62 has a solid support surface having immobilized thereon a first sequencing primer, at least one end of the nucleic acid molecule comprises at least a portion of a sequence that is capable of complementary pairing with at least a portion of the first sequencing primer, and the first reaction comprises complementary pairing of at least a portion of the nucleic acid molecule with the first sequencing primer for ligation into flow cell 62.

The so-called "first sequencing primer" is an oligonucleotide (a short nucleic acid sequence of known sequence) of known sequence immobilized on the chip surface, often also called a "probe".

In some embodiments, the second reaction solution comprises a first nucleotide, a first polymerase, and a cleavage reagent, and step S140 comprises: (a) controlling the pump 14 to operate to bring a first nucleotide and a first polymerase into the flow cell 62, and subjecting the flow cell 62 to conditions suitable for a polymerization reaction to bind the first nucleotide to a nucleic acid molecule by extending a first sequencing primer, the first nucleotide comprising a base, a sugar unit, a cleavable blocking group, and a detectable label; (b) exciting the detectable label and collecting a signal from the detectable label; (c) controlling the pump 14 to operate to cause the cleavage reagent to enter the flow cell 62 to remove the cleavable blocking group and the detectable label of the first nucleotide; (d) repeating (a) - (c) at least once.

In particular, by "subjecting to conditions suitable for a polymerization reaction", temperature conditions are generally involved in addition to the components/reagents required for the polymerization reaction (e.g., polymerase, reaction substrates, i.e., nucleotides, and/or sequencing primers). For example, the nucleic acid sequencing system 90 may further comprise a temperature control system that controls the temperature of the flow cell 62/reaction chamber to achieve "conditions suitable for polymerization".

The detectable label is for example an optically detectable label, for example a fluorescent molecule. The cleavable blocking group can prevent/inhibit the binding of other nucleotides (first nucleotide) in the reaction system to the next position of the nucleic acid molecule to be detected, and can be a physical block such as a sugar group with an azido group (-N) at the 3' -position₃) It may be a non-physical block (virtual block) such as a steric conformation in which the blocking group can form a barrier to the progress of the extension reaction in the extension reaction solution system.

In certain embodiments, the second reaction solution further comprises a second nucleotide, and step S140 further comprises performing the following after (a): controlling the pump 14 to operate to bring the second nucleotide and the first polymerase into the flow cell and to place the flow cell under conditions suitable for a polymerization reaction to bind the second nucleotide to the nucleic acid molecule by continuing to extend the product after (a), the second nucleotide comprising a base, a sugar unit and a cleavable blocking group. This step is advantageously performed to synchronize the reaction of multiple nucleic acid molecules in a cluster, i.e., to eliminate or reduce to some extent the reaction of leading (preceding) or lagging (phasing) nucleic acid molecules in a cluster, which facilitates the sequencing reaction.

Referring to fig. 19, in some embodiments, the second reaction solution comprises a third nucleotide, a fourth nucleotide, a second polymerase, a third polymerase, a cleavage reagent, and a second sequencing primer, at least one end of the nucleic acid molecule comprises at least a portion of a sequence that is complementary to at least a portion of the second sequencing primer, and the operation of the pump 14 is controlled such that the second reaction solution enters the flow cell 62 through the second flow channel to perform the second reaction comprises: (i) controlling the pump 14 to operate to bring a third nucleotide and a second polymerase into the flow cell 62, and subjecting the flow cell 62 to conditions suitable for a polymerization reaction to bind the third nucleotide to the nucleic acid molecule by extending the first sequencing primer to obtain a nascent strand, the third nucleotide being a nucleotide that carries neither a cleavable blocking group nor a detectable label; (ii) controlling pump 14 to operate to bring a fourth nucleotide, a third polymerase and a second sequencing primer into flow cell 62, and subjecting flow cell 62 to conditions suitable for a polymerization reaction to cause the second sequencing primer to bind to the nascent strand and the fourth nucleotide to bind to the nascent strand by extending the second sequencing primer, the fourth nucleotide comprising a base, a sugar unit, a cleavable blocking group and a detectable label; (iii) exciting the detectable label and collecting a signal from the detectable label; (iv) controlling the pump 14 to operate to cause the cleavage reagent to enter the flow cell 62 to remove the cleavable blocking group and the detectable label of the fourth nucleotide; (v) (iii) repeating (ii) - (iv) at least once. Thus, a second reaction can be achieved by steps (ii) to (v) to achieve sequencing of the nucleic acid molecule.

The third nucleotide can be, for example, a natural nucleotide; the fourth nucleotide may be identical to the first nucleotide; the first to third polymerases can be the same or different, e.g., different types of DNA polymerases or different mutants of the same type of DNA polymerase, each independently capable of binding to a specified nucleotide effective to catalyze the performance of a specified extension/polymerization reaction.

Compared with the sequencing method in the previous embodiment, the method can be used for sequencing the sequence of the other end of the nucleic acid molecule to be detected by synthesizing the complementary strand of the nucleic acid molecule to be detected, so as to realize sequencing of the other end of the nucleic acid molecule to be detected.

Similarly, in certain embodiments, the second reaction solution further comprises a fifth nucleotide, the method further comprising after (ii), controlling the pump 14 to operate so that the fifth nucleotide and the third polymerase enter the flow cell, and subjecting the flow cell to conditions suitable for a polymerization reaction to bind the fifth nucleotide to the nascent strand by continuing to extend the product after (ii), the fifth nucleotide comprising a base, a sugar unit, and a cleavable blocking group. The fifth nucleotide, for example, the second nucleotide, is a reversible terminator without a detectable label, and this step is performed to facilitate simultaneous reaction of multiple nucleic acid molecules in a cluster, i.e., to eliminate or reduce a certain amount of reaction-leading (preceding) or-lagging (phasing) nucleic acid molecules in a cluster, to facilitate the sequencing reaction, and to obtain longer reads (reads).

It should be noted that the explanations of the technical features of the flow path selection valve 10 and/or the fluid path system 12, such as the structure, connection relationship and operation control, in the above embodiments are also applicable to the method for implementing the sequencing by the control system of any of the embodiments, how to implement the method for implementing the sequencing by the control system of any of the embodiments by using the flow path selection valve 10 and the related elements/structural components, not expanded herein, a person skilled in the art can understand how to utilize and control the flow path selection valve 10 and/or the liquid path system 12 and/or the sequencing system 90 in the above embodiments to implement the corresponding sequencing method by the exemplary descriptions of the structure, connection relationship, function and operation manner of the flow path selection valve 10 and/or the liquid path system 12 in the above embodiments and the current sequencing method.

The present application also provides a method, which includes a method for implementing sequencing by the control system according to any of the above embodiments.

In the description herein, reference to the description of the terms "certain embodiments," "one embodiment," "some embodiments," "illustrative embodiments," "examples," "specific examples," or "some examples" means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the application. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

The logic and/or steps represented in the flowcharts or otherwise described herein, such as an ordered listing of executable instructions that can be considered to implement logical functions, can be embodied in any computer-readable storage medium for use by or in connection with an instruction execution system, apparatus, or device, such as a computer-based system, processor-containing system, or other system that can fetch the instructions from the instruction execution system, apparatus, or device and execute the instructions. For the purposes of this description, a "computer-readable storage medium" can be any means that can contain, store, communicate, propagate, or transport the program for use by or in connection with the instruction execution system, apparatus, or device.

In addition, functional units in the embodiments of the present invention may be integrated into one processing module, or each unit may exist alone physically, or two or more units are integrated into one module. The integrated module can be realized in a hardware mode, and can also be realized in a software functional module mode. The integrated module, if implemented in the form of a software functional module and sold or used as a separate product, may also be stored in a computer-readable storage medium.

Although embodiments of the present application have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present application, and that variations, modifications, substitutions and alterations of the above embodiments may be made by those of ordinary skill in the art within the scope of the present application, which is defined by the claims and their equivalents.

Claims

1. A fluid path system, comprising:

a plurality of flow paths fluidly connected with the flow cell to support a target analyte when the flow cell is installed in the fluidic circuit system;

a flow path selector valve connected to the flow path, the flow path selector valve including a manifold provided with a common port, a plurality of first ports, and a plurality of second ports, and a first spool provided with a communication groove, the first spool being rotatable or slidable with respect to the manifold to switch the flow path selector valve between a first valve position, through which the first ports communicate with the second ports, and a second valve position, through which the first ports communicate with the common port when the flow path selector valve is in the first valve position, and the second valve position, through which the first ports communicate with the common port;

a pump fluidly connected to the flow cell when the flow cell is installed in the fluid path system and flowing liquid through the flow path selected by the flow path selection valve during an analysis operation; and

a control circuit operatively coupled to the flow path selection valve, the control circuit for controlling the flow path selection valve to select a specified flow path.

2. The fluid path system of claim 1, wherein the flow path selector valve includes a second spool disposed on the manifold, the first spool disposed on the second spool, the second spool disposed with a first passage in communication with the first port, a second passage in communication with the second port, and a third passage in communication with the common port,

the communication groove communicates the first passage and the second passage to communicate the first port and the second port with the flow path selector valve in the first valve position;

the communication groove communicates the first passage and the third passage to communicate the first port with the common port with the flow path selection valve in the second valve position; optionally, the manifold is provided with a cavity, the second valve spool being at least partially received in the cavity; optionally, a first connection port, a second connection port and a third connection port are arranged on the bottom surface of the cavity, the first connection port is communicated with the first port and the first channel, the second connection port is communicated with the second port and the second channel, and the third connection port is communicated with the common port and the third channel; optionally, at least two of the first port, the second port and the common port are located on different sides of the manifold, respectively; optionally, the communication groove includes a first communication groove and a second communication groove which are provided at intervals, the first port and the second port communicate through the first communication groove with the flow path selector valve in the first valve position, and the first port and the common port communicate through the second communication groove with the flow path selector valve in the second valve position; optionally, the flow path selector valve comprises a drive member for driving the first spool to rotate or slide; optionally, the flow path selector valve comprises a slider connected to the first spool, the slider being connected to the drive member, the drive member driving the first spool to slide via the slider; optionally, the drive component provides an electromagnetic drive; optionally, the flow path selector valve comprises a housing removably connected to the manifold, the first spool and the slider being housed in the housing; optionally, the housing is provided with a guide groove, the slider includes a connecting portion and a guide rail portion connected to the connecting portion, the connecting portion is connected to the first valve element, and the guide rail portion is matched with the guide groove to guide the slider to drive the first valve element to slide.

3. The fluid path system of claim 1, further comprising:

a reagent selection valve to select a reagent from a plurality of reagents according to an analysis protocol; and

a flow cell carrying a target analyte;

the flow path selection valve is fluidly connected between the reagent selection valve and the flow cell, the flow path selection valve selecting a flow path of a flow-through flow cell from a plurality of flow paths through the flow cell according to an analytical protocol and directing the selected reagent through the flow cell, the pump flowing the selected reagent through the selected flow path according to the analytical protocol; optionally, the flow cell comprises a plurality of channels arranged in parallel, one end of the channels communicating with the first port; optionally, one end of the channel is un-piped to the first port.

4. The fluid path system of claim 1, further comprising a first memory and a second memory,

the first storage stores a sample solution to be tested, the first storage is connected with the second port,

the second memory stores a reaction reagent, and the second memory is connected with the common port.

5. A sequencing system comprising the fluid path system of any one of claims 1-4.

6. A method of controlling a system to effect sequencing, the system comprising a plurality of flow paths, a flow cell connected to the plurality of flow paths, a flow path selection valve and pump, a first reservoir, and a second reservoir,

the flow path selection valve includes a manifold provided with a common port, a plurality of first ports and a plurality of second ports, and a first spool provided with a communication groove, the first spool being rotatable or slidable with respect to the manifold to selectively communicate the common port with the first port or the first port with the second port, thereby selecting different flow paths,

the first memory is connected with the second port,

the second memory is connected to the common port,

the first memory bears a first reaction liquid, the first reaction liquid contains nucleic acid molecules,

the second memory carries a second reaction solution containing a component necessary for nucleic acid sequencing,

the pump is used to flow liquid through the flow path selected by the flow path selection valve,

the method comprises the following steps:

switching the flow path selector valve to a first valve position to cause the communication groove to communicate the first port and the second port to communicate the first reservoir and the flow cell;

controlling the pump to operate to cause the first reaction solution to enter the flow cell to perform a first reaction including attaching at least a portion of the nucleic acid molecule to the flow cell with the flow path selection valve in the first valve position;

switching the flow path selector valve to a second valve position to cause the communication groove to communicate the first port and the common port to communicate a second reservoir and a flow cell;

controlling the pump to operate to cause the second reaction solution to enter the flow cell to perform a second reaction including causing a nucleic acid molecule in the flow cell after the first reaction is performed to interact with the second reaction solution to undergo a polymerization reaction and detecting a signal from the reaction with the flow cell in a state where the flow path selection valve is in the second valve position to effect sequencing of the nucleic acid molecule.

7. The method of claim 6, wherein the system further comprises a third reservoir connected to the common port, the third reservoir carrying a third reaction solution comprising components required for performing amplification, the method further comprising, after performing the first reaction and before performing the second reaction:

controlling the pump to work so that a third reaction solution in a third storage enters the flow cell to perform a third reaction, wherein the third reaction comprises the interaction of the nucleic acid molecules in the flow cell after the first reaction and the third reaction solution so as to realize the amplification of the nucleic acid molecules; optionally, the system further comprises a fourth reservoir connected to the common port, the fourth reservoir carrying a wash solution, the method further comprising, prior to performing the first reaction:

controlling the pump to operate to cause wash solution in a fourth reservoir to enter the flow cell with the flow path selection valve in the first valve position; optionally, further comprising, prior to performing the first reaction:

controlling the pump to operate to pump air to the flow cell with the flow path selection valve in the first valve position; optionally, the system further comprises a fourth reservoir connected to the common port, the fourth reservoir carrying a wash solution, further comprising, prior to performing the first reaction and/or prior to performing the second reaction:

controlling the pump to work so that the washing solution in the fourth storage tank enters the flow cell.

8. The method of claim 6, wherein the flow cell has a solid support surface having a first sequencing primer immobilized thereon, at least one end of the nucleic acid molecule comprises at least a portion of a sequence capable of complementary pairing with at least a portion of the first sequencing primer, and the first reaction comprises complementary pairing of at least a portion of the nucleic acid molecule with the first sequencing primer for ligation into the flow cell; optionally, the second reaction solution comprises a first nucleotide, a first polymerase and a cleavage reagent, and the controlling the pump to operate to cause the second reaction solution to enter the flow cell to perform a second reaction comprises:

(a) controlling the pump to operate so that the first nucleotide and the first polymerase enter the flow cell and the flow cell is under conditions suitable for a polymerization reaction to bind the first nucleotide to the nucleic acid molecule by extending the first sequencing primer, the first nucleotide comprising a base, a sugar unit, a cleavable blocking group, and a detectable label;

(b) exciting the detectable label and collecting a signal from the detectable label;

(c) controlling the pump to operate to cause the cleavage reagent to enter the flow cell to remove the cleavable blocking group and detectable label of the first nucleotide;

(d) repeating (a) - (c) at least once; optionally, the second reaction solution further comprises a second nucleotide, the controlling the pump to operate to cause the second reaction solution to enter the flow cell for a second reaction, further comprising after (a),

controlling the pump to operate so that the second nucleotide and the first polymerase enter the flow cell and the flow cell is subjected to conditions suitable for a polymerization reaction to bind the second nucleotide to the nucleic acid molecule by continuing to extend the product after (a), the second nucleotide comprising a base, a sugar unit and a cleavable blocking group; optionally, the second reaction solution comprises a first nucleotide, a second nucleotide, a first polymerase, a second polymerase, a cleavage reagent, and a second sequencing primer, at least one end of the nucleic acid molecule comprises at least a portion of a sequence capable of complementary pairing with at least a portion of the second sequencing primer, and the controlling the pump to operate to cause the second reaction solution to enter the flow cell for a second reaction comprises:

(i) controlling the pump to operate so that the first nucleotide and the first polymerase enter the flow cell and the flow cell is under conditions suitable for a polymerization reaction to bind the first nucleotide to the nucleic acid molecule by extending the first sequencing primer to obtain a nascent strand, the first nucleotide being a nucleotide bearing neither a cleavable blocking group nor a detectable label;

(ii) controlling the pump to operate so that the second nucleotide, the second polymerase and the second sequencing primer enter the flow cell and the flow cell is under conditions suitable for a polymerization reaction so that the second sequencing primer binds to the nascent strand and the second nucleotide binds to the nascent strand by extending the second sequencing primer, the second nucleotide comprising a base, a sugar unit, a cleavable blocking group and a detectable label;

(iii) exciting the detectable label and collecting a signal from the detectable label;

(iv) controlling the pump to operate to cause the cleavage reagent to enter the flow cell to remove the cleavable blocking group and detectable label of the second nucleotide;

(v) (iii) repeating (ii) - (iv) at least once; optionally, the second reaction solution further comprises a third nucleotide, the controlling the pump to operate to cause the second reaction solution to enter the flow cell for a second reaction, further comprising after (ii),

(iii) controlling the pump to operate so that the third nucleotide and the second polymerase enter the flow cell and the flow cell is subjected to conditions suitable for a polymerisation reaction to bind the third nucleotide to the nascent strand by continuing to extend the product after (ii), the third nucleotide comprising a base, a sugar unit and a cleavable blocking group.

9. The method of claim 6, wherein the flow path selection valve includes a second spool disposed on the manifold, the first spool being disposed on the second spool, the second spool being provided with a first passage communicating with the first port, a second passage communicating with the second port, and a third passage communicating with the common port,

the communication groove communicates the first passage and the third passage to communicate the first port with the common port with the flow path selection valve in the second valve position; optionally, the manifold is provided with a cavity, the second valve spool being at least partially received in the cavity; optionally, a first connection port, a second connection port and a third connection port are arranged on the bottom surface of the cavity, the first connection port is communicated with the first port and the first channel, the second connection port is communicated with the second port and the second channel, and the third connection port is communicated with the common port and the third channel; optionally, at least two of the first port, the second port and the common port are respectively located on different sides of the manifold; optionally, the communication groove includes a first communication groove and a second communication groove which are provided at intervals, the first port and the second port communicate through the first communication groove with the flow path selector valve in the first valve position, and the first port and the common port communicate through the second communication groove with the flow path selector valve in the second valve position; optionally, the flow path selector valve comprises a drive member for driving the first spool to rotate or slide; optionally, the flow path selection valve comprises a slider connected to the first valve core, the slider is connected to the driving part, and the driving part drives the first valve core to slide through the slider; optionally, the drive component provides an electromagnetic drive; optionally, the flow path selector valve comprises a housing removably connected to the manifold, the first spool and the slider being housed in the housing; optionally, the housing is provided with a guide groove, the slider includes a connecting portion and a guide rail portion connected to the connecting portion, the connecting portion is connected to the first valve element, and the guide rail portion is matched with the guide groove to guide the slider to drive the first valve element to slide.

10. The method of claim 6, wherein the system further comprises:

a reagent selection valve to select a reagent from a plurality of reagents according to an analysis protocol;

the flow path selection valve is fluidly connected between the reagent selection valve and the flow cell, the flow path selection valve selects a flow path of a flow-through flow cell from a plurality of flow paths through the flow cell according to an analytical protocol and directs the selected reagent through the flow cell, the pump flows the selected reagent through the selected flow path according to the analytical protocol; optionally, the flow cell comprises a plurality of channels arranged in parallel, one end of the channels communicating with the first port; optionally, one end of the channel is un-piped to the first port.