CN110904206A - Liquid path system, biomolecule analysis system and nucleic acid sequence measuring system - Google Patents

Abstract

Translated fromChinese

本申请公开了一种液路系统及生物分子分析系统。液路系统用于为分析生物分子提供溶液环境，第一反应包括利用第一试剂使生物分子连接至反应装置中，第二反应包括利用第二试剂对连接至反应装置中的生物分子进行检测。液路系统包括阀体组件及驱动组件。阀体组件包括第一多通阀及第二多通阀。驱动组件包括第一泵及第二泵。在第一多通阀连通反应装置和第一泵，且第二多通阀连通反应装置和第一试剂的状况下，第一泵用于驱动使第一试剂沿第一方向进入反应装置，以进行第一反应；在第二多通阀连通反应装置和第二泵，且第一多通阀连通反应装置和第二试剂的状况下，第二泵用于驱动使第二试剂沿第二方向进入反应装置，以进行第二反应。

The present application discloses a liquid circuit system and a biomolecule analysis system. The liquid circuit system is used to provide a solution environment for analyzing biomolecules. The first reaction includes using the first reagent to connect the biomolecules to the reaction device, and the second reaction includes using the second reagent to detect the biomolecules connected to the reaction device. The hydraulic system includes valve body components and drive components. The valve body assembly includes a first multi-way valve and a second multi-way valve. The drive assembly includes a first pump and a second pump. Under the condition that the first multi-port valve communicates with the reaction device and the first pump, and the second multi-port valve communicates with the reaction device and the first reagent, the first pump is used to drive the first reagent to enter the reaction device along the first direction, so as to Carry out the first reaction; under the condition that the second multi-port valve communicates with the reaction device and the second pump, and the first multi-port valve communicates with the reaction device and the second reagent, the second pump is used to drive the second reagent along the second direction Enter the reaction device for the second reaction.

Description

Liquid path system, biomolecule analysis system and nucleic acid sequence measuring system

Technical Field

The present application relates to the field of biomolecule analysis technology, and more particularly, to a fluid path system, a biomolecule analysis system, and a nucleic acid sequence determination system.

Background

In the related art, apparatuses for performing nucleic acid analysis on a nucleic acid sample placed on a solid substrate, such as nucleic acid sequencing (sequencing) based on chip detection, generally involve sample processing before loading a sample to be tested so as to meet the requirements of the loading on a computer, such as connection to a chip, and loading the processed sample into a sequencer for sequencing.

With the development of automated related technologies, related commercially available sequencing platforms often include two or more separate or functionally linked devices, which are typically sold or purchased in sets when a certain sequencing platform is sold or purchased.

For a sequencing platform comprising two associated devices, typically one of the devices for processing a nucleic acid sample to be tested comprises attaching it to a reaction device, such as a chip, the device often being referred to as a hybridization apparatus, a sample introduction apparatus or a sample processing apparatus; the other device detects the chip connected with the nucleic acid sample output by the last device, thereby realizing the determination of the nucleic acid sequence, and the device is the most main hardware of a sequencing platform and is often called a sequencer.

A commercial sequencing platform, which comprises a plurality of automated devices for processing and detecting samples, is at least partly due to the complexity of the fluid control involved in processing samples before computer and in computer sequencing, for example, sample processing before computer often includes a plurality of steps and a plurality of reactions, involving a plurality of reagents and a plurality of fluid inlet and outlet sequences, and some of the reagents or reactions may have special requirements on the material, flow rate, pressure, etc. of the pipeline, and further, sequencing itself generally involves a plurality of reagents and a plurality of reactions; still another part is the susceptibility of biochemical reactions to interference, including the susceptibility of reagents to contamination.

Therefore, the liquid path system for sample processing before the computer and the liquid path system for sequencing are integrated, a set of liquid path system which can realize the processing of the sample before the computer and the sequencing of the processed sample can be constructed, the interference or the pollution can not be caused, and the method is also suitable for industrialization and is very difficult.

Disclosure of Invention

The embodiment of the application provides a liquid path system, a biomolecule analysis system and a nucleic acid sequence measuring system.

The liquid path system of this application embodiment is used for providing solution environment for the analysis biomolecule, the analysis biomolecule includes and carries out first reaction and second reaction on reaction unit, first reaction includes that to utilize first reagent to make the biomolecule be connected to in the reaction unit, the second reaction includes to utilize the second reagent to be connected to biomolecule among the reaction unit detects, the liquid path system includes valve body subassembly and drive assembly, the valve body subassembly includes first multi-way valve and second multi-way valve, first multi-way valve can communicate the second reagent with reaction unit, the second multi-way valve can communicate the first reagent with reaction unit; the driving assembly comprises a first pump and a second pump, the first pump is connected with the first multi-way valve, and the second pump is connected with the second multi-way valve; under the condition that the first multi-way valve is communicated with the reaction device and the first pump, and the second multi-way valve is communicated with the reaction device and the first reagent, the first pump is used for driving the first reagent to enter the reaction device along a first direction so as to perform the first reaction; and under the condition that the second multi-way valve is communicated with the reaction device and the second pump, and the first multi-way valve is communicated with the reaction device and the second reagent, the second pump is used for driving the second reagent to enter the reaction device along a second direction so as to perform the second reaction.

The biological molecules are referred to as biological macromolecules, such as proteins or nucleic acids. In certain embodiments, the biomolecule is a nucleic acid, the first reaction comprises a hybridization reaction, and/or the second reaction comprises a sequencing reaction.

In certain embodiments, the valve body assembly further comprises a third multi-way valve that can communicate the second reagent and the first multi-way valve.

In certain embodiments, the third multi-way valve includes a stator and a rotor that are communicable, the third multi-way valve includes a common port, the stator includes a plurality of ports, the rotor includes a communication groove, and the rotor is rotatable to communicate the common port with at least one of the ports through the communication groove.

In some embodiments, the first multi-way valve includes a plurality of ports, the plurality of ports are connected to at least the second reagent, the reaction device and the first pump, respectively, and any two ports on the first multi-way valve can be communicated.

In some embodiments, the reaction apparatus includes a first unit and a second unit, the first multi-way valve is a four-way valve, and four ports of the four-way valve are respectively connected to the second reagent, the first unit, the second unit, and the first pump.

In some embodiments, the second multi-way valve includes three ports, three of the ports are respectively connected to the first reagent, the reaction device and the second pump, and any two of the ports of the second multi-way valve can be communicated.

In certain embodiments, the second multi-way valve is a three-way valve.

In some embodiments, the reaction device comprises a plurality of channels, the number of the second multi-way valves and the number of the second pumps are not less than the number of the channels, and one second multi-way valve can communicate one channel and one second pump.

In certain embodiments, the fluid path system further comprises a manifold assembly in communication with the first pump, the manifold assembly configured to collect the first reagent after the first reaction; and/or the collecting assembly is communicated with the second pump and is used for collecting the second reagent after the second reaction.

In certain embodiments, the manifold assembly is further configured to collect the second reagent driven by the first pump when the first pump and the second reagent are in communication through the first multi-way valve.

In certain embodiments, the manifold assembly includes a fluid trap including a first port in communication with a plurality of the second ports and a plurality of second ports, a first pump in communication with the second ports, a second pump in communication with the second ports, and a waste bottle in communication with the first ports.

In certain embodiments, after performing the first reaction or before beginning the second reaction, the first multi-way valve can communicate the first pump and the second reagent, and/or communicate the second pump and the second reagent, and the first pump and/or the second pump can be used to drive the second reagent to fill the flow path of the first multi-way valve with the second reagent.

In certain embodiments, the first direction is opposite the second direction.

The biomolecule analysis system of the embodiments of the present application includes the liquid path system of any of the embodiments of the present application.

In certain embodiments, the biomolecule analysis system further includes a reaction device connecting the first and second multi-way valves.

In some embodiments, the biomolecule is a nucleic acid, the reaction device includes a first unit and a second unit, the biomolecule analysis system further includes a signal acquisition device for acquiring a signal; the second reaction is a nucleic acid sequencing reaction comprising a plurality of repetitive reactions, one of the repetitive reactions comprising a base extension reaction, signal acquisition, and radical cleavage; and (b) performing the base extension reaction and/or the radical cleavage in one of the first unit and the second unit by using the liquid path system, and simultaneously performing the signal acquisition in the other of the first unit and the second unit by using the signal acquisition device. Thus, the efficiency of measuring the nucleic acid molecule can be improved.

The nucleic acid sequencing system of the embodiments of the present application includes the fluid path system of any of the embodiments of the present application.

In certain embodiments, the nucleic acid sequence determination system further comprises a reaction device connecting the first multi-way valve and the second multi-way valve.

In the fluid path system, biomolecule analytic system and nucleic acid sequence measurement system of any embodiment of this application, through switching first multi-way valve intercommunication reaction unit and first pump, and switch second multi-way valve intercommunication reaction unit and first reagent, can carry out first reaction in the reaction unit, through switching second multi-way valve intercommunication reaction unit and second pump, and switch first multi-way valve intercommunication reaction unit and second reagent, can carry out the second reaction in the reaction unit, can realize carrying out first reaction and second reaction on the reaction unit through a fluid path system, the structure of the fluid path system of realization first reaction and second reaction is simpler, and, the cost of setting up this integrated fluid path system is far less than the sum of the cost of setting up the fluid path system of realizing first reaction alone and the fluid path system of realizing the second reaction alone.

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 a block diagram of a biomolecule analysis system in one state according to an embodiment of the present application;

FIG. 2 is a schematic block diagram of a biomolecule analysis system in another state according to an embodiment of the present application;

FIG. 3 is a schematic illustration of the third and first multi-way valves of an embodiment of the present application;

fig. 4 is a block diagram of a biomolecule analysis system according to an embodiment of the present application in still another state.

Description of the main element symbols:

the system comprises abiomolecule analysis system 1000, aliquid path system 100, avalve body assembly 10, a firstmulti-way valve 11,ports 111, 112, 113, 114, a secondmulti-way valve 12,ports 121, 122, 123, a thirdmulti-way valve 13, astator 131, arotor 132, acommon port 133, aport 134, acommunication groove 135, a driving assembly 20, afirst pump 21, asecond pump 22, aflow collecting assembly 30, aliquid collecting piece 31, asecond port 311, afirst port 312, awaste liquid bottle 32, areaction device 200, afirst unit 201, asecond unit 202, achannel 203, asecond reagent 300 and afirst reagent 400.

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 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.

Referring to fig. 1, the present embodiment provides abiomolecule analysis system 1000, and thebiomolecule analysis system 1000 includes afluid path system 100 and adetachable reaction device 200 connected to thefluid path system 100. Thebiomolecule analysis system 1000 may be specifically a nucleic acid sequence determination system, and thefluid path system 100 is used to provide a solution environment for analyzing biomolecules including performing a first reaction and a second reaction in thereaction device 200. Wherein the first reaction includes coupling the biomolecule into thereaction device 200 using thefirst reagent 400, and the second reaction includes detecting the biomolecule coupled into thereaction device 200 using thesecond reagent 300.

Thefluid circuit system 100 includes avalve body assembly 10 and a drive assembly 20. Thevalve body assembly 10 includes a firstmulti-way valve 11 and a secondmulti-way valve 12. The firstmulti-way valve 11 may communicate thesecond reagent 300 with thereaction device 200, and the secondmulti-way valve 12 may communicate thefirst reagent 400 with thereaction device 200. The driving assembly 20 includes afirst pump 21 and asecond pump 22, thefirst pump 21 is connected to the firstmulti-way valve 11, and thesecond pump 22 is connected to the secondmulti-way valve 12. Thereaction device 200 is connected between the first and secondmulti-way valves 11 and 12.

In a state where the firstmulti-way valve 11 communicates with thereaction device 200 and thefirst pump 21, and the secondmulti-way valve 12 communicates with thereaction device 200 and thefirst reagent 400, thefirst pump 21 is used to drive thefirst reagent 400 to enter thereaction device 200 in the first direction to perform the first reaction. Under the condition that the secondmulti-way valve 12 communicates with thereaction device 200 and thesecond pump 22, and the firstmulti-way valve 11 communicates with thereaction device 200 and thesecond reagent 300, thesecond pump 22 is used for driving thesecond reagent 300 to enter thereaction device 200 along the second direction to perform the second reaction.

In thebiomolecule analysis system 1000 according to the embodiment of the present application, thereaction apparatus 200 and thefirst pump 21 are communicated by switching the firstmulti-way valve 11, and thereaction apparatus 200 and thefirst reagent 400 are communicated by switching the secondmulti-way valve 12, a first reaction may be carried out in thereaction apparatus 200 by switching the secondmulti-way valve 12 to communicate thereaction apparatus 200 with thesecond pump 22, and switching the firstmulti-way valve 11 to communicate thereaction apparatus 200 with thesecond reagent 300, the second reaction can be carried out in thereaction device 200, the first reaction and the second reaction can be carried out in thereaction device 200 through oneliquid path system 100, the structure of theliquid path system 100 for realizing the first reaction and the second reaction is simpler, the first reaction and the second reaction do not need to be carried out in different systems separately, the operation of a user is simpler and more convenient, moreover, the cost of building this integratedfluid circuit system 100 is much lower than the sum of the costs of building a fluid circuit system that implements the first reaction alone and a fluid circuit system that implements the second reaction alone.

Specifically, referring to fig. 1 and 2, fig. 1 is a schematic block diagram illustrating a first reaction performed on areaction apparatus 200, and fig. 2 is a schematic block diagram illustrating a second reaction performed on thereaction apparatus 200. The first reaction of the present embodiment includes a reaction of attaching a biomolecule into thereaction device 200, or is referred to as a hybridization reaction. The term biomolecule includes 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. Thereaction apparatus 200 may be a reaction site including a substrate, and thereaction apparatus 200 may be in the form of a chip, and thereaction apparatus 200 may be detachably connected to theliquid path system 100. 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, so that the biomolecule is attached to thereaction device 200. Thefirst reagent 400 may include a solution for performing a first reaction, for example, a hybridization solution including the above-described biomolecules.

The second reaction includes a reaction for detecting a biomolecule, for example, a nucleic acid, attached to thereaction device 200, and the second reaction may be a sequencing reaction, so-called sequencing, 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 comprise one or more sub-reactions, in one example, sequencing the DNA, the second reaction being sequencing, based on sequencing by synthesis or sequencing by ligation, the sequencing comprising a plurality of sub-reactions including base extension reaction, signal acquisition and radical excision; 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).

Wherein the base extension reaction comprises binding nucleotide glycosides (including modified nucleotides) to nucleic acid molecules by polymerase or ligase based on the base complementary principle for Sequencing By Synthesis (SBS) or Sequencing By Ligation (SBL) on thereaction apparatus 200 on which the nucleic acid molecules are immobilized, and collecting corresponding reaction signals. For example, an engineered nucleotide may refer to a nucleotide with a label that allows the engineered nucleotide to be detected under certain circumstances, e.g., a nucleotide with a fluorescent molecular label that fluoresces when excited by a laser of a particular wavelength; in general, for SBS, an engineered nucleotide typically 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, e.g., a blocking group such as an azide (-N) group attached at the 3' position of the sugar moiety of the nucleotide₃)。

Thesecond reagent 300 comprises a polymerase and an engineered nucleotide, and in one example, thesecond reagents 300 are each loaded in five separate containers, each of which is loaded with a polymerase and four engineered nucleotides, the polymerase and one or more nucleotides are mixed in thereaction apparatus 200, and the controlled polymerase chain reaction is performed under suitable conditions, i.e., the base extension reaction is achieved. In another example, thesecond reagent 300 further includes other solutions, such as imaging reagents, washing reagents, etc., that are independently supported.

Collecting the signal includes collecting the signal emitted from the modified nucleotide bound to the nucleic acid molecule, for example, by irradiating a specific region of thereaction apparatus 200 after the base extension reaction, in which the nucleotide labeled with the fluorescent molecule fluoresces, optionally, thesecond reagent 300 further includes an information collecting reagent such as an imaging reagent, for example, an antioxidant, to facilitate collection of the fluorescent signal, and the information collecting reagent can be carried independently in a container, and such second reagent 300 (hereinafter also referred to as the information collecting reagent) can be added to thereaction apparatus 200 to facilitate collection of information about the modified nucleotide, for example, to facilitate acquisition of the light emitted from the modified nucleotide or the fluorescent molecule thereon by the imaging apparatus into an image.

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. In one example, thesecond reagent 300 further comprises a cleavage reagent separately carried in a container that is passed to simultaneously remove the detectable label and blocking group on the engineered nucleotide upon the performance of the radical cleavage sub-reaction.

Thesecond reagent 300 introduced into thereaction apparatus 200 may be different depending on the different sub-reactions being performed, for example, thesecond reagent 300 including the modified nucleotide may be introduced when the base extension reaction is performed, the second reagent 300 (i.e., one of the information collecting reagents) advantageous for performing imaging may be introduced when the signal collection is performed, and thesecond reagent 300 for removing the detectable label and the blocking group on the modified nucleotide (hereinafter referred to as a cleavage reagent) may be introduced when the group cleavage is performed. Further, after the completion of the previous sub-reaction and before the start of the next sub-reaction, a washing reagent may be introduced to remove unreacted substances remaining in thereaction apparatus 200 or in thechannel system 100, substances interfering with the reaction or signal collection, the washing reagent may be one of thesecond reagents 300, and the washing reagent may be a buffer solution not interfering with the base extension reaction.

Unless otherwise specified, the second reaction described below is sequencing, including any one or more of the sub-reactions of the second reaction described above, and a washing procedure between the two sub-reactions.

With continued reference to fig. 1 and 2, thereaction apparatus 200 can provide reaction sites for the first reaction and the second reaction. Specifically, thereaction device 200 may include one ormore channels 203, and thefirst reagent 400 may enter thechannel 203 or flow through thechannel 203 in a first direction (e.g., a first direction X shown in fig. 1) when performing the first reaction, and thesecond reagent 300 may enter thechannel 203 or flow through thechannel 203 in a second direction (e.g., a second direction Y shown in fig. 1, which may be opposite to the first direction X) when performing the second reaction.

In the embodiment of the present application, thereaction apparatus 200 includes afirst unit 201 and asecond unit 202. Thefirst unit 201 comprises one ormore channels 203 and thesecond unit 202 comprises one ormore channels 203. The same reaction may be performed in thefirst cell 201 and thesecond cell 202, or different reactions may be performed in the first cell and the second cell. For example, when the first reaction is performed in thefirst unit 201, the first reaction or the second reaction may be performed in thesecond unit 202, or no reaction may be performed in thesecond unit 202; when the second reaction is performed in thefirst unit 201, the first reaction or the second reaction may be performed in thesecond unit 202, or no reaction may be performed in thesecond unit 202; when a sub-reaction (e.g., base extension) of the second reaction is performed in thefirst unit 201, another sub-reaction (e.g., signal acquisition) of the second reaction may be performed in thesecond unit 202, which is not limited herein.

In the example shown in fig. 1 and 2, thefirst unit 201 includes a plurality ofchannels 203. One end of the plurality ofchannels 203 of thefirst unit 201 adjacent to the firstmulti-way valve 11 may be commonly connected to oneport 112 of the firstmulti-way valve 11. One end of eachchannel 203 of thefirst unit 201 close to the secondmulti-way valve 12 can be connected with oneport 121 of one secondmulti-way valve 12, so that differentfirst reagents 400 can be introduced intodifferent channels 203 during the first reaction, different biomolecules can be connected intodifferent channels 203, and the differentfirst reagents 400 cannot be cross-contaminated before the first reaction. The arrangement of thechannels 203 in thesecond unit 202 may be the same as the arrangement of thechannels 203 in thefirst unit 201, and will not be described herein again.

With continued reference to fig. 1 and 2, the firstmulti-way valve 11 may communicate thefirst pump 21 with thereaction device 200, the firstmulti-way valve 11 may further communicate thesecond reagent 300 with thereaction device 200, and the firstmulti-way valve 11 may further communicate thesecond reagent 300 with thefirst pump 21. Specifically, the firstmulti-way valve 11 includes a plurality ofports 111, 112, 113, and 114, the plurality ofports 111, 112, 113, and 114 are connected to at least thesecond reagent 300, thereaction device 200, and thefirst pump 21, respectively, and any two of the plurality ofports 111, 112, 113, and 114 can communicate with the firstmulti-way valve 11. The firstmulti-way valve 11 may be a four-way valve, a five-way valve, a six-way valve, etc., and is not limited herein. When thereaction apparatus 200 includes only one of thefirst unit 201 and thesecond unit 202, the firstmulti-way valve 11 may also be a three-way valve. In the example shown in fig. 1 and 2, theport 111 is connected to thesecond reagent 300, theport 112 is connected to thefirst unit 201, theport 113 is connected to thesecond unit 202, and theport 114 is connected to thefirst pump 21.

The firstmulti-way valve 11 may communicate with any two of the plurality ofports 111, 112, 113, 114 to communicate a flow passage between any two of theports 111, 112, 113, 114. Theport 112 may be made to communicate with theport 114, for example, by switching the state of the firstmulti-way valve 11, to communicate thefirst pump 21 with thefirst unit 201; theport 113 may be made to communicate with theport 114 to communicate thefirst pump 21 with thesecond unit 202;port 111 may be brought into communication withport 112 to communicate thesecond reagent 300 with thefirst cell 201;port 111 may be put in communication withport connection 113 to communicate thesecond reagent 300 with thesecond cell 202;port 111 may be brought into communication withport 114 to communicate thesecond reagent 300 with thefirst pump 21.

With continued reference to fig. 1 and 2, the secondmulti-way valve 12 may communicate thefirst reagent 400 with thereaction apparatus 200, the secondmulti-way valve 12 may communicate thesecond pump 22 with thereaction apparatus 200, and the secondmulti-way valve 12 may further communicate thefirst reagent 400 with thesecond pump 22. Specifically, the secondmulti-way valve 12 includes threeports 121, 122, 123, the threeports 121, 122, 123 are respectively connected to thereaction device 200, thefirst reagent 400, and thesecond pump 22, and the secondmulti-way valve 12 can communicate any two of the threeports 121, 122, 123. The secondmulti-way valve 12 may be a three-way valve, a four-way valve, a five-way valve, etc., and is not limited herein.

The secondmulti-way valve 12 may communicate any two of the threeports 121, 122, 123 to communicate the flow passages to which any twoports 121, 122, 123 are connected. For example, by switching the state of the secondmulti-way valve 12, theport 121 can be made to communicate with theport 122 to communicate thefirst reagent 400 with thereaction device 200; theport 121 and theport 123 may be made to communicate thesecond pump 22 with the reaction device 200 (thefirst unit 201 or the second unit 202); by switching the state of the secondmulti-way valve 12, theport 122 may be brought into communication with theport 123 to communicate thefirst reagent 400 with thesecond pump 22.

When the number of thepassages 203 is plural, the number of the secondmulti-way valves 12 may also be plural, the number of the secondmulti-way valves 12 is not less than the number of thepassages 203, and one secondmulti-way valve 12 communicates with onepassage 203. At this time, the communication states of the plurality of secondmulti-way valves 12 may not be affected, for example, a part of the secondmulti-way valves 12 may communicate thefirst reagent 400 with thereaction apparatus 200, and another part of the secondmulti-way valves 12 may communicate thesecond pump 22 with thereaction apparatus 200, so thatdifferent channels 203 of thereaction apparatus 200 may perform different reactions.

With continued reference to fig. 1 and 2, thefirst pump 21 is connected to the firstmulti-way valve 11. When the firstmulti-way valve 11 communicates thefirst pump 21 with thesecond reagent 300, thefirst pump 21 may be used to drive thesecond reagent 300 into thefirst pump 21; when the firstmulti-way valve 11 communicates thefirst pump 21 with thereaction device 200, thefirst pump 21 can be used to drive the reagent in thereaction device 200 into thefirst pump 21. Thesecond pump 22 is connected to the secondmulti-way valve 12. When the secondmulti-way valve 12 communicates thefirst reagent 400 with thesecond pump 22, thesecond pump 22 may be used to drive thefirst reagent 400 into thesecond pump 22; when the secondmulti-way valve 12 communicates thereaction device 200 with thesecond pump 22, thesecond pump 22 may be used to drive the reagent in thereaction device 200 into thesecond pump 22. The number of the second pumps 22 may be plural, and the number of the second pumps 22 may be the same as the number of the secondmulti-way valves 12, and each of the second pumps 22 is connected to one of the secondmulti-way valves 12.

In one example, the firstmulti-way valve 11 can be directly connected to thesecond reagent 300, and in another example, the firstmulti-way valve 11 can be indirectly connected to thesecond reagent 300, for example, a valve body can be disposed between the firstmulti-way valve 11 and thesecond reagent 300. Referring to fig. 3, in the embodiment of the present application, thevalve body assembly 10 further includes a thirdmulti-way valve 13. The thirdmulti-way valve 13 may communicate thesecond reagent 300 with the firstmulti-way valve 11. The thirdmulti-way valve 13 may in particular be a rotary valve.

Specifically, the thirdmulti-way valve 13 includes astator 131 and arotor 132 that are communicable, the thirdmulti-way valve 13 includes acommon port 133, and thestator 131 includes a plurality ofports 134. Therotor 132 includes acommunication groove 135, and therotor 132 is rotatable to communicate thecommon port 133 with at least oneport 134 through thecommunication groove 135.

Thecommon port 133 may be connected to the firstmulti-way valve 11, and in particular, may be connected to theport 111 of the firstmulti-way valve 11. The number ofports 134 is plural, and may be six, seven, eight, ten, eleven, twelve, thirteen, fourteen, sixteen, etc., and eachport 134 may communicate with a different composition ofsecond reagent 300, such as oneport 134 communicating with reagent I, anotherport 134 communicating with reagent II, yet anotherport 134 communicating with reagent III, etc. Thesecond reagent 300 may be contained in reagent tubes, thesecond reagents 300 of different compositions may be contained in different reagent tubes, respectively, the number of theports 134 may be the same as the number of the reagent tubes, and oneport 134 may be connected to thesecond reagent 300 in one reagent tube through a pipe. Thecommon port 133 can be made to communicate with thedifferent ports 134 through thecommunication groove 135 by rotating therotor 132 to communicate the firstmulti-way valve 11 with thesecond reagent 300 of different composition to meet the demand for the different types ofsecond reagents 300 in the second reaction currently in progress.

Referring to fig. 1 and 2, in some embodiments, thefluid path system 100 further includes a collectingassembly 30, the collectingassembly 30 is in communication with thefirst pump 21, and the collectingassembly 30 is configured to collect thefirst reagent 400 after the first reaction. When thefirst pump 21 and thesecond reagent 300 are communicated through the firstmulti-way valve 11, the collectingmodule 30 may also be used to collect thesecond reagent 300 driven by thefirst pump 21, and at this time, thesecond reagent 300 may not pass through thereaction device 200, and thesecond reagent 300 enters the collectingmodule 30 after passing through thefirst pump 21.Manifold assembly 30 may also be in communication withsecond pump 22, andmanifold assembly 30 is configured to collectsecond reagent 300 after the second reaction.

Therefore, the collectingassembly 30 can be used for collecting the waste liquid after the first reaction and the second reaction, so that the waste liquid can be conveniently treated in a centralized manner. Specifically, themanifold assembly 30 includes aliquid trap 31 and awaste bottle 32. Theliquid trap 31 includes afirst port 312 and a plurality ofsecond ports 311, and thefirst port 312 communicates with the plurality ofsecond ports 311. Thefirst pump 21 communicates with thesecond port 311, thesecond pump 22 communicates with thesecond port 311, and thewaste bottle 32 communicates with thefirst port 312. Thefirst reagent 400 after the first reaction or thesecond reagent 300 after the second reaction can enter theliquid trap 31 through thesecond port 311, and the reagent entering theliquid trap 31 flows out of thefirst port 312 into thewaste liquid bottle 32.

The following will illustrate the process of performing the first reaction and the second reaction by taking thebiomolecule analysis system 1000 in the example shown in FIGS. 1, 2 and 4 as an example:

as shown in fig. 1, when the first reaction is required, theport 121 of the secondmulti-way valve 12 communicates with theport 122 to communicate the secondmulti-way valve 12 with thereaction device 200 and thefirst reagent 400, and theport 112 or 113 of the firstmulti-way valve 11 communicates with theport 114 to communicate the firstmulti-way valve 11 with thereaction device 200 and thefirst pump 21. Thefirst pump 21 is turned on so that thefirst pump 21 drives thefirst reagent 400 into thereaction apparatus 200 along the first direction X to perform the first reaction. Under the driving of thefirst pump 21, thefirst reagent 400 can also enter thefirst pump 21 and further enter theliquid trap 31 and thewaste liquid bottle 32. Because the plurality ofchannels 203 of thereaction apparatus 200 are connected to the plurality of secondmulti-way valves 12 in a one-to-one correspondence manner, and the plurality of secondmulti-way valves 12 are connected to the plurality offirst reagents 400 in a one-to-one correspondence manner, differentfirst reagents 400 can flow intodifferent channels 203, different biomolecules can be connected todifferent channels 203, and cross contamination is not generated whendifferent channels 203 perform the first reaction. In addition, thefirst reagent 400 enters thereaction device 200 along the first direction X, and before thefirst reagent 400 enters thereaction device 200 for the first reaction, the first reagent does not need to pass through the pipelines such as the firstmulti-way valve 11 and the thirdmulti-way valve 13, so that the wall hanging of thefirst reagent 400 on the pipelines is reduced, the liquid passing of the microfirst reagent 400 can be realized, the loss of thefirst reagent 400 is reduced, and the cost is saved.

As shown in fig. 2 and 3, after the first reaction is completed, theliquid path system 100 may be cleaned using the cleaning reagent in thesecond reagent 300. Specifically, by rotating therotor 132 of the thirdmulti-way valve 13, theport 134 of the thirdmulti-way valve 13, which communicates with the cleaning reagent, is made to communicate with thecommon port 133; switching theport 111 of the firstmulti-way valve 11 to communicate with theport 112 or 113, so that the firstmulti-way valve 11 communicates with thereaction device 200 and thesecond reagent 300; theport 121 of the secondmulti-way valve 12 is switched to communicate with theport 123 to communicate the secondmulti-way valve 12 with thereaction device 200 and thesecond pump 22. Thesecond pump 22 is turned on, so that thesecond pump 22 drives the cleaning reagent to enter thereaction apparatus 200 along the second direction Y, and the cleaning reagent cleans thefirst reagent 400 remained in thereaction apparatus 200. The cleaning reagent may also enter thesecond pump 22, and further enter theliquid trap 31 and thewaste liquid bottle 32, driven by thesecond pump 22.

As shown in fig. 4, before the second reaction is started or after the first reaction is performed, the type of thesecond reagent 300, for example, reagent I, to be introduced into thereaction device 200 for the second reaction may be determined, and theport 134 of the thirdmulti-way valve 13, which is communicated with the reagent I, is communicated with thecommon port 133 by rotating therotor 132 of the thirdmulti-way valve 13; theport 111 of the firstmulti-way valve 11 is switched to communicate with theport 114, so that the firstmulti-way valve 11 communicates with the common port 133 (at this time, thecommon port 133 communicates with the reagent I) and thefirst pump 21. Thefirst pump 21 is started, so that thefirst pump 21 drives thesecond reagent 300 to quickly fill the flow path of the firstmulti-way valve 11 with thesecond reagent 300, and the flow speed of thesecond reagent 300 is increased, specifically, the reagent I is filled in the pipelines of thesecond reagent 300 and theport 134, the pipeline inside the thirdmulti-way valve 13, the pipeline between thecommon port 133 and the firstmulti-way valve 11, and the pipeline inside the firstmulti-way valve 11, so that in the subsequent second reaction, the reagent I does not need to be filled in the flow path of thesecond reagent 300 to the firstmulti-way valve 11 again, the reagent I can quickly enter thereaction device 200 to perform the second reaction, and meanwhile, bubbles are prevented from entering thereaction device 200 to influence the performance of the second reaction. Under the driving of thefirst pump 21, the reagent I can also fill the pipeline between the firstmulti-way valve 11 and thefirst pump 21, the pipeline inside thefirst pump 21, and the pipeline between thefirst pump 21 and the liquid collecting assembly, so as to facilitate the next reaction or the quick cleaning of thebiomolecule analysis system 1000.

Before the second reaction is started, after the first reaction is performed, or when thebiomolecule analysis system 1000 is cleaned, theport 111 of the firstmulti-way valve 11 may be switched to communicate with theport 112 or 113, and theport 121 of the secondmulti-way valve 12 may be switched to communicate with theport 123, so that the firstmulti-way valve 11 and the secondmulti-way valve 12 communicate with the common port 133 (in this case, thecommon port 133 may communicate with the reagent I) and thesecond pump 22. Thesecond pump 22 is turned on so that thesecond pump 22 drives thesecond reagent 300 to quickly fill the flow path of thesecond reagent 300 to the firstmulti-way valve 11. Further, optionally, theport 111 of the firstmulti-way valve 11 may be communicated with theport 114, while theport 111 of the firstmulti-way valve 11 is communicated with theport 112 or 113, theport 121 of the secondmulti-way valve 12 is communicated with theport 123, so that the firstmulti-way valve 11 is communicated with the common port 133 (at this time, thecommon port 133 may be communicated with the reagent I) and thefirst pump 21, while the firstmulti-way valve 11 and the secondmulti-way valve 12 are communicated with thecommon port 133 and thesecond pump 22, and thefirst pump 21 and thesecond pump 22 are simultaneously turned on, so that thefirst pump 21 and thesecond pump 22 simultaneously drive thesecond reagent 300 to quickly fill thesecond reagent 300 into the flow path of the firstmulti-way valve 11, thereby increasing the flow rate of thesecond reagent 300.

As shown in fig. 2, when the second reaction is performed, theport 111 of the firstmulti-way valve 11 communicates with theport 112 or 113 to allow the firstmulti-way valve 11 to communicate with thereaction device 200 and thesecond reagent 300, and theport 121 of the secondmulti-way valve 12 communicates with theport 123 to allow the secondmulti-way valve 12 to communicate with thereaction device 200 and thesecond pump 22. Thesecond pump 22 is turned on, so that thesecond pump 22 drives thesecond reagent 300 into thereaction device 200 along the second direction Y to perform the second reaction.

In combination with the above, the second reaction includes a plurality of sub-reactions, the process of performing the second reaction in thereaction apparatus 200 may be sub-reactions such as base extension reaction, signal acquisition, and radical cleavage in thereaction apparatus 200, and a cleaning process may be performed between the two sub-reactions. Specifically, the firstmulti-way valve 11 communicates thecommon port 133 with thereaction apparatus 200, the secondmulti-way valve 12 communicates thereaction apparatus 200 with thesecond pump 22, and therotor 132 of the thirdmulti-way valve 13 is rotated to communicate theport 134 of the thirdmulti-way valve 13 communicating with the modified nucleotide with thecommon port 133 during the base extension reaction; when signals are collected, therotor 132 of the thirdmulti-way valve 13 is rotated, so that aport 134 in the thirdmulti-way valve 13, which is communicated with the information collecting reagent, is communicated with thecommon port 133; when radical excision is performed, therotor 132 of the thirdmulti-way valve 13 is rotated, so that theport 134 of the thirdmulti-way valve 13, which is communicated with the excision reagent, is communicated with thecommon port 133; when the cleaning process is performed, therotor 132 of the thirdmulti-way valve 13 is rotated so that theport 134 of the thirdmulti-way valve 13, through which the cleaning agent is communicated, is communicated with thecommon port 133.

In one example, when thereaction apparatus 200 includes only one reaction unit, the sub-reactions of the second reaction and the cleaning process between the two sub-reactions may be sequentially performed on the reaction unit.

In another example, when thereaction apparatus 200 includes a plurality of reaction units, different sub-reactions in the second reaction may be performed on different reaction units, respectively. In the embodiment of the present application, thereaction apparatus 200 includes afirst unit 201 and asecond unit 202, thefirst unit 201 and thesecond unit 202 are respectively communicated with theport 112 and theport 113 of the firstmulti-way valve 11, so that differentsecond reagents 300 can be respectively introduced into thefirst unit 201 and thesecond unit 202, and thefirst unit 201 and thesecond unit 202 can respectively perform different sub-reactions, thereby improving the overall efficiency of performing the second reaction.

For example, the second reaction includes a nucleic acid sequencing reaction including a plurality of repetitive reactions, one repetitive reaction including a base extension reaction, signal acquisition, and radical cleavage. While one of thefirst unit 201 and thesecond unit 202 is undergoing a base extension reaction or radical cleavage, a signal is collected from the other of thefirst unit 201 and thesecond unit 202. In one example, the sum of the reaction time required for the group cleavage and the reaction time required for the base extension reaction is approximately equal to the time required for signal acquisition.

Thebiomolecule analysis system 1000 may further include a signal acquisition device that may be used for signal acquisition, and in particular, the signal acquisition device may be an imaging device that may perform signal acquisition on thefirst unit 201 or thesecond unit 202 that is performing signal acquisition. Because only one of thefirst unit 201 and thesecond unit 202 can collect signals at the same time, the number of the signal collecting devices can be set to one set, and the signal collecting devices collect signals of thefirst unit 201 or thesecond unit 202 which is collecting signals, so that two sets of signal collecting devices are not needed, and the cost is saved.

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.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of the feature. In the description of the present application, "a plurality" means at least two, e.g., two, three, unless specifically limited otherwise.

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 (10)

1. A fluid path system for providing a solution environment for analyzing biomolecules, the analyzing biomolecules including performing a first reaction and a second reaction on a reaction device, the first reaction including coupling the biomolecules into the reaction device using a first reagent, the second reaction including detecting the biomolecules coupled into the reaction device using a second reagent, the fluid path system comprising:

the valve body assembly comprises a first multi-way valve and a second multi-way valve, the first multi-way valve can be communicated with the second reagent and the reaction device, and the second multi-way valve can be communicated with the first reagent and the reaction device; and

the driving assembly comprises a first pump and a second pump, the first pump is connected with the first multi-way valve, and the second pump is connected with the second multi-way valve;

under the condition that the first multi-way valve is communicated with the reaction device and the first pump, and the second multi-way valve is communicated with the reaction device and the first reagent, the first pump is used for driving the first reagent to enter the reaction device along a first direction so as to perform the first reaction;

and under the condition that the second multi-way valve is communicated with the reaction device and the second pump, and the first multi-way valve is communicated with the reaction device and the second reagent, the second pump is used for driving the second reagent to enter the reaction device along a second direction so as to perform the second reaction.

2. The fluid path system of claim 1, wherein the biomolecule is a nucleic acid, the first reaction comprises a hybridization reaction, and/or the second reaction comprises a sequencing reaction;

optionally, the valve body assembly further comprises a third multi-way valve, the third multi-way valve being communicable with the second reagent and the first multi-way valve;

optionally, the third multi-way valve comprises a stator and a rotor that are communicable, the third multi-way valve comprises a common port, the stator comprises a plurality of ports, the rotor comprises a communication groove, and the rotor is rotatable to communicate the common port with at least one of the ports through the communication groove.

3. The fluid path system of claim 1, wherein the first multi-way valve comprises a plurality of ports, the plurality of ports are connected to at least the second reagent, the reaction device and the first pump, respectively, and any two ports on the first multi-way valve can be communicated;

optionally, the reaction device comprises a first unit and a second unit, the first multi-way valve is a four-way valve, and four ports of the four-way valve are respectively connected with the second reagent, the first unit, the second unit and the first pump.

4. The fluid path system of claim 1, wherein the second multi-way valve comprises three ports, the three ports are respectively connected with the first reagent, the reaction device and the second pump, and any two ports of the second multi-way valve can be communicated;

optionally, the second multi-way valve is a three-way valve;

optionally, the reaction device comprises a plurality of channels, the number of the second multi-way valves and the number of the second pumps are not less than the number of the channels, and one second multi-way valve can be communicated with one channel and one second pump.

5. The fluid path system of claim 1, further comprising a manifold assembly in communication with the first pump, the manifold assembly configured to collect the first reagent after the first reaction; and/or

The collecting assembly is communicated with the second pump and is used for collecting the second reagent after the second reaction;

optionally, the manifold assembly is further configured to collect the second reagent driven by the first pump when the first pump and the second reagent are in communication through the first multi-way valve;

optionally, the manifold assembly includes a fluid collection piece and a waste fluid bottle, the fluid collection piece includes a first port and a plurality of second ports, the first port communicates with the plurality of second ports, the first pump communicates with the second ports, the second pump communicates with the second ports, and the waste fluid bottle communicates with the first port.

6. The fluid path system according to claim 1, wherein the first multi-way valve is capable of communicating the first pump and the second reagent and/or communicating the second pump and the second reagent after the first reaction is performed or before the second reaction is started, and the first pump and/or the second pump is configured to drive the second reagent to fill the flow path of the first multi-way valve with the second reagent;

optionally, the first direction is opposite the second direction.

7. A biomolecule analysis system, comprising the liquid path system according to any one of claims 1 to 6.

8. The biomolecule analysis system of claim 7, further comprising a reaction device connecting the first and second multi-way valves;

optionally, the biomolecule is a nucleic acid, the reaction device comprises a first unit and a second unit, and the biomolecule analysis system further comprises a signal acquisition device for acquiring a signal;

the second reaction is a nucleic acid sequencing reaction comprising a plurality of repetitive reactions, one of the repetitive reactions comprising a base extension reaction, signal acquisition, and radical cleavage;

and (b) performing the base extension reaction and/or the radical cleavage in one of the first unit and the second unit, and simultaneously performing the signal acquisition in the other of the first unit and the second unit by using the signal acquisition device.

9. A nucleic acid sequencing system comprising the fluid path system of any one of claims 1 to 6.

10. The nucleic acid sequence determination system of claim 9, further comprising a reaction device that removably connects the first and second multi-way valves.

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