CN115197830A - Nucleic acid amplification detection device and nucleic acid amplification method - Google Patents

Nucleic acid amplification detection device and nucleic acid amplification method Download PDF

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Publication number
CN115197830A
CN115197830A CN202110388554.3A CN202110388554A CN115197830A CN 115197830 A CN115197830 A CN 115197830A CN 202110388554 A CN202110388554 A CN 202110388554A CN 115197830 A CN115197830 A CN 115197830A
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CN
China
Prior art keywords
reaction tube
reaction
temperature
nucleic acid
tube fixing
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Pending
Application number
CN202110388554.3A
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Chinese (zh)
Inventor
解亚平
赵一帆
范旭
戴立忠
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Sansure Biotech Inc
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Sansure Biotech Inc
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Priority to CN202110388554.3A priority Critical patent/CN115197830A/en
Priority to PCT/CN2022/085485 priority patent/WO2022218199A1/en
Publication of CN115197830A publication Critical patent/CN115197830A/en
Pending legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • B01L9/06—Test-tube stands; Test-tube holders
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • B01L7/5255—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones by moving sample containers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B01L2200/147—Employing temperature sensors
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0654—Lenses; Optical fibres
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04—Details of the conveyor system
    • G01N2035/0401—Sample carriers, cuvettes or reaction vessels
    • G01N2035/0412—Block or rack elements with a single row of samples
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations
    • G01N35/04—Details of the conveyor system
    • G01N2035/0439—Rotary sample carriers, i.e. carousels
    • G01N2035/0441—Rotary sample carriers, i.e. carousels for samples

Abstract

The present invention relates to a nucleic acid amplification detection apparatus and a nucleic acid amplification method, the nucleic acid amplification detection apparatus including: the reaction tube fixing module comprises a reaction tube fixing support, and the reaction tube fixing support is used for fixing the reaction tube; the reaction detection module comprises a fixed disc and at least two temperature control optical modules, and the at least two temperature control optical modules are arranged on the fixed disc; the reaction tube fixing support and the temperature control optical module are controlled to relatively lift along the first direction and relatively rotate around a rotating shaft extending along the first direction, so that the reaction tube fixed on the reaction tube fixing support can be selectively inserted into any one of the temperature control optical modules. According to the nucleic acid amplification detection device, the reaction tubes can be rapidly switched in different temperature areas formed by different temperature control optical modules in a circulating manner to carry out amplification reaction through the relative rotation and relative lifting of the temperature control optical modules and the reaction tube fixing supports for fixing the reaction tubes, so that the detection efficiency of the amplification reaction is improved.

Description

Nucleic acid amplification detection device and nucleic acid amplification method
Technical Field
The invention relates to the technical field of in-vitro diagnosis, in particular to a nucleic acid amplification detection device and a nucleic acid amplification method.
Background
The PCR (Polymerase Chain Reaction) technique is a molecular biology technique for amplifying and amplifying specific DNA (deoxyribonucleic acid) sequences in vitro. The PCR technology has the characteristics of strong specificity, high sensitivity, low purity requirement, simplicity, convenience and rapidness, so that the PCR technology is widely applied to molecular biological detection and analysis.
Generally, the process of amplifying DNA by PCR technology is: in a mixed system of a proper buffer solution and heat-resistant DNA polymerase, the DNA undergoes a high-temperature melting reaction at the temperature of about 95 ℃, namely hydrogen bonds between double strands of the DNA are broken to form 2 complementary single-stranded DNAs; the single-stranded DNA is subjected to annealing (renaturation) reaction under the mediation of the directional and specific oligonucleotide chain as a primer, namely, the temperature of the single-stranded DNA is rapidly reduced to the range of the design temperature value (generally about 50-65 ℃) of the primer, and the single-stranded DNA and the primer are combined along the base complementary pairing principle; the temperature is quickly increased to about 72 ℃, the extension reaction is carried out after the single-stranded DNA is combined with the primer, DNA polymerase starts to combine with the deoxyribonucleoside triphosphate from the 3' end of the primer, and the DNA polymerase is sequentially extended according to corresponding bases on the template and the complementary pairing principle, so that a new DNA fragment complementary with the template is formed. After PCR amplification, the number of initial DNA molecules is doubled, which is a cycle; the multiplied DNA molecules then become the template for the next cycle, and so on, after 30-40 cycles, the number of DNA molecules will be amplified to approximately 10 of the initial value 9 And (4) doubling.
Therefore, in the process of amplifying DNA by the PCR detector, the reaction temperature needs to be adjusted by continuously and rapidly increasing and decreasing the temperature, and the time for increasing and decreasing the temperature is long, which greatly prolongs the time of an amplification cycle, thereby reducing the amplification efficiency.
Disclosure of Invention
Accordingly, it is necessary to provide a nucleic acid amplification detecting apparatus and a nucleic acid amplification method which can achieve the technical effect of reducing the time for temperature change.
According to one aspect of the present application, there is provided a nucleic acid amplification detecting apparatus including:
the reaction tube fixing module comprises a reaction tube fixing support, and the reaction tube fixing support is used for fixing a reaction tube; and
the reaction detection module is positioned on one side of the reaction tube fixing module and comprises a fixed disc and at least two temperature control optical modules, and the at least two temperature control optical modules are arranged on the fixed disc;
the reaction tube fixing support and the temperature control optical module are controlled to relatively lift along a first direction and relatively rotate around a rotating shaft extending along the first direction, so that the reaction tube fixed on the reaction tube fixing support can be selectively inserted into any one of the temperature control optical modules.
In one embodiment, the reaction detection module includes three temperature control optical modules, each temperature control optical module forms an independent temperature control temperature zone, the three temperature control optical modules are arranged at intervals around the rotating shaft, and an included angle between a central point of each two adjacent temperature control optical modules and a connecting line of the rotating shaft is 120 °.
In one embodiment, the reaction tube fixing module is provided with one or more reaction holes arranged along a linear direction, and each of the temperature control optical modules can simultaneously detect samples in the one or more reaction holes arranged along the linear direction.
In one embodiment, the reaction tube fixing module comprises a lifting driving assembly, the lifting driving assembly is in transmission connection with the reaction tube fixing support, and the lifting driving assembly is used for driving the reaction tube fixing support to reciprocate in the first direction;
the reaction detection module comprises a rotary driving assembly, the rotary driving assembly is in transmission connection with the fixed disc, and the rotary driving assembly is used for driving the fixed disc to rotate in the first direction.
In one embodiment, the lifting driving assembly includes a mounting seat, a lifting driving member and a screw transmission mechanism, the lifting driving member and the lifting transmission mechanism are both mounted on the mounting seat, the lifting driving member is in transmission connection with the reaction tube fixing bracket through the screw transmission mechanism, and the lifting driving member drives the reaction tube fixing bracket to reciprocate along the first direction through the screw transmission mechanism.
In one embodiment, the screw rod transmission mechanism includes a guide rail and a transmission screw rod, one end of the transmission screw rod is in transmission connection with the lifting driving member, the other end of the transmission screw rod extends along the first direction and is rotatably connected to the mounting base, the guide rail is mounted on the mounting base and extends along the first direction, the reaction tube fixing frame is in transmission connection with the transmission screw rod and is limited on the guide rail, and the lifting driving member can drive the reaction tube fixing member to reciprocate in the first direction along the guide rail through the transmission screw rod.
In one embodiment, the rotary driving assembly includes a rotary driving member and a synchronous belt transmission mechanism, the rotary driving member is connected to the fixed disc through the synchronous belt transmission mechanism, and the rotary driving member drives the fixed disc to rotate in the first direction through the synchronous belt transmission mechanism.
In one embodiment, the reaction detection module further includes a bearing disc, the bearing disc covers the fixed disc and defines an accommodating space together with the fixed disc, the temperature control optical module is accommodated in the accommodating space, and the bearing disc is provided with a bearing hole correspondingly communicating the temperature control optical module with an external environment;
in the process that the reaction tube fixing support and the temperature control optical module rotate around the rotating shaft relatively, each reaction hole and one of the bearing holes correspondingly communicated with any one of the temperature control optical modules are coaxial in the first direction.
In one embodiment, the reaction detection module further comprises a heat preservation medium, and the heat preservation medium is located in the accommodating space.
In one embodiment, the temperature control optical module includes a temperature control optical module casing, a heating element, and a fluorescence signal detection optical element, the heating element and the fluorescence signal detection optical element are both contained in the temperature control optical module casing, the heating element is used for generating heat to form a temperature zone, and the fluorescence signal detection optical element is used for detecting a fluorescence signal in the reaction tube.
According to one aspect of the present invention, there is provided a nucleic acid amplification method using the nucleic acid amplification detecting apparatus, comprising:
providing a reaction detection module having at least two independently controlled temperature zones;
and controlling the reaction tube to be switched in different temperature zones.
In one embodiment, the step of controlling the reaction tube to switch among different temperature zones comprises the following specific steps:
and controlling the reaction tube and the reaction detection module to relatively lift along a first direction or relatively rotate around a rotating shaft extending along the first direction so as to enable the reaction tube to be positioned in any one of the temperature zones.
The invention has the following technical effects:
1. the nucleic acid amplification detection device of the invention is different from the prior art that the temperature requirement of the amplification reaction is achieved by controlling the temperature rise and fall of a single temperature zone, and the amplification reaction can be carried out by rapidly and circularly switching the reaction tube in different temperature zones formed by different temperature control optical modules through the relative rotation and relative rise and fall of the temperature control optical module and the reaction tube fixing bracket for fixing the reaction tube on the space, thereby reducing the time of temperature change and improving the detection efficiency of the amplification reaction.
2. The nucleic acid amplification detection device of the present invention can process one sample at a time, and can also process a plurality of samples, and the processing efficiency is high.
3. The traditional amplification device has deviation in temperature rise and fall, and cannot realize accurate temperature control, and each temperature zone of the nucleic acid amplification detection device is independently controlled and is constant and accurate in temperature, so that the reaction environment of a sample is more stable, and more accurate detection can be realized. The nucleic acid amplification detection device is provided with the bearing plate, the heat preservation medium is arranged in the bearing plate, the effects of preventing heat loss and saving energy are achieved, and the temperature control is more accurate, so that the detection accuracy is further improved.
Drawings
FIG. 1 is a schematic structural diagram of a nucleic acid amplification detecting apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a reaction detection module of the nucleic acid amplification detection apparatus shown in FIG. 1;
FIG. 3 is a schematic diagram showing a partial structure of a reaction detection module of the nucleic acid amplification detection apparatus shown in FIG. 1;
FIG. 4 is a schematic diagram of a nucleic acid amplification method according to an embodiment of the present invention.
The reference numbers illustrate:
100. a nucleic acid amplification detection device; 120. a reaction tube fixing module; 121. a lift drive assembly; 1212a, a first mounting portion; 1212b, a second mounting portion; 1214. a lifting drive member; 1216. a screw drive mechanism; 1216a, a coupler; 1216b, bearings; 1216c, a drive screw; 1216d, guide rails; 123. a reaction tube fixing bracket; 1231. a first fixed part; 1234. a second fixed part; 140. a reaction detection module; 141. a rotary drive assembly; 1412. a rotary drive member; 1414. a synchronous belt transmission mechanism; 1414a, a first timing pulley; 1414b, a second timing pulley; 1414c, a synchronous belt; 1414d, a connecting shaft; 143. fixing the disc; 145. a temperature-controlled optical module; 147. a carrier tray; 1472. a bearing hole; 200. and (3) a reaction tube.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced device or element must have a particular orientation, be constructed and operated in a particular orientation, and are not to be considered limiting of the invention.
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 such feature. In the description of the present invention, "a plurality" means at least two, e.g., two, three, etc., unless specifically limited otherwise.
In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; they may be directly connected or indirectly connected through intervening media, or they may be connected internally or in any other suitable relationship, unless expressly stated otherwise. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.
In the present invention, unless otherwise expressly stated or limited, 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 an intermediate. 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 "under," "beneath," and "under" a second feature may be directly under or obliquely under the second feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
It will be understood that when an element is referred to as being "secured to" or "disposed on" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "upper," "lower," "left," "right," and the like as used herein are for illustrative purposes only and do not denote a unique embodiment.
FIG. 1 is a schematic diagram showing a structure of a nucleic acid amplification detecting apparatus 100 according to an embodiment of the present invention.
Referring to fig. 1, an embodiment of the present invention provides a nucleic acid amplification detecting apparatus 100 for amplifying a sample (e.g., DNA fragment) in a reaction tube 200. The nucleic acid amplification detecting apparatus 100 includes a reaction tube fixing module 120 and a reaction detecting module 140, wherein the reaction tube fixing module 120 is used for fixing the reaction tube 200, and the reaction detecting module 140 is used for heating and fluorescence detecting the reaction tube 200.
Referring to fig. 1, the reaction tube fixing module 120 includes a lifting driving assembly 121 and a reaction tube fixing bracket 123. The lifting driving assembly 121 is in transmission connection with the reaction tube fixing support 123, the reaction tube fixing support 123 is used for fixing the reaction tube 200, and the lifting driving assembly 121 is used for driving the reaction tube fixing support 123 to lift in a reciprocating manner in the first direction, so as to drive the reaction tube 200 to lift in a reciprocating manner in the first direction.
Specifically, the lift drive assembly 121 includes a mount, a lift drive 1214, and a lead screw drive 1216. The lifting driving member 1214 and the lifting transmission mechanism are both mounted on the mounting base, the lifting driving member 1214 is connected to the reaction tube fixing bracket 123 through the screw transmission mechanism 1216 in a transmission manner, and the lifting driving member 1214 drives the reaction tube fixing bracket 123 to reciprocate along the first direction through the screw transmission mechanism 1216. It is understood that the specific configuration of the elevating driving assembly 121 is not limited thereto, and the elevating driving member 1214 may be drivingly connected to the reaction tube fixing bracket 123 by various driving methods.
Specifically, in an embodiment, the mounting seat includes a first mounting portion 1212a and a second mounting portion 1212b, the first mounting portion 1212a and the second mounting portion 1212b are disposed at an interval in the first direction, and both the first mounting portion 1212a and the second mounting portion 1212b are rectangular plate-mounted structures. The length directions of the first and second mounting portions 1212a and 1212b are the second direction in the following embodiments, the width directions of the first and second mounting portions 1212a and 1212b are the third direction in the following embodiments, and the first, second, and third directions are perpendicular to each other.
The lifting driving member 1214 is a driving motor, and includes a main body and an output shaft extending out of one end of the main body, the main body is mounted on a side of the first mounting portion 1212a away from the second mounting portion 1212b, and the output shaft extends through the first mounting portion 1212a toward the second mounting portion 1212b along the first direction.
The lead screw transmission 1216 includes a coupler 1216a, a bearing 1216b, a transmission lead screw 1216c, and two guide rails 1216d. One end of the transmission screw 1216c is in transmission connection with an output shaft of the lifting driving part 1214 through a coupler 1216a, so that the transmission screw 1216c can synchronously rotate along with the output shaft of the lifting driving part 1214; the other end of the drive screw 1216c extends in the first direction and is rotatably coupled to the second mounting portion 1212b by a bearing 1216 b. Two guide rails 1216d are spaced apart along the second direction and are located on opposite sides of the driving screw 1216c in the second direction, one end of each guide rail 1216d is fixed to the second mounting portion 1212b, and the other end of each guide rail 1216d extends along the first direction toward the first mounting portion 1212 a.
The reaction tube fixing bracket 123 includes a first fixing portion 1232 and a second fixing portion 1234. The first fixing portion 1232 is located on one side of the two guide rails 1216d in the third direction, two sides of the first fixing portion 1232 in the second direction are respectively limited on the two guide rails 1216d, and the middle portion of the first fixing portion 1232 is in transmission connection with the transmission screw 1216 c. The second fixing portion 1234 is disposed on a side of the first fixing portion 1232 facing the second mounting portion 1212b in the first direction, and the second fixing portion 1234 is bent and extended along the third direction from an end connected to the first fixing portion 1232.
The second fixing portion 1234 has one or more reaction holes, the reaction holes are linearly arranged along the second direction, and each reaction hole is used for fixing one reaction tube 200. Thus, the simultaneous detection of one sample or a plurality of samples can be realized by using different numbers of reaction holes according to the number of the detected samples. It can be understood that the number and arrangement of the reaction holes are not limited, and the reaction holes can be arranged as required to meet different detection requirements.
Thus, the lifting driving member 1214 drives the transmission screw 1216c to rotate through the output shaft, and the transmission screw 1216c converts the torque output by the lifting driving member 1214 into a linear motion along the first direction, so as to drive the reaction tube fixing bracket 123 to reciprocate along the guide rail 1216d in the first direction, thereby realizing the position adjustment of the reaction tube 200 in the first direction.
Referring to FIGS. 2 and 3, FIG. 2 is a schematic diagram illustrating a partial structure of a reaction detecting module of the nucleic acid amplification detecting apparatus 100 according to an embodiment of the present invention; FIG. 3 is a side view showing a partial structure of a reaction detecting module 140 of the nucleic acid amplification detecting apparatus according to the embodiment of the present invention.
In some embodiments, the reaction detecting module 140 is located at one side of the reaction tube fixing module 120, and includes a rotation driving assembly 141, a fixing plate 143, and at least two temperature-controlled optical modules 145. The rotation driving assembly 141 is in transmission connection with the fixed disk 143, the temperature control optical module 145 is disposed on the fixed disk 143, and the rotation driving assembly 141 is configured to drive the fixed disk 143 to rotate around a rotation axis extending along the first direction, so as to drive the temperature control optical module 145 located on the fixed disk 143 to rotate around the rotation axis, thereby heating different temperature control optical modules 145 for the reaction tube 200.
Specifically, the rotary driving assembly 141 includes a rotary driving element 1412 and a timing belt transmission mechanism 1414, the rotary driving element 1412 and the fixed tray 143 are disposed at an interval in the second direction, the rotary driving element 1412 is connected to the fixed tray 143 by the timing belt transmission mechanism 1414 in a transmission manner, and the rotary driving element 1412 drives the fixed tray 143 to rotate around the first direction by the timing belt transmission mechanism 1414. The rotary driving component 141 can be connected, the specific configuration is not limited, and the rotary driving component 1412 can be in transmission connection with the fixed disc 143 through other transmission mechanisms.
Specifically, in one embodiment, the rotary drive 1412 is a drive motor and includes a rotary drive body and an output shaft extending out of one end of the rotary drive body. The synchronous belt transmission mechanism 1414 comprises a first synchronous belt pulley 1414a, a second synchronous belt pulley 1414b, a synchronous belt 1414c and a connecting shaft 1414d, the first synchronous belt pulley 1414a is fixedly connected to the output shaft of the rotary driving member 1412, the second synchronous belt pulley 1414b is connected to the fixed disc 143 through the connecting shaft 1414d in a transmission manner, and the synchronous belt 1414c is wound on the first synchronous belt pulley 1414a and the second synchronous belt pulley 1414b.
Thus, the rotary driving member 1412 can drive the first synchronous pulley 1414a to rotate, and further drive the synchronous pulley 1414c to move forward and rotate with the second synchronous pulley 1414b, and finally drive the fixed tray 143 to rotate around the rotating shaft, which coincides with the central axis of the fixed tray 13.
In some embodiments, the reaction detecting module 140 includes three temperature control optical modules 145, each temperature control optical module 145 is substantially cubic, the three temperature control optical modules 145 are spaced around the rotation axis of the fixing plate 143, and an angle between a central point of each adjacent temperature control optical module 145 and a line connecting the rotation axes is 120 °.
Each temperature control optical module 145 includes a temperature control optical module case, and a heating element, a fluorescence signal detection optical element, and a control circuit board accommodated in the temperature control optical module case, wherein the heating element is used to generate heat to form a temperature zone, and the fluorescence signal detection optical element is used to collect a fluorescence signal in the reaction tube 200 under the control of the control circuit board.
Thus, each temperature control optical module 145 forms an independent temperature control temperature zone, the three temperature control optical modules 145 form three temperature zones with independent temperature control, such as a low temperature zone, a medium temperature zone, and a high temperature zone, and the temperature of each temperature zone is independently controlled without affecting each other. During the relative rotation of the reaction tube fixing bracket 123 and the temperature control optical module 145 about the rotation axis, the reaction tube 200 fixed to the reaction tube fixing bracket 123 is selectively inserted into any one of the temperature control optical modules 145 to be in different temperature environments.
In some embodiments, the reaction detecting module 140 includes two temperature control optical modules 145, the two temperature control optical modules 145 are disposed opposite to each other in the radial direction of the fixed disk 143, the two temperature control optical modules 145 form two low temperature regions and a high temperature region with independent temperature control, and the reaction tube leaving the high temperature region can be cooled by standing outside the temperature control optical modules 145 to cool and then enter the low temperature region.
Further, the reaction detecting module 140 further includes a bearing plate 147, the bearing plate 147 covers the fixing plate 143 and defines an accommodating space together with the fixing plate 143, the temperature control optical module 145 is accommodated in the accommodating space, a bearing hole 1472 for communicating the temperature control optical module 145 with an external environment is formed in the bearing plate 147, and the reaction hole of the reaction tube fixing support 123 and the bearing hole 1472 of any one of the temperature control optical modules 145 are coaxially arranged in the first direction. Thus, the reaction tube 200 can correspondingly extend into the corresponding temperature control optical module 145 through one of the bearing holes 1472, and the arrangement of the bearing plate 147 creates a dark light environment for the temperature control optical module 145, thereby facilitating fluorescence detection.
In some embodiments, the reaction detecting module 140 further includes a thermal insulation medium formed by thermal insulation materials such as thermal insulation cotton, the thermal insulation medium is located in the accommodating space, and the thermal insulation medium is used for preventing heat dissipation of the temperature control optical module 145, so as to save energy and enable temperature control to be more accurate. It is understood that the material forming the insulating medium is not limited and can be set as required to meet different requirements.
It will be appreciated that in other embodiments, the fixed plate 143 may be held stationary, and may be moved up and down in the first direction and rotated about a rotation axis extending in the first direction by the reaction tube fixing bracket 123. In this way, the reaction tube fixing bracket 123 and the temperature control optical module 145 of the present application are controlled to be relatively lifted and lowered along the first direction and relatively rotated around a rotation axis extending along the first direction, so that the reaction tube 200 fixed to the reaction tube fixing bracket 123 is selectively inserted into any one of the temperature control optical modules 145.
The present application also provides a nucleic acid amplification method comprising the steps of:
s110: a reaction detection module 140 is provided, the reaction detection module 140 having at least two independently controlled temperature zones.
S120: the reaction tube 200 is controlled to switch in different temperature zones.
Specifically, the reaction tube 200 and the reaction detecting module 140 are controlled to be relatively lifted or rotated along a first direction or a rotational axis extending along the first direction, so that the reaction tube 200 is located in any one of the temperature zones.
The nucleic acid amplification method is described in detail below by taking the example that the nucleic acid amplification detecting apparatus 100 has three temperature-controlled optical modules 145:
the three temperature control optical modules 145 form three temperature zones of independent temperature control, a low temperature zone, a medium and high temperature zone, respectively.
The fixing plate 143 may be rotated by the rotation driving assembly until one of the temperature-controlled optical modules 145 is aligned with the reaction tube 200 fixed to the reaction tube fixing bracket 123 in the first direction. Then, the elevating driving assembly 121 drives the reaction tube fixing bracket 123 to descend in the first direction until the reaction tube 200 is inserted into one of the temperature control optical modules 145 through the bearing hole 1472 of the bearing plate 147.
After the reaction in the current temperature zone is completed, the elevating driving unit 121 drives the reaction tube fixing bracket 123 to be elevated in the first direction to detach the reaction tube 200 from the temperature-controlled optical module 145. Then, the rotational driving unit 141 drives the temperature-controlled optical module 145 to rotate by 120 ° so that the other temperature-controlled optical module 145 is aligned with the reaction tube 200 fixed to the reaction-tube fixing bracket 123 in the first direction. Thereafter, the elevating driving assembly 121 drives the reaction tube fixing bracket 123 to descend again in the first direction until the reaction tube 200 is inserted into the temperature control optical module 145 through the bearing hole 1472 of the bearing plate 147.
It can be understood that, when the nucleic acid amplification detecting apparatus 100 has only two temperature control optical modules 145, in the process of switching the reaction tube 200 from the high temperature region to the low temperature region, after the reaction tube 200 is completely separated from the temperature control optical module 145 forming the middle temperature region, the elevation driving assembly 121 may control the reaction tube 200 to stand in the air until the temperature of the sample in the reaction tube 200 is reduced to a preset value, and then adjust the height of the reaction tube fixing support 123 to make the reaction tube 200 enter the low temperature region.
The nucleic acid amplification detection device 100 can rapidly perform amplification reaction through the relative rotation and relative lifting of the temperature control optical module 145 and the reaction tube fixing support 123 for fixing the reaction tube 200 in space, thereby realizing rapid circulation of a sample in the reaction tube 200 among three temperature zones, being different from the change of the time control heating temperature in the prior art, obviously reducing the time of temperature change and the time required by amplification reaction, independently controlling each temperature zone, keeping the temperature constant and accurate, enabling the reaction environment of the sample to be more stable, and realizing more accurate detection. In addition, the three temperature control optical modules 145 can detect the fluorescent signal in real time while controlling the temperature to be constant, so that the detection sensitivity and convenience are higher. In addition, since only the rotation of the temperature control optical module 145 can be controlled without rotating the reaction tube fixing holder 123 for fixing the reaction tube 200, the reaction environment of the sample is more stable, and one sample or a plurality of samples can be processed at a time, thereby having a high processing effect.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent should be subject to the appended claims.

Claims (12)

1. A nucleic acid amplification detection apparatus comprising:
the reaction tube fixing module comprises a reaction tube fixing support, and the reaction tube fixing support is used for fixing a reaction tube; and
the reaction detection module is positioned on one side of the reaction tube fixing module and comprises a fixed disc and at least two temperature control optical modules, and the at least two temperature control optical modules are arranged on the fixed disc;
the reaction tube fixing support and the temperature control optical module are controlled to relatively lift along a first direction and relatively rotate around a rotating shaft extending along the first direction, so that the reaction tube fixed on the reaction tube fixing support can be selectively inserted into any one of the temperature control optical modules.
2. The apparatus according to claim 1, wherein the reaction detecting module comprises three temperature-controlled optical modules, each of the three temperature-controlled optical modules forms an independently temperature-controlled temperature zone, the three temperature-controlled optical modules are arranged at intervals around the rotation axis, and an included angle between a central point of each of two adjacent temperature-controlled optical modules and a connecting line of the rotation axis is 120 °.
3. The apparatus of claim 1, wherein the reaction tube fixing module has one or more reaction wells arranged in a linear direction, and each of the temperature-controlled optical modules is capable of simultaneously detecting one or more samples in the reaction wells arranged in the linear direction.
4. The nucleic acid amplification detecting device according to claim 1, wherein the reaction tube fixing module includes a lifting driving component, the lifting driving component is in transmission connection with the reaction tube fixing support, and the lifting driving component is configured to drive the reaction tube fixing support to reciprocate in the first direction;
the reaction detection module comprises a rotary driving assembly, the rotary driving assembly is in transmission connection with the fixed disc, and the rotary driving assembly is used for driving the fixed disc to rotate in the first direction.
5. The nucleic acid amplification detecting device according to claim 4, wherein the lifting driving assembly includes a mounting base, a lifting driving member and a screw transmission mechanism, the lifting driving member and the lifting transmission mechanism are both mounted on the mounting base, the lifting driving member is connected to the reaction tube fixing bracket through the screw transmission mechanism, and the lifting driving member drives the reaction tube fixing bracket to reciprocate along the first direction through the screw transmission mechanism.
6. The nucleic acid amplification detecting apparatus according to claim 5, wherein the screw transmission mechanism includes a guide rail and a transmission screw, one end of the transmission screw is drivingly connected to the elevating driving member, the other end of the transmission screw extends in the first direction and is rotatably connected to the mounting base, the guide rail is mounted on the mounting base and extends in the first direction, the reaction tube fixing frame is drivingly connected to the transmission screw and is retained on the guide rail, and the elevating driving member can drive the reaction tube fixing member to reciprocate in the first direction along the guide rail through the transmission screw.
7. The nucleic acid amplification detecting device of claim 4, wherein the rotary driving component comprises a rotary driving element and a synchronous belt transmission mechanism, the rotary driving element is connected to the fixing disc through the synchronous belt transmission mechanism, and the rotary driving element drives the fixing disc to rotate in the first direction through the synchronous belt transmission mechanism.
8. The nucleic acid amplification detection device of claim 1, wherein the reaction detection module further comprises a bearing tray, the bearing tray covers the fixed tray and defines an accommodating space together with the fixed tray, the temperature-controlled optical module is accommodated in the accommodating space, and bearing holes correspondingly communicating the temperature-controlled optical module with the external environment are formed in the bearing tray;
in the process that the reaction tube fixing support and the temperature control optical module rotate around the rotating shaft relatively, each reaction hole and one of the bearing holes correspondingly communicated with any one of the temperature control optical modules are coaxial in the first direction.
9. The nucleic acid amplification detection apparatus of claim 8, wherein the reaction detection module further comprises an incubation medium, and the incubation medium is located in the accommodating space.
10. The apparatus of claim 1, wherein the temperature-controlled optical module comprises a housing, a heating element and a fluorescence-signal-detecting optical element, the heating element and the fluorescence-signal-detecting optical element are accommodated in the housing, the heating element is configured to generate heat to form a temperature zone, and the fluorescence-signal-detecting optical element is configured to detect a fluorescence signal in the reaction tube.
11. A nucleic acid amplification method using the nucleic acid amplification detecting apparatus according to any one of claims 1 to 10, comprising the steps of:
providing a reaction detection module having at least two independently controlled temperature zones; and controlling the reaction tube to be switched in different temperature zones.
12. The nucleic acid amplification method according to claim 11, wherein controlling the reaction tubes to switch between different temperature zones comprises the following steps:
and controlling the reaction tube and the reaction detection module to relatively lift along a first direction or relatively rotate around a rotating shaft extending along the first direction so as to enable the reaction tube to be positioned in any one of the temperature zones.
CN202110388554.3A 2021-04-12 2021-04-12 Nucleic acid amplification detection device and nucleic acid amplification method Pending CN115197830A (en)

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PCT/CN2022/085485 WO2022218199A1 (en) 2021-04-12 2022-04-07 Nucleic acid amplification test device and nucleic acid amplification method

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AU2006259239A1 (en) * 2005-06-16 2006-12-28 Stratagene California Heat blocks and heating
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