CN211847961U

CN211847961U - Nucleic acid detection device and system

Info

Publication number: CN211847961U
Application number: CN201921974672.7U
Authority: CN
Inventors: 方雪恩; 杨波; 朱美娇; 王耀华; 孔继烈
Original assignee: Shanghai Superchip Biological Science & Technology Co ltd; Shanghai Suchuang Diagnostic Products Co ltd
Current assignee: Shanghai Superchip Biological Science & Technology Co ltd; Shanghai Suchuang Diagnostic Products Co ltd
Priority date: 2019-11-15
Filing date: 2019-11-15
Publication date: 2020-11-03
Anticipated expiration: 2029-11-15

Abstract

The utility model relates to a nucleic acid detection device and system. The device comprises: a microfluidic module including a microfluidic chip; the temperature control area is arranged below the microfluidic chip and comprises a first temperature area, a second temperature area, a temperature insulation area and a detection hole; the rotating module is connected with the microfluidic chip; the temperature control module is connected to the temperature control area; and the detection module is used for detecting the fluorescent signals generated by the samples in the reaction holes through the detection holes and outputting detection results. Through the cooperation of each module of nucleic acid detecting device, according to the utility model discloses nucleic acid detecting device can realize the integration of nucleic acid purification, nucleic acid detection, reduces operation flow to improve work efficiency.

Description

Nucleic acid detection device and system

Technical Field

The utility model relates to a detect technical field, especially relate to a nucleic acid detection device and system.

Background

The microfluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes into a micron-scale chip, and automatically completes the whole analysis process. Microfluidic chips (Microfluidic chips) are a hot spot area for the development of current micro Total analysis systems (minidesigned Total analysis systems).

The traditional nucleic acid detection technology is based on PCR (Polymerase Chain Reaction), and needs to be rapidly switched among different temperatures to realize the cycle of denaturation, annealing and extension functions. The switching between the temperatures has high requirements on equipment, takes long time, leads to long whole detection time, and one test usually needs one hour or two hours. The constant temperature amplification technology which is started in the last decade only needs to carry out amplification reaction at a fixed temperature, thereby greatly improving the amplification efficiency and shortening the test time.

However, the prior art does not mention an integrated scheme of nucleic acid extraction, isothermal amplification reaction, and melting test.

Therefore, the integration of nucleic acid extraction, isothermal amplification reaction and melting test is significant for reducing operation flow and improving work efficiency.

SUMMERY OF THE UTILITY MODEL

Technical problem

In view of this, the technical problem to be solved by the present invention is how to realize the integration of nucleic acid extraction, isothermal amplification reaction, and melting test, so as to reduce the operation procedures and improve the work efficiency.

Solution scheme

In order to solve the above technical problem, according to an embodiment of the present invention, there is provided a nucleic acid detecting apparatus including: the microfluidic module comprises a microfluidic chip, wherein the microfluidic chip comprises a plurality of sample adding holes, a plurality of liquid dividing areas and a plurality of reaction holes, the sample adding holes are used for adding samples, the reaction holes are used for carrying out amplification reaction and melting reaction on the samples, and the liquid dividing areas are connected with the plurality of reaction holes through microchannels and microvalves;

the temperature control area is arranged below the microfluidic chip and comprises a first temperature area, a second temperature area, a temperature insulation area and a detection hole;

the rotating module is connected with the microfluidic chip and used for driving the microfluidic chip to rotate at a first rotating speed for a first time and at a second rotating speed for a second time, so that a sample in the sample adding hole enters the reaction hole through the liquid separating area, the microchannel and the microvalve under the action of centrifugal force;

the temperature control module is connected to the temperature control area, the temperature control module is used for:

controlling the temperature of the first temperature region to stabilize at a first temperature when the sample is located in the well, so that the sample releases nucleic acids;

controlling the temperature of the second temperature region to be stabilized at a second temperature when the sample is positioned in the reaction hole, so that the sample is subjected to an amplification reaction;

when the amplification reaction is finished, controlling the temperature of the second temperature area to rise from the second temperature to a third temperature at a constant speed, so that the sample is subjected to a melting reaction;

the temperature control module further comprises a temperature auxiliary area, and the temperature auxiliary area is used for forming a relatively sealed space with the temperature control area;

and the detection module is used for detecting the fluorescent signals generated by the samples in the reaction holes through the detection holes and outputting detection results.

In one possible embodiment, the first temperature zone, the second temperature zone, and the thermal insulation zone are annular, the thermal insulation zone is disposed between the first temperature zone and the second temperature zone to isolate the first temperature zone from the second temperature zone,

wherein the sample application well is covered by the first temperature zone and the reaction well is covered by the second temperature zone.

In a possible embodiment, the driving the microfluidic chip to rotate at a first rotation speed for a first time and at a second rotation speed for a second time includes:

driving the microfluidic chip to rotate at a first rotation speed for a first time, and repeating the rotation for a first time, wherein each rotation is separated by a first preset time, so that the sample enters the liquid separation area from the sample adding hole;

and after the sample enters the liquid distribution area, driving the microfluidic chip to rotate at a second rotating speed for a second time, and repeating the rotation for a second time, wherein the rotation is separated by a second preset time every time, so that the sample enters the reaction hole from the liquid distribution area through the microchannel and the microvalve.

In one possible embodiment, the first rotation speed is 1400rpm to 1800rpm, the first time is 7s to 14s, the first number of times is 2 to 5 times, the second rotation speed is 4200rpm to 4600rpm, the second time is 7s to 14s, and the second number of times is 2 to 5 times.

In one possible embodiment, the detecting the fluorescent signal generated by the sample in the reaction well through the detection well includes:

and detecting the fluorescence signal in each reaction hole on the microfluidic chip in real time in the amplification reaction and the melting reaction.

In a possible embodiment, the diameter of the detection hole is 3.5mm to 5.5mm, the first temperature is 90 to 100 ℃, the second temperature is 60 to 65 ℃, and the third temperature is 90 to 110 ℃, wherein a light guide tube with the length of 4 to 15mm is further arranged in the detection module.

In one possible embodiment, the detection module comprises:

the light-emitting unit is used for emitting first light;

a light propagation unit for propagating the first light to the detection hole; when the rotating module drives the reaction hole of the microfluidic chip to rotate above the detection hole, and the first light irradiates the sample of the reaction hole through the detection hole, the light transmission unit transmits second light which is emitted by the sample and is excited by the first light; and

the detection unit is used for detecting the second light and generating a first signal according to the second light, the first signal is an electric signal, the detection unit is further used for carrying out amplification, filtering and analog-to-digital conversion processing on the first signal to obtain a digital signal serving as the detection result, and the detection result is used for analyzing the sample.

In order to solve the above technical problem, according to another embodiment of the present invention, there is provided a nucleic acid detecting system including:

one or more of said nucleic acid detection devices;

and the control device is electrically connected with the one or more nucleic acid detection devices and is used for controlling the one or more nucleic acid detection devices.

Advantageous effects

Through above device, the utility model discloses each aspect of embodiment can realize that nucleic acid extraction, amplification reaction, melting reaction integration can be accomplished the complete detection to nucleic acid in a device, can reduce operation flow, improve detection efficiency, owing to adopt the integrated design, the utility model provides a detection result that nucleic acid detecting device obtains has higher accuracy. And, the embodiment of the utility model provides an in the rotation module drive micro-fluidic chip with first rotational speed rotate the very first time, rotate the second time with the second rotational speed, make sample in the application of sample hole passes through under the effect of centrifugal force divide the liquid zone the microchannel the micro-valve gets into the reaction hole can the flow direction and the position of sample among the accurate distributed control micro-fluidic chip, realize evenly dividing liquid, broken valve flow to realize nucleic acid detecting device's stable detection.

Other features and aspects of the present invention will become apparent from the following detailed description of exemplary embodiments, which proceeds with reference to the accompanying drawings.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments, features, and aspects of the present invention and, together with the description, serve to explain the principles of the invention.

FIG. 1 is a block diagram showing a nucleic acid detecting apparatus according to an embodiment of the present invention.

Fig. 2 shows a schematic diagram of a microfluidic chip according to an embodiment of the present invention.

FIG. 3 is a schematic view showing a nucleic acid detecting apparatus according to an embodiment of the present invention.

FIG. 4 is a block diagram showing a nucleic acid detecting apparatus according to an embodiment of the present invention.

Fig. 5 is a schematic structural diagram of a light propagation unit according to an embodiment of the present invention.

FIG. 6 is a schematic structural diagram of a nucleic acid detecting apparatus according to an embodiment of the present invention.

FIG. 7 is a schematic structural diagram of a nucleic acid detecting apparatus according to an embodiment of the present invention.

Fig. 8a shows a perspective view of a nucleic acid detecting apparatus according to an embodiment of the present invention, and fig. 8b shows a front view of the nucleic acid detecting apparatus according to an embodiment of the present invention.

Fig. 9 shows a flow chart of a nucleic acid detection method according to an embodiment of the present invention.

Fig. 10 shows a block diagram of a nucleic acid detection system according to an embodiment of the present invention.

Detailed Description

Various exemplary embodiments, features and aspects of the present invention will be described in detail below with reference to the accompanying drawings. In the drawings, like reference numbers can indicate functionally identical or similar elements. While the various aspects of the embodiments are presented in drawings, the drawings are not necessarily drawn to scale unless specifically indicated.

The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.

Furthermore, in the following detailed description, numerous specific details are set forth in order to provide a better understanding of the present invention. It will be understood by those skilled in the art that the present invention may be practiced without some of these specific details. In some instances, methods, means, elements and circuits that are well known to those skilled in the art have not been described in detail so as not to obscure the present invention.

Referring to FIG. 1, FIG. 1 shows a block diagram of a nucleic acid detecting apparatus 10 according to an embodiment of the present invention.

Referring to fig. 2, fig. 2 is a schematic diagram of a microfluidic chip according to an embodiment of the present invention.

Referring to FIG. 3, FIG. 3 is a schematic diagram of a nucleic acid detecting apparatus 10 according to an embodiment of the present invention.

As shown in fig. 1, 2, and 3, a microfluidic module 100 includes a microfluidic chip, the microfluidic chip includes a plurality of sample wells 101, a plurality of liquid distribution areas 102, and a plurality of reaction wells 104, the sample wells 101 are used for adding a sample, the reaction wells 104 are used for performing an amplification reaction and a melting reaction on the sample, and the liquid distribution areas 102 are connected to the plurality of reaction wells 104 through micro channels and micro valves 103;

the temperature control area 130 is arranged below the microfluidic chip and comprises a first temperature area 131, a second temperature area 132, a temperature insulation area 134 and a detection hole 133;

the rotating module 110 is connected to the microfluidic chip, and is configured to drive the microfluidic chip to rotate at a first rotation speed for a first time and at a second rotation speed for a second time, so that the sample in the sample addition hole 101 enters the reaction hole 104 through the liquid separation region 102, the microchannel, and the microvalve under the action of centrifugal force;

a temperature control module 140 connected to the temperature control zone 130, the temperature control module 140 being configured to:

controlling the temperature of the first temperature region 131 to be stabilized at a first temperature when the sample is located in the well 101, so that the sample releases nucleic acid;

controlling the temperature of the second temperature region 132 to be stabilized at a second temperature when the sample is located in the reaction well 104, so that the sample performs an amplification reaction;

when the amplification reaction is finished, controlling the temperature of the second temperature region 132 to increase from the second temperature to a third temperature at a uniform speed, so that the sample undergoes a melting reaction, an

The detection module 120 is configured to detect a fluorescence signal generated by the sample in the reaction well 104 through the detection well 133 and output a detection result.

Through above device, the embodiment of the utility model provides a can realize that nucleic acid draws, amplification reaction, melting reaction integration, can accomplish the complete detection to nucleic acid in a device, can reduce operation flow, improve detection efficiency, owing to adopt the integrated design, the utility model provides a detection result that the nucleic acid detection device that provides obtained has higher accuracy. And, the embodiment of the utility model provides an in rotate module 110 and drive micro-fluidic chip rotates the very first time with first rotational speed, rotates the second time with the second rotational speed, makes the sample in the application of sample hole 101 passes through under the effect of centrifugal force divide liquid district 102 the microchannel the microvalve gets into reaction hole 104, the flow direction and the position of sample among the control micro-fluidic chip of can accurate distribution realize evenly dividing liquid, broken valve flow to realize nucleic acid detecting device's stable detection.

In one possible embodiment, as shown in fig. 2, the microfluidic chip may be divided into 4 identical regions (A, B, C, D), each region comprising at least one sample well 101, a plurality of reaction wells 104, and a liquid separation region 102, a microchannel, and a microvalve therebetween.

In one possible embodiment, as shown in fig. 3, the first temperature region 131, the second temperature region 132, and the thermal insulation region may be annular, and the thermal insulation region is disposed between the first temperature region 131 and the second temperature region 132 to isolate the first temperature region 131 and the second temperature region 132.

In one possible embodiment, the loading well 101 is covered by the first temperature zone 131 and the reaction well 104 is covered by the second temperature zone 132.

In one example, a sample (e.g., a liquid sample to be tested) can be added to the well 101 and the well 101 is closed, and when the sample is in the well 101, the temperature control module 140 of the embodiment of the present invention can control the temperature of the first temperature region 131, so that the sample is cracked to achieve the purpose of nucleic acid extraction. After nucleic acid extraction is accomplished, the rotation module 110 of the embodiment of the present invention can control the rotation of the microfluidic module 100, so that the sample enters the reaction well 104 through the separation region 102, the micro-channel and the micro-valve under the action of centrifugal force. When the sample enters the reaction hole 104, the temperature control module 140 of the embodiment of the present invention can control the second temperature region 132 to reach the temperature required by the amplification reaction, because the reaction hole 104 is covered by the second temperature region 132, the sample can perform the amplification reaction in the reaction hole 104, and when the sample performs the amplification reaction, the detection module 120 of the embodiment of the present invention can monitor the amplification reaction through the detection hole 133; after the amplification reaction is accomplished, the utility model discloses temperature control module 140 can control the temperature of the regional 132 of second temperature and reach the required temperature of melting reaction, and the sample carries out melting reaction in reaction hole 104, and detection module 120 can real-time detection melting process's signal through inspection hole 133 to obtain the melting curve through analysis and processing, obtain the melting temperature of amplification product.

The process of detecting nucleic acid using the nucleic acid detecting apparatus according to the embodiment of the present invention is described above, and specific parameters will be described below.

In a possible embodiment, the driving the microfluidic chip to rotate at a first rotation speed for a first time and at a second rotation speed for a second time may include:

driving the microfluidic chip to rotate at a first rotation speed for a first time, and repeating for a first time, wherein each rotation is separated by a first preset time, so that the sample enters the liquid separation area 102 from the sample adding hole 101;

when the sample enters the liquid separation area 102, the microfluidic chip is driven to rotate at a second rotation speed for a second time, and the rotation is repeated for a second time, wherein each time the microfluidic chip rotates for a second preset time, so that the sample enters the reaction hole 104 from the liquid separation area 102 through the microchannel and the microvalve.

In this way, the utility model discloses rotation module 110 can realize the hierarchical control of rotational speed, and rotation module 110 rotates with first rotational speed control micro-fluidic module 100, can realize that the sample evenly distributed divides liquid zone 102 (micro-fluidic entry) to second rotational speed control micro-fluidic module 100 rotates, can realize that the sample is quick, evenly get into reaction hole 104 to realize that nucleic acid stably detects.

In one possible embodiment, the first rotation speed may be 1400rpm to 1800rpm, the first time may be 7s to 14s, the first number of times may be 2 to 5 times, the second rotation speed may be 4200rpm to 4600rpm, the second time may be 7s to 14s, and the second number of times may be 2 to 5 times.

In one example, the first time and the second time may be equal, for example, both may be 10s, and the first number and the second number may also be equal, for example, both may be 3.

It should be understood that the above-mentioned first rotation speed, second rotation speed, first time, second time, first number of times, and second number of times are exemplary descriptions, the present invention does not limit the rotation speed, the rotation number, and the rotation time of the rotation module 110, the rotation module 110 can rotate at any time and any number of times within the allowable speed, and the first rotation speed, the second rotation speed, the first time, the second time, the first number of times, and the second number of times can be set according to actual situations.

Through the setting, the embodiment of the utility model provides a can realize the flow direction and the position of sample among the accurate control micro-fluidic chip, realize evenly dividing liquid, broken valve flow.

In one possible embodiment, the rotation module 110 may include a stepper motor, which may be, for example, a high-speed micro stepper motor. In one example, the rotation module 110 may rotate the microfluidic module 100 step by step at a fixed angle. The rotating module 110 can be freely switched between high-speed and low-speed rotation, for example, between 100 rpm and 6000 rpm. Of course, the rotating module 110 may also include other types of motors, and the present invention is not limited thereto.

The embodiment of the utility model provides an in the rotation module 110 can realize high-speed rotation, low-speed rotation, forward rotation, reverse rotation, rotational speed can linear increase or nonlinear increase, to the control of rotation module 110, the utility model discloses do not limit, the operation mode of rotation module 110 can be confirmed as required to the technical staff in the field.

In one possible embodiment, the first temperature may be 90 to 100 ℃, the second temperature may be 60 to 65 ℃, and the third temperature may be 90 to 110 ℃. It should be understood that the temperatures of the first temperature region 131 and the second temperature region 132 may be other temperatures, and the present invention is not limited thereto.

In one example, the temperature control module 140 can control the temperature of the first temperature region 131 within ± 0.5 ℃ of a certain temperature in a range of 90-100 ℃, for example, between 94.5 ℃ and 95.5 ℃.

When the temperature of the first temperature region 131 is 90-100 ℃, the sample can be subjected to nucleic acid cracking, thereby realizing nucleic acid extraction. When the temperature of the first temperature region 131 is stabilized within a certain temperature range of 90 to 100 ℃ plus or minus 0.5 ℃, the conditions for performing nucleic acid lysis on the sample are optimal.

In a possible embodiment, when the added sample is a mixture solution including a clinical sample and impurities, after the sample is added to the sample adding hole 101 in the microfluidic chip, the temperature control module 140 controls the temperature of the first temperature region 131 to be a certain temperature of 90-100 ℃, and when the sample in the sample adding hole 101 is at a high temperature (e.g., 90-100 ℃), the sample in the sample adding hole 101 is pyrolyzed, thereby releasing nucleic acid. The temperature control module 140 may further control the temperature of the well 101 to be maintained at a certain temperature for a certain time, which may be determined according to practical situations, for example, the time may be 3-8 minutes, such as 5 minutes, and by controlling the temperature of the first temperature region 131 to be maintained for the certain time, the sample in the well 101 may be pyrolyzed, thereby releasing the nucleic acid.

In one example, the temperature control module 140 can control the temperature of the second temperature region 132 to be within ± 0.5 ℃ of a certain temperature within a range of 60-65 ℃ so that the sample in the reaction well 104 can smoothly perform the amplification reaction, for example, the temperature control module 140 can control the temperature of the second temperature region 132 to be between 62.5 ℃ and 63.5 ℃.

After the sample in the sample adding hole 101 is cracked, the micro-fluidic module 100 may be driven to rotate by the rotation module 110, so as to uniformly distribute the sample in the sample adding hole 101 to the reaction hole 104 through centrifugal force, and when an amplification reaction needs to be performed, the temperature control module 140 may control the temperature of the second temperature region 132 to be within ± 0.5 ℃ of a certain temperature within a range of 60 to 65 ℃, so that the sample in the reaction hole 104 may be subjected to the amplification reaction. The embodiment of the utility model provides a temperature through control second temperature region 132 is within a certain temperature +/-0.5 ℃ of 60 ~ 65 ℃ within range, can realize going on smoothly of amplification reaction.

Of course, the embodiment of the present invention can keep the temperature of the second temperature region 132 to be within a certain temperature range of 60-65 ℃ by the temperature control module 140 within ± 0.5 ℃ for the preset time, so that the amplification reaction can be completed by the sample, of course, the embodiment of the present invention does not limit the specific value of the preset time, and the skilled in the art can determine according to the actual situation.

In one possible embodiment, the detecting the fluorescent signal generated by the sample in the reaction well through the detection well may include:

In one possible embodiment, the third temperature is greater than the second temperature.

When the sample in the reaction well 104 completes the amplification reaction, the temperature control module 140 may control the temperature of the second temperature region 132 to linearly increase (increase at a constant rate) from the second temperature to the third temperature, for example, from 63 ℃ to 100 ℃, so that the melting reaction may proceed smoothly.

In a possible embodiment, the detecting the sample in the reaction well 104 through the detection well 133 may include:

and detecting a fluorescence signal in the melting reaction process in real time in the process that the temperature of the second temperature region 132 linearly rises from the second temperature to the third temperature, and analyzing to obtain the melting temperature of the amplification product.

After the amplification reaction is completed, the temperature control module 140 gradually increases the temperature of the second temperature region 132, and simultaneously monitors the fluorescence signal of each reaction well in the microfluidic chip in real time through the detection module 120 to obtain a curve of the fluorescence signal of each reaction well with respect to the temperature change, a melting curve is generated through analysis and calculation, a characteristic peak (Tm, the temperature at which 50% of the DNA double strand is melted) is present on the melting curve, the specific product can be distinguished from other products such as primer dimer by using the characteristic peak, whether the amplification product is the target product is examined through the melting curve, and the melting temperature of the amplification product is obtained. Of course, the above description is exemplary and should not be construed as limiting the invention.

Referring to fig. 3, as shown in fig. 3, the temperature control module may further include a temperature auxiliary region 135 for forming a relatively sealed space with the temperature control region, and during the amplification reaction and the melting reaction, the temperature is set to be the same as or slightly higher than the second temperature region to improve the temperature control effect, and the temperature can be controlled more precisely through the temperature auxiliary region 135.

Through above device, the utility model discloses can realize nucleic acid extraction, amplification reaction, three main functions in an organic whole of fusion test, let the product integrate, simplify the operation flow, improve detection efficiency. The temperature control module 140 controls each temperature area of the temperature control area 130, so that the requirements of three functions of nucleic acid extraction, amplification reaction and melting test on temperature are met, and accurate temperature control of each temperature area is realized in a compact mechanism. And, the embodiment of the utility model provides a carry out the integrated design with fluid control and optical detection in the micro-fluidic chip, make equipment miniaturization as far as possible, reduce micro-fluidic chip's transfer process, improve detection efficiency. The flow direction and the position of a liquid sample of the microfluidic chip are precisely and step-by-step controlled by intelligently and precisely controlling the centrifugal rotating speed, so that different requirements of uniform liquid separation, valve breaking flow, stable detection and the like are met.

The detection module 120 is described in an exemplary manner below.

The detection module 120 may be configured to detect the sample in the reaction well 104 and output a detection result.

In one possible embodiment, when the sample enters each reaction well 104 under the action of centrifugal force, the detection module 120 may perform detection on the sample in each reaction well 104 and output the detection result.

Referring to FIG. 4, FIG. 4 is a block diagram of a nucleic acid detecting apparatus according to an embodiment of the present invention.

In one possible implementation, as shown in fig. 4, the detection module 120 may include a light emitting unit 1201, a light propagating unit 1202 and a detection unit 1203.

In a possible embodiment, the light emitting unit 1201 may emit a first light, the first light may irradiate a sample of the reaction well 104 after propagating through the light propagating unit 1202, the sample may emit a second light when being excited by the first light, the second light may be received by the detecting unit 1203 after propagating through the light propagating unit, the detecting unit 1203 detects the second light, and may generate a first signal according to the second light, the first signal is an electrical signal, the detecting unit 1203 is further configured to amplify, filter, and perform analog-to-digital conversion on the first signal, so as to obtain a digital signal as the detection result, and the detection result may be used to analyze the sample.

By matching the units in the detection module 120, the detection module 120 can output a detection result, which can be used to analyze the concentration of the sample in the reaction well 104, the negative and positive properties, the melting temperature of the amplification product, and the like.

It should be noted that, when the sample is subjected to an amplification reaction or a melting reaction in the reaction well 104, the rotation module may control the rotation of the microfluidic module, so that the detection module may monitor the reaction process in the reaction well 104 through the detection well. Of course, the rotation speed of the rotation module in the amplification reaction or the melting reaction of the present invention is not limited, and can be determined as needed by those skilled in the art.

In one possible embodiment, the light emitting unit 1201 may be a maintenance-free high power LED light source or other form of light source.

In one possible embodiment, the light propagation unit 1202 may include a plurality of optical components.

Referring to fig. 5, fig. 5 is a schematic structural diagram of a light propagation unit 1202 according to an embodiment of the present invention.

In one possible embodiment, the light propagation unit may include a first collimating lens 301, a first optical filter 302, a dichroic mirror 303, a second collimating lens 304, a second optical filter 305, and a condensing lens 306.

In one possible embodiment, the light emitting unit 1201 emits a first light, the first light irradiates a sample 307 (the sample in the reaction well 104) through the first collimating lens 301, the first optical filter 302, the dichroic mirror 303 and the second collimating lens 304, the sample 307 is excited by the first light to emit a second light, and the second light is received by the detecting unit 1203 after propagating through the second collimating lens 304, the dichroic mirror 303, the second optical filter 305 and the converging lens 306.

By the cooperation of the optical components of the light transmission unit, the first light emitted from the light emitting unit 1201 can be irradiated to the sample 307, and the light emitted from the sample 307 can also be transmitted to the detection unit 1203.

In one possible implementation, the detection unit 1203 may include a photodiode, or other device or apparatus that can convert an optical signal into an electrical signal. The detection unit 1203 may further include a signal amplification function and/or an a/D conversion function, for example, may further include a plurality of signal amplifiers to achieve high amplification of weak signals, and may further include an a/D converter to convert analog signals into digital signals for convenient transmission.

In one possible embodiment, the detection result output by the detection module 120 can be used to detect the concentration, negative or positive, of the sample in the reaction well 104.

In a possible implementation mode, a light guide pipe with the length of 4-15 mm can be further arranged in the detection module, and the light guide pipe is used for improving the light collection efficiency and reducing the loss of the fluorescence propagating in the air, so that the detection sensitivity is improved. The material of the light guide pipe can be, for example, glass material, optical fiber, etc. When setting up the light pipe, first light with the second light does not scatter and disappear, can not take place the light leak, sets up the light pipe and helps improving the efficiency that detecting element detected, promotes nucleic acid detection device nucleic acid detection's effect.

In one possible embodiment, a light pipe (not shown) may be disposed between the dichroic mirror 303 and the second collimating lens 304.

In one example, the light guide tube may be cylindrical, rectangular parallelepiped, or the like, and when the sample 307 in the reaction well 104 of the microfluidic chip is excited to emit fluorescence, the light may be transmitted through the second collimating lens 304, the light guide tube, the dichroic mirror 303, the second filter 305, and the converging lens 306 and then received by the detection unit 1203.

The above description has been introduced to the detection module, and it should be understood that the present invention is not limited thereto, and in other embodiments, the embodiments of the present invention can also utilize components and parts such as phase-locked loops to perform phase-locked amplification on the collected dual-optical-path signal, and perform signal integration through integrators and the like, so that the processing of the optical signal is more accurate.

Please refer to fig. 1-5.

In one possible embodiment, the light emitting unit 1201 may emit a first light, which may be transmitted through the detection hole 133 while passing through the light transmitting unit 1202, when the rotation module 110 drives the microfluidic module 100 to step at a low speed, the reaction wells 104 in the microfluidic chip pass through the detection wells 133, the first light irradiates the sample in the reaction hole 104, the sample can emit a second light when excited by the first light, the second light is transmitted by the light transmission unit and the light guide tube and then can be received by the detection unit 1203, the detection unit 1203 detects the second light, a first signal can be generated according to the second light, where the first signal is an electrical signal, and the detection unit 1203 amplifies, filters, and performs analog-to-digital conversion on the first signal to obtain a corresponding digital signal as the detection result, where the digital signal is used for analyzing the sample. By arranging the detection hole 133 in the temperature control region 130, the detection module 120 detects the sample on the microfluidic chip through the detection hole 133, so that the nucleic acid detection device 10 has a compact structure, breaks through the limitation on space, and can save space and materials, thereby saving cost.

In a possible embodiment, the diameter of the detection hole 133 may be set to be slightly larger than the diameter of the reaction hole 104 of the microfluidic chip, and of course, in order to achieve smooth signal collection and transmission, the diameter of the detection hole 133 may also be set to be slightly larger than the diameter of the reaction hole 104 of the microfluidic chip, and at the same time: the diameter < (reaction well 104 diameter +2 × reaction well spacing) of the detection well 133. For example, when the diameter of the reaction hole 104 of the microfluidic chip is 2.7mm, the diameter of the detection hole 133 may be set to be 2.5-5.0 mm (e.g., 4.5 mm). By the above arrangement, when a sample in a certain reaction well 104 is detected through the detection well 133, the influence of fluorescence emitted from samples in other adjacent reaction wells 104 can be eliminated, and the fluorescence collection rate and the detection sensitivity can be improved.

In one possible embodiment, the concentration, positivity, amplification result, etc. of the sample can be obtained by analyzing the first signal from which an amplification curve can be drawn.

After the amplification reaction is completed, the sample in the reaction well 104 may be subjected to a melting reaction, and a melting curve may be drawn by analyzing a fluorescence signal of the sample during the melting reaction and performing an analysis process.

It should be noted that, in the melting reaction, the workflow of the detection unit is similar to that of the amplification unit, and thus the detailed description is omitted.

For the detailed description of other modules, please refer to the foregoing, which is not described herein again.

Although the present invention has been described above by way of example with reference to the nucleic acid detecting apparatus 10, it will be understood by those skilled in the art that the present invention is not limited thereto. In fact, the user can flexibly set the functions of each module according to personal preference and/or practical application scenes, or increase and decrease the number of the modules, as long as the modules are matched to realize integration of sample adding, nucleic acid purification and nucleic acid detection.

Thus, by matching the respective modules of the nucleic acid detecting apparatus 10, the nucleic acid detecting apparatus 10 according to the present invention can realize integration of nucleic acid purification, amplification reaction, and melting reaction, and reduce the operation flow, thereby improving the work efficiency.

Referring to FIG. 6, FIG. 6 shows a schematic structural diagram of a nucleic acid detecting device 10 according to an embodiment of the present invention.

As shown in FIG. 6, the nucleic acid detecting apparatus 10 may further include a transmission module 150.

The transmission module 150 may be electrically connected to the detection module 120, and may be configured to transmit the detection result output by the detection module 120.

In a possible implementation manner, the transmission module 150 may be a wired transmission module or a wireless transmission module, and the detection result is transmitted in a wired or wireless manner. For example, the wireless mode may include WiFi, bluetooth, ZigBee, 3G, 4G, 5G, and the like.

It should be noted that although the present invention has been described with reference to the transmission module 150, the transmission module 150 may be integrated into other modules, for example, a main control chip (e.g., MCU/CPU/DSP/FPGA, etc.) may be disposed in the main control circuit to control the operation of the apparatus, i.e., the transmission module 150 may be integrated into the main control circuit. The master control circuit may be implemented in combination with executable logic instructions to perform the working process of the master control circuit, wherein the executable logic instructions may be implemented based on prior art means. The utility model discloses do not limit to master control circuit's concrete implementation. Similarly, the utility model provides a control to rotation module, temperature control module etc. can realize through correlation technique, the utility model discloses do not restrict to how to control the motor rotation, how to control temperature's embodiment.

In one example, the transmission module 150 can transmit the detection result to an analysis module (not shown), which can be disposed outside the nucleic acid detecting device.

The analysis module may receive the detection result, analyze the sample according to the detection result, and output an analysis result.

In one possible embodiment, the analysis result may include the concentration of the sample, the negative or positive, the status of the amplification reaction, and the like.

In a possible implementation manner, the analysis module may be an analysis module composed of any one or more chips such as MCU/CPU/DSP/FPGA.

In a possible implementation manner, the analysis module may also be a terminal (e.g., a mobile phone, a tablet computer, a host, etc.), a server, or other devices with a computing function.

Of course, although the present invention has been described in a manner in which the analysis module is independent from the nucleic acid detecting apparatus, it should be understood that the present invention is not limited thereto, and in other embodiments, the analysis module may be integrated into the nucleic acid detecting apparatus to realize the analysis function of nucleic acid.

Although the present invention has been described above by way of example with reference to the nucleic acid detecting apparatus 10, it will be understood by those skilled in the art that the present invention is not limited thereto. In fact, the user can flexibly set the functions of each module according to personal preference and/or practical application scenes, or increase and decrease the number of modules, as long as the cooperation of each module can realize integration of nucleic acid release, sample distribution, nucleic acid purification, nucleic acid detection and data transmission.

Like this, through the cooperation of each module of nucleic acid detecting device 10, according to the utility model discloses the integration of release nucleic acid, distribution sample, nucleic acid purification, nucleic acid detection, amplification reaction, melting reaction, data transmission can be realized to nucleic acid detecting device 10 of above-mentioned embodiment, reduces operation flow to improve work efficiency.

Referring to FIG. 7, FIG. 7 is a schematic structural diagram of a nucleic acid detecting device 10 according to an embodiment of the present invention.

As shown in FIG. 7, the nucleic acid detecting apparatus 10 may further include an in-out cartridge module 180.

The inlet and outlet module 180 can be arranged in a cuboid structure or a cube structure, the inlet and outlet module 180 can comprise a first space, the microfluidic module 100 can be arranged in the first space, the microfluidic module 100 passes through the inlet and outlet module 180 to enter and exit the device, and when the inlet and outlet module 180 is driven to enter the device, the first space is a closed space. Wherein, the in-out module 180 may comprise a motor.

Referring to fig. 8a and 8b, fig. 8a shows a perspective view of a nucleic acid detecting device 10 according to an embodiment of the present invention, and fig. 8b shows a front view of the nucleic acid detecting device 10 according to an embodiment of the present invention.

As shown in FIGS. 8a and 8b, the nucleic acid detecting apparatus 10 may include: the device comprises a microfluidic module 100, a rotating module 110, a detection module 120, an in-out bin module 180, a lifting module 200, a temperature auxiliary area 135 and a control module 300.

The in-out module 180 can comprise a motor and can be used for automatic in and out of the microfluidic module 100 to take and place the microfluidic chip, so that the reaction process is completely isolated from the operator, the stability of nucleic acid detection can be improved, and the safety of the operator can be ensured.

Wherein, the lifting module 200 may comprise a motor, which may be used for automatic lifting of the temperature auxiliary zone 135.

The temperature auxiliary region 135 may be disposed on the top of the microfluidic module, and may be used for auxiliary heating to improve heating efficiency, or cooling the entire reaction chamber and assisting in preventing over-temperature.

Wherein, control module 300 can include general purpose chips such as central processing unit CPU, microprocessor MCU, also can include special chip, to this, the utility model discloses do not restrict.

The following is an exemplary description of the nucleic acid detection procedure.

In one example, the microfluidic chip may be placed on a chip tray (in the in-out module 180), and the chip tray is driven by a horizontal motion motor of the in-out module 180 to enter a detection position (i.e., directly below the temperature auxiliary region 135) in the instrument chamber. When the sample in the sample adding hole of the microfluidic chip is to be subjected to high-temperature cracking, the lifting module 200 can drive the temperature auxiliary area 135 to lift, so that the temperature auxiliary area 135 is close to the microfluidic module 100 to form a relatively closed space, and the temperature control module can control the temperature of the first temperature area to reach the cracking temperature, so that the sample in the microfluidic chip releases nucleic acid. After a period of time (which may be set according to actual conditions), the sample lysis is completed, and the temperature auxiliary region 135 may be cooled or the sample may be naturally cooled to a certain temperature to prepare for centrifugation. Then, the lifting module 200 drives the temperature auxiliary area 135 to rise to a suitable distance, the rotating module 110 drives the microfluidic module 100 to rotate at a low speed and a high speed for centrifugation, so that the sample nucleic acid solution enters the reaction hole, the lifting module 200 drives the temperature auxiliary area 135 to press down to be close to but not tightly attached to the microfluidic chip, the temperature of the second temperature area is controlled to reach the temperature of the amplification reaction, or the temperature auxiliary area 135 can also be synchronously heated in an auxiliary manner, so as to improve the stability of the temperature of the reaction hole. So that a constant temperature space of 63 ℃ is formed in the enclosed space for amplification reaction, the rotation module 110 drives the microfluidic module 100 to rotate during the amplification reaction, and the detection module 120 monitors the fluorescence intensity of each reaction hole in real time and transmits the fluorescence intensity to the analysis module (current PC). At the end of amplification, the second temperature region is heated, or the temperature auxiliary region 135 is heated simultaneously, so that the temperature of the second temperature region is raised at a constant speed, and the amplification product is subjected to a melting reaction. The rotation module 110 drives the microfluidic module 100 to rotate, and the detection module 120 monitors the fluorescence intensity of each reaction well in real time and transmits the fluorescence intensity to the analysis module (current PC). After the melting reaction is finished, the temperature of the temperature control area is waited for natural cooling or the temperature is assisted by the temperature auxiliary area to be controlled by 135 ℃ to reach the safe temperature, and the inlet and outlet module 180 controls the micro-fluidic module 100 to be taken out of the warehouse after the safe temperature is reached.

Although the present invention has been described above by way of example with reference to the nucleic acid detecting apparatus 10, it will be understood by those skilled in the art that the present invention is not limited thereto. In fact, the user can flexibly set the functions of each component or increase or decrease the number of components according to personal preference and/or practical application scenarios, as long as the cooperation of each component can realize integration of nucleic acid release, sample distribution, nucleic acid purification, nucleic acid detection (including amplification reaction monitoring and melting reaction monitoring), data transmission and data analysis.

Thus, by matching the respective parts of the nucleic acid detecting apparatus 10, the nucleic acid detecting apparatus 10 according to the present invention can realize integration of nucleic acid release, sample distribution, nucleic acid purification, nucleic acid detection, amplification reaction, melting reaction, data transmission, and analysis, and reduce the operation flow, thereby improving the work efficiency.

Referring to fig. 9, fig. 9 is a flow chart illustrating a method for detecting nucleic acid according to an embodiment of the present invention.

As shown in fig. 9, the method may include:

step S110, controlling the temperature of the first temperature region 131 to be stabilized at a first temperature when the sample is located in the well 101, so that the sample releases nucleic acid.

And step S120, controlling the micro-fluidic chip to rotate at the first rotation speed for the first time, so that the micro-fluidic chip rotates at the first rotation speed for the first time and at the second rotation speed for the second time, and the sample in the sample adding hole 101 enters the reaction hole 104 through the liquid separating area 102, the micro-channel and the micro-valve under the action of centrifugal force.

Step S130, controlling the temperature of the second temperature region 132 to be stabilized at a second temperature when the sample is located in the reaction well 104, so that the sample performs an amplification reaction; when the amplification reaction is finished, controlling the temperature of the second temperature region 132 to rise from the second temperature to a third temperature at a constant speed, so that the sample is subjected to a melting reaction;

step S140, detecting the fluorescence signal generated by the sample in the reaction well 104 through the detection well 133 disposed in the temperature control region 130, and outputting the detection result.

For the detailed description of steps S110 to S140, please refer to the description of the nucleic acid detecting device 10 before, which is not repeated herein.

It should be noted that the nucleic acid detection method is a method corresponding to the nucleic acid detection device, and for specific description, reference is made to the description of the device before, which is not repeated herein.

It should be noted that, although the present invention has been described above by way of example of a nucleic acid detection method, those skilled in the art will appreciate that the present invention is not limited thereto. In fact, the user can flexibly set each step according to personal preference and/or practical application scene, or increase or decrease the number of steps, or change the sequence of steps, as long as the integration of sample adding, nucleic acid purifying and nucleic acid detecting can be realized by the cooperation of each step.

Through above method, the embodiment of the utility model provides a can realize nucleic acid extraction, amplification reaction, melting reaction integration, can reduce operation flow, improve detection efficiency, owing to adopt the integrated design, the embodiment of the utility model provides a detection result that the nucleic acid testing method that provides obtained has higher accuracy. And, the embodiment of the utility model provides an in rotate module 110 and drive micro-fluidic chip rotates the very first time with first rotational speed, rotates the second time with the second rotational speed, makes the sample in the application of sample hole 101 passes through under the effect of centrifugal force divide liquid district 102 the microchannel the microvalve gets into reaction hole 104, the flow direction and the position of sample among the control micro-fluidic chip of can accurate distribution realize evenly dividing liquid, broken valve flow to realize nucleic acid detecting device's stable detection.

Referring to fig. 10, fig. 10 is a block diagram of a nucleic acid detecting system according to an embodiment of the present invention.

As shown in FIG. 10, the nucleic acid detecting system may include at least one nucleic acid detecting apparatus 10 (FIG. 10 is an example, showing that one nucleic acid detecting apparatus 10 is included, but the nucleic acid detecting system may include a plurality of nucleic acid detecting apparatuses 10) and a control apparatus 20.

And a control unit 20 electrically connected to the one or more nucleic acid detecting units 10 for controlling the one or more nucleic acid detecting units 10.

In one possible embodiment, the control device 20 may include a router, and the plurality of nucleic acid detecting devices 10 are connected to the router and control the plurality of nucleic acid detecting devices 10.

In one possible embodiment, the control device 20 may be a terminal (including but not limited to a mobile phone, a computer, etc.), a server, or other devices capable of implementing control and operation functions.

In one possible embodiment, the control device 20 may control the plurality of nucleic acid detecting devices 10 to operate synchronously, thereby realizing synchronous control, and in another embodiment, the control device 20 may control the plurality of nucleic acid detecting devices 10 to operate asynchronously, thereby realizing independent control of the plurality of nucleic acid detecting devices 10.

For a detailed description of the nucleic acid detecting device 10, reference is made to the foregoing description and no further description is made herein.

It should be noted that, although the present invention has been described above by taking a nucleic acid detecting system as an example, those skilled in the art will understand that the present invention should not be limited thereto. In fact, the user can flexibly set the functions of the modules or increase or decrease the number of modules according to personal preference and/or practical application scenarios, as long as the cooperation of the modules can realize the control of one or more nucleic acid detecting devices 10.

Thus, the nucleic acid detecting system according to the above-described embodiment of the present invention can realize the control of one or more nucleic acid detecting apparatuses 10 by the cooperation of the respective modules of the nucleic acid detecting system.

The above description is only for the specific embodiments of the present invention, but the protection scope of the present invention is not limited thereto, and any person skilled in the art can easily think of the changes or substitutions within the technical scope of the present invention, and all should be covered within the protection scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims

1. A nucleic acid detecting device, comprising:

the microfluidic module comprises a microfluidic chip, wherein the microfluidic chip comprises a plurality of sample adding holes, a plurality of liquid dividing areas and a plurality of reaction holes, the sample adding holes are used for adding samples, the reaction holes are used for carrying out amplification reaction and melting reaction on the samples, and the liquid dividing areas are connected with the plurality of reaction holes through microchannels and microvalves;

2. The nucleic acid detecting apparatus according to claim 1, wherein the first temperature region, the second temperature region, and a temperature insulating region are annular, and the temperature insulating region is provided between the first temperature region and the second temperature region to isolate the first temperature region and the second temperature region,

3. The nucleic acid detecting apparatus according to claim 1, wherein the first rotational speed is 1400 to 1800rpm, the first time is 7 to 14s, the second rotational speed is 4200 to 4600rpm, and the second time is 7 to 14 s.

4. The nucleic acid detecting apparatus according to claim 1, wherein the detecting of the fluorescent signal generated from the sample in the reaction well through the detection well includes:

5. The nucleic acid detecting apparatus according to claim 1, wherein the diameter of the detection hole is 3.5mm to 5.5mm, the first temperature is 90 to 100 ℃, the second temperature is 60 to 65 ℃, and the third temperature is 90 to 110 ℃, and wherein a light guide tube having a length of 4 to 15mm is further provided in the detection module.

6. The nucleic acid detecting apparatus according to claim 1, wherein the detecting module includes:

the light-emitting unit is used for emitting first light;

7. A nucleic acid detection system, comprising:

one or more nucleic acid detecting devices according to any one of claims 1 to 6;