CN112694972A - Reaction module and corresponding multiple multi-channel real-time fluorescent nucleic acid detector - Google Patents
Reaction module and corresponding multiple multi-channel real-time fluorescent nucleic acid detector Download PDFInfo
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Abstract
The invention relates to a reaction module for a multiplex multi-channel real-time fluorescent nucleic acid detector, which is of a compact integral structure and consists of a heating body assembly and an optical fiber pair; the heating body assembly comprises a heating module, a cooling module and a temperature control plate; the temperature raising module comprises a heating sheet for inserting and heating the reaction tube; the cooling module comprises a fan for cooling; the optical fiber pair comprises an incident optical fiber and an emergent optical fiber which run parallel to each other; the invention also relates to a multiplex multi-channel real-time fluorescent nucleic acid detector adopting the reaction module, which further comprises: the device comprises a light source excitation and fluorescence detection assembly, a movement assembly, a computer and an external power supply; the heating plate, the fan and the optical fiber pairs are all positioned in a space surrounded by the shell plate of the reaction module. Therefore, the requirement that a small amount of samples are detected at any time can be met, and conditions are created for the nucleic acid detector to enter a basic level inspection mechanism, a small and medium-sized hospital and be used for bedside detection.
Description
Technical Field
The invention relates to the technical field of nucleic acid amplification detection, in particular to a reaction module and a corresponding multiplex real-time fluorescent nucleic acid detector.
Background
The nucleic acid detecting instrument is an instrument for amplifying and detecting nucleic acid by a PCR technique or a constant temperature amplification method, and the real-time fluorescent nucleic acid detecting instrument is an instrument for reflecting the amount of an amplified product by detecting the accumulation amount of a fluorescent signal generated by a fluorescent dye or a specific probe added to a reaction system.
During PCR amplification and isothermal amplification, the real-time fluorescent nucleic acid detector needs to use a light source to irradiate the reaction solution in the reaction tube, and then detect fluorescence generated by a fluorescent dye or a fluorescent probe in the reaction solution through the detector. The traditional real-time fluorescent nucleic acid detector is characterized in that each reaction tube corresponds to one set of light source and one set of detector, one or more excitation channels are arranged in one set of light source, one or more detection channels are arranged in one set of detector, the number of the channels of the real-time fluorescent nucleic acid detector is limited to 4-6 channels due to the influence of the overlapping of fluorescent dye spectrums, only 4-6 target genes can be detected corresponding to each reaction tube, and the traditional real-time fluorescent nucleic acid detector is obviously not enough for the application requiring more detection, such as the detection of mixed infection of respiratory viruses or the identification of infection sources of diseases with similar symptoms.
Chinese patent: a multiplex PCR reaction tube, its assembling method and application are disclosed in application publication No. CN110564584A, which describes a multiplex reaction tube having a plurality of reaction chambers in the lower reaction chamber 221 for carrying out multiplex reactions. However, this patent does not provide a real-time fluorescent nucleic acid detector that can perform multiplex fluorescence detection using such a multiplex PCR reaction tube.
In the conventional nucleic acid detecting instrument and the real-time fluorescent nucleic acid detecting instrument, a plurality of reaction holes are formed in one metal module, a plurality of reaction tubes can be accommodated, the number of the reaction holes is 8, 16, 32, 48 or 96, and a user can select a suitable nucleic acid detecting instrument and a suitable real-time fluorescent nucleic acid detecting instrument according to the amount of a sample to be detected at one time. The more the reaction holes of the instrument are, the more the requirement for batch sample detection is suitable, but the more the holes of the instrument are, the better the hole number of the instrument is, the more the instrument structure is possibly complicated, the volume, the weight and the cost of the instrument are also increased, the less the sample amount is, but the user who needs follow-up examination needs to collect a plurality of samples to be installed on the machine together for reducing the starting-up cost and the depreciation cost of the instrument, the time for giving the detection result is prolonged, the diagnosis of the disease state is delayed, even the infectious diseases are accelerated to spread in susceptible people, if only a small amount of samples are detected at one time, the hole position and the electric quantity are wasted, the utilization efficiency and the service life of the instrument are reduced, and the instrument which can follow-up examination is urgently needed in the market.
The existing nucleic acid detector and the real-time fluorescent nucleic acid detector have the advantages of high flux and strong functions, but have the defects of high price, large volume, heaviness and limited number of detection channels, and are difficult to enter basic level inspection institutions, small and medium-sized hospitals and be used for bedside detection in batches.
Disclosure of Invention
The invention aims to provide a reaction module and a corresponding multiplex real-time fluorescent nucleic acid detector aiming at the defects in the prior art.
In order to achieve the purpose, the invention adopts the technical scheme that:
the invention provides a reaction module for a multiplex real-time fluorescent nucleic acid detector, which is a compact integral structure and consists of a heating body assembly and an optical fiber pair;
the heating body assembly comprises a heating module, a cooling module and a temperature control plate; the heating module comprises a heating sheet for inserting and heating the reaction tube, the cross section of the heating sheet is the same as the cross section of the reaction bin positioned at the lower end of the reaction tube in shape and size, and the edge of the heating sheet is flush with the edge of the reaction bin; the cooling module comprises a fan for cooling; the temperature control plate is used for controlling a temperature rising and reducing process and is respectively connected with the temperature rising module and the temperature reducing module;
the optical fiber pair comprises an incident optical fiber and an emergent optical fiber which run in parallel, one end of the optical fiber pair is abutted against one side of the reaction bin, the other end of the incident optical fiber is communicated with the light source excitation module, and the other end of the emergent optical fiber is communicated with the fluorescence detection module;
the heating plate, the fan and the optical fiber pairs are all positioned in a space surrounded by the shell plate of the reaction module; the multiple reaction modules are connected in parallel, and the computer controls the temperature rise and fall, light source excitation and fluorescence detection processes of the single reaction module, so that the multiple reaction modules connected in parallel can carry out multi-path temperature reaction and multi-path fluorescence detection; each instrument can be provided with one, two, three, four or more reaction modules, preferably, each instrument is provided with four reaction modules, and four temperature reactions and four fluorescence detections can be simultaneously carried out.
The heating module of the heating body assembly further comprises a sleeve plate which is embedded above the shell plate of the reaction module, the sleeve plate is used for fixing and supporting the heating sheet and enabling the reaction tube to keep upright, the sleeve plate and the optical fiber orientation plate are of a separated structure or are integrally formed, the heating module is not limited by the invention, and the sleeve plate is positioned in a space which is defined by the cover plate and the opening above the front shell plate and can contain the reaction tube.
By adopting the technical scheme, each reaction module of the instrument is relatively independent and compact in structure, and cannot interfere with each other during use, so that the same reaction can be carried out simultaneously, different reactions can be carried out simultaneously, and the reactions can not be carried out simultaneously, so that the use flexibility is improved, and the instrument is suitable for being inspected at any time. When one reaction module breaks down (for example, a fan does not rotate or a heating sheet does not heat), the normal work of other reaction modules cannot be influenced, the durability of the instrument is improved, and even parts of the fault reaction module only need to be replaced without returning the whole instrument to the factory for maintenance when the on-site maintenance is needed in an emergency situation, so that the express cost is reduced, the maintenance speed is increased, and the utilization rate of the instrument is further improved.
Preferably, the heating body assembly further comprises an optical fiber orientation plate and a fixing frame which determine the trend of the optical fiber and fix the optical fiber; the optical fiber orientation plate and the fixing frame are positioned in a space surrounded by the shell plate of the reaction module; the optical fiber directional plate is provided with at least one optical fiber directional groove, each optical fiber directional groove can be paved with an optical fiber pair or one or a plurality of optical fibers, which are called as optical fibers (pairs) hereinafter, one side or two sides of the optical fiber directional plate are provided with the optical fiber directional grooves, the optical fiber directional plate is stacked in a single layer or a plurality of layers and is used for placing the single layer or the plurality of layers of optical fibers (pairs), the fixing frame is one or a plurality of fixed frames, the shape of the fixing frame can be one or a plurality of shapes such as I shape, L shape, V shape, Y shape, X shape, T shape, W shape, C shape, U shape or O shape, the fixing frame is made of rigid or flexible bendable material, two ends of the fixing frame and/or the inside of the fixing frame are provided with one or two or a plurality of threaded holes, the threaded holes can correspond to the threaded holes on the optical fiber directional plate, the optical fibers (pairs) paved with the optical fibers are fixed by the fixing, the fixing method of the optical fiber orientation plate and the fixing frame can also adopt other methods and/or structures, and the orientation and the positioning of the optical fiber (pair) can also adopt other methods and/or structures, for example, two foldable optical fiber orientation plates similar to a hinge are used for fixing, for example, the optical fiber (pair) is bundled by using a wire arranging tape, and part of the fixing frame can also be used for fixing components such as a heating sheet, and the invention is not limited to this.
By adopting the technical scheme, the trend and the position of the optical fiber are fixed, the shock resistance strength is improved, the instrument is suitable for being used in a bumpy environment, and the failure rate and the maintenance cost of the instrument are reduced.
Preferably, the included angle θ e (0, π) between one end of the optical fiber pair and one side of the reaction chamber is set.
Preferably, the included angle θ is pi/2, so that the incident light is perpendicular to the edge and the emergent light is perpendicular to the edge, thereby reducing interference.
Preferably, each optical fiber pair abutting against each side of the reaction chamber corresponds to at most one independent reaction chamber of the reaction chamber, and the reaction chambers are far apart from each other, so that optical interference between the reaction chambers is reduced, and optical isolation can be performed between the reaction chambers by adopting a proper mode and/or materials; the number of reaction chambers independent near each side of the reaction chamber can be zero, one, two or more, the number of optical fiber pairs abutted against the corresponding side of the reaction chamber can be zero, one, two or more, and each reaction tube at least comprises one reaction chamber, so that each reaction chamber has at least one corresponding optical fiber pair.
The working principle of the invention is that the sample to be measured in each reaction chamber of the reaction chambers of the reaction tube reacts at a constant temperature or a plurality of temperatures controlled by the temperature rising module and the temperature lowering module, after the light emitted by the light source irradiates the sample to be measured in one reaction chamber of the reaction chamber through the incident optical fiber of the optical fiber pair which is abutted against one edge of the reaction chamber, if the amplification product is generated, the accumulated fluorescence enters a detector through the emergent optical fiber of the same optical fiber pair which is abutted with the same side of the reaction bin to form a fluorescence signal, and the fluorescence signal is displayed through a screen after being analyzed by a computer, and similarly, the incident optical fibers corresponding to the reaction chambers respectively conduct light of the light source to illuminate the reaction chambers, fluorescence generated by the reaction chambers is conducted to the detector through the corresponding emergent optical fibers to form fluorescence signals, and the fluorescence signals of the reaction chambers are analyzed by the computer and then are uniformly displayed through the screen.
The second aspect of the present invention provides a multiplex real-time fluorescent nucleic acid detector using the reaction module, further comprising: the device comprises a light source excitation and fluorescence detection assembly, a movement assembly, a computer and an external power supply;
the light source excitation and fluorescence detection assembly comprises a light source excitation module, a fluorescence detection module and a daylighting panel; the light source excitation module comprises a light source, an optical filter and a first group of optical elements for condensation, collimation and the like; the fluorescence detection module comprises a detector, an optical filter and a second group of optical elements for condensation, collimation and the like; the daylighting panel is used for controlling light source excitation and fluorescence detection and is respectively connected with the light source and the detector;
the motion assembly comprises a motor and a motor driver;
the temperature control plate, the daylighting panel, the motor driver and the computer are respectively connected with the external power supply.
Preferably, the method further comprises the following steps: the device comprises a power interface, a USB interface, a power switch, a screen and a status indicator lamp;
the power interface is connected with the external power supply; the USB interface is connected with the computer; the screen is integrated with the computer; the number of the status indicator lamps is the same as or multiple of the number of the reaction modules, so that the working status of the corresponding reaction module is indicated.
Preferably, the method further comprises the following steps: a housing;
structural components of the multiplex real-time fluorescent nucleic acid detector are all positioned in the shell, and the shell comprises a cover plate and an integrally formed shell plate; the shell plates comprise a front shell plate, a rear shell plate and a bottom shell plate; the middle part of the upper part of the front shell plate is provided with an opening, a sunken groove is formed at the opening, the cover plate is positioned at the opening, the cover plate is connected with the front shell plate or the cover plate is connected with the reaction module shell plate through a rotating shaft at one side, and any feasible mode can be adopted for the opening/closing mode of the cover plate, which is not limited by the invention; the number of the cover plates is the same as that of the reaction modules and is positioned above the corresponding reaction modules, a reaction tube can be accommodated in a space defined by the cover plates and the opening above the front shell plate, the space above the reaction tube can be sealed by each cover plate, so that external dust and ambient light can be prevented from entering, sealing gaskets or other sealing modes for enhancing sealing can be arranged at the opening above the front shell plate and/or in the circumferential direction below the cover plates, and the invention is not limited to the above; the cover plate can be made of heat preservation and insulation materials, or the heat preservation and insulation materials are arranged in or outside the cover plate and are used for preserving heat on the upper part of the reaction tube and insulating heat among the reaction modules, and the heat insulation effect can be strengthened by enlarging the distance between the reaction modules, which is not limited in the invention; the front shell plate is connected with the bottom shell plate through a first side plate extending downwards from two sides of the front shell plate, and is connected with the rear shell plate through a second side plate extending downwards from the rear side of the front shell plate, the screen and the USB interface are arranged on the side plate in front of the front shell plate, and the USB interface is arranged on one side close to the screen; the rear shell plate is provided with the power interface and the power switch; the bottom shell plate is fixed with a lower base plate, the temperature control plate, the daylighting plate and the motor driver are fixed on the lower base plate, a support rod is further installed on the lower base plate, a middle base plate is installed on the support rod, and the reaction module is fixed on the middle base plate through the reaction module shell plate.
By adopting the technical scheme, the multiplex multi-channel real-time fluorescent nucleic acid detector has a sealed integral structure, is not easily influenced by external dust, water mist and the like, has a small volume and is convenient to carry and move. The cover plate can be conveniently opened or closed, and the reaction tube is convenient to take. The instrument is provided with a data interface and a display screen, so that the operation of an external computer is not needed, and the portability and the use convenience of the whole instrument are further improved.
By adopting the technical scheme, compared with the prior art, the invention has the following technical effects:
the invention can break through the quantity limitation of multiple detection target genes of the existing real-time fluorescent nucleic acid detector, each reaction chamber in the multiple reaction tube is equivalent to a traditional complete reaction tube, if the quantity of the multiple detection target genes of the existing real-time fluorescent nucleic acid detector is n, and the quantity of the reaction chambers in the multiple reaction tube is m, the quantity of the multiple detection target genes of the multiple real-time fluorescent nucleic acid detector is nxm, thereby realizing the improvement of m times. The multi-channel reaction module provided by the invention can change the limitation that the samples of the existing nucleic acid detector and the real-time fluorescent nucleic acid detector need to be detected in batch and cannot be detected at any time, and provides a better choice for users, particularly users who have a small single detection quantity of the samples and need to be detected at any time. The problems of false negative caused by nucleic acid degradation due to long-time storage of samples caused by overstocked large amount of samples and delay of diagnosis and treatment due to long-time waiting of detection results are solved.
The multi-path reaction module provided by the multi-path real-time fluorescence detector can simultaneously carry out a plurality of different reactions, so that the optimization time can be reduced by times for users who often need to optimize reaction conditions and develop new detection kits, the working intensity of kit developers is reduced, and the marketing speed of the detection kits is accelerated.
By using the multiple multi-channel real-time fluorescent nucleic acid detector provided by the invention, a basic inspection mechanism and a small and medium-sized hospital can realize the purpose that samples can be inspected at any time, a plurality of reagent kits with different reaction conditions and used for detecting different pathogens can be simultaneously used on a plurality of reaction modules to carry out multi-tube multi-detection, or specific primers and probes for detecting different pathogens are fixed in different reaction chambers of a reaction tube in advance, and the same reagent kit is used for carrying out single-tube multi-detection, so that a diagnosis and treatment scheme is provided for determining an infection source as soon as possible, examining infectious disease infected patients and suspected patients as soon as possible, and a diagnosis and treatment scheme is provided for confirmed patients as soon as possible, thereby providing possibility for restraining the. The nucleic acid detector can further expand the number of detected samples by adding the reaction modules, and the number of the reaction modules of the nucleic acid detector is not limited by the invention.
Drawings
FIG. 1 is a perspective view of the multiplex real-time fluorescent nucleic acid detecting apparatus of the present invention, in which FIG. 1A is a lid-closed view, FIG. 1B is a lid-opened view, and FIG. 1B is a view showing the positions of reaction tubes;
FIG. 2 is a diagram showing the relationship between the reaction module structure and the reaction tube of the multiplex real-time fluorescent nucleic acid detecting apparatus of the present invention, in which FIG. 2A is an exploded view and FIG. 2B is an assembled view;
FIG. 3 is an assembly diagram of the internal structure and reaction tubes of the multiplex real-time fluorescent nucleic acid detecting apparatus according to the present invention;
FIG. 4 is an assembly diagram of the outer casing of the multiplex real-time fluorescent nucleic acid detector of the present invention;
FIG. 5 is a schematic front view and a schematic back view of a reaction tube having five reaction chambers used in example 2 of the present invention;
wherein the reference numerals include: heat patch 1a 1; race plate 1a 3; fan 1B 1; a temperature control plate 1C; an optical fiber orientation plate 1D; fiber orientation groove 1D 1; a fixed frame 1E; a pair of optical fibers 2; a reaction bin 221; a light source excitation and fluorescence detection assembly 3; a daylighting panel 38; a moving assembly 4; a motor 41; a motor driver 42; an external power supply 5; a power supply interface 51; a USB interface 52; a power switch 53; a screen 6; a reaction module 7; the reaction module shell 71; a status indicator light 72; a cover plate 81; a front shell plate 82; a first side plate 821; a front side plate 822; a second side panel 823; a rear shell plate 83; a bottom shell plate 84; a lower substrate 841; a support bar 842; a middle substrate 843.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
It should be noted that the embodiments and features of the embodiments may be combined with each other without conflict.
The invention is further described with reference to the following drawings and specific examples, which are not intended to be limiting.
Example 1
This example provides a multiplex real-time fluorescent nucleic acid detector, as shown in fig. 1-4, comprising: the device comprises a reaction bin 221, a light source excitation and fluorescence detection component 3, a movement component 4, a computer, an external power supply 5 and four mutually independent reaction modules 7;
the reaction bin 221 is positioned at the lower end of the reaction tube;
the light source excitation and fluorescence detection component 3 comprises a light source excitation module, a fluorescence detection module and a daylighting panel 38; the light source excitation module comprises a light source, an optical filter and a first group of optical elements for condensation, collimation and the like; the fluorescence detection module comprises a detector, an optical filter and a second group of optical elements for condensation, collimation and the like; the lighting panel 38 is used for controlling light source excitation and fluorescence detection, and is respectively connected with the light source and the detector;
the moving assembly 4 comprises a motor 41 and a motor driver 42;
as shown in fig. 2 and 3, the reaction module 7 is a compact integral structure, and is composed of a heating body assembly and an optical fiber pair 2; the heating body assembly comprises a heating module, a cooling module and a temperature control plate 1C; the heating module comprises a heating plate 1A1 for inserting and heating a reaction tube, the cross section of the heating plate 1A1 is polygonal, the shape and the size of the cross section of the heating plate are the same as those of the cross section of the reaction bin 221, and the edges of the heating plate 1A1 are flush with the edges of the reaction bin 221; the cooling module comprises a fan 1B1 for cooling; the temperature control plate 1C is used for controlling a temperature rise and reduction process and is respectively connected with the temperature rise module and the temperature reduction module; the optical fiber pair 2 comprises an incident optical fiber and an emergent optical fiber which run in parallel, one end of the optical fiber pair 2 is abutted to one side of the reaction bin 221, the other end of the incident optical fiber is communicated with the light source excitation module, and the other end of the emergent optical fiber is communicated with the fluorescence detection module; the heating plate 1A1, the fan 1B1 and the optical fiber pair 2 are all positioned in a space surrounded by the reaction module shell plate 71; the four reaction modules 7 are connected in parallel, and the computer controls the temperature rise and fall, the light source excitation and the fluorescence detection processes of the single reaction module 7, so that the four reaction modules 7 connected in parallel can carry out four-way temperature reaction and four-way fluorescence detection; by adopting the technical scheme, the whole structure of the reaction module is compact, and the fault of one reaction module does not influence the normal use of other reaction modules;
the temperature control plate 1C, the lighting plate 38, the motor driver 42 and the computer are respectively connected with the external power supply 5.
The four-way reaction module 7 provided by the multi-path real-time fluorescence detector can avoid the limitation that samples of the existing nucleic acid detector and the real-time fluorescence nucleic acid detector need to be detected in batches, when a user acquires a new sample to be detected immediately, the user does not need to wait for the detector in use to finish operation or wait for the whole batch of samples to be accumulated, and only needs to have the idle reaction module 7 to start reaction immediately, so that convenience is provided for the user needing frequent sampling and detection, and the problems of false negative caused by degradation of nucleic acid due to long-time storage and the problem of lag of detection results caused by overstock of a large number of samples are avoided. Assuming that 1.5 hours is required for one detection experiment, 5 detections can be performed during 8 hours a day using the existing nucleic acid detector and real-time fluorescent nucleic acid detector, and 20 detections can be performed during 8 hours a day using the real-time fluorescent nucleic acid detector provided with the four-way reaction module of the present invention. When a new detection kit is developed, reaction conditions including parameters such as denaturation time, annealing/extension temperature and the like need to be optimized on a detector, on the assumption that each experiment needs 1.5 hours and 2 times of repetition, the optimization process of using the existing nucleic acid detector and the real-time fluorescent nucleic acid detector needs 90 hours, and if the real-time fluorescent nucleic acid detector provided with the four-way reaction module is adopted, the experiment time can be reduced to 22.5 hours, so that the working intensity of kit developers is reduced, and the marketing speed of the detection kit is accelerated.
Preferably, the method further comprises the following steps: a power interface 51, a USB interface 52, a power switch 53, a screen 6 and a status indicator lamp 72;
the power interface 51 is connected with the external power supply 5; the USB interface 52 is connected with the computer; the screen 6 is integrated with the computer; the number of the status indicator lamps 72 is the same as the number of the reaction modules 7, thereby indicating the operation status of the corresponding reaction module 7.
The working principle of the invention is that samples to be measured in each reaction chamber in a reaction tube in which one or more reaction modules 7 are inserted react at a constant temperature or a plurality of temperatures controlled by a heating module and a cooling module, when light emitted by a light source irradiates the samples to be measured in each reaction chamber through an incident optical fiber abutted against one side of a reaction chamber 221 at the lower end of the reaction tube, if an amplification product is generated, accumulated fluorescence enters a detector through an emergent optical fiber abutted against the side of the reaction chamber 221 of the reaction tube to form a fluorescence signal, and the fluorescence signal is acquired by a computer and displayed through a screen 6.
Preferably, as shown in fig. 1B and 4, the method further includes: a housing;
the structural components of the multiplex real-time fluorescent nucleic acid detector are all positioned in the shell, and the shell comprises a cover plate 81 and an integrally formed shell plate; the shell plates include a front shell plate 82, a rear shell plate 83, a bottom shell plate 84; the middle part of the upper part of the front shell plate 82 is opened, a sunken groove is formed at the opening, the cover plate 81 is positioned at the opening, the cover plate 81 is connected with the front shell plate 82 through a rotating shaft at one side, and any feasible mode can be adopted for the opening/closing mode of the cover plate 81, which is not limited in the invention; the number of the cover plates 81 is one or more, preferably, each instrument is provided with four cover plates 81, each cover plate 81 is positioned above the corresponding reaction module 7, a reaction tube can be accommodated in a space defined by the openings above each cover plate 81 and the front shell plate 82, and each cover plate 81 can seal the space above the reaction tube to prevent external dust and ambient light from entering; the front shell 82 is connected to the bottom shell 84 through a first side plate 821 with two sides extending downward, and is connected to the rear shell 83 through a second side plate 823 with a rear extending downward, the front side plate 822 of the front shell 82 is provided with the screen 6 and the USB interface 52, and the USB interface 52 is arranged on a side close to the screen 6; the rear shell plate 83 is provided with the power interface 51 and the power switch 53; a lower substrate 841 is fixed on the bottom shell plate 84, the temperature control plate 1C, the daylighting plate 38 and the motor driver 42 are fixed on the lower substrate 841, a support rod 842 is further installed on the lower substrate 841, a middle substrate 843 is installed on the support rod 842, and the reaction module 7 is fixed on the middle substrate 843 through the reaction module shell plate 71.
By adopting the technical scheme, the multiplex multi-channel real-time fluorescent nucleic acid detector has a sealed integral structure, is not easily influenced by external dust, water mist and the like, has a small volume and is convenient to carry and move. The cover plate can be conveniently opened or closed, and the reaction tube is convenient to take. The instrument is provided with a data interface and a display screen, so that the operation of an external computer is not needed, and the portability and the use convenience of the whole instrument are further improved.
Preferably, as shown in fig. 2A, the heating body assembly further includes an optical fiber orientation plate 1D and a fixing frame 1E for determining the orientation and fixing of the optical fiber; the optical fiber orientation plate 1D and the fixing frame 1E are positioned in a space surrounded by the reaction module shell plate 71; two optical fiber orientation grooves 1D1 are respectively arranged on the left wing and the right wing of the optical fiber orientation plate 1D, and an optical fiber pair 2 can be laid in each optical fiber orientation groove 1D1, so that the optical fiber pair 2 is bent and then guided to two longer vertical side edges of the reaction bin 221. The optical fiber positioning device is characterized in that the fixing frames 1E are multiple, threaded holes are formed in the fixing frames 1E and can correspond to threaded hole positions on the optical fiber positioning plates 1D, and the fixing frames 1E are used for fixing four optical fiber pairs 2 with bent trends and other three optical fiber pairs 2 with unbent trends in the optical fiber positioning grooves 1D1 with optical fibers laid.
The invention uses the optical fiber orientation plate 1D with the optical fiber orientation groove 1D1 to determine the trend of the optical fiber pair 2, and uses a plurality of fixing frames 1E to fix each optical fiber, thereby improving the shock resistance of the system, enabling the instrument to be suitable for being used in a bumpy environment, and reducing the failure rate and the maintenance cost of the instrument.
Preferably, an included angle θ between one end of the optical fiber pair 2 and one side of the reaction chamber is pi/2, so that incident light is perpendicular to the side, and emergent light is perpendicular to the side, thereby reducing interference.
The edge of the lower end of the reaction bin 221 of the reaction tube which is abutted against the optical fiber pair 2 is made of transparent material;
the lower end of the reaction bin 221 is provided with five sides, the optical fiber pairs 2 abutted with the five sides at the lower end of the reaction bin 221 are seven, and the seven optical fiber pairs 2 abutted with the five sides at the lower end of the reaction bin 221 can correspond to an independent reaction bin chamber close to a certain side at the lower end of the reaction bin 221. The number of the reaction chambers close to the bottom edge of the reaction chamber 221 is one, the number of the reaction chambers close to the two oblique side edges of the reaction chamber 221 is one, the number of the reaction chambers close to the two longer vertical side edges of the reaction chamber 221 is two, the number of the optical fiber pairs 2 abutted against the bottom edge, the two oblique side edges and the two vertical side edges of the lower end of the corresponding reaction chamber 221 is one, two or two, and each reaction tube comprises seven reaction chambers.
By adopting the technical scheme, the multiple real-time fluorescence detector can simultaneously detect the fluorescence signals of the samples to be detected in the seven reaction chambers, and compared with the existing products on the market, the multiple detection capability is improved by seven times.
Example 2
This example is an alternative configuration of the multiplex reaction tube used in the multiplex real-time fluorescent nucleic acid detecting apparatus described in example 1, and is mainly different in that the reaction tube used has five reaction chambers (as shown in FIG. 5);
five of the seven optical fiber pairs 2 abutting against five sides of the lower end of the reaction bin 221 may respectively correspond to an independent reaction bin chamber near a certain side of the lower end of the reaction bin 221. The number of the reaction chambers close to the bottom edge of the reaction chamber 221 is one, the number of the reaction chambers close to two oblique side edges of the reaction chamber 221 is one, the number of the reaction chambers close to two longer vertical side edges of the reaction chamber 221 is also one, the number of the optical fiber pairs 2 abutted against the bottom edge, the two oblique side edges and the two vertical side edges of the lower end of the corresponding reaction chamber 221 is one, two or two, each reaction tube comprises five reaction chambers, and two optical fiber pairs 2 in seven optical fiber pairs 2 abutted against five edges of the reaction chamber 221 do not correspond to the reaction chambers.
When the reaction tube with five reaction chambers is used, under the command of a computer of the detector, the light collecting plate controls five of the seven optical fiber pairs 2 to respectively collect illumination and fluorescence signals of a sample to be detected of the five reaction chambers, and controls the other two optical fiber pairs 2 not to perform illumination and/or fluorescence signal collection, so that the detector is compatible with the reaction tube types with less reaction chambers downwards.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention.
Claims (8)
1. The reaction module for the multiplex multi-channel real-time fluorescent nucleic acid detector is characterized in that the reaction module (7) is of a compact integral structure and consists of a heating body assembly and an optical fiber pair (2);
the heating body assembly comprises a heating module, a cooling module and a temperature control plate (1C); the heating module comprises a heating sheet (1A1) used for inserting and heating a reaction tube, the cross section of the heating sheet (1A1) is the same as the cross section of a reaction bin (221) positioned at the lower end of the reaction tube in shape and size, and the edge of the heating sheet (1A1) is flush with the edge of the reaction bin (221); the cooling module comprises a fan (1B1) for cooling; the temperature control plate (1C) is used for controlling a temperature rising and reducing process and is respectively connected with the temperature rising module and the temperature reducing module;
the optical fiber pair (2) comprises an incident optical fiber and an emergent optical fiber which run in parallel, one end of the optical fiber pair (2) is abutted against one side of the reaction bin (221), the other end of the incident optical fiber is communicated with the light source excitation module, and the other end of the emergent optical fiber is communicated with the fluorescence detection module;
the heating plate (1A1), the fan (1B1) and the optical fiber pair (2) are all positioned in a space surrounded by a reaction module shell plate (71); the reaction modules (7) are connected in parallel, and the computer controls the temperature rise and fall, the light source excitation and the fluorescence detection process of the single reaction module (7), so that the reaction modules (7) connected in parallel can carry out multi-path temperature reaction and multi-path fluorescence detection.
2. The reaction module according to claim 1, wherein the heating body assembly further comprises a fiber orientation plate (1D) and a fixing frame (1E) which determine the orientation and fixing of the optical fibers; the optical fiber orientation plate (1D) and the fixing frame (1E) are positioned in a space surrounded by the reaction module shell plate (71).
3. The reaction module according to claim 1, wherein the angle θ e (0, π) between one end of said pair of fibers (2) and one side of said reaction chamber (221) is in contact.
4. A reaction module according to claim 3, wherein the angle θ ═ pi/2.
5. A reaction module according to claim 3, wherein each of said pairs of optical fibers (2) abutting each side of said reaction chamber (221) corresponds to at most one independent reaction chamber of said reaction chamber (221), and each reaction tube comprises at least one reaction chamber, so that each of said reaction chambers has at least one corresponding pair of optical fibers (2).
6. A multiplex real-time fluorescent nucleic acid detector employing the reaction module of any one of claims 1-5, further comprising: the device comprises a light source excitation and fluorescence detection assembly (3), a movement assembly (4), a computer and an external power supply (5);
the light source excitation and fluorescence detection assembly (3) comprises a light source excitation module, a fluorescence detection module and a daylighting panel (38); the light source excitation module comprises a light source, an optical filter and a first group of optical elements for condensation, collimation and the like; the fluorescence detection module comprises a detector, an optical filter and a second group of optical elements for light gathering and collimation; the daylighting panel (38) is used for controlling light source excitation and fluorescence detection and is respectively connected with the light source and the detector;
the motion assembly (4) comprises a motor (41) and a motor driver (42);
the temperature control plate (1C), the daylighting plate (38), the motor driver (42) and the computer are respectively connected with the external power supply (5).
7. The multiplex real-time fluorescent nucleic acid detector of claim 6, further comprising: the device comprises a power interface (51), a USB interface (52), a power switch (53), a screen (6) and a status indicator lamp (72);
the power supply interface (51) is connected with the external power supply (5); the USB interface (52) is connected with the computer; the screen (6) is integrated with the computer; the number of the status indicator lamps (72) is the same as the number of the reaction modules (7) or is a multiple of the number of the reaction modules (7), so that the operating status of the corresponding reaction modules (7) is indicated.
8. The multiplex real-time fluorescent nucleic acid detector of claim 6, further comprising: a housing;
structural components of the multiplex real-time fluorescent nucleic acid detector are all positioned in the shell, and the shell comprises a cover plate (81) and an integrally formed shell plate; the shell plates comprise a front shell plate (82), a rear shell plate (83) and a bottom shell plate (84); the middle part of the upper part of the front shell plate (82) is provided with an opening, a sunken groove is formed at the opening, the cover plate (81) is positioned at the opening, the cover plate (81) is connected with the front shell plate (82) through a rotating shaft at one side or the cover plate (81) is connected with the reaction module shell plate (71), the number of the cover plates (81) is the same as that of the reaction modules (7), and the cover plates are positioned above the corresponding reaction modules (7); the front shell plate (82) is connected with the bottom shell plate (84) through a first side plate (821) with two sides extending downwards, and is connected with the rear shell plate (83) through a second side plate (823) with a rear side extending downwards, the screen (6) and the USB interface (52) are arranged on a front side plate (822) of the front shell plate (82), and the USB interface (52) is arranged on one side close to the screen (6); the rear shell plate (83) is provided with the power interface (51) and the power switch (53); be fixed with infrabasal plate (841) on bottom shell plate (84), be fixed with on infrabasal plate (841) temperature control plate (1C), plane skylight (38) and motor drive (42), still install bracing piece (842) on infrabasal plate (841), base plate (843) in installing on bracing piece (842), pass through on well base plate (843) reaction module shell plate (71) are fixed with reaction module (7).
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117402730A (en) * | 2023-12-14 | 2024-01-16 | 鲲鹏基因（北京）科技有限责任公司 | Temperature control device for PCR detection and PCR instrument |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6369893B1 (en) * | 1998-05-19 | 2002-04-09 | Cepheid | Multi-channel optical detection system |
| CN105675574A (en) * | 2016-03-17 | 2016-06-15 | 苏州天隆生物科技有限公司 | Multi-fluorescence channel detection system for real-time fluorescence quantitative PCR |
| CN110564584A (en) * | 2019-08-23 | 2019-12-13 | 默礼生物(杭州)有限公司 | Multiple PCR reaction tube and assembling method and application thereof |
| CN111394235A (en) * | 2020-01-17 | 2020-07-10 | 杭州柏恒科技有限公司 | Fluorescence detection device and detection method thereof |
| CN211856359U (en) * | 2020-01-19 | 2020-11-03 | 杭州优米仪器有限公司 | Double scanning formula fluorescence detection device |
| CN215668001U (en) * | 2020-11-23 | 2022-01-28 | 默礼生物(杭州)有限公司 | Reaction module and corresponding multiple multi-channel real-time fluorescent nucleic acid detector |
-
2020
- 2020-11-23 CN CN202011333261.7A patent/CN112694972A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6369893B1 (en) * | 1998-05-19 | 2002-04-09 | Cepheid | Multi-channel optical detection system |
| CN105675574A (en) * | 2016-03-17 | 2016-06-15 | 苏州天隆生物科技有限公司 | Multi-fluorescence channel detection system for real-time fluorescence quantitative PCR |
| CN110564584A (en) * | 2019-08-23 | 2019-12-13 | 默礼生物(杭州)有限公司 | Multiple PCR reaction tube and assembling method and application thereof |
| CN111394235A (en) * | 2020-01-17 | 2020-07-10 | 杭州柏恒科技有限公司 | Fluorescence detection device and detection method thereof |
| CN211856359U (en) * | 2020-01-19 | 2020-11-03 | 杭州优米仪器有限公司 | Double scanning formula fluorescence detection device |
| CN215668001U (en) * | 2020-11-23 | 2022-01-28 | 默礼生物(杭州)有限公司 | Reaction module and corresponding multiple multi-channel real-time fluorescent nucleic acid detector |
Non-Patent Citations (1)
| Title |
|---|
| 王伟平;: "实时荧光定量PCR检测系统的研究", 硅谷, no. 22, 23 November 2010 (2010-11-23), pages 89 - 90 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117402730A (en) * | 2023-12-14 | 2024-01-16 | 鲲鹏基因（北京）科技有限责任公司 | Temperature control device for PCR detection and PCR instrument |
| CN117402730B (en) * | 2023-12-14 | 2024-03-05 | 鲲鹏基因（北京）科技有限责任公司 | Temperature control device for PCR detection and PCR instrument |
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