CN219010300U

CN219010300U - Real-time fluorescence quantitative PCR detection device

Info

Publication number: CN219010300U
Application number: CN202221316380.6U
Authority: CN
Inventors: 付沈斌; 唐安华; 张键
Original assignee: Guangzhou Fuyi Medical Technology Co ltd
Current assignee: Guangzhou Fuyi Medical Technology Co ltd
Priority date: 2022-05-27
Filing date: 2022-05-27
Publication date: 2023-05-12
Anticipated expiration: 2032-05-27

Abstract

The utility model relates to the technical field of PCR detection, in particular to a real-time fluorescence quantitative PCR detection device, which comprises a PCR reaction tube, wherein a sealing film is arranged at the opening of the PCR reaction tube; the device also comprises a detection seat, an optical detection assembly for detecting the sample in the PCR reaction tube and a membrane opening assembly for puncturing the sealing membrane; the optical detection assembly and the membrane opening assembly are both arranged on the detection seat. The utility model has simple structure and arrangement, and can puncture the sealing film when the sample in the PCR reaction tube is required to be detected through the arrangement of the film opening assembly, thereby effectively preventing the PCR reaction tube from being in an open state all the time, and causing the sample in the tube to be polluted to influence the subsequent detection.

Description

Real-time fluorescence quantitative PCR detection device

Technical Field

The utility model relates to the technical field of PCR instrument detection, in particular to a real-time fluorescent quantitative PCR detection device.

Background

The real-time fluorescent quantitative PCR instrument consists of fluorescent quantitative system and computer for monitoring the fluorescence of the circulation process, and the computer connected to the real-time equipment collects fluorescent data, which is displayed in the form of graph via developed real-time analysis software.

The optical detection element of the existing real-time fluorescent quantitative PCR instrument detection device is convenient to integrate into the instrument, the volume is usually small, samples in reaction tubes at different positions are detected by moving the optical detection element, and the reaction tubes are generally required to be opened in advance, so that reagents in the reaction tubes can be exposed to the outside in advance, the reagents are easy to be polluted, and the preservation and subsequent detection of the samples are not facilitated.

The utility model discloses a portable real-time fluorescence ration PCR appearance, including box, heating cooling unit, light source emission receiving unit, display element and power, the box is provided with year dish and slide rail, is provided with a plurality of standing grooves of arranging in a row on the year dish, places the test bottle in the standing groove, opens heating cooling unit and carries out the circulation heating to the test bottle, can make the DNA fragment in the test bottle obtain the rapid amplification, simultaneously carries out the mark control with light source emission receiving unit alignment test bottle to the product through the slide rail to obtain real-time fluorescence quantity. The portable real-time fluorescence quantitative PCR instrument aims at a light source transmitting and receiving unit at a test bottle through sliding adjustment of the sliding rail, but the reagent bottle is inconvenient to open, and if the reagent bottle is opened completely in advance, a sample is polluted.

Disclosure of Invention

The utility model provides a real-time fluorescence quantitative PCR detection device which is simple in structure and can perform membrane opening operation on a reaction tube, and aims to solve the problems that the cover of the reaction tube in the prior art is inconvenient or the reaction tube needs to be opened in advance to cause sample pollution.

In order to solve the technical problems, the utility model adopts the following technical scheme: the real-time fluorescence quantitative PCR detection device comprises a PCR reaction tube, wherein a sealing film is arranged at the opening of the PCR reaction tube; the device also comprises a detection seat, an optical detection assembly for detecting a sample in the PCR reaction tube and a membrane opening assembly for puncturing the sealing membrane; the optical detection assembly and the membrane opening assembly are both arranged on the detection seat.

The PCR reaction tube is used for placing the sample to be detected, the sealing film is sealed after the sample to be detected is placed in the PCR reaction tube, the sealing film is punctured by the film opening component on the detection seat when the sample in the PCR reaction tube needs to be detected, and the optical detection component can detect the sample in the PCR reaction tube with the film opening. The utility model has simple structure and arrangement, and can puncture the sealing film of the PCR reaction tube when the sample in the PCR reaction tube is required to be detected through the arrangement of the film opening assembly, thereby effectively preventing the PCR reaction tube from being in an open state all the time, and causing the sample in the tube to be polluted to influence the subsequent detection.

Preferably, a slot is arranged at the bottom of the detection seat; the membrane opening assembly is arranged in the groove.

Preferably, the membrane opening assembly comprises a piercing member capable of moving up and down in the slot and a driving structure for driving the piercing member to move; the driving structure is connected with the piercing member; the driving structure is arranged in the slot.

Preferably, the driving structure comprises a cylinder and a connecting piece; the air cylinder is arranged at the bottom of the slot, and the connecting piece is connected with a piston rod of the air cylinder; the piercing member is arranged on the connecting member.

Preferably, the side wall of the slot is provided with a chute; the connecting piece is provided with a sliding block; the sliding block is in sliding connection with the sliding groove; and a hemispheroid is arranged on one side of the sliding block, which faces the sliding groove.

Preferably, the piercing member comprises an opening member and a piercing head; one end of the opening part is connected with the connecting part, and the other end of the opening part is connected with the puncture head.

Preferably, the puncture head is arranged at the middle position of the bottom of the spreading piece.

Preferably, the opening piece is a hollow structure with two open ends.

Preferably, a second channel extending to the outside of the detection seat is communicated with the center of the bottom of the groove; the connecting piece is arranged as a ring piece; one end of the opening member is connected with the connecting member, and the piercing head is arranged at the bottom edge of the other end of the opening member.

Preferably, the detection seat is provided with a first channel; the optical detection assembly comprises an excitation assembly for emitting excitation light and an emission assembly for receiving the emitted light, and the excitation assembly and the emission assembly are both arranged in the first channel.

Compared with the prior art, the utility model has the beneficial effects that:

1) The utility model has simple structure and arrangement, can perform membrane opening operation in the PCR reaction tube through the arrangement of the membrane opening assembly, and puncture the sealing membrane before the sample in the PCR reaction tube is required to be detected, thereby effectively preventing the PCR reaction tube from being in an open state all the time, and causing the sample in the tube to be polluted to influence the subsequent detection;

2) The second channel and the membrane opening assembly are matched, so that liquid can be discharged or absorbed into the PCR reaction tube when needed, and the PCR reaction tube is very convenient to use and high in practicability.

Drawings

FIG. 1 is a schematic diagram of the structure of a real-time fluorescent quantitative PCR detection device of the present utility model;

FIG. 2 is an enlarged view at A in FIG. 1;

FIG. 3 is an enlarged view at B in FIG. 1;

FIG. 4 is a schematic view of the structure of the spreader of the present utility model;

FIG. 5 is a schematic diagram showing the structure of another embodiment of the real-time fluorescent quantitative PCR detection apparatus according to the present utility model;

FIG. 6 is an enlarged view at C in FIG. 5;

FIG. 7 is a schematic top view of the test seat of the present utility model in a separated state.

In the accompanying drawings: 1-PCR reaction tube; 11-sealing film; 12-extension; 2-a detection seat; 21-a first channel; 211-excitation light channels; 212-an emission light channel; 22-slotting; 23-a second channel; 24-a first half-seat; 25-a second half-seat; 3-an optical detection assembly; 31-an excitation assembly; 311-TIR collimator lens; 312-a collimating cylindrical lens; 313-a first narrowband filter; 314—single color LED lamp; a 32-transmitting assembly; 321-a first plano-convex lens; 322-a second narrowband filter; 323-silicon photomultiplier; 324-a second plano-convex lens; 33-two-phase color separation mirror; 4-opening a membrane assembly; 6-piercing member; 61-a spreader; 611-a first cylindrical portion; 612—a diameter-variable portion; 613-a second cylindrical portion; 62-piercing head; 42-a drive structure; 421-cylinder; 422-connection; 51-sliding grooves; 52-sliding blocks; 521-hemispheres; 7-pipe frames; 8-side plates; 9-top cap.

Detailed Description

The drawings are for illustrative purposes only and are not to be construed as limiting the present patent; for the purpose of better illustrating the embodiments, certain elements of the drawings may be omitted, enlarged or reduced and do not represent the actual product dimensions; it will be appreciated by those skilled in the art that certain well-known structures in the drawings and descriptions thereof may be omitted. The positional relationship depicted in the drawings is for illustrative purposes only and is not to be construed as limiting the present patent.

The same or similar reference numbers in the drawings of embodiments of the utility model correspond to the same or similar components; in the description of the present utility model, it should be understood that, if there are orientations or positional relationships indicated by terms "upper", "lower", "left", "right", "long", "short", etc., based on the orientations or positional relationships shown in the drawings, this is merely for convenience in describing the present utility model and simplifying the description, and is not an indication or suggestion that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, so that the terms describing the positional relationships in the drawings are merely for exemplary illustration and are not to be construed as limitations of the present patent, and that it is possible for those of ordinary skill in the art to understand the specific meaning of the terms described above according to specific circumstances.

The technical scheme of the utility model is further specifically described by the following specific embodiments with reference to the accompanying drawings:

example 1

Referring to fig. 1-4, an embodiment 1 of a real-time fluorescent quantitative PCR detection device is shown, which comprises a PCR reaction tube 1, a detection seat 2, an optical detection assembly 3 and a membrane opening assembly 4, wherein the PCR reaction tube 1 is a universal uncovered PCR reaction tube, a sealing membrane 11 is sealed at the opening of the PCR reaction tube after a sample to be detected is filled into the PCR reaction tube 1, and the sealing membrane 11 is a thin membrane made of aluminum foil or silica gel.

The PCR reaction tube 1 is placed on the pipe frame 7, holes are uniformly distributed on the pipe frame 7, an extension portion 12 is arranged at the opening end of the PCR reaction tube 1, and when the PCR reaction tube 1 is placed on the pipe frame 7, the extension portion 12 is lapped on the pipe frame 7.

The detection seat 2 is located above the pipe frame 7, the detection seat 2 can move above the pipe frame 7, so that samples in PCR reaction pipes at different positions can be conveniently detected, the structure for driving the detection seat 2 to move is in the prior art, corresponding structures are not drawn in the drawings, and redundant description is omitted.

Wherein optical detection subassembly 3 and division membrane assembly 4 all set up on detecting seat 2, wherein the bottom of detecting seat 2 is equipped with fluting 22, fluting 22 is the cylinder cell body in this embodiment, division membrane assembly 4 sets up in fluting 22, specifically, division membrane assembly 4 includes piercing member 6 and drive structure 42, piercing member 6 can reciprocate in fluting 22, drive structure 42 is used for driving piercing member 6 and removes, piercing member 6 is located directly over sealing membrane 11, drive structure 42 drives piercing member 6 and moves down, piercing member 6 contacts with sealing membrane 11 and punctures sealing membrane 11, the sample in the follow-up optical detection subassembly 3 to PCR reaction tube 1 of being convenient for detects.

Specifically, the driving structure 42 includes a cylinder 421 and a connecting member 422, wherein the cylinder 421 is a general cylinder, the cylinder 421 is disposed at the bottom of the slot 22, a piston rod of the cylinder 421 is connected with the connecting member 422, the connecting member 422 is a circular plate, and the piercing member 6 is connected at a central position of the connecting member 422.

Wherein the piercing member 6 comprises a spreading member 61 and a piercing head 62, wherein the piercing head 62 is conical, when contacting with the sealing film 11, the sealing film 11 is conveniently pierced, wherein one end of the spreading member 61 is connected with the piercing head 62, the piercing head 62 is arranged at the middle position of the bottom of the spreading member 61, the other end of the spreading member 61 is connected with the connecting member 422, the opening of the sealing film 11 pierced by the piercing head 62 can be further enlarged by the arrangement of the spreading member 61, so that light rays emitted by the subsequent optical detection assembly 3 can smoothly enter and exit the PCR reaction tube 1.

Specifically, the spreader 61 includes a first cylindrical portion 611, a reducing portion 612, and a second cylindrical portion 613, where one end of the reducing portion 612 is connected to the first cylindrical portion 611, and the other end is connected to the second cylindrical portion 613, and the first cylindrical portion 611, the reducing portion 612, and the second cylindrical portion 613 are integrally formed, where the first cylindrical portion 611 is connected to the connector 422, and the second cylindrical portion 613 is connected to the piercing head 62, where the cross section of the reducing portion 612 gradually decreases from the first cylindrical portion 611 to the second cylindrical portion 613, where the cross section of the first cylindrical portion 611 is larger than the cross section of the second cylindrical portion 613, specifically, the cross section of the first cylindrical portion 611 is the same as the largest cross section of the reducing portion 612, and the cross section of the second cylindrical portion 612 is the same as the smallest cross section of the reducing portion 612.

In order to further improve the stability of the connecting piece 422 driving the puncturing piece 6 to move in the slot 22, the side wall of the slot 22 is provided with a sliding groove 51, the connecting piece 422 is provided with a sliding block 52, the sliding block 52 is in sliding connection with the sliding groove 51, namely the sliding block 52 can move in the sliding groove 51, wherein the sliding groove 51 is provided with two groups which are respectively positioned on two opposite side walls of the slot 22, and the sliding block 52 is provided with two groups which are respectively positioned on opposite side sides of the connecting piece 422.

Wherein the detection seat 2 is provided with a first channel 21, the optical detection component 3 is arranged in the first channel 21, and the first channel 21 is in the shape of

The first channel 21 includes an excitation light channel 211 and an emission light channel 212, and the excitation light channel 211 and the emission light channel 212 are disposed perpendicular to each other.

Wherein the optical detection assembly 3 comprises an excitation assembly 31 and an emission assembly 32, wherein the emission assembly 32 is arranged in the emission light channel 212, wherein the excitation assembly 31 is arranged in the excitation light channel 211, wherein a two-phase color splitter 33 is arranged at the intersection of the excitation light channel 211 and the emission light channel 212, wherein the two-phase color splitter 33 divides the emission light channel 212 into an upper channel and a lower channel, wherein the emission assembly 32 is arranged in the upper channel in the emission light channel 212.

Specifically, the excitation assembly 31 includes a TIR collimating lens 311, a collimating cylindrical lens 312 and a first narrow band filter 313, where the TIR collimating lens 311 is disposed at a port of the excitation light channel 211, and the TIR collimating lens 311 and the two-phase color splitter 33 are sequentially disposed along a direction from the TIR collimating lens 311 to the two-phase color splitter 33, the excitation assembly 31 further includes a single-color LED lamp 314 disposed on a side plate 8, the side plate 8 is detachably disposed on a side wall of the detection seat 2, in this embodiment, the side plate 8 is mounted on the detection seat 2 through a bolt assembly, and when the side plate 8 is mounted on the detection seat 2, the single-color LED lamp 314 is just located in the excitation light channel 211 and faces the TIR collimating lens 311.

The emission component 32 includes a first plano-convex lens 321, a second narrowband filter 322, and a silicon photomultiplier 323, where the silicon photomultiplier 323 is disposed on the top cover 9, and the top cover 9 is detachably connected to the detection seat 1, and in this embodiment, the top cover 9 is mounted on the detection seat 2 through a bolt component. When the top cover 9 is mounted on the detecting seat 1, the silicon photomultiplier 323 is just positioned in the emission light channel 212, and the first plano-convex lens 321 and the second narrow-band filter 322 are sequentially arranged between the silicon photomultiplier 323 and the two-phase color splitter 33 along the direction from the silicon photomultiplier 323 to the two-phase color splitter 33.

When the optical detection component 3 detects the sample in the PCR reaction tube, the emission light channel 212 is opposite to the open end of the PCR reaction tube 1, and a second plano-convex lens 324 is arranged in the lower channel of the emission light channel 212.

As a preferable technical scheme, the detection seat 2 is arranged to be in a two-half structure and comprises a first half seat 24 and a second half seat 25, the first half seat 24 and the second half seat 25 are detachably connected, the specific connection mode can be realized by the prior art, for example, the detection seat is connected by bolts or realized by means of buckling, plugging and the like, and the arrangement can facilitate the assembly, disassembly and maintenance of the optical detection assembly 3 and the membrane opening assembly 4, and the use is very convenient.

The distance between the piercing member 6 and the first through hole 21 is adjusted according to the distance between the holes of the PCR reaction tubes 1 placed on the tube frame 7, when the device is used, the piercing member 6 can be aligned with one of the PCR reaction tubes 1, the first channel 21 is just aligned with the adjacent PCR reaction tube 1, so that the film opening and the detection operation can be simultaneously carried out, and the improvement of the working efficiency is facilitated.

The specific working principle of the utility model is as follows:

the detection seat 2 is moved to the position of the PCR reaction tube 1 at the required position, the puncturing piece 6 is positioned right above the PCR reaction tube 1, the air cylinder 421 is started, the air cylinder 421 drives the connecting plate 422 to move downwards, the connecting plate 422 drives the puncturing piece 6 to move downwards and contact the sealing film 11 to puncture the sealing film 11, then the air cylinder 421 is started again, and the air cylinder 421 drives the puncturing piece 6 to move upwards to the original position by driving the connecting plate 422, so that the film opening operation of the PCR reaction tube 1 at the position is finished.

The optical detection assembly 3 can detect the sample in the PCR reaction tube 1 where the sealing film 11 has been pierced, the specific detection principle is a conventional technical means, specifically, the excitation light is emitted by the monochromatic LED lamp 314 with a specific wavelength, the excitation light is scattered at a certain emission angle to form scattered light, after the scattered light is collimated by the TIR collimating lens 311 and the collimating cylindrical lens 312, most of the scattered light is collimated into parallel light, the parallel light passes through the first narrowband filter 313 with a specific wavelength range, the spectrum outside the required wavelength range is filtered, and then is incident into the dichroic mirror 33 placed at an included angle of 45 ° with the parallel light, the parallel light is refracted by 90 °, and the refracted parallel light is focused by the second planoconvex lens 324, and the parallel light is located under the liquid surface of the sample to be detected in the PCR reaction tube 1. Because the sample to be detected in the PCR reaction tube 1 is provided with fluorescent substances, the fluorescent substances emit other specific wavelength spectrums at the positions under excitation, the emitted light is collimated by the second plano-convex lens 324, then the emitted light is transmitted by the dichroic mirror 33, the transmitted light is filtered by the second narrow-band filter 322 in the specific wavelength range to remove spectrums outside the required wavelength range, the filtered spectrums are subjected to light focusing by the first plano-convex lens 321, the focusing point is the silicon photomultiplier 323, and the silicon photomultiplier 323 transmits data to a computer for data analysis.

Example 2

This embodiment is an embodiment 2 of a real-time fluorescent quantitative PCR detection device, which is different from embodiment 1 in that: as shown in fig. 2, in the present embodiment, a hemisphere 521 is disposed on a side of the sliding block 52 facing the sliding slot 51, and the hemisphere 521 can reduce friction with the sliding slot 51 and improve the moving smoothness of the connecting piece 422.

Example 3

This embodiment is an embodiment 3 of a real-time fluorescent quantitative PCR detection device, which is different from embodiment 2 in that: as shown in fig. 5-7, in this embodiment, the spreader 61 is a hollow structure with two open ends, wherein the center of the bottom of the slot 22 is communicated with the second channel 23 extending to the outside of the detection seat 2, wherein the connector 422 is a ring, wherein the inner diameter of the connector 422 is the same as the inner diameter of the first cylindrical member 611, and wherein the piercing head 62 is disposed at the edge of the bottom of the spreader 61.

Wherein be equipped with through-hole 91 on top cap 9, when installing on detecting seat 2 on top cap 9, through-hole 91 is just to second passageway 23, wherein is provided with the lid 92 that can cover on the through-hole 91, and lid 92 passes through the elastic strip to be connected on top cap 9 in this embodiment.

In this embodiment, the membrane opening assembly 4 not only can puncture the sealing membrane 11 on the PCR reaction tube 1, but also can realize the operations of adding and absorbing liquid to the PCR reaction tube under the cooperation of the second channel 23.

It is to be understood that the above examples of the present utility model are provided by way of illustration only and not by way of limitation of the embodiments of the present utility model. Other variations or modifications of the above teachings will be apparent to those of ordinary skill in the art. It is not necessary here nor is it exhaustive of all embodiments. Any modification, equivalent replacement, improvement, etc. which come within the spirit and principles of the utility model are desired to be protected by the following claims.

Claims

1. The real-time fluorescence quantitative PCR detection device comprises a PCR reaction tube (1), wherein a sealing film (11) is arranged at the opening of the PCR reaction tube (1); the method is characterized in that: the device also comprises a detection seat (2), an optical detection assembly (3) for detecting the sample in the PCR reaction tube (1) and a membrane opening assembly (4) for puncturing the sealing membrane (11); the optical detection assembly (3) and the membrane opening assembly (4) are both arranged on the detection seat (2).

2. The real-time fluorescent quantitative PCR detection device as set forth in claim 1, wherein: a slot (22) is formed in the bottom of the detection seat (2); the membrane opening assembly (4) is arranged in the groove (22).

3. The real-time fluorescent quantitative PCR detection device as set forth in claim 2, wherein: the membrane opening assembly (4) comprises a piercing member (6) capable of moving up and down in the slot (22) and a driving structure (42) for driving the piercing member (6) to move; the drive structure (42) is connected to the lancing element (6); the driving structure (42) is arranged in the slot (22).

4. A real-time fluorescent quantitative PCR detection device in accordance with claim 3 wherein: the driving structure (42) comprises a cylinder (421) and a connecting piece (422); the air cylinder (421) is arranged at the bottom of the slot (22), and the connecting piece (422) is connected with a piston rod of the air cylinder (421); the piercing member (6) is provided on the connecting member (422).

5. The real-time fluorescent quantitative PCR detection apparatus as set forth in claim 4, wherein: a chute (51) is arranged on the side wall of the slot (22); the connecting piece (422) is provided with a sliding block (52); the sliding block (52) is in sliding connection with the sliding groove (51); a hemispheroid (521) is arranged on one side of the sliding block (52) facing the sliding groove (51).

6. The real-time fluorescent quantitative PCR detection apparatus as set forth in claim 4, wherein: the piercing member (6) comprises a spreader (61) and a piercing head (62); one end of the spreading piece (61) is connected with the connecting piece (422), and the other end is connected with the piercing head (62).

7. The real-time fluorescent quantitative PCR detection apparatus as set forth in claim 6, wherein: the piercing head (62) is provided with a middle position of the bottom of the opening piece (61).

8. The real-time fluorescent quantitative PCR detection apparatus as set forth in claim 6, wherein: the opening piece (61) is a hollow structure with two open ends.

9. The real-time fluorescent quantitative PCR detection device as set forth in claim 8, wherein: a second channel (23) extending to the outside of the detection seat (2) is communicated with the bottom center of the slot (22); the connection piece (422) is arranged as a ring piece; the piercing head (62) is arranged at the bottom edge of the other end of the opening piece (61).

10. The real-time fluorescent quantitative PCR detection apparatus as set forth in any one of claims 1-9 wherein: the detection seat (2) is provided with a first channel (21); the optical detection assembly (3) comprises an excitation assembly (31) for emitting excitation light and an emission assembly (32) for receiving the emitted light, and the excitation assembly (31) and the emission assembly (32) are both arranged in the first channel (21).