CN112501013A

CN112501013A - PCR instrument with movable heating device

Info

Publication number: CN112501013A
Application number: CN202011578021.3A
Authority: CN
Inventors: 颜菁; 翟峰; 王开林
Original assignee: Jiangsu Huixian Pharmaceutical Technology Co ltd
Current assignee: Jiangsu Huixian Pharmaceutical Technology Co ltd
Priority date: 2020-12-28
Filing date: 2020-12-28
Publication date: 2021-03-16

Abstract

The invention discloses a PCR instrument with a movable heating device. The PCR instrument with a movable heating device comprises: the heating device is movably arranged on the bracket along the y direction; a first slider movably disposed on the bracket along the x-direction; a first connecting rod having a first end and a second end, the first end of the first connecting rod being rotatably connected to the heating device, the lower end of the first connecting rod being rotatably connected to the first slider; and the limiting piece is used for limiting the sliding stroke of the first sliding block so that the heating device moves to the same position of the sample area each time, and the limiting piece is connected to the bracket. The moving device can drive the heating device to reciprocate so as to match the temperature cycle required by the PCR instrument; the first connecting rod can controllably move the heating device to the same position of the sample area through the matching of the mechanism consisting of the first connecting rod and the first slide block and the limiting piece.

Description

PCR instrument with movable heating device

Technical Field

The invention belongs to the field of PCR equipment, and relates to a PCR instrument with a movable heating device.

Background

Microfluidics is a technology for precisely controlling and controlling microscale fluids, and particularly relates to a technology for automatically completing the whole analysis process by integrating basic operation units of sample preparation, reaction, separation, detection and the like in the processes of biological, chemical and medical analysis on a microfluidic chip with the square centimeter. Due to the huge potential in the fields of biology, chemistry, medicine and the like, the PCR amplification detection method has been developed into a new research field crossing the subjects of biology, chemistry, medicine, fluid, electronics, materials, machinery and the like, when PCR (polymerase chain reaction) amplification is used for detecting nucleic acid on a microfluidic chip, the PCR is formed by three steps of denaturation, annealing and extension, and the control of the temperature of the PCR process is the key for success or failure of detection. At present, the PCR appearance that generally on the market is the disconnect-type, needs the manual work to take and put the kit during the heating, and this mode is comparatively loaded down with trivial details inconvenient, needs independent firing equipment to do moreover, and reagent detects with high costs.

Disclosure of Invention

The invention aims to provide a PCR instrument with a movable heating device, which can controllably move the heating device to a preset position to realize the purpose of circularly heating a sample.

In order to achieve the purpose, the invention adopts the following technical scheme:

a heating device-movable PCR instrument, comprising a heating device for heating a sample, wherein a sample area for placing the sample is arranged in the PCR instrument, the PCR instrument further comprises a moving device for moving the heating device into and out of the sample area, the moving device comprises: the heating device is movably arranged on the bracket along the y direction; a first slider movably disposed on the bracket in the x direction; a first link having a first end and a second end, the first end of the first link being rotatably connected to the heating device, and the lower end of the first link being rotatably connected to the first slider; and a stopper for limiting a sliding stroke of the first slider to move the heating device to the same position of the sample area each time, the stopper being connected to the rack.

In an embodiment, the moving device further includes a second connecting rod having a first end and a second end, the first end of the second connecting rod is rotatably connected to the first sliding block, the second end of the second connecting rod is rotatably connected to the limiting member, and the limiting member is movably connected to the bracket along a predetermined stroke.

In one embodiment, the bracket has a first position-limiting portion and a second position-limiting portion, and the position-limiting member has a pin movably disposed between the first position-limiting portion and the second position-limiting portion.

In an embodiment, the bracket is provided with a long hole extending along the y direction, the first limiting portion and the second limiting portion are respectively formed on hole walls at two end portions of the long hole, and the pin is slidably inserted into the long hole along the y direction.

In an embodiment, the moving device further includes a second slider movably disposed on the support in the y direction, and the heating device is disposed on the second slider.

In one embodiment, the heating device includes a fixing plate and one or more heat-conducting fins disposed on the fixing plate.

In one embodiment, the heating device further comprises heat preservation cotton wrapped around the heat conducting sheet.

In one embodiment, the heat conducting sheet has a front surface for contacting with a sample and a back surface opposite to the front surface, and the heating device further includes a refrigerating sheet disposed at the back surface of the heat conducting sheet, wherein the refrigerating sheet is attached to the back surface of the heat conducting sheet or has a certain gap with the back surface.

In one embodiment, the refrigeration plate is a TEC refrigeration plate.

In an embodiment, the heating device further includes a heat sink, and the fixing plate is disposed on the heat sink.

In an embodiment, the heating device further comprises a fan.

Compared with the prior art, the invention has the following advantages by adopting the scheme:

according to the PCR instrument with the movable heating device, the moving device can drive the heating device to reciprocate so as to match the temperature cycle required by the PCR instrument; the first connecting rod can controllably move the heating device to the same position of the sample area through the matching of the mechanism consisting of the first connecting rod and the first slide block and the limiting piece.

Drawings

In order to more clearly illustrate the technical solution of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.

FIG. 1 is a block diagram showing a PCR apparatus with a movable heating unit according to an embodiment of the present invention;

FIG. 2 is a front view of the heating device and the moving device;

FIG. 3 is a left side view of the heating device and the moving device shown in FIG. 2;

FIG. 4 is a cross-sectional view taken along line B-B of FIG. 3;

fig. 5 is a top view of the heating device and the moving device shown in fig. 2.

Wherein the content of the first and second substances,

2. a mobile device; 20. a support; 201. a first guide rail; 202. a second guide rail; 203. a long hole; 21. a first slider; 22. a second slider; 23. a first link; 24. a second link; 25. a limiting member; 251. a pin;

3. a heating device; 30. a mounting seat; 31. a heat conductive sheet; 32. heat preservation cotton; 33. a fixing plate; 34. a heat sink; 35. a fan; 36. a refrigeration plate;

4. a sample area;

5. a fluorescence detection device;

6. and a chip fixing device.

Detailed Description

Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings so that the advantages and features of the invention may be more readily understood by those skilled in the art. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto.

As used in this specification and the appended claims, the terms "comprises" and "comprising" are intended to only encompass the explicitly identified steps and elements, which do not constitute an exclusive list, and that a method or apparatus may include other steps or elements. As used herein, the term "and/or" includes any combination of one or more of the associated listed items.

It should be noted that, unless otherwise specified, when a feature is referred to as being "fixed" or "connected" to another feature, it may be directly fixed or connected to the other feature or indirectly fixed or connected to the other feature. Further, the description of the upper, lower, left, right, etc. used in the present invention is only with respect to the positional relationship of the respective components of the present invention with respect to each other in the drawings.

It is further understood that the use of "a plurality" in this disclosure means two or more, as other terms are analogous.

The terms of orientation such as x-direction and y-direction are defined herein based on fig. 1 and the orientation of the PCR instrument in actual use, and the x-direction and the y-direction are perpendicular to each other and respectively indicate the left-right direction and the up-down direction.

PCR consists of three steps, denaturation-annealing-extension, the specificity of which depends on oligonucleotide primers complementary to both ends of the target sequence, similar to the natural replication process of DNA. Different reaction stages of PCR require different holding temperatures, DNA is denatured into single strands at 95 ℃, primers and single strands are combined according to the base complementary pairing principle at about 60 ℃, the optimal reaction temperature of DNA polymerase is reached when the temperature is adjusted to about 72 ℃, and the DNA polymerase catalyzes and synthesizes complementary strands from a hydroxyl end to a phosphate group direction along a template according to the base complementary pairing principle.

The present embodiment provides a PCR instrument with a movable heating device, which can heat and amplify DNA by combining the above features of heating and melting DNA and cooling. Referring to fig. 1, the PCR instrument includes a sample area 4, a heating device 3, a moving device 2, a chip fixing device 6, and a fluorescence detecting device 5. The sample area 4 is used for placing a biological sample to be tested, such as a biological sample in the microfluidic chip, and the sample area is particularly used for carrying the microfluidic chip. The chip fixing device 6 is used for positioning and fixing the microfluidic chip. The heating device 3 is used for heating or cooling the microfluidic chip in the sample area 4. The moving device 2 is used for driving and controlling the movement of the heating device 3. The fluorescence detection device 6 is used for detecting fluorescence signals in the microfluidic chip. Wherein the heating device 3 and the moving device 2 will be described in detail with reference to fig. 2 to 5; the fluorescence detecting means 5 and the chip holding means 6 are known means, and will not be described in detail.

The moving device 2 includes a bracket 20, a first slider 21, a second slider 22, a first link 23, a second link 24, and a stopper 25. The bracket 201 is fixed on the casing of the PCR instrument. The bracket 20 is a U-shaped structure formed by two side plates and a bottom plate. The first slider 21 is movably disposed on the bracket 20 in the x direction; specifically, a first guide rail 201 extending in the x direction is fixedly provided on the bottom plate of the bracket 20, and the first slider 21 is slidably fitted to the first guide rail 201. The second slide block 22 is arranged on the bracket 20 in a sliding way along the y direction; specifically, a second guide rail 202 extending in the y direction is fixedly arranged on the left side plate of the bracket 20, and the second slider 22 is arranged on the second guide rail 202 in a sliding fit manner; the heating device 3 is arranged on the second slide 22. The first link 23 has a first end and a second end, the first end of the first link 23 is rotatably connected to the heating device 3 by a pivot, and the lower end of the first link 23 is rotatably connected to the first slider 21 by a pivot. The second link 24 has a first end and a second end, the first end of the second link 24 is rotatably connected to the first slider 21 via a pivot, and the second end of the second link 24 is rotatably connected to the limiting member 25 via a pivot. The limiting member 25 is movably connected to the rack 20 along a predetermined stroke, and the limiting member 25 limits the rotation of the second connecting rod 24, so as to limit the sliding stroke of the first sliding block 21, so that the first connecting rod 23 can move the heating device 3 to the same position of the sample area each time, thereby satisfying the thermal cycling program of the PCR reaction. The axes of the pivot shafts for pivotal connection extend horizontally in the front-rear direction and are parallel to each other.

Specifically, the right side plate of the bracket 20 has a first position-limiting portion and a second position-limiting portion disposed up and down, and the limiting member 25 has a pin 251 movably disposed between the first position-limiting portion and the second position-limiting portion. Further, a long hole 203 extending along the y direction is formed in the right side plate of the bracket 20, an upper hole wall of the long hole 203 is a first limiting portion, a lower hole wall of the long hole 203 is a second limiting portion, and the pin 251 is slidably inserted into the long hole 203 along the y direction.

The movement process of each part when the moving device 2 moves the heating device 3 is as follows: when the first slider 21 slides leftwards along the first guide rail 201, the first slider 21 drives the first connecting rod 23 to move leftwards and simultaneously rotate clockwise, the trend of circular motion is converted into linear motion, and the first connecting rod 23 drives the heating device 3 to move upwards along the second guide rail 202; meanwhile, the second connecting rod 24 connected to the first slider 21 moves clockwise while moving, the second end of the second connecting rod 24 drives the limiting member 25 to move relative to the bracket 20, the pin 251 on the limiting member 25 slides upwards in the long hole 203 on the bracket 20, and when the pin slides to the upper end of the long hole 203, the pin receives the upper side hole wall to stop moving, so that the first slider 21 can be always stopped at a preset position by moving the heating device 3 on each time, and the heating device 3 can reach the position of the microfluidic chip every time without colliding with the microfluidic chip. Conversely, when the first slider 21 moves to the right, the heating device 3 will move downward with a similar movement principle.

The heating device 3 includes a mounting seat 30, a heat conducting sheet 31, heat insulating cotton 32, a fixing plate 33, a radiator 34, a fan 35 and a refrigerating sheet 36. The mounting seat 30 is fixedly disposed on the second slider 22 and rotatably connected to the first end of the first link 23 by a pivot. The heat sink 34 is provided on the mount 30, and the fan 35 is provided below the heat sink 34. The fixing plate 33 is disposed on the heat sink 34, and the heat-conducting sheet 31 is disposed on the fixing plate 33 with its upper surface exposed to contact the microfluidic chip. The number of the heat-conducting fins 31 is one, and a refrigerating fin 36 is arranged below each heat-conducting fin 31. The heat-insulating cotton 32 is wrapped around the heat-conducting fin 31.

The heat dissipation fan 35 is fixed below the heat sink 34, and cools the heat sink 34. The cooling plate 36 is a TEC cooling plate 36(Thermo Electric Cooler). The cooling plate 36 has a fast cooling speed and a short cooling period, so that the rapid amplification of molecules can be realized. The heat insulation cotton 32 reduces the heat in the heating area from diffusing and losing to the periphery.

In the present embodiment, the heat conductive sheet 31 has a front surface for contacting the sample and a back surface opposite to the front surface, and the cooling sheet 36 is attached to the back surface of the heat conductive sheet 31. In other embodiments, there may be a gap between the back surface of the heat-conducting sheet 31 and the cooling sheet 36.

The PCR instrument with the movable heating device 3 has the following advantages: in the integrated machine, the connecting rod sliding block can flexibly control the heating device 3 at the same position and quickly and tightly attach the heating position to the microfluidic chip. The heating device 3 can be attached at any height in a certain range, and can fix the microfluidic chip in the sample area. The heat sink 34 is provided with a plurality of heat conductive sheets 31 to form a plurality of spot heating, thereby providing flexibility. By adopting a TEC heating mode, the temperature rise and fall speed is high, the period is short, and the rapid molecular amplification can be realized.

The above embodiments are merely illustrative of the technical ideas and features of the present invention, and are preferred embodiments, which are intended to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the scope of the present invention. All equivalent changes or modifications made according to the principles of the present invention should be covered within the protection scope of the present invention.

Claims

1. A PCR instrument with a movable heating device, comprising a heating device for heating a sample, wherein a sample area for placing the sample is provided in the PCR instrument, the PCR instrument further comprises a moving device for moving the heating device into and out of the sample area, the moving device comprises:

the heating device is movably arranged on the bracket along the y direction;

a first slider movably disposed on the bracket in the x direction;

a first link having a first end and a second end, the first end of the first link being rotatably connected to the heating device, and the lower end of the first link being rotatably connected to the first slider; and

a stopper for limiting a sliding stroke of the first slider to move the heating device to the same position of the sample area each time, the stopper being connected to the rack.

2. The PCR instrument as claimed in claim 1, wherein the moving device further comprises a second link having a first end and a second end, the first end of the second link is rotatably connected to the first slide, the second end of the second link is rotatably connected to the stopper, and the stopper is movably connected to the frame along a predetermined stroke.

3. The PCR instrument of claim 2, wherein the frame has a first position-limiting portion and a second position-limiting portion, and the position-limiting member has a pin movably disposed between the first position-limiting portion and the second position-limiting portion.

4. The PCR instrument of claim 3, wherein the holder has a slot extending in the y-direction, the first and second stoppers are formed on the walls of the slot at both ends of the slot, and the pin is slidably inserted into the slot in the y-direction.

5. The PCR instrument of claim 1, wherein the moving means further comprises a second slide movably disposed on the support in the y-direction, the heating means being disposed on the second slide.

6. The PCR instrument of claim 1, wherein the heating device comprises a fixing plate and one or more heat-conducting fins disposed on the fixing plate.

7. The PCR instrument of claim 6, wherein the heating device further comprises heat-insulating cotton wrapped around the heat-conducting strip.

8. The PCR instrument of claim 6, wherein the heat-conducting plate has a front surface for contacting the sample and a back surface opposite to the front surface, and the heating device further comprises a cooling plate disposed at the back surface of the heat-conducting plate, the cooling plate being attached to the back surface of the heat-conducting plate or having a gap with the back surface.

9. The PCR instrument of claim 8, wherein the refrigeration plate is a TEC refrigeration plate.

10. The PCR instrument of claim 6, wherein the heating device further comprises a heat sink, the fixing plate being disposed on the heat sink; and/or the heating device further comprises a fan.