CN116179333A - Temperature-controlled amplification device and PCR instrument - Google Patents
Temperature-controlled amplification device and PCR instrument Download PDFInfo
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- CN116179333A CN116179333A CN202310423353.1A CN202310423353A CN116179333A CN 116179333 A CN116179333 A CN 116179333A CN 202310423353 A CN202310423353 A CN 202310423353A CN 116179333 A CN116179333 A CN 116179333A
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- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 46
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- 238000001816 cooling Methods 0.000 claims description 20
- 230000004308 accommodation Effects 0.000 claims description 12
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- 238000003752 polymerase chain reaction Methods 0.000 description 12
- 238000001514 detection method Methods 0.000 description 11
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- 239000000463 material Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 230000004544 DNA amplification Effects 0.000 description 1
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
- B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
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Abstract
Embodiments of the present disclosure provide a temperature controlled amplification device and a PCR instrument. The temperature-controlled amplification device includes: heating and refrigerating piece; the temperature control part can be heated or refrigerated by the heating and refrigerating part, and a plurality of accommodating cavities are formed on one side of the temperature control part, which is away from the heating and refrigerating part, and are respectively used for accommodating the lower areas of corresponding sample tubes in the plurality of sample tubes; the heat insulation pad is arranged above the temperature control piece and comprises a plurality of first through holes corresponding to the plurality of accommodating cavities; a constant temperature member disposed above the heat insulating mat and including a first top plate, and a plurality of protrusions extending from the first top plate toward the heat insulating mat, a plurality of second through holes being provided in the plurality of protrusions and the first top plate, the plurality of first through holes and the plurality of second through holes being disposed in correspondence, walls of the plurality of second through holes being capable of surrounding and contacting upper regions of the respective sample tubes, respectively, for keeping the upper regions of the respective sample tubes constant temperature; and a thermal cover disposed over the thermostatic element.
Description
Technical Field
Embodiments of the present disclosure relate generally to the field of PCR instrument technology, and more particularly, to a temperature controlled amplification device and a PCR instrument.
Background
Molecular diagnosis techniques are techniques for diagnosing human states and diseases by detecting the presence, defects, or abnormal expression of genes using deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) as diagnostic materials and using molecular biology techniques. The basic principle of the diagnosis technology is to detect whether the structure of DNA or RNA is changed, the quantity of DNA or RNA is more or less and the expression function is abnormal, so as to determine whether the abnormal change of the gene level of a tested person exists, and the diagnosis technology has important significance for preventing, predicting, diagnosing, treating and prognosing diseases.
Polymerase Chain Reaction (PCR) is a molecular biological technique for amplifying specific DNA fragments, the basic principle of which is similar to the natural replication process of DNA, consisting of three basic reaction steps, denaturation, annealing and extension, i.e., denaturation of template DNA, annealing (renaturation) of template DNA with primers, extension of primers. By repeating the three processes of denaturation, annealing and extension, more "semi-reserved replication chains" are obtained, and the new chains can be used as templates for the next cycle. In this way, the target gene to be amplified can be amplified by a desired multiple. Finally, the target gene in the copied sample can be detected by a real-time fluorescent quantitative PCR technology. Real-time fluorescent quantitative PCR instruments are used to perform such detection.
In PCR instruments, it is often necessary to perform multiple cycles of heating and cooling the sample to complete the amplification and replication of the sample. The faster the temperature rise and the temperature drop, the time required by sample amplification and replication can be shortened, and the detection efficiency of the instrument can be improved. However, the temperature control module of the instrument is rapidly switched between high temperature and low temperature, so that the sample is easily condensed in the sample tube, and the condensed liquid is contaminated on the wall of the sample tube or other pollutants at other positions, so that the sample is polluted, and the accuracy of the detection result is further affected. In addition, the condensation phenomenon can reduce the speed of sample amplification and replication, and the detection efficiency of the instrument is affected. Therefore, how to further improve the detection efficiency of the apparatus and ensure the accuracy of the detection result is a problem to be solved.
Disclosure of Invention
It is an object of the present disclosure to provide a temperature controlled amplification device and a PCR instrument to at least partially solve the above-mentioned problems.
In a first aspect of the present disclosure, there is provided a temperature-controlled amplification device comprising: heating and refrigerating piece; the temperature control part is arranged above the heating and refrigerating part and can be heated or refrigerated by the heating and refrigerating part, one side of the temperature control part, which is away from the heating and refrigerating part, is provided with a plurality of accommodating cavities which are respectively used for accommodating lower areas of corresponding sample tubes in the plurality of sample tubes; the heat insulation pad is arranged above the temperature control piece and comprises a plurality of first through holes corresponding to the plurality of accommodating cavities; a thermostat provided above the heat insulating mat and including a first top plate, and a plurality of protrusions extending from the first top plate toward the heat insulating mat, the plurality of protrusions and the first top plate having a plurality of second through holes provided therein, the plurality of first through holes and the plurality of second through holes being provided in correspondence, walls of the plurality of second through holes being capable of encircling and contacting upper regions of the respective sample tubes, respectively, for keeping the upper regions of the respective sample tubes at a constant temperature; and a thermal cover disposed over the thermostatic element.
In the embodiment according to the disclosure, when the heating and refrigerating member is rapidly changed from low temperature to high temperature, the temperature control member correspondingly and rapidly heats up the lower region of the sample tube, and the vaporized sample is not condensed after contacting the inner wall of the lower region of the sample tube; meanwhile, the constant temperature piece keeps the upper area of the sample tube between the heat cover and the heat insulation pad constant temperature, and the vaporized sample is not condensed after contacting the inner wall of the upper area of the sample tube; finally, the thermal cover maintains the top opening of the sample tube in contact therewith at a high temperature, and the vaporized sample is not condensed after contacting the sample tube in contact with the thermal cover and the thermal cover.
Therefore, when the heating and refrigerating piece is rapidly changed from low temperature to high temperature, the vaporized sample is contacted with the inner wall of the sample tube, and no condensation phenomenon occurs. The temperature control amplification device can prevent pollution to the sample, ensure the accuracy of a detection result, and further improve the detection efficiency of the instrument.
In some embodiments, the thermostat further includes a support portion disposed along an edge of the first top plate and extending toward the heat insulation pad, the first top plate, the support portion, and the heat insulation pad enclosing a first enclosed space.
In some embodiments, the temperature control further comprises a bottom plate adjacent the heating and cooling element, a second top plate opposite the bottom plate, and a side plate extending from an edge of the second top plate to the bottom plate, the plurality of receiving cavities extending from the second top plate in a direction toward the bottom plate.
In some embodiments, the second top panel, the bottom panel, and the side panels enclose a receiving space, and the plurality of receiving cavities extend from the second top panel into the receiving space.
In some embodiments, the accommodating space is filled with a heat-conducting medium.
In some embodiments, ends of the plurality of receiving cavities proximate the floor are disposed in contact with the floor.
In some embodiments, the temperature controlled amplification device further comprises a sample container comprising the plurality of sample tubes and a body, the plurality of sample tubes extending from a top surface of the body toward the temperature control member, the top surface of the body in contact with the thermal cap.
In some embodiments, a peripheral portion of the body around the top surface is in contact with the heat insulating pad, the body, the heat insulating pad, and the thermostat enclosing a second enclosed space.
In some embodiments, an outer surface of each of the plurality of sample tubes is surrounded by the first through hole, the second through hole, and the receiving cavity.
In some embodiments, each of the plurality of receiving cavities is disposed about the same axis as the respective first and second through holes, and the axis is perpendicular to the direction of extension of the insulation blanket.
In some embodiments, the temperature value of the thermal cover is greater than the temperature value of the thermostat.
In a second aspect of the present disclosure, there is provided a PCR instrument comprising any one of the temperature controlled amplification devices according to the first aspect of the present disclosure.
It should be understood that what is described in this section is not intended to limit the key features or essential features of the embodiments of the present disclosure, nor is it intended to limit the scope of the present disclosure. Other features of the present disclosure will become apparent from the following description.
Drawings
The above and other features, advantages and aspects of embodiments of the present disclosure will become more apparent by reference to the following detailed description when taken in conjunction with the accompanying drawings. In the drawings, wherein like or similar reference numerals denote like or similar elements, in which:
FIG. 1 shows a schematic perspective cross-sectional structure of a temperature controlled amplification device according to one embodiment of the present disclosure;
FIG. 2 shows a schematic perspective cross-sectional structure of a temperature controlled amplification device according to another embodiment of the present disclosure, in which a sample container is not shown;
FIG. 3 illustrates a schematic perspective cross-sectional structure of a temperature controlled amplification device according to another embodiment of the present disclosure, in which a sample container is shown;
FIG. 4 illustrates a schematic structural view of a thermostatic element according to one embodiment of the present disclosure when viewed in one direction;
fig. 5 shows a schematic view of the thermostatic element of fig. 4 viewed in another direction.
Reference numerals illustrate:
100 is a temperature controlled amplification device; 1 is a heating and refrigerating piece; 2 is a temperature control piece, 21 is a second top plate, 22 is a bottom plate, 23 is a side plate, and 24 is a containing cavity; 3 is a heat insulation pad, 31 is a first through hole; 4 is a constant temperature piece, 41 is a first top plate, 42 is a protruding part, 43 is a second through hole, and 44 is a supporting part; 5 is a sample container, 51 is a sample tube, and 52 is a main body; 6 is a thermal cover; 10 is a first enclosed space; 20 is an accommodating space; 30 is a second enclosed space.
Detailed Description
Preferred embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While the preferred embodiments of the present disclosure are illustrated in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
The term "comprising" and variations thereof as used herein means open ended, i.e., "including but not limited to. The term "or" means "and/or" unless specifically stated otherwise. The term "based on" means "based at least in part on". The terms "one example embodiment" and "one embodiment" mean "at least one example embodiment. The term "another embodiment" means "at least one additional embodiment". The terms "first," "second," and the like, may refer to different or the same object.
As described above, in the process of amplifying and copying a sample by a PCR apparatus, a temperature control module of the apparatus is rapidly switched between high and low temperatures, so that a condensation phenomenon of the sample in a sample tube is very easy to occur, which may affect the accuracy of a detection result and the detection efficiency of the apparatus. Embodiments of the present disclosure provide a temperature-controlled amplification device and a PCR instrument to avoid condensation when a vaporized sample contacts an inner wall of a sample tube. Hereinafter, the principles of the present disclosure will be described with reference to fig. 1 to 5.
Fig. 1 shows a schematic perspective cross-sectional structure of a temperature-controlled amplification apparatus 100 according to one embodiment of the present disclosure. As shown in fig. 1, the temperature controlled amplification device 100 described herein generally includes a heating and cooling element 1, a temperature control element 2, a sample container 5, and a thermal cover 6.
The heating and refrigerating element 1 can heat and refrigerate the temperature control element 2, for example, it can raise the temperature of the temperature control element 2, and it can also lower the temperature of the temperature control element 2. The heating and cooling element 1 according to the embodiment of the present disclosure may be various types of temperature raising and lowering elements currently known or available in the future, and the embodiment of the present disclosure is not limited thereto. For example, in some embodiments, the heating and cooling element 1 may be a peltier element.
The temperature control 2 is arranged above the heating and refrigerating element 1 to raise and lower the temperature under the control of the heating and refrigerating element 1, thereby realizing the temperature control of the sample in the sample container 5. As shown in fig. 1, the temperature control element 2 is formed with a plurality of receiving chambers 24 on the side facing away from the heating and cooling element 1, the receiving chambers 24 being used for receiving the sample containers 5.
As shown in fig. 1, in some embodiments, the sample container 5 includes a body 52 and a plurality of sample tubes 51. A plurality of sample tubes 51 extend from the top surface of the body 52 toward the temperature control 2. The sample tube 51 is used to contain the sample to be amplified and replicated. The lower regions of the plurality of sample tubes 51 may be embedded within the respective receiving cavities 24 of the temperature control 2 for heating and cooling under the control of the temperature control 2. The top surface of the body 52 may be in contact with the thermal cover 6 so that the contact position of the sample tube 51 with the thermal cover 6 is maintained at a constant temperature. The contact position of the sample tube 51 and the heat cover 6 is always maintained in a constant temperature state, so that the vaporized sample is not condensed in the case of contacting the top opening of the sample tube 51 and the heat cover 6.
It should be appreciated that the sample container 5 according to embodiments of the present disclosure may be various types of sample containers. For example, in some embodiments, the sample container 5 may contain 96 sample tubes 51, e.g., a 96-well full skirt. In other embodiments, the sample container 5 may be a 96-well semi-skirt or 96-well non-skirt. It should be noted that the numbers, values, numbers, etc. mentioned above and as may be referred to elsewhere in the disclosure are exemplary and are not intended to limit the scope of the disclosure in any way. Any other suitable numbers, values, numbers are possible. For example, the sample container 5 may include more or fewer sample tubes 51 depending on the particular application scenario and requirements.
As described above, the side of the temperature control member 2 facing away from the heating and cooling member 1 is formed with a plurality of accommodation chambers 24. The accommodation chamber 24 is for accommodating the sample tube 51 of the sample container 5, and wraps the lower region of the sample tube 51. Since the temperature control 2 is provided above the heating and cooling element 1, the temperature control 2 can correspondingly realize rapid transition of low temperature and high temperature according to the adjustment of the heating and cooling element 1. With this configuration, when the heating and cooling member 1 is rapidly changed from low temperature to high temperature, the temperature controller 2 can rapidly raise the temperature of the lower region of the sample tube 51 located in the accommodation chamber 24 accordingly, and the vaporized sample does not come into contact with the inner wall of the lower region of the sample tube 51.
It will be appreciated that in the embodiment according to the present disclosure shown in fig. 1, when the heating and cooling member 1 is rapidly changed from low temperature to high temperature, the temperature control member 2 rapidly heats up the lower region of the sample tube 51 located in the accommodation chamber 24 accordingly, and the vaporized sample is not condensed after contacting the inner wall of the lower region of the sample tube 51 located in the accommodation chamber 24; at the same time, the thermal cover 6 maintains the position of the top opening of the sample tube 51 in contact therewith in a high temperature state, and the vaporized sample is not condensed as well after contacting the top opening of the sample tube 51 in contact with the thermal cover 6 and the thermal cover 6.
Referring again to fig. 1, the inventors noted that in the case where the sample tube 51 has just undergone the cooling process, the temperature of the portion of the sample tube 51 located between the bottom surface of the heat cover 6 and the top surface of the temperature control 2 (i.e., the upper region of the sample tube 51) is low, and thus the inner wall temperature of the upper region of the sample tube 51 is low. Then, when the sample tube 51 is heated again by the temperature controller 2, the temperature of the upper region of the sample tube 51 is raised slowly due to the fact that the upper region is not wrapped by the accommodating cavity 24 of the temperature controller 2, so that the vaporized sample contacts the inner wall of the sample tube 51 in the region and then is condensed, and the condensed liquid is stained on the inner wall of the upper region of the sample tube 51, so that sample pollution is caused, and the accuracy of the detection result is affected.
To solve the above-described problems, according to an embodiment of the present disclosure, there is provided a temperature-controlled amplification apparatus 100, which includes a thermostat 4 for maintaining an upper region of a sample tube 51 at a relatively high constant temperature, thereby preventing an inner wall of the upper region of the sample tube 51 from generating a condensation phenomenon. Such a temperature-controlled amplification apparatus 100 will be described below with reference to fig. 2 to 5. Referring first to fig. 2 and 3, fig. 2 shows a schematic perspective cross-sectional structure of a temperature-controlled amplification apparatus 100 according to another embodiment of the present disclosure, in which a sample container 5 is not shown; fig. 3 illustrates a schematic perspective cross-sectional structure of a temperature-controlled amplification apparatus 100 according to another embodiment of the present disclosure, in which a sample container 5 is shown.
As shown in fig. 2 and 3, the temperature-controlled amplification device 100 includes a heating and cooling member 1, a temperature control member 2, a heat insulating pad 3, a constant temperature member 4, and a heat cover 5. The structures of the heating and cooling element 1, the temperature control element 2, and the heat cover 5 are similar to those of the heating and cooling element 1, the temperature control element 2, and the heat cover 5 in the temperature-controlled amplification apparatus 100 shown in fig. 1. Hereinafter, differences between the temperature-controlled amplification apparatus 100 shown in fig. 2 and 3 and the temperature-controlled amplification apparatus 100 shown in fig. 1 will be described in detail, and will not be described in detail for the same portions.
As shown in fig. 2 and 3, the heat insulation pad 3 is disposed above the temperature control 2, and a plurality of first through holes 31 corresponding to the plurality of accommodating chambers 24 are disposed on the heat insulation pad 3 contacting with the top surface of the temperature control 2, so as to facilitate the corresponding sample tubes 51 to pass through the heat insulation pad 3.
As shown in fig. 2 and 3, the thermostat 4 is provided above the heat insulating mat 3 in contact with the top surface of the heat insulating mat 3. The thermostatic element 4 is provided with a plurality of second through holes 43 corresponding to the plurality of first through holes 31, respectively for facilitating the passage of the corresponding sample tube 51 through the thermostatic element 4.
In particular, fig. 4 and 5 show schematic structural views of the thermostat 4 as viewed in different directions according to one embodiment of the present disclosure. As shown in fig. 4 and 5, the thermostat 4 includes a first top plate 41 and a plurality of protruding portions 42. The plurality of protruding portions 42 extend from the first top plate 41 toward the heat insulating mat 3, and a plurality of second through holes 43 are provided in the plurality of protruding portions 42 and the first top plate 41. In other words, the plurality of through holes are provided on the plurality of protruding portions 42, the plurality of through holes are provided on the first top plate 41 correspondingly, and the plurality of through holes on the plurality of protruding portions 42 and the plurality of through holes on the first top plate 41 communicate one by one, thereby forming the plurality of second through holes 43. In this way, the walls of the plurality of second through holes 43 can respectively surround and contact the upper regions of the respective sample tubes 51 for keeping the upper regions of the respective sample tubes 51 constant. With this arrangement, the vaporized sample does not undergo condensation in the case of contacting the inner wall of the sample tube 51 in this region.
The thermostat 4 according to the embodiment of the present disclosure may be maintained at a constant temperature in various types of ways. For example, in some embodiments, the thermostat 4 is electrically heated. In other embodiments, a liquid flow path may be formed inside the thermostat 4, and the liquid flow path of the liquid flow path may be circulated to supply the high-temperature liquid to maintain the thermostat 4 in a constant temperature state.
As further shown in fig. 4 and 5, in some embodiments, the thermostat 4 further includes a support 44, the support 44 being disposed along an edge of the first top plate 41 and extending toward the heat insulating mat 3. Referring to fig. 2 to 5, the first top plate 41, the supporting portion 44 and the heat insulation pad 3 define a first enclosed space 10, and the protruding portion 42 is located in the first enclosed space 10. In the case where the thermostat 4 is operated for a while, the temperature of the air in the first enclosed space 10 reaches or approaches the temperature of the thermostat 4, so that the heat transfer amount of the protruding portion 42 to the first enclosed space 10 can be reduced. In this way, the temperature of the protruding portion 42 can be maintained at a high state, thereby keeping the upper region of the sample tube 51 wrapped by the protruding portion 42 at a constant temperature, and finally solving the problem of condensation occurring when the vaporized sample contacts the inner wall of the sample tube 51 in this region.
In some embodiments, the temperature value of the thermal cover 6 may be set to be greater than the temperature value of the thermostatic element 4, and the temperature value of the thermostatic element 4 may be set to be higher than the highest temperature of the sample tube 51. For example, in one embodiment, the temperature of the thermostatic element 4 may be maintained between 95-105 ℃, and the maximum temperature of the sample tube 51 may be 95 ℃. As an example, in case the thermal cover 6 is set to 105 ℃, the thermostat 4 may be set to 100 ℃, of course, these temperature values may be additionally set according to specific application scenarios and requirements, e.g. the thermal cover 6 may be set to 104 ℃ and the thermostat 4 may be set to 102 ℃. In other embodiments, the temperature value of the thermal cover 6 may also be set equal to the temperature value of the thermostatic element 4.
In some embodiments, as shown in fig. 2 and 3, a peripheral portion of the body 52 around its top surface is in contact with the insulation pad 3. The main body 52, the heat insulating mat 3, and the thermostat 4 can enclose the second closed space 30. The second sealed space 30 is located outside the first sealed space 10. In the case where the thermostat 4 is operated for a certain period of time, the temperature of the air in the second closed space 30 can also reach or approach the temperature of the thermostat 4, thereby reducing the amount of heat transfer from the thermostat 4 to the external environment, and thus maintaining the temperature of the inside of the thermostat 4 at a higher level. In this way, the upper region of the sample tube 51 wrapped by the protrusion 42 can be kept constant, and the problem of condensation occurring when the vaporized sample contacts the inner wall of the sample tube 51 in this region is also solved.
With the above arrangement, the heat insulating mat 3 encloses the first closed space 10 with the first top plate 41 and the supporting portion 44 and the second closed space 30 with the main body 52 and the thermostat 4 on the one hand, so that a closed space is formed in the upper region of the heat insulating mat 3, which can prevent the vaporized sample from condensing when contacting the inner wall of the sample tube 51 in this region, and the detailed description has been discussed above, and will not be repeated here. On the other hand, the heat insulating pad 3 can also function to block heat transfer from the region of the temperature control 2 to the region of the thermostatic element 4, thereby enabling the temperature control 2 to transfer heat efficiently to the lower region of the sample tube 51 located within the accommodation chamber 24. Since the sample is usually located in the lower region of the sample tube 51, this can avoid waste of the internal heat of the temperature control 2, and eventually increase the amplification replication speed of the sample inside the sample tube 51.
The insulation mat 3 according to the embodiments of the present disclosure may be various types of insulation mats 3, and the embodiments of the present disclosure are not limited thereto. For example, in some embodiments, the heat insulation pad 3 is made of silica gel. In other embodiments, the heat insulation pad 3 may be a nano-base heat insulation pad or the material of the heat insulation pad 3 is foamed plastic or polyurethane, etc.
In some embodiments, as shown in fig. 1 and 2, temperature control member 2 further includes a bottom plate 22, a second top plate 21, and a side plate 23 extending from an edge of second top plate 21 to bottom plate 22. The bottom plate 22 is disposed adjacent to the heating-cooling member 1 and in contact with the heating-cooling member 1. The second top plate 21 is disposed opposite to the bottom plate 22. A plurality of receiving cavities 24 extend from the second top plate 21 in a direction toward the bottom plate 22. That is, the second top plate 21, the bottom plate 22, the side plates 23 and the accommodating cavity 24 are in an integrated structure, which also ensures that the whole structure of the temperature control 2 is firm and reliable.
In some embodiments, the second top plate 21, the side plate 23, and the bottom plate 22 collectively enclose the accommodation space 20. That is, the temperature control member 2 is hollow. A plurality of receiving cavities 24 extend from the second top plate 21 into the receiving space 20. It will be appreciated that, since the temperature control 2 is hollow, conditions are created for the present disclosure to fill the heat transfer medium in the accommodating space 20. The heat transfer medium may be a fluid that is vaporizable by heat, such as water. In the case where the heating and cooling element 1 is heated, the bottom plate 22 of the temperature control element 2 is correspondingly heated, so that the heat transfer medium located in the receiving space 20 absorbs a large amount of heat energy and evaporates into a gaseous substance. The gaseous substance can rapidly fill the entire accommodation space 20, and in the case where the gaseous substance contacts the outer wall of the accommodation chamber 24, the gaseous substance condenses to liquefy and release heat and the heat is transferred to the inner and outer walls of the sample tube 51 wrapped by the accommodation chamber 24. In this way, the inner and outer walls of the sample tube 51 can be rapidly warmed up, thereby avoiding condensation of the vaporized sample after contacting the inner wall of the sample tube 51 having a low temperature. Meanwhile, the heat generated by the heating and refrigerating element 1 is rapidly transmitted to the whole accommodating space 20 through the gaseous substances in a state of not being accumulated locally, and the sample tube 51 can be heated uniformly by the heat released by the gaseous substances in the condensation process.
With the above configuration, the sample tube 51 wrapped by the accommodation chamber 24 can be heated up quickly, and the phenomenon that the vaporized sample is condensed after contacting the inner wall of the sample tube 51 with low temperature can be avoided. Meanwhile, since the sample tube 51 is uniformly heated, the efficiency of sample amplification and replication can be improved.
In some embodiments, as shown in fig. 2, the ends of the plurality of receiving cavities 24 proximate to the base 22 may be disposed in contact with the base 22. In the case where the end of the receiving chamber 24 is in contact with the bottom plate 22, the bottom plate 22 can rapidly transfer heat to the receiving chamber 24 to more rapidly heat up the sample tube 51 in the receiving chamber 24. In other embodiments, the ends of the plurality of receiving cavities 24 proximate the base 22 may also be disposed a distance from the base 22.
In some embodiments, the outer surface of each sample tube 51 of the plurality of sample tubes 51 is surrounded by the first through hole 31, the second through hole 43, and the receiving cavity 24. With this configuration, on the one hand, the temperature control member 2 and the temperature control member 4 can be enabled to rapidly transfer heat to the sample tube 51, so that no condensation phenomenon occurs at any portion where the vaporized sample contacts the sample tube 51 in the case where the sample is vaporized. On the other hand, because the clearance between the outer surface of the sample tube 51 and the first through hole 31 is very small, the heat of the temperature control 2 can be prevented from being transferred to the constant temperature element 4, and then the temperature control 2 can be efficiently transferred to the lower area of the sample tube 51 in the accommodating cavity 24, so that the waste of the internal heat of the temperature control 2 is avoided.
In some embodiments, as shown in fig. 3, each of the plurality of receiving cavities 24 is disposed about the same axis as the respective first through hole 31 and second through hole 43, and the axis is perpendicular to the direction of extension of the insulation pad 3. That is, the sample tube 51 is inserted in the temperature control 2 substantially perpendicular to the extending direction of the heat insulating mat 3.
Embodiments of the present disclosure also provide a PCR instrument comprising any of the temperature controlled amplification devices 100 described above.
The temperature-controlled amplification device 100 according to the embodiment of the present disclosure may be applied to various PCR instruments in order to avoid a condensation phenomenon of a sample within the sample tube 51 when gene amplification is performed. It should be appreciated that the temperature controlled amplification device 100 according to embodiments of the present disclosure may also be applied to other biochemical reactions, as well, embodiments of the present disclosure are not limited in this regard.
The foregoing description of the embodiments of the present disclosure has been presented for purposes of illustration and description, and is not intended to be exhaustive or limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the various embodiments described. The terminology used herein was chosen in order to best explain the principles of the embodiments, the practical application, or the technical improvement in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.
Claims (12)
1. A temperature-controlled amplification device (100), characterized in that the temperature-controlled amplification device (100) comprises:
a heating and refrigerating element (1);
a temperature control element (2) which is arranged above the heating and refrigerating element (1) and can be heated or refrigerated by the heating and refrigerating element (1), wherein a plurality of accommodating cavities (24) are formed on one side of the temperature control element (2) which is away from the heating and refrigerating element (1), and the plurality of accommodating cavities (24) are respectively used for accommodating lower areas of corresponding sample tubes (51) in a plurality of sample tubes (51);
a heat insulation pad (3) which is arranged above the temperature control piece (2) and comprises a plurality of first through holes (31) corresponding to the plurality of accommodating cavities (24);
a thermostat (4) provided above the heat insulating mat (3) and including a first top plate (41), and a plurality of protrusions (42) extending from the first top plate (41) toward the heat insulating mat (3), the plurality of protrusions (42) and the first top plate (41) being provided therein with a plurality of second through holes (43), the plurality of first through holes (31) and the plurality of second through holes (43) being provided in correspondence, the walls of the plurality of second through holes (43) being capable of encircling and contacting respectively the upper regions of the respective sample tubes (51) for keeping the upper regions of the respective sample tubes (51) at a constant temperature; and
and a thermal cover (6) arranged above the constant temperature piece (4).
2. The temperature controlled amplification device (100) according to claim 1, wherein the thermostat (4) further comprises a support (44), the support (44) being arranged along an edge of the first top plate (41) and extending towards the heat insulation pad (3), the first top plate (41), the support (44) and the heat insulation pad (3) enclosing a first closed space (10).
3. The temperature controlled amplification device (100) according to claim 1 or 2, wherein the temperature control member (2) further comprises a bottom plate (22) adjacent to the heating and cooling member (1), a second top plate (21) opposite to the bottom plate (22), and a side plate (23) extending from an edge of the second top plate (21) to the bottom plate (22), the plurality of accommodating chambers (24) extending from the second top plate (21) in a direction toward the bottom plate (22).
4. A temperature controlled amplification device (100) according to claim 3, wherein the second top plate (21), the bottom plate (22) and the side plates (23) enclose a receiving space (20), and the plurality of receiving cavities (24) extend from the second top plate (21) into the receiving space (20).
5. The temperature-controlled amplification device (100) according to claim 4, wherein the accommodating space (20) is filled with a heat-conducting medium.
6. The temperature-controlled amplification device (100) of claim 5, wherein ends of the plurality of receiving chambers (24) proximate the bottom plate (22) are disposed in contact with the bottom plate (22).
7. The temperature controlled amplification device (100) according to claim 1 or 2, wherein the temperature controlled amplification device (100) further comprises a sample container (5), the sample container (5) comprising the plurality of sample tubes (51) and a body (52), the plurality of sample tubes (51) extending from a top surface of the body (52) towards the temperature control member (2), the top surface of the body (52) being in contact with the thermal cover (6).
8. The temperature controlled amplification device (100) of claim 7, wherein a peripheral portion of the body (52) around the top surface is in contact with the heat insulating pad (3), the body (52), the heat insulating pad (3) and the thermostatic element (4) enclosing a second enclosed space (30).
9. The temperature controlled amplification device (100) according to claim 1 or 2, wherein an outer surface of each sample tube (51) of the plurality of sample tubes (51) is surrounded by the first through hole (31), the second through hole (43) and the accommodation chamber (24).
10. The temperature controlled amplifying device (100) according to claim 1 or 2, wherein each receiving cavity (24) of the plurality of receiving cavities (24) is arranged around the same axis as the respective first through hole (31) and the second through hole (43), and the axis is perpendicular to the direction of extension of the heat insulating mat (3).
11. The temperature controlled amplification device (100) according to claim 1 or 2, wherein the temperature value of the thermal cover (6) is greater than the temperature value of the thermostatic element (4).
12. A PCR instrument, characterized in that it comprises a temperature controlled amplification device (100) according to any one of claims 1 to 11.
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Address after: Building 1, No. 42 Qibei Road, Changping District, Beijing Patentee after: Kunpeng Gene (Beijing) Scientific Instrument Co.,Ltd. Country or region after: China Address before: 2202, 19th Floor, Building 9, No. 8 Ronghua Middle Road, Daxing District, Beijing Patentee before: ROCGENE TECNOLOGY Co. Country or region before: China |