CN213037766U - Quick PCR reaction tube and instrument thereof - Google Patents

Abstract

The utility model discloses a quick PCR reaction tube, PCR reaction tube includes at least: the container comprises a container tube, a tube cover, a piston and a piston rod, wherein one end of the container tube is provided with the tube cover used for realizing tube body sealing, the piston is arranged inside the tube body at the other end, and the piston is connected with the piston rod. The position adjustment of the reaction liquid in the tube is realized through the PCR reaction tube, so that the reaction liquid is in different temperature areas, and the rapid temperature adjustment of the reaction liquid is completed. The temperature rise and fall speed in the matched instrument can be considered to be at least more than 15 ℃ per second. In addition, the temperature rise and drop requirements are not met, so that the instantaneous power is small, and no energy is wasted.

Description

Quick PCR reaction tube and instrument thereof

Technical Field

The utility model belongs to a molecular biology device, in particular to a rapid PCR reaction tube and an instrument thereof.

Background

PCR polymerase chain reaction is a reaction for DNA/RNA replication. In addition to isothermal PCR, most of the other reactions require constant switching among several different temperatures, so the reaction is usually performed in a temperature cycling device. The temperature cycling device needs to have the capability of rapid temperature rise and fall to provide different temperatures needed for PCR reaction.

Generally, the current PCR reaction mostly adopts two temperature step cycles, namely high temperature denaturation, low temperature annealing and extension. Most PCR requires high temperatures between 90-95 degrees and low temperatures between 50-60 degrees. The speed of temperature change becomes an important performance index of the PCR instrument. It is currently generally accepted that a temperature rise and fall speed of 5 degrees/second is a faster PCR instrument.

Meanwhile, the faster the PCR instrument, the greater the instantaneous power is generally, under otherwise substantially identical conditions. And the power is used for overcoming the influence caused by the last opposite temperature change process and is basically wasted energy.

SUMMERY OF THE UTILITY MODEL

The utility model aims to provide a: in order to overcome the problems of the prior art, the rapid PCR reaction tube is provided, and the position adjustment of the reaction liquid in the tube is realized through the PCR reaction tube, so that the reaction liquid is in different temperature areas, and the rapid temperature adjustment of the reaction liquid is completed.

The purpose of the utility model is realized through the following technical scheme:

a rapid PCR reaction tube, the PCR reaction tube comprising at least: the container comprises a container tube, a tube cover, a piston and a piston rod, wherein one end of the container tube is provided with the tube cover used for realizing tube body sealing, the piston is arranged inside the tube body at the other end, and the piston is connected with the piston rod.

According to a preferred embodiment, the containment tube comprises a bullet-shaped body portion and an elongated body portion; the elongated tube body part is connected with the pointed end of the bullet-shaped tube body part, and the other end of the bullet-shaped tube body part is connected with the tube cover; the piston is assembled in the slender pipe body, and a piston rod is connected in the direction back to the bullet type pipe body and can slide in the slender pipe body.

According to a preferred embodiment, the end of the piston rod is further provided with a recessed pull rod.

According to a preferred embodiment, the tube cover and the container tube are integrally injection-molded and are connected by a connecting rib.

According to a preferred embodiment, the elongated body portion has an internal pipe diameter of 2mm and a length of 25mm to 30 mm; the length of the pointed cone of the bullet type pipe body is 8-10mm, and the length of the equal-diameter part of the bullet type pipe body is 8-10 mm; the wall thickness of the PCR reaction tube is 0.25mm-0.35 mm.

A rapid PCR reaction instrument at least comprises a PCR reaction tube, a thermal cover, a first heating block and a second heating block, wherein the PCR reaction tube at least comprises: the container comprises a container tube, a tube cover, a piston and a piston rod, wherein the tube cover used for realizing tube body sealing is arranged at one end of the container tube, the piston is arranged in the tube body at the other end of the container tube, and the piston is connected with the piston rod; the heat cover is arranged on the top of the tube cover and is in contact with the tube cover; the first heating block and the second heating block are respectively sleeved with the outer side of the container tube and are attached to the container tube, and a gap is formed between the first heating block and the second heating block; the first heating block and a corresponding accommodating area in the container tube form a first temperature adjusting area; and the second heating block and the corresponding accommodating area in the container pipe form a second temperature adjusting area.

According to a preferred embodiment, the thermal cover is provided with a hole body structure, and an optical assembly for imaging the reaction liquid in the PCR reaction tube is arranged in the hole body structure.

According to a preferred embodiment, the containment tube comprises a bullet-shaped body portion and an elongated body portion; the elongated tube body part is connected with the pointed end of the bullet-shaped tube body part, and the other end of the bullet-shaped tube body part is connected with the tube cover; the piston is assembled in the elongated tube body, and a piston rod is connected in the direction back to the bullet-shaped tube body and can slide in the elongated tube body; the first heating block is sleeved outside the pointed conical section of the bullet type pipe body part; the second heating block is sleeved outside the elongated tube body.

According to a preferred embodiment, a gap space between the first heating block and the second heating block is provided with a heat insulation block.

According to a preferred embodiment, the reaction device further comprises a catch configured for completing the grasping of the trailing end of the piston rod for effecting the driving up and down movement of the piston rod.

The main scheme and the further selection schemes of the utility model can be freely combined to form a plurality of schemes, which are the schemes that can be adopted and claimed by the utility model; and the utility model discloses also can the independent assortment between (each non-conflict selection) selection and between other choices. The technical solutions to be protected by the present invention, which are various combinations that can be known to those skilled in the art based on the prior art and the common general knowledge after understanding the present invention, are not exhaustive herein.

The utility model has the advantages that: the position adjustment of the reaction liquid in the tube is realized through the PCR reaction tube, so that the reaction liquid is in different temperature areas, and the rapid temperature adjustment of the reaction liquid is completed. The temperature rise and fall speed in the matched instrument can be considered to be at least more than 15 ℃ per second. In addition, the temperature rise and drop requirements are not met, so that the instantaneous power is small, and no energy is wasted.

Drawings

FIG. 1 is a schematic perspective view of a PCR reaction tube according to the present invention;

FIG. 2 is a cross-sectional view of a PCR reaction tube of the present invention;

FIG. 3 is a schematic structural view of the PCR reaction apparatus of the present invention;

FIG. 4 is a schematic structural view of a device for setting a reaction solution in a first temperature control region in a PCR reaction apparatus of the present invention;

FIG. 5 is a schematic structural view of a device for setting a reaction solution in a second temperature control zone in the PCR reaction apparatus of the present invention;

the device comprises a container tube 1, a piston 2, a piston 3, a piston rod 4, a tube cover 10, a PCR reaction tube 11, a heat cover 12, a first heating block 13, a heat insulation block 14, a second heating block 15 and an optical assembly.

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the disclosure herein. The present invention can also be implemented or applied through other different specific embodiments, and various details in the present specification can be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict.

It should be noted that, in order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, not all embodiments. The components of embodiments of the present invention, as generally described and illustrated in the figures herein, may be arranged and designed in a wide variety of different configurations.

Thus, the following detailed description of the embodiments of the present invention, presented in the accompanying drawings, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", and the like indicate the position or positional relationship based on the position or positional relationship shown in the drawings, or the position or positional relationship which is usually placed when the product of the present invention is used, and are only for convenience of description and simplification of the description, but do not indicate or imply that the device or element referred to must have a specific position, be constructed and operated in a specific orientation, and thus, should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used solely to distinguish one from another and are not to be construed as indicating or implying relative importance.

Furthermore, the terms "horizontal", "vertical", "overhang" and the like do not imply that the components are required to be absolutely horizontal or overhang, but may be slightly inclined. For example, "horizontal" merely means that the direction is more horizontal than "vertical" and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.

In the description of the present invention, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meaning of the above terms in the present invention can be understood in specific cases to those skilled in the art.

Additionally, the utility model discloses it is pointed out that, in the utility model, if do not write out structure, connection relation, positional relationship, power source relation etc. that concretely relates to very much, then the utility model relates to a structure, connection relation, positional relationship, power source relation etc. are technical personnel in the field on prior art's basis, can not learn through creative work.

Example 1:

referring to fig. 1 and 2, there is shown a rapid PCR reaction tube 10 including at least: a container tube 1, a tube cover 4, a piston 2 and a piston rod 3.

The container tube comprises a container tube 1 and is characterized in that one end of the container tube 1 is provided with a tube cover 4 used for achieving tube body sealing, a piston 2 is arranged inside the tube body at the other end, and the piston 2 is connected with a piston rod 3.

Preferably, the tube cap 4 is sealed to the tail end of the bullet-shaped tube after adding the reagent, and together with the piston 2, seals the reagent in the receptacle tube 1. Preferably, the optical detection instrument uses upper excitation/collection light, and the tube cover is a transparent tube cover.

Preferably, the containment tube 1 comprises a bullet-type body portion and an elongated body portion.

Preferably, the elongated body portion is connected to a pointed end of the bullet-shaped body portion, and the other end of the bullet-shaped body portion is connected to the cap 4.

Preferably, the piston 2 is fitted inside the elongated tubular body, and a rod of the piston 2 is attached in a direction facing away from the bullet-shaped tubular body and is slidable inside the elongated tubular body.

Further, the piston 2 is made of a soft material that is resistant to high temperature and has a small amount of elastic deformation, and is pre-pressed and sealed in consideration of different hardness. Preferably, the piston 2 can be pre-pressed by 0.2mm by using 60-degree silica gel or fluorine gel.

Preferably, the end of the piston rod 3 is further provided with a concave pull rod. So that the pulling of the piston rod 3 can be smoothly performed.

Preferably, the container tube 1 is made of high-temperature-resistant plastic. And the wall thickness of the PCR reaction tube or the container tube 1 is 0.25mm-0.35 mm.

Preferably, the pipe cover 4 and the container pipe 1 are integrally formed by injection molding, and are connected through a connecting rib in the middle.

Preferably, the elongated body portion has an internal tube diameter of 2mm and a length of 25mm to 30 mm. The length of the pointed cone of the bullet type pipe body is 8-10mm, and the length of the equal-diameter part of the bullet type pipe body is 8-10 mm.

The PCR reaction tube is vertically arranged as shown in FIG. 2, and when the piston 2 is moved to the uppermost position, the reaction solution 20-A is in the bullet type tube body portion. When the piston 2 moves to the lowermost portion, the reaction liquid 20-B is in the elongated tubular body portion. Therefore, when different heating devices are arranged on the outer sides of the bullet type tube body part and the elongated tube body part respectively, the position adjustment of the reaction liquid in the tube can be realized through the PCR reaction tube, so that the reaction liquid is in different temperature areas, and the rapid temperature adjustment of the reaction liquid is completed.

Example 2

On the basis of embodiment 1, reference is also made to fig. 3 to 5. The utility model also discloses a quick PCR reaction appearance. The reaction instrument at least comprises a PCR reaction tube 10, a thermal cover 11, a first heating block 12 and a second heating block 14.

Preferably, the thermal cover 11 is disposed on top of the tube cover 4 and contacts the tube cover 4.

Furthermore, a hole structure is arranged on the thermal cover 11, and an optical assembly 15 for fluorescence excitation and detection of the reaction solution in the PCR reaction tube 10 is arranged in the hole structure.

Preferably, the apparatus may be provided with an optical probe at a location where an optical reading is required. In principle, the optical assembly 15 can be arranged at any location, but preferably the instrument can be arranged to excite/receive light upwards, i.e. in the thermal cover 11. The thermal cover thus has a hole in the middle, in which the optical component 15 is arranged, of which only the interface part of the optical component 15 is shown.

Preferably, a heating assembly is further disposed in the thermal cover 11 for heating the tube cover 4 to prevent condensation in the tube cover 4 from affecting the PCR reaction. Specifically, if condensation occurs in the PCR reaction, the concentration of the fluorescent substance will be drastically changed, which affects the PCR reaction.

Preferably, the material of the thermal cover 11 is pure aluminum or red copper. Which is generally provided on the cover of the instrument and is separated from the instrument body when the PCR reaction tube 10 is inserted into the instrument and is snapped to the instrument body before operation.

Preferably, the first heating block 12 and the second heating block 14 are respectively sleeved on the outer side of the containment tube 1 and attached to the containment tube 1. And a gap is arranged between the first heating block 12 and the second heating block 14.

Preferably, the thickness of the first heating block 12 is as high as that of the tapered section of the bullet-shaped body of the reaction tube, and a tapered through hole having the same shape as that of the tapered section is formed in the middle. The material of the first heating block 12 is preferably red copper or pure aluminum which has high heat conductivity coefficient and is economical in cost.

Preferably, the combined thickness of the second heating block 14 and the insulating block 13 does not exceed the length of the elongated tubular body minus the length of the piston. The second heating block 14 is provided with a hole in the middle with the same outer diameter as the elongate tubular body portion. Preferably, the thickness of the second heating block 14 is 18 mm. The heating block is preferably made of red copper or pure aluminum which has high heat conductivity coefficient and is economical in cost.

Preferably, the first heating block 12 and the corresponding accommodating area in the container tube 1 form a first temperature adjusting area. The second heating block 14 and the corresponding accommodating area in the container pipe 1 form a second temperature adjusting area.

Further, the first heating block 12 is sleeved outside the pointed conical section of the bullet-shaped pipe body. The second heating block 14 is sleeved outside the elongated pipe body part.

Preferably, a heat insulation block 13 is disposed in a gap space between the first heating block 12 and the second heating block 14.

Preferably, the heat insulating block 13 has a hole in the middle of the heat insulating material, the hole having the same outer diameter as that of the elongated tubular body portion having the same diameter as that of the reaction tube. The insulation is preferably silica aerogel blanket. Preferably, the insulating material of the insulating block 13 has a thickness of 3mm to 5 mm.

Preferably, the reaction apparatus further comprises a grapple configured for completing the grasping of the rear end of the rod of the piston 2 to effect the driving of the rod of the piston 2 up and down.

The specific implementation process of the reactor is as follows:

the method comprises the following steps: the piston 2 of the PCR reaction tube 10 is pushed to the uppermost position, and the reagent is added from the nozzle, and the tube cap 4 is closed.

Step two: the instrument is uncapped (i.e., the thermal cap 11), the reaction tube is inserted into the well in the instrument, and the instrument is capped, with the optical interface in the thermal cap aligned with the reaction tube cap 4, as shown in FIG. 4.

Step three: the hot lid 11 is heated to 105 degrees and is kept at 105 degrees from the beginning to the end of the reaction, and is used for heating the reaction tube cap 4 and preventing condensation.

Step four: if the template in the reaction solution is RNA, reverse transcription of RNA may occur. At this time, the first heating block 12 is directly heated to a reverse transcription required temperature, typically 50 to 60 degrees. If there is no RNA reverse transcription process, step four can be skipped.

Step five: the grapple pulls the piston 2 in the reaction tube and the reaction solution enters the elongated body portion (shown in fig. 5). At this point the second heating block 14 is warmed to the PCR pre-denaturation/hot start temperature, typically 90-95 degrees. Typically, this temperature is also the step seven temperature. The first heating block 12 may now be adjusted to the temperature required in step six.

Step six: prior to this step, the first heating block 12 should have been heated to the annealing/extension temperature required for PCR, typically 50-60 degrees. After this step, the piston 2 in the reaction tube is pushed by the grapple, and the reaction liquid enters the tapered portion in the bullet type tube body portion, as shown in fig. 4. The temperature of the reagent in the reaction tube is immediately reduced to the set temperature. After the annealing/extension is completed, the fluorescence information is read by the optical assembly. In this step, the second heating block 14 maintains the PCR denaturation temperature.

Step seven: prior to this step, the second heating block 14 should have been heated to the denaturation temperature required for PCR, typically 90-95 degrees. After this step, the grapple pushes the piston in the reaction tube and the reaction solution enters the elongated tube portion as shown in FIG. 5. The temperature of the reagent in the reaction tube was immediately raised to the set temperature. In this step, the first heating block 12 should maintain the PCR annealing/extension temperature.

Step eight: repeating the sixth step and the seventh step for a plurality of times to complete the amplification and obtain real-time fluorescence amplification data.

Step nine: if a melting curve response is required, the instrument grapple can push the push rod of the reaction tube to the uppermost part. At this time, the first heating block 12 is gradually raised from the annealing temperature (50-60 degrees) required for PCR to the denaturation temperature (90-95 degrees), and the optical assembly is used to collect optical information to complete the melting curve. There is generally no additional reaction after the melting profile and the second heating block 14 can be turned off early to reduce the temperature.

Step ten: turning on the fan to cool the heating head

Step eleven: the instrument was closed and the reaction tube removed.

The instrument of the invention is characterized in that:

1. the temperatures of the first heating block and the second heating block are not changed in the whole reaction process, and the temperature rising and falling speed only depends on the speed of the piston rod 3. According to the test, if the size of the reaction tube is designed according to the optimal size, the push rod can push the reaction liquid from the elongated tube body to the bullet type tube body without causing spray splash, the equivalent temperature reduction speed is more than 15 degrees/second and can reach 22.5 degrees/second at most, the process that the piston rod 3 is pulled downwards to allow the reaction liquid to enter the elongated tube body from the bullet type tube body can be completed in less than one second, the equivalent temperature increase speed is more than 30 degrees/second and can reach 45 degrees/second at most.

2. In the reaction process, the first heating block 12 and the second heating block 14 do not need to be heated or cooled, and only need small power to maintain the temperature after the temperature is stable, so that the power consumption of the instrument is very small.

3. The first heating block 12 and the second heating block 14 are not heated or cooled, all the power of the first heating block is used for maintaining the reaction temperature, the negative effect caused by overcoming the last temperature is not generated, and the energy consumption waste is less.

4. If all the sizes of the reaction tube are designed according to the optimal size, the system formed by the reaction tube and the instrument can be compatible with a PCR system of 10uL to 50uL only by slightly adjusting the distance of the grapple.

5. The instrument can complete PCR reaction. And based on the corresponding collected data, the melting curve analysis after the reaction can be carried out.

The aforesaid the utility model discloses basic embodiment and each further alternative can the independent assortment in order to form a plurality of embodiments, is the utility model discloses can adopt and claim the embodiment of protection. In the scheme of the utility model, each selection example can be combined with any other basic examples and selection examples at will.

The above description is only exemplary of the present invention and should not be taken as limiting the scope of the present invention, as any modifications, equivalents, improvements and the like made within the spirit and principles of the present invention are intended to be included within the scope of the present invention.

Claims (10)

1. A rapid PCR reaction tube, comprising at least: a container tube (1), a tube cover (4), a piston (2) and a piston rod (3),

the container comprises a container tube (1), a tube cover (4) used for sealing the tube body is arranged at one end of the container tube (1), a piston (2) is arranged inside the tube body at the other end of the container tube, and the piston (2) is connected with a piston rod (3).

2. The rapid PCR reaction tube according to claim 1, wherein the containment tube (1) comprises a bullet-shaped tube portion and an elongated tube portion;

the elongated tube body part is connected with the pointed end of the bullet-shaped tube body part, and the other end of the bullet-shaped tube body part is connected with the tube cover (4);

the piston (2) is assembled in the elongated tube body, is connected with a piston rod (3) in the direction back to the bullet-shaped tube body and can slide in the elongated tube body.

3. The rapid PCR reaction tube according to claim 2, wherein the end of the piston rod (3) is further provided with a concave pull rod.

4. The rapid PCR reaction tube of claim 1, wherein the tube cover (4) and the vessel tube (1) are integrally formed by injection molding and connected by a connecting rib therebetween.

5. The rapid PCR reaction tube of claim 3, wherein the elongated body portion has an inner diameter of 2mm and a length of 25mm to 30 mm;

the length of the pointed cone of the bullet type pipe body is 8-10mm, and the length of the equal-diameter part of the bullet type pipe body is 8-10 mm;

the wall thickness of the PCR reaction tube is 0.25mm-0.35 mm.

6. A rapid PCR reaction instrument is characterized in that the reaction instrument at least comprises a PCR reaction tube (10), a hot cover (11), a first heating block (12) and a second heating block (14),

the PCR reaction tube (10) comprises at least: the container comprises a container tube (1), a tube cover (4), a piston (2) and a piston rod (3), wherein one end of the container tube (1) is provided with the tube cover (4) for realizing tube body sealing, the piston (2) is arranged in the tube body at the other end, and the piston (2) is connected with the piston rod (3);

the hot cover (11) is arranged on the top of the tube cover (4) and is in contact with the tube cover (4);

the first heating block (12) and the second heating block (14) are respectively sleeved with the outer side of the container pipe (1) and attached to the container pipe (1), and a gap is formed between the first heating block (12) and the second heating block (14);

moreover, the first heating block (12) and a corresponding accommodating area in the container pipe (1) form a first temperature adjusting area;

the second heating block (14) and a corresponding containing area in the container pipe (1) form a second temperature adjusting area.

7. A rapid PCR reaction apparatus according to claim 6, wherein a well structure is provided on the container tube (1), and an optical assembly (15) for fluorescence excitation and detection is provided in the well;

a hole body structure is arranged on the hot cover (11), and an optical assembly (15) used for fluorescence excitation and detection of reaction liquid in the PCR reaction tube (10) is arranged in the hole body structure.

8. The rapid PCR reaction instrument according to claim 6, wherein the containment tube (1) comprises a bullet-shaped tube portion and an elongated tube portion;

the elongated tube body part is connected with the pointed end of the bullet-shaped tube body part, and the other end of the bullet-shaped tube body part is connected with the tube cover (4);

the piston (2) is assembled in the elongated tube body, is connected with a piston rod (3) in the direction back to the bullet-shaped tube body and can slide in the elongated tube body;

the first heating block (12) is sleeved outside the pointed conical section of the bullet type pipe body part;

the second heating block (14) is sleeved outside the elongated pipe body part.

9. A rapid PCR reaction apparatus according to claim 8, wherein the gap space between the first heating block (12) and the second heating block (14) is provided with a heat insulating block (13).

10. A rapid PCR reaction instrument according to claim 9, characterized in that the instrument further comprises a grapple configured for completing the grasping of the trailing end of the piston rod (3) for driving the piston rod (3) up and down.

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