Background
In recent years, with the rapid development of molecular biology techniques, diagnostic methods based on nucleic acid detection have been widely established and widely applied to laboratory detection of human diseases, isothermal amplification technology is one of them, and isothermal amplification has the advantages of rapidness, high efficiency and specificity compared with other nucleic acid amplification technologies and does not need special equipment, so that it is considered by many scholars as a detection method possibly comparable to PCR once it appears. 2019-when the new coronavirus is in a big outbreak, the PCR molecular diagnosis technology is set as the gold standard for detection, but the requirement on a thermal cycler is high, and the detection in the field and the primary hospitals is not facilitated. Thus, scientists have attempted to use alternative exponential amplification techniques, such as loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification, hyperbranched rolling circle amplification, exponential amplification reactions, and exponential strand displacement amplification, among others. The exponential amplification technology reacts under the constant temperature condition without thermal cycle, thereby not only expanding the application scene of the nucleic acid detection technology, but also exciting the potential of POCT application. The molecular diagnosis has the characteristics of high sensitivity and strong specificity, and if the advantages of quick POCT are further integrated, the molecular diagnosis inevitably occupies an important position in the detection field.
POCT refers to a detection mode which is carried out on a sampling site and utilizes a portable analysis instrument and a matched reagent to quickly obtain a detection result. POCT meaning can be understood from two aspects: spatially, tests performed at the patient's side, i.e. "bedside tests"; temporally, a "point-of-care" test can be performed.
The nucleic acid amplification product produced based on the constant temperature amplification technology can perform single molecule nucleic acid detection at a constant and low temperature within 30 min. The current nucleic acid amplification products in China have low requirements on environment and hardware facilities, and have good application in the aspects of biological protection, water body inspection, food inspection, medical diagnosis, microfluid, veterinarian and the like; however, the current nucleic acid amplification products consider the problems of application scenes and cost, and the colloidal gold test strip is used for terminal detection, so that the integration of amplification and test strip detection is realized, the general scene requirements are met, and the cost is reduced, but the product has the defects when being applied to the nucleic acid detection of the new coronavirus: the amplified product needs to be uncovered by a worker, the worker puts the product into a corresponding portable instrument, false positive of detection can be increased, the nucleic acid amplification sample can also produce aerosol pollution to pollute a laboratory, and therefore the pollution problem caused by the amplified product is solved by generally using a sealing device, the complexity of operation is obviously increased, and the risk of pollution still exists in the operation process.
Namely, the nucleic acid amplification products in the prior art have the problems of complicated operation and aerosol pollution.
SUMMERY OF THE UTILITY MODEL
The utility model aims to provide a detection device for a nucleic acid amplification product, which solves the problems of complicated operation and aerosol pollution of the existing nucleic acid amplification products.
In order to achieve the purpose, the utility model adopts the following technical scheme:
a detection apparatus for a nucleic acid amplification product, comprising an amplification part, a dilution part and a detection part; an amplification cavity is formed in the amplification part, a dilution cavity is formed in the dilution part, a detection cavity is formed in the detection part, and a detection unit is arranged in the detection cavity;
the amplification part is detachably connected with the dilution part; when a sample is amplified, the amplification part is connected with the dilution part, and the amplification cavity is not communicated with the dilution cavity; when a sample is diluted, the amplification part is connected with the dilution part, and the amplification cavity is communicated with the dilution cavity;
the diluting part is detachably connected with the detecting part, when a sample is detected, the diluting part is connected with the detecting part, and the diluting cavity is communicated with the detecting cavity.
Optionally, a third opening is formed in the diluting cavity on the wall of the diluting cavity close to the detecting part, and a sealing film is arranged on the third opening;
the detection part is provided with a fourth opening on the wall of the detection cavity close to the dilution part, and the detection part is provided with a puncture part protruding out of the detection cavity;
when a sample is detected, the puncture part penetrates through the sealing film, and the third opening is communicated with the fourth opening.
Optionally, the puncturing portion includes a first tip portion and a second tip portion, and the first tip portion protrudes from the second tip portion relative to the detecting portion.
Optionally, the number of the second tip portions is two, and the second tip portions are respectively disposed on two sides of the first tip portion.
Optionally, the length direction of the first tip portion is arranged to extend along a first direction, the length direction of the second tip portion is arranged to extend along a second direction, and the first direction is perpendicular to the second direction.
Optionally, the puncturing part further comprises a puncturing base, and a puncturing baffle is convexly arranged on the puncturing base;
the first tip portion and the second tip portion are both arranged on one side, far away from the puncture base, of the puncture baffle.
Optionally, the detection portion includes a drainage unit, one end of the drainage unit is located in the detection cavity, and the other end of the drainage unit passes through the fourth opening and is connected with the puncture portion.
Optionally, the detection cavity comprises a detection pool and a buffer pool which are separated by a partition plate, and one end of the drainage unit is located in the buffer pool;
the baffle corresponds detection element's position has been seted up and has been dodged the hole, detection element is located the detection pond, detection element's one end is passed dodge the hole and extend to in the buffer pool.
Optionally, the bottom wall area of the puncture base is smaller than the top surface area of the drainage unit.
Optionally, a first opening is formed in the wall of the amplification cavity, and a second opening is formed in the wall of the dilution cavity;
the amplification part is rotatable relative to the dilution part when the amplification part is connected to the dilution part; and when the amplification part rotates to a preset first angle relative to the dilution part, the first opening is communicated with the second opening.
Compared with the prior art, the utility model has the following beneficial effects:
when the detection device for the nucleic acid amplification product provided by the utility model is used for detecting nucleic acid, a nucleic acid sample and an amplification reagent are placed in an amplification cavity, then, an amplification part is connected with a dilution part, and a nucleic acid amplification sample is obtained after the amplification reaction is finished; then, communicating the amplification cavity with the dilution cavity, and mixing the nucleic acid amplification sample with the dilution reagent in the dilution cavity to obtain a nucleic acid dilution sample; then, the diluting part is connected with the detecting part, so that the diluting cavity is communicated with the detecting cavity, the nucleic acid diluting sample in the diluting cavity flows into the detecting cavity and contacts with the detecting unit, and the detecting unit can feed back a result to an operator after the reaction to be detected is finished; in the detection process, the amplification, dilution and reaction processes of the nucleic acid sample are in a sealed state, so that aerosol pollution of nucleic acid pollutants to the environment is prevented, complicated-operation equipment such as a sealing device and the like is omitted, and the detection device is convenient to operate.
Detailed Description
In order to make the objects, features and advantages of the present invention more apparent and understandable, the embodiments of the present invention will be described in detail and completely with reference to the accompanying drawings, and it is to be understood that the embodiments described below are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and thus, are not to be construed as limiting the present invention. It should be noted that when one component is referred to as being "connected" to another component, it can be directly connected to the other component or intervening components may also be present.
The technical scheme of the utility model is further explained by the specific implementation mode in combination with the attached drawings.
Referring to fig. 1 to 8, fig. 1 is a schematic overall structure diagram of a detection apparatus for a nucleic acid amplification product according to an embodiment of the present invention, fig. 2 is a schematic bottom structure diagram of an amplification part according to an embodiment of the present invention, fig. 3 is a schematic top structure diagram of a dilution part according to an embodiment of the present invention, fig. 4 is a schematic partial cross-sectional structure diagram of the detection apparatus for a nucleic acid amplification product according to an embodiment of the present invention, fig. 5 is a schematic partially enlarged structure diagram of fig. 4 at a, fig. 6 is a schematic front structure diagram of a puncture part according to an embodiment of the present invention, fig. 7 is a schematic side structure diagram of the puncture part according to an embodiment of the present invention, and fig. 8 is a schematic top structure diagram of the puncture part according to an embodiment of the present invention.
The detection device for the nucleic acid amplification product is applied to the nucleic acid detection scene, completes the amplification, dilution and detection of the nucleic acid sample under the condition of keeping the interior closed by improving the structure of the detection device, and has simple and convenient operation and high safety.
As shown in FIGS. 1 and 4, the detection apparatus for a nucleic acid amplification product used in the present embodiment includes an amplification part 1, a dilution part 2, and a detection part 3; the amplification part 1 is internally provided with an amplification cavity, the dilution part 2 is internally provided with a dilution cavity, the detection part 3 is internally provided with a detection cavity, and the detection cavity is internally provided with a detection unit 4. Wherein, the diluting cavity is pre-stored with a diluting reagent which can dilute the nucleic acid amplification sample, thereby improving the detection sensitivity and the detection efficiency; an observation window is arranged on the wall of the detection cavity and is made of transparent material; the detection part 3 comprises an upper detection shell and a lower detection shell, and the upper detection shell and the lower detection shell are hermetically connected and are butted and encircled to form a detection cavity; the detection unit 4 comprises detection test paper, the detection test paper can react with the nucleic acid diluted sample, and the result is fed back through color change; alternatively, the detection unit 4 includes a test tube filled with a detection solution, and the nucleic acid dilution sample can enter the test tube to be mixed with the detection solution to perform a detection reaction.
The amplification part 1 and the dilution part 2 are detachably connected; when a sample is amplified, the amplification part 1 is connected with the dilution part 2, and the amplification cavity is not communicated with the dilution cavity; when the sample is diluted, the amplification part 1 is connected with the dilution part 2, and the amplification cavity is communicated with the dilution cavity.
Dilution portion 2 and detection portion 3 detachably are connected, and when detecting the sample, dilution portion 2 is connected with detection portion 3, dilutes the cavity and communicates with the detection cavity.
Specifically, when nucleic acid detection is performed, a nucleic acid sample and an amplification reagent are placed in an amplification cavity, then, the amplification part 1 is connected with the dilution part 2, and a nucleic acid amplification sample is obtained after an amplification reaction is finished; then, communicating the amplification cavity with the dilution cavity, and mixing the nucleic acid amplification sample with the dilution reagent in the dilution cavity to obtain a nucleic acid dilution sample; then, the diluting part 2 is connected with the detecting part 3, so that the diluting cavity is communicated with the detecting cavity, the nucleic acid diluting sample in the diluting cavity flows into the detecting cavity and contacts with the detecting unit 4, and after the reaction to be detected is finished, the detecting unit 4 can feed back a result to an operator; in the detection process, the amplification, dilution and reaction processes of the nucleic acid sample are in a sealed state, so that aerosol pollution of nucleic acid pollutants to the environment is prevented, complicated-operation equipment such as a sealing device and the like is omitted, and the detection device is convenient to operate.
Further, as shown in fig. 2 to 3, a first opening 51 is opened on the wall of the amplification chamber, and a second opening 52 is opened on the wall of the dilution chamber.
When the amplification part 1 is connected to the dilution part 2, the amplification part 1 can rotate relative to the dilution part 2; and when the amplification part 1 is rotated to a predetermined first angle with respect to the dilution part 2, the first opening 51 communicates with the second opening 52. Illustratively, when performing nucleic acid detection, a nucleic acid sample and amplification reagents are added to the amplification chamber through the first opening 51; then, the amplification part 1 and the dilution part 2 are connected, at this time, the angle of the amplification part 1 relative to the dilution part 2 is not the first angle, namely, the first opening 51 and the second opening 52 are staggered, at this time, the first opening 51 is blocked by the outer wall of the dilution part 2, the second opening 52 is blocked by the outer wall of the amplification part 1, and the amplification cavity is isolated from the dilution cavity; after the amplification reaction is finished, the operator rotates the amplification part 1 to a preset first angle relative to the dilution part 2, so that the first opening 51 is communicated with the second opening 52, and the nucleic acid amplification sample is mixed with the dilution reagent to obtain the nucleic acid dilution sample.
It should be added that a sealing ring is arranged at the joint of the amplification part 1 and the dilution part 2, and when the amplification part 1 is connected with the dilution part 2, the sealing ring plays a role of sealing the gap between the amplification part 1 and the dilution part 2.
Further, as shown in fig. 4 and 5, the diluting part 2 has a third opening 53 formed on the wall of the diluting cavity near the detecting part 3, and a sealing film 55 is provided on the third opening 53.
The detecting part 3 is provided with a fourth opening 54 on the wall of the detecting cavity close to the diluting part 2, and the detecting part 3 is provided with a puncturing part 6 protruding from the detecting cavity. When a sample is tested, the punctured portion 6 passes through the sealing film 55, and the third opening 53 communicates with the fourth opening 54. More specifically, when a sample needs to be detected, the diluting part 2 is connected to the detecting part 3, and during the connection process, the puncturing part 6 punctures the sealing film 55, so that the nucleic acid diluted sample in the diluting cavity flows into the detecting cavity and contacts with the detection test paper.
Further, as shown in fig. 6 to 8, the puncturing part 6 includes a first tip part 61 and a second tip part 62, and the first tip part 61 protrudes from the second tip part 62 relative to the detecting part 3. In the process of connecting the diluting part 2 and the detecting part 3, the first tip part 61 pierces the sealing membrane 55 first, and the second tip part 62 enlarges the opening of the sealing membrane 55, so that the sealing membrane 55 is expanded for the second time, and the efficiency of the nucleic acid diluted sample flowing into the detection cavity is improved.
In a specific embodiment, the number of the second pointed portions 62 is two, and the second pointed portions are respectively disposed at both sides of the first pointed portion 61. It should be understood that the number of the second tip portions 62 can also be four, six, eight, etc., and are evenly distributed on both sides of the first tip portion 61.
Further, the first tip portion 61 is disposed to extend in a first direction in a longitudinal direction thereof, and the second tip portion 62 is disposed to extend in a second direction in a longitudinal direction thereof, the first direction being perpendicular to the second direction. At this time, as shown in fig. 8, the first pointed portion 61 and the second pointed portion 62 cross each other in a cross shape.
Further, the puncturing part 6 further comprises a puncturing base 63, and a puncturing baffle 64 is convexly arranged on the puncturing base 63. The first and second pointed portions 61, 62 are disposed on a side of the puncture barrier 64 remote from the puncture base 63.
Specifically, when the diluting part 2 is connected to the detecting part 3, the first tip part 61 first pierces the sealing film 55, and the second tip part 62 forms a larger opening at the pierced sealing film 55; with the penetration of the piercing part 6, the piercing baffle 64 further enlarges the opening, thereby effectively piercing the sealing membrane 55, avoiding the piercing part 6 from being stuck on the opening of the sealing membrane 55, avoiding the occurrence of blocking, and ensuring the flow rate of the nucleic acid diluted sample.
In one embodiment, first tip portion 61 is formed by four piercing arms extending from piercing barrier 64 in a direction away from piercing base 63, with the tips of the four piercing arms converging at an apex to form first tip portion 61.
Further, as shown in fig. 5, the detecting portion 3 includes a drainage unit 31, one end of the drainage unit 31 is located in the detecting cavity, and the other end of the drainage unit 31 passes through the fourth opening 54 and is connected to the puncturing portion 6. The drainage unit 31 is made of sponge, cotton and other water-absorbing materials, is equivalent to a capacity pool capable of temporarily storing liquid, can make the nucleic acid diluted sample move directionally to prevent the nucleic acid diluted sample from diffusing to other places, and can adjust the speed of the nucleic acid diluted sample reaching the detection unit 4 to prevent the sample washing phenomenon; in addition, it enables the nucleic acid diluted sample to overcome the internal pressure difference due to the sealing of the detection device and flow into the detection chamber smoothly.
In this embodiment, the one end of drainage unit 31 extends to and detects the casing down, and the diapire of detection cavity, and the other end of drainage unit 31 passes fourth opening 54 and overlaps and be equipped with the sealing washer, and this sealing washer sets up outside detecting the cavity to offset with the border of fourth opening 54.
Further, the detection chamber comprises a detection cell 33 and a buffer cell 32 which are separated by a partition 35, and one end of the drainage unit 31 is located in the buffer cell 32. The buffer pool 32 plays a role in buffering the diluted nucleic acid sample, so as to prevent the diluted nucleic acid sample from directly impacting the detection unit 4.
The position that baffle 35 corresponds detecting element 4 has seted up and has dodged the hole, and detecting element 4 is located detection pond 33, and the one end of detecting element 4 passes and dodges the hole and extend to in the buffer pool 32. The joint of the avoiding hole and the detection unit 4 is provided with a sealing ring, the sealing ring respectively seals the avoiding hole and the detection unit 4, and the sealing ring fills the gap between the detection unit 4 and the partition plate 35 and does not press the detection unit 4. The detecting unit 4 is not deformed thereby, and normal operation of the detection reaction is ensured.
In addition, the surrounding portion 34 is protruded from the detecting portion 3 outside the buffer tank 32, the surrounding portion 34 surrounds the fourth opening 54, and a gap for inserting the diluting portion 2 is left between the fourth opening 54 and the surrounding portion 34, so as to prevent the diluted nucleic acid sample from being splashed into the external environment. Meanwhile, a plurality of sealing rings are arranged on the surrounding part 34 for sealing and connecting the diluting part 2 and the detecting part 3.
Further, the bottom wall area of the puncture base 63 is smaller than the top surface area of the drainage unit 31. So that the nucleic acid diluted sample can flow down the puncture base 63 to the drainage cell 31.
In summary, the detection device for a nucleic acid amplification product provided in this embodiment has the advantages of simple operation and high safety, and further has the characteristics of sample washout prevention, high detection efficiency, high detection accuracy, high stability, and the like.
The above-mentioned embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.