CN113980797B - Detection device and detection method for nucleic acid amplification product - Google Patents
Detection device and detection method for nucleic acid amplification product Download PDFInfo
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- CN113980797B CN113980797B CN202111277311.9A CN202111277311A CN113980797B CN 113980797 B CN113980797 B CN 113980797B CN 202111277311 A CN202111277311 A CN 202111277311A CN 113980797 B CN113980797 B CN 113980797B
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- 230000003321 amplification Effects 0.000 title claims abstract description 110
- 238000003199 nucleic acid amplification method Methods 0.000 title claims abstract description 110
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- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 74
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 74
- 238000010790 dilution Methods 0.000 claims abstract description 67
- 239000012895 dilution Substances 0.000 claims abstract description 67
- 238000007789 sealing Methods 0.000 claims abstract description 55
- 238000012360 testing method Methods 0.000 claims abstract description 43
- 238000006243 chemical reaction Methods 0.000 claims abstract description 18
- 238000007865 diluting Methods 0.000 claims abstract description 18
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 14
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- 238000012123 point-of-care testing Methods 0.000 description 5
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- 230000000694 effects Effects 0.000 description 3
- 239000003344 environmental pollutant Substances 0.000 description 3
- 238000011010 flushing procedure Methods 0.000 description 3
- 238000011901 isothermal amplification Methods 0.000 description 3
- 231100000719 pollutant Toxicity 0.000 description 3
- 238000007397 LAMP assay Methods 0.000 description 2
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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Abstract
The invention discloses a detection device and a detection method for a nucleic acid amplification product, wherein when nucleic acid detection is carried out, a nucleic acid sample and an amplification reagent are added into an amplification part, the amplification part is connected with a dilution part to form a sample storage unit, and the sample storage unit is uniformly shaken to complete nucleic acid amplification reaction; the puncturing part punctures the sealing film, the amplifying cavity is communicated with the diluting cavity to form a closed cavity, and the sealed cavity is uniformly shaken to finish dilution; and connecting the sample storage unit with the detection part, enabling the sample in the closed cavity to flow into the detection cavity, and carrying out detection reaction with the detection test paper to finish detection. The nucleic acid sample is always in the airtight cavity after being added into the amplification part, so that leakage can not be caused when the nucleic acid sample is amplified and diluted, the safety is high, and the whole operation process of the detection device only needs to carry out two connection actions and one puncturing action, so that the action of uncovering is omitted, the operation complexity of nucleic acid detection is reduced, the leakage risk is further avoided, and the advantages of simplicity and convenience in operation and high safety are achieved.
Description
Technical Field
The invention relates to the field of nucleic acid amplification detection, in particular to a detection device and a detection method for a nucleic acid amplification product.
Background
In recent years, with the rapid development of molecular biology techniques, diagnostic methods based on nucleic acid detection have been established in large numbers and widely used in laboratory detection of human diseases, and isothermal amplification technology is one of them, and compared with other nucleic acid amplification technologies, isothermal amplification has the advantages of being rapid, efficient, specific and free from special equipment, so that once it has appeared, it has been considered by many students as a detection method that is likely to be comparable to PCR. In the virus detection, the PCR molecular diagnosis technology is defined as a gold standard for detection, but the PCR molecular diagnosis technology has high requirements on a thermal cycler, and is unfavorable for field and primary hospital detection. Accordingly, 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, exponential strand displacement amplification, and the like. The index amplification technology reacts under the constant temperature condition without thermal cycling, so that the application scene of the nucleic acid detection technology is expanded, and the potential of POCT application is stimulated. The molecular diagnosis has the characteristics of high sensitivity and strong specificity, and if the molecular diagnosis is integrated with the advantage of POCT (point of care testing) rapidly, the molecular diagnosis will certainly occupy important position in the detection field.
POCT refers to a detection mode which is performed on a sampling site and can rapidly obtain a detection result by using a portable analysis instrument and a matched reagent. POCT meaning can be understood from two aspects: spatially, the test is performed at the patient's side, i.e. "bedside test"; in time, a "point-in-time test" may be performed.
The nucleic acid amplification product based on isothermal amplification technology can detect single-molecule nucleic acid at a constant and low temperature in only 30 min minutes. The current nucleic acid amplification products in China have low requirements on environment and hardware facilities, and have good application in aspects of biological protection, water body inspection, food inspection, medical diagnosis, microfluidics, veterinarian and the like; however, the existing nucleic acid amplification products consider the problems of application scenes and cost, a colloidal gold test strip is used for terminal detection, the integration of amplification and test strip detection is realized, the general scene requirement is met, the cost is reduced, and the defects are exposed when the product is applied to the nucleic acid detection of the novel coronavirus: the amplified product needs to be uncapped by a worker, and the product is put into a corresponding portable instrument by the worker, so that false positives of detection can be increased, a laboratory can be polluted, and therefore, the problem of pollution caused by the amplified product is solved by commonly 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 complex operation and low safety.
Disclosure of Invention
The invention aims to provide a detection device and a detection method for a nucleic acid amplification product, which are used for solving the problems of complicated operation and low safety of the nucleic acid amplification product in the prior art.
To achieve the purpose, the invention adopts the following technical scheme:
The detection device for the nucleic acid amplification product comprises a sample storage unit and a detection part which are detachably connected, wherein a closed cavity is formed in the sample storage unit, a detection cavity provided with a fourth opening is formed in the detection part, and a detection unit is arranged in the detection cavity;
When the sample storage unit is connected with the detection part, one end of the sample storage unit is sleeved in the fourth opening, and the closed cavity is communicated with the detection cavity;
When the puncture rod body is not pushed, the sample storage unit comprises an amplifying part and a diluting part which are detachably connected, an amplifying cavity with a first opening is formed in the amplifying part, and a diluting cavity with a second opening and a third opening is formed in the diluting part; a sealing film for separating the amplification cavity from the dilution cavity is arranged on the second opening, and a puncturing part for puncturing the sealing film is movably sleeved on the third opening;
When the amplifying part is connected with the diluting part, the puncturing part can move along the direction close to the second opening so as to puncture the sealing film, and the amplifying cavity is communicated with the diluting cavity to form the closed cavity.
Optionally, the piercing part comprises a piercing rod body, and a piercing cone, a first limiting part and a second limiting part are sequentially arranged on the piercing rod body along the direction away from the sealing film;
The puncture cone is positioned in the dilution cavity and used for puncturing the sealing film; the first limiting part is positioned in the dilution cavity and used for preventing the puncture rod body from sliding out of the closed cavity; the second limiting part is positioned outside the dilution cavity and used for preventing the puncture cone from being contacted with the sealing film before dilution.
Optionally, the first limiting part comprises a limiting plate protruding on the puncture rod body, and the length of the limiting plate is greater than the caliber of the third opening;
The second limiting part comprises a first pin hole formed in the puncture rod body and a first bayonet lock movably inserted into the first pin hole;
When the sealing film is not punctured, the limiting plate is arranged in the dilution cavity and is abutted to the edge of the third opening, and the first bayonet lock is arranged outside the dilution cavity and is abutted to the edge of the third opening.
Optionally, the length of the limiting plate is smaller than the caliber of the second opening.
Optionally, a third limiting portion is further formed on the puncture rod body at a side, far away from the sealing film, of the second limiting portion, and the third limiting portion includes a second pin hole formed in the puncture rod body and a second bayonet lock movably inserted into the second pin hole.
Optionally, the detection unit comprises detection test paper, a baffle is arranged in the detection part, and the baffle separates the detection cavity to form a buffer pool and a detection pool;
the baffle is provided with an avoidance hole, the detection test paper passes through the avoidance hole and extends from the buffer pool to the detection pool;
the detection part comprises a communication part, and when the sample storage unit is connected with the detection part, the sample storage unit is communicated with the buffer pool through the communication part.
Optionally, the communicating portion extends inwards from a cavity wall of the fourth opening, a protruding portion is further protruding on one side, close to the piercing portion, of the communicating portion, a flow hole is formed in the protruding portion, corresponding to the third opening, and the flow hole is communicated with the buffer tank;
When the sample storage unit is connected with the detection part, the dilution part is inserted into the fourth opening, the puncture part is lifted by the protruding part, and the flow hole is communicated with the closed cavity.
Optionally, a drainage unit is further disposed in the detection portion, one end of the drainage unit is located in the detection tank, and the other end of the drainage unit is inserted into the flow hole.
Optionally, the communication part comprises a puncture unit arranged in the detection cavity;
When the sample storage unit is connected with the detection part, the amplification part is inserted into the fourth opening, the puncture unit is inserted into the amplification cavity, and the buffer pool is communicated with the closed cavity.
Optionally, the dodging hole is formed in the end portion, away from the bottom wall of the detection cavity, of the baffle, the detection test paper is located at a first interval between one end portion of the detection tank and the bottom wall of the detection cavity is greater than a second interval between the other end portion of the buffer tank and the bottom wall of the detection cavity.
Optionally, in the detection tank, the detection part is further provided with a test paper rack and a flange part; the test paper rack bears the test paper, flange portion set up in the both ends of test paper.
A detection method applied to the detection device for the nucleic acid amplification product comprises the following steps:
Adding a nucleic acid sample and an amplification reagent into the amplification part, connecting the amplification part with the dilution part to form the sample storage unit, and shaking uniformly to complete a nucleic acid amplification reaction;
The puncture part moves along the direction close to the second opening, so that the puncture part punctures the sealing film, the amplification cavity is communicated with the dilution cavity to form the closed cavity, and the sealing film is uniformly shaken to finish dilution;
and connecting the sample storage unit with the detection part, communicating the closed cavity with the detection cavity, and enabling the sample in the closed cavity to flow into the detection cavity and perform detection reaction with the detection unit to finish detection.
The detection method is applied to the detection device for the nucleic acid amplification product, wherein the amplification part is pre-stored with an amplification reagent, and the detection method comprises the following steps:
releasing the connection between the amplifying part and the diluting part, adding a nucleic acid sample into the amplifying part, connecting the amplifying part and the diluting part to form a sample storage unit, and shaking uniformly to complete the nucleic acid amplification reaction;
The puncture part moves along the direction close to the second opening, so that the puncture part punctures the sealing film, the amplification cavity is communicated with the dilution cavity to form a closed cavity, and the sealing film is uniformly shaken to finish dilution;
And connecting the sample storage unit with the detection part, wherein the closed cavity is communicated with the detection cavity, and the sample in the closed cavity flows into the detection cavity and performs detection reaction with the detection unit to finish detection.
Compared with the prior art, the invention has the following beneficial effects:
The invention provides a detection device and a detection method for a nucleic acid amplification product, in particular to a detection device and a detection method for the nucleic acid amplification product, which are characterized in that a nucleic acid sample and an amplification reagent are added into an amplification part, the amplification part and a dilution part are connected to form a sample storage unit, and the sample storage unit is uniformly shaken to complete a nucleic acid amplification reaction; the puncture part is moved along the direction close to the second opening, so that the puncture part punctures the sealing film, the amplification cavity is communicated with the dilution cavity to form a closed cavity, and the dilution is completed by shaking; the sample storage unit is connected with the detection part, the closed cavity is communicated with the detection cavity, and the sample in the closed cavity flows into the detection cavity and is subjected to detection reaction with the detection test paper to finish detection. The nucleic acid sample is always in the airtight cavity after being added into the amplifying part, so that leakage can not be caused when the nucleic acid sample is amplified and diluted, the safety is high, and the whole operation process of the detecting device after the nucleic acid sample is added is only required to carry out two connecting actions and one puncturing action, so that the action of uncovering is omitted, the operation complexity of nucleic acid detection is reduced, the leakage risk is further avoided, and the detecting device and the detecting method for the nucleic acid amplified product are simple and convenient to operate and high in safety.
Drawings
In order to more clearly illustrate the embodiments of the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only some embodiments of the invention, and that other drawings can be obtained from these drawings without inventive faculty for a person skilled in the art.
The structures, proportions, sizes, etc. shown in the drawings are shown only in connection with the present disclosure, and are not intended to limit the scope of the invention, since any modification, variation in proportions, or adjustment of the size, etc. of the structures, proportions, etc. should be considered as falling within the spirit and scope of the invention, without affecting the effect or achievement of the objective.
Fig. 1 is a schematic diagram of the overall structure of a detection device according to an embodiment of the present invention;
Fig. 2 is a schematic diagram of a front view cross-section structure of a detection device according to an embodiment of the present invention;
FIG. 3 is a schematic view of the partial enlarged structure of FIG. 2 at A;
fig. 4 is a schematic diagram of a right-side cross-sectional structure of a detection device according to an embodiment of the present invention;
FIG. 5 is a schematic diagram illustrating a first connection status structure of a sample storage unit according to an embodiment of the present invention;
FIG. 6 is a schematic diagram showing a second connection status structure of a sample storage unit according to an embodiment of the present invention;
FIG. 7 is a schematic diagram of the structure of an amplifying section according to an embodiment of the present invention;
FIG. 8 is a schematic view of a dilution unit according to an embodiment of the invention;
fig. 9 is a schematic front view of another detecting device according to an embodiment of the present invention;
FIG. 10 is a schematic diagram illustrating a front sectional structure of another detecting device according to an embodiment of the present invention;
fig. 11 is a schematic right-view cross-sectional structure of another detecting device according to an embodiment of the present invention.
Illustration of: 1. an amplification unit; 11. a first opening; 12. an amplification chamber;
2. a dilution section; 21. a second opening; 22. a third opening; 23. a dilution chamber;
3. A puncturing portion; 31. a puncture cone; 32. a first limit part; 33. a second limit part; 4. a sealing film;
5. A detection unit; 51. a buffer pool; 52. a detection pool; 53. a baffle; 54. detecting test paper; 55. a boss; 56. a drainage unit; 57. a puncturing unit; 6. and (3) sealing rings.
Detailed Description
In order to make the objects, features and advantages of the present invention more comprehensible, the technical solutions in the embodiments of the present invention are described in detail below with reference to the accompanying drawings, and it is apparent that the embodiments described below are only some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the description of the present invention, it should be understood that the directions or positional relationships indicated by the terms "upper", "lower", "top", "bottom", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as limiting the present invention. It is 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 invention is further described below by the specific embodiments with reference to the accompanying drawings.
Referring to fig. 1 to 11, fig. 1 is a schematic overall structure of a detection device according to an embodiment of the present invention, fig. 2 is a schematic front view cross-sectional structure of a detection device according to an embodiment of the present invention, fig. 3 is a schematic partial enlarged structure of a portion of fig. 2 at a, fig. 4 is a schematic right view cross-sectional structure of a detection device according to an embodiment of the present invention, fig. 5 is a schematic first connection state structure of a sample storage unit according to an embodiment of the present invention, fig. 6 is a schematic second connection state structure of a sample storage unit according to an embodiment of the present invention, fig. 7 is a schematic structure of an amplifying portion according to an embodiment of the present invention, fig. 8 is a schematic structure of a dilution portion according to an embodiment of the present invention, fig. 9 is a schematic front view cross-sectional structure of another detection device according to an embodiment of the present invention, fig. 10 is a schematic front view cross-sectional structure of another detection device according to an embodiment of the present invention, and fig. 11 is a schematic right view cross-sectional structure of another detection device according to an embodiment of the present invention.
Example 1
The detection device for the nucleic acid amplification product provided by the embodiment adopts a constant-temperature nucleic acid amplification technology, can amplify, dilute and detect a nucleic acid product in a normal-temperature environment, reduces the risk of pollutant diffusion and simplifies the operation steps by improving the detection device.
As shown in fig. 1 to 4, 7 and 8, the detection device for nucleic acid amplification product of the present embodiment includes a sample storage unit and a detection unit 5 detachably connected, a closed cavity is formed in the sample storage unit, a detection cavity configured with a fourth opening is formed in the detection unit 5, and a detection unit is disposed in the detection cavity. The sample storage unit can store a nucleic acid sample and amplify and dilute the nucleic acid sample; the detection unit refers to a unit capable of performing a detection reaction on a sample, and specifically includes a detection strip 54 or a test tube filled with a detection reagent.
As shown in fig. 2 and 9, when the sample storage unit is connected to the detection portion 5, one end of the sample storage unit is sleeved in the fourth opening, and the closed cavity is communicated with the detection cavity. The joint of the sample storage unit and the detection part 5 is provided with a sealing ring 6, so that the tightness of the detection device can be further improved; the detection part 5 is provided with an observation window which is made of transparent materials, the observation window is in sealing connection with the detection part 5, and workers can observe the change of the test paper from the outside.
As shown in fig. 5 to 8, the sample storage unit includes an amplification section 1 and a dilution section 2 detachably connected, an amplification chamber 12 in which a first opening 11 is arranged is formed in the amplification section 1, and a dilution chamber 23 in which a second opening 21 and a third opening 22 are arranged is formed in the dilution section 2; the second opening 21 is provided with a sealing membrane 4 for separating the amplification chamber 12 from the dilution chamber 23, and the third opening 22 is movably sleeved with a puncturing part 3 for puncturing the sealing membrane 4. The first opening 11 is matched with the second opening 21, that is, the first opening 11 can be directly abutted with the second opening 21, in this embodiment, the cavity wall of the amplifying part 1 in the first opening 11 can be sleeved in the cavity wall of the diluting part 2 in the second opening 21, so that the stability of connection is improved; the amplification reagent is arranged in the amplification cavity 12, the buffer solution is arranged in the dilution cavity 23, the sample is obtained after the buffer solution is mixed with the amplification product, and the sensitivity and the detection rate of the buffer solution can be improved.
When the amplification part 1 is connected to the dilution part 2, the puncturing part 3 can move in a direction approaching the second opening 21 to puncture the sealing membrane 4, and the amplification chamber 12 communicates with the dilution chamber 23 to form a closed chamber. It should be noted that the sealing ring 6 is provided at the joint between the amplification unit 1 and the dilution unit 2, so that the sealability of the sample storage unit can be further improved.
Specifically, when nucleic acid detection is performed, a nucleic acid sample and an amplification reagent are added into an amplification part 1, the amplification part 1 and a dilution part 2 are connected to form a sample storage unit, and the sample storage unit is uniformly shaken to complete a nucleic acid amplification reaction; the puncture part 3 is moved along the direction close to the second opening 21, so that the puncture part 3 punctures the sealing membrane 4, the amplification cavity 12 is communicated with the dilution cavity 23 to form a closed cavity, and the closed cavity is uniformly shaken to finish dilution; the sample storage unit is connected with the detection part 5, the closed cavity is communicated with the detection cavity, and the sample in the closed cavity flows into the detection cavity and performs detection reaction with the detection test paper 54 to complete detection. The nucleic acid sample is always in the airtight cavity after being added into the amplifying part 1, so that leakage can not be caused when the nucleic acid sample is amplified and diluted, the safety is high, and the whole operation process of the detecting device after the nucleic acid sample is added only needs to carry out two connecting actions and one puncturing action, so that the action of uncovering is omitted, the operation complexity of nucleic acid detection is reduced, the leakage risk is further avoided, and the detecting device and the detecting method for the nucleic acid amplified product are simple and convenient to operate and high in safety.
Further, as shown in fig. 2, 5 and 6, the piercing portion 3 includes a piercing rod body, and a piercing cone 31, a first limiting portion 32 and a second limiting portion 33 are sequentially disposed on the piercing rod body along a direction away from the sealing film 4.
When the puncture rod body is not pushed, the puncture cone 31 is positioned in the dilution cavity 23 and is used for puncturing the sealing membrane 4; the first limiting part 32 is positioned in the dilution cavity 23 and is used for preventing the puncture rod body from sliding out of the closed cavity; the second stopper 33 is located outside the dilution chamber 23 for preventing the puncture cone 31 from contacting the sealing membrane 4 before diluting the amplified product (amplified nucleic acid sample).
Specifically, after the amplification part 1 and the dilution part 2 are connected to form a sample storage unit, the first limiting part 32 prevents the puncture rod body from sliding out of the closed cavity, so that liquid in the dilution cavity 23 is prevented from leaking, the second limiting part 33 prevents the puncture rod body from sliding into the closed cavity completely, and also prevents the puncture cone 31 from puncturing the sealing film 4 in advance before the nucleic acid sample is amplified, so that the stability of the sample storage unit is improved; when the amplified product (amplified nucleic acid sample) needs to be diluted, the worker releases the locking of the second limiting part 33 and pushes the puncture push rod, so that the puncture cone 31 punctures the sealing membrane 4, the amplified cavity 12 is communicated with the dilution cavity 23 to form a closed cavity, and the amplified product is mixed with the buffer solution to dilute the amplified product.
Further, the first limiting portion 32 includes a limiting plate protruding on the piercing rod body, and the length of the limiting plate is greater than the caliber of the third opening 22.
The second limiting portion 33 includes a first pin hole formed in the piercing rod body and a first bayonet lock movably inserted into the first pin hole.
When the sealing film 4 is not pierced, the limiting plate is arranged in the dilution cavity 23 and is abutted with the edge of the third opening 22, and the first bayonet lock is arranged outside the dilution cavity 23 and is abutted with the edge of the third opening 22.
Specifically, the limiting plate cooperates with the first bayonet to limit the puncturing part 3 to a fixed position before the amplified product (amplified nucleic acid sample) needs to be diluted, i.e., when the sealing membrane 4 is not punctured. Meanwhile, the limiting plate can be tightly attached to the third opening 22, so that the tightness between the third opening 22 and the puncturing part 3 is enhanced, and the liquid in the dilution cavity 23 is prevented from leaking to the outside or being polluted by the outside in the transportation process before use.
Further, the length of the limiting plate is smaller than the caliber of the second opening 21. Specifically, when the worker pushes the puncture push rod, the puncture cone 31 and the limiting plate are contacted with the sealing membrane 4 in sequence, so that the first limiting part 32 can prevent the puncture cone 31 from sliding from the dilution part 2 to the outside, and can further advance after the puncture cone 31 punctures the sealing membrane 4, so that the notch on the sealing membrane 4 becomes larger, the buffer solution is contacted with the amplified product more easily, and the mixing speed is accelerated. It will also be appreciated that if the size of the puncture cone 31 is merely increased to increase the gap in the sealing membrane 4, it may happen that the puncture cone 31 is caught by the second opening 21, and by the above arrangement, the rate of mixing is increased while ensuring a stable operation of the detection device.
Further, a third limiting portion (not shown) is further formed on the piercing rod body at a side of the second limiting portion 33 away from the sealing film 4, and the third limiting portion includes a second pin hole formed on the piercing rod body and a second bayonet lock movably inserted into the second pin hole. Specifically, the second bayonet lock can prevent the puncture part 3 from completely falling into the dilution cavity 23 after the sealing film 4 is punctured, and the second bayonet lock can also enable one end of the puncture rod part to be always located outside the dilution part 2, so that sealing connection is always formed between the puncture rod body and the third opening 22, and the sealing performance and stability of the detection device are improved. When there is a case where it is necessary to take out the content of the detecting device, the second bayonet is pulled out, and the end of the piercing rod is pushed to enter the diluting chamber 23 entirely, so that the third opening 22 of the diluting part 2 is opened, and the content is taken out.
Further, as shown in fig. 2 and 3, a baffle 53 is provided in the detection portion 5, and the baffle 53 partitions the detection chamber into a buffer tank 51 and a detection tank 52.
The baffle 53 is provided with an avoidance hole, and the test paper 54 passes through the avoidance hole and extends from the buffer tank 51 to the detection tank 52. It is to be added that a sealing ring is arranged between the avoidance hole and the detection test paper 54, and the sealing ring can fill the residual gap of the avoidance hole, but can not press the detection test paper 54, so that the tightness and stability of the detection device are improved.
The detection section 5 includes a communication section through which the sample storage unit communicates with the buffer pool 51 when the sample storage unit is connected to the detection section 5.
Specifically, after the sample storage unit is connected with the detection part 5, the communication part enables the closed cavity of the sample storage unit to be communicated with the buffer pool 51, so that samples in the closed cavity flow to the buffer pool 51 and then flow into the detection pool 52, and the phenomenon of sample flushing is effectively prevented through the arrangement of the buffer pool 51.
In a specific embodiment, as shown in fig. 1 to 8, the communicating portion extends inward from the cavity wall of the fourth opening, a protruding portion 55 is further protruding from a side of the communicating portion near the piercing portion 3, and a through hole is formed in the protruding portion 55 at a position corresponding to the third opening 22, and the through hole is in communication with the buffer pool 51.
When the sample storage unit is connected with the detection part 5, the dilution part 2 is inserted into the fourth opening, the puncture part 3 is lifted up by the bulge part 55, and the circulation hole is communicated with the closed cavity.
Specifically, when the sample storage unit is connected to the detecting portion 5, the protruding portion 55 can push the piercing portion 3 into the closed cavity, so that the third opening 22 is no longer plugged by the piercing rod, and the sample in the closed cavity flows out of the third opening 22 and flows into the buffer cell 51 through the flow hole. Meanwhile, the sealing ring 6 is sleeved outside the boss 55, and the sealing ring 6 is connected with the dilution part 2, so that the tightness between the detection part 5 and the dilution part 2 can be improved. In the process, the step of uncovering by a worker is omitted, the risk of leakage of pollutants is reduced, the steps are saved, and the safety is improved.
Further, a drainage unit 56 is further provided in the detection portion 5, one end of the drainage unit 56 is located in the detection cell 52, and the other end is inserted into the flow hole. The drainage unit 56 may be a sponge, cotton, or other absorbent material. The drainage unit 56 is equivalent to a "capacity pool", on one hand, through the water absorption capacity, the sample is moved directionally to prevent the leakage of pollutants, and on the other hand, the rate of reaching the detection part 5 of the sample can be regulated and controlled to prevent the phenomenon of sample flushing; and the sample storage unit is in a sealing state when being connected with the detection part 5, and by arranging the drainage unit 56, the problem that liquid in the amplification part 1 is not easy to flow into the detection part 5 due to atmospheric pressure can be overcome, so that the drainage unit 56 can continuously absorb samples in the closed cavity and drain the samples to the buffer pool 51, and the detection efficiency of the detection device is improved.
In another specific embodiment, as shown in fig. 9 to 11, the communicating portion includes a puncturing unit 57 provided in the detection chamber.
When the sample storage unit is connected to the detection unit 5, the amplification unit 1 is inserted into the fourth opening, the puncture unit 57 is inserted into the amplification chamber 12, and the buffer tank 51 communicates with the closed chamber. Wherein, one end part of the amplifying part 1 far away from the diluting part 2 is matched with the size of the fourth opening, so that the amplifying part 1 can be inserted into the fourth opening; after the amplification part 1 is inserted into the fourth opening, the puncturing unit 57 punctures the amplification part 1 so that the sample in the amplification chamber 12 flows into the closed chamber.
Further, as shown in fig. 2 and 3, the avoidance hole is formed on the end portion of the baffle 53 away from the bottom wall of the detection cavity, and a first distance between one end portion of the detection tank 52 and the bottom wall of the detection cavity is larger than a second distance between the other end portion of the detection tank 51 and the bottom wall of the detection cavity, where the detection test paper 54 is located.
In the detection tank 52, the detection part 5 is also provided with a test paper rack and a flange part; the test paper rack carries test paper 54, and the flange parts are arranged at two ends of the test paper 54.
It should be understood that the test paper 54 can be avoided Kong Taisheng to form a certain angle with the horizontal plane, so that the phenomenon of flushing can be effectively prevented. The test strip holder is formed by a bottom plate or boss that can hold the test strip overhead and prevent the test strip 54 from contacting the bottom wall of the test cell 52. And be formed with the test paper groove between flange portion and the test paper frame at test paper 54 both ends, this test paper groove can effectively restrict the orbit of sample liquid, prevents on the one hand that the sample from contacting with the four walls of detection pond 52, can prevent that the sample from leaking from the four walls of detection pond 52, on the other hand forms capillary action, makes the sample flow on test paper 54 fast to the detection rate has been accelerated, thereby has improved detection device's security and detection efficiency.
In summary, the detection device provided in this embodiment has the advantages of simple operation and high safety, and simultaneously has the characteristics of high detection efficiency, high stability, simple operation and high precision.
Example two
The embodiment provides a detection method applied to a detection device for a nucleic acid amplification product in an implementation, which comprises the following steps:
S11, adding a nucleic acid sample and an amplification reagent into the amplification part 1, connecting the amplification part 1 with the dilution part 2 to form a sample storage unit, and shaking uniformly to complete a nucleic acid amplification reaction; wherein, before adding the nucleic acid sample and the amplification reagent to the amplification part 1, the amplification part 1 can be pre-connected with the dilution part 2 to form a sample storage unit, or can be stored separately;
s12, enabling the puncture part 3 to move along the direction close to the second opening 21, enabling the puncture part 3 to puncture the sealing film 4, enabling the amplification cavity 12 to be communicated with the dilution cavity 23 to form a closed cavity, shaking uniformly, and completing dilution;
And S13, connecting the sample storage unit with the detection part 5, communicating the closed cavity with the detection cavity, enabling the sample in the closed cavity to flow into the detection cavity, and carrying out detection reaction with the detection unit to finish detection.
In the first embodiment, the specific structure and technical effects of the detection device for a nucleic acid amplification product are described, and the detection method is realized by the detection device for a nucleic acid amplification product, which also has the technical effects.
Additionally, another way exists for the detection method:
The amplification part 1 of the sample storage unit is pre-stored with the amplification reagent in advance, and at this time, before detection, the amplification part 1 and the dilution part 2 are required to be connected to form the sample storage unit, so that the amplification reagent is prevented from being polluted;
s21, disconnecting the amplification part 1 from the dilution part 2, adding a nucleic acid sample into the amplification part 1, connecting the amplification part 1 with the dilution part 2 to form a sample storage unit, and shaking uniformly to complete the nucleic acid amplification reaction;
S22, enabling the puncture part 3 to move along the direction close to the second opening 21, enabling the puncture part 3 to puncture the sealing film 4, enabling the amplification cavity 12 to be communicated with the dilution cavity 23 to form a closed cavity, shaking uniformly, and completing dilution;
S23, connecting the sample storage unit with the detection part 5, communicating the closed cavity with the detection cavity, enabling the sample in the closed cavity to flow into the detection cavity, and carrying out detection reaction with the detection unit to finish detection.
In summary, the detection device provided in this embodiment has the advantages of simple operation and high safety, and simultaneously has the characteristics of high detection efficiency, high stability, simple operation and high precision.
The above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the 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 scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (6)
1. The detection device for the nucleic acid amplification product is characterized by comprising a sample storage unit and a detection part (5) which are detachably connected, wherein a closed cavity is formed in the sample storage unit, a detection cavity provided with a fourth opening is formed in the detection part (5), and a detection unit is arranged in the detection cavity;
when the sample storage unit is connected with the detection part (5), one end of the sample storage unit is sleeved in the fourth opening, and the closed cavity is communicated with the detection cavity;
the sample storage unit comprises an amplifying part (1) and a diluting part (2) which are detachably connected, an amplifying cavity (12) for configuring a first opening (11) is formed in the amplifying part (1), and a diluting cavity (23) for configuring a second opening (21) and a third opening (22) is formed in the diluting part (2); the second opening (21) is provided with a sealing film (4) for separating the amplification cavity (12) and the dilution cavity (23), and the third opening (22) is movably sleeved with a puncturing part (3) for puncturing the sealing film (4);
The puncture part (3) comprises a puncture rod body, and a puncture cone (31), a first limit part (32) and a second limit part (33) are sequentially arranged on the puncture rod body along the direction away from the sealing film (4);
When the puncture rod body is not pushed, the puncture cone (31) is positioned in the dilution cavity (23) and used for puncturing the sealing membrane (4); the first limiting part (32) is positioned in the diluting cavity (23) and is used for preventing the puncture rod body from sliding out of the closed cavity; the second limiting part (33) is positioned outside the diluting cavity (23) and is used for preventing the puncture cone (31) from contacting with the sealing film (4) before dilution;
the first limiting part (32) comprises a limiting plate which is convexly arranged on the puncture rod body, and the length of the limiting plate is larger than the caliber of the third opening (22);
The second limiting part (33) comprises a first pin hole formed in the piercing rod body and a first bayonet lock movably inserted into the first pin hole;
When the sealing film (4) is not pierced, the limiting plate is arranged in the dilution cavity (23) and is abutted against the edge of the third opening (22), and the first bayonet lock is arranged outside the dilution cavity (23) and is abutted against the edge of the third opening (22);
The length of the limiting plate is smaller than the caliber of the second opening (21);
A third limiting part is further formed on the puncture rod body at one side of the second limiting part (33) away from the sealing film (4), and comprises a second pin hole formed on the puncture rod body and a second bayonet lock movably inserted into the second pin hole;
the detection unit comprises detection test paper (54), a baffle (53) is arranged in the detection part (5), and the baffle (53) separates the detection cavity to form a buffer pool (51) and a detection pool (52);
The baffle plate (53) is provided with an avoidance hole, the detection test paper (54) passes through the avoidance hole and extends from the buffer tank (51) to the detection tank (52);
The detection part (5) comprises a communication part, and when the sample storage unit is connected with the detection part (5), the sample storage unit is communicated with the buffer pool (51) through the communication part;
The communication part extends inwards from the cavity wall of the fourth opening, a protruding part (55) is further protruding from one side of the communication part, which is close to the puncture part (3), a flow hole is formed in the protruding part (55) at a position corresponding to the third opening (22), and the flow hole is communicated with the buffer tank (51);
when the sample storage unit is connected with the detection part (5), the dilution part (2) is inserted into the fourth opening, the puncture part (3) is lifted by the bulge part (55), and the flow hole is communicated with the closed cavity;
the avoidance hole is formed in the end portion, away from the bottom wall of the detection cavity, of the baffle plate (53), the first interval between one end portion of the detection pool (52) and the bottom wall of the detection cavity is larger than the second interval between the other end portion of the buffer pool (51) and the bottom wall of the detection cavity, of the detection test paper (54).
2. The device for detecting a nucleic acid amplification product according to claim 1, wherein a drainage unit (56) is further provided in the detection section (5), one end of the drainage unit (56) is positioned in the detection cell (52), and the other end is inserted into the flow hole.
3. The detection apparatus for a nucleic acid amplification product according to claim 1, wherein the communication section includes a puncturing unit (57) provided in the detection chamber;
When the sample storage unit is connected with the detection part (5), the amplification part (1) is inserted into the fourth opening, the puncture unit (57) is inserted into the amplification cavity (12), and the buffer pool (51) is communicated with the closed cavity.
4. The detection apparatus for a nucleic acid amplification product according to claim 1, wherein the detection section (5) is further provided with a test strip holder and a flange section in the detection cell (52); the test paper rack bears the test paper (54), and the flange parts are arranged at two ends of the test paper (54).
5. A method for detecting a non-therapeutic target nucleic acid, which is applied to the detection device for a nucleic acid amplification product according to any one of claims 1 to 4, comprising:
Adding a nucleic acid sample and an amplification reagent into the amplification part, connecting the amplification part with the dilution part to form the sample storage unit, and shaking uniformly to complete a nucleic acid amplification reaction;
The puncture part moves along the direction close to the second opening, so that the puncture part punctures the sealing film, the amplification cavity is communicated with the dilution cavity to form the closed cavity, and the sealing film is uniformly shaken to finish dilution;
and connecting the sample storage unit with the detection part, communicating the closed cavity with the detection cavity, and enabling the sample in the closed cavity to flow into the detection cavity and perform detection reaction with the detection unit to finish detection.
6. The method for detecting a non-therapeutic target nucleic acid according to claim 5, wherein the amplification unit has an amplification reagent pre-stored therein,
And disconnecting the amplification part from the dilution part, and adding a nucleic acid sample into the amplification part.
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| CN115466671B (en) * | 2022-09-28 | 2023-12-15 | 广纳达康(广州)生物科技有限公司 | Anti-flushing liquid nucleic acid amplification product detection device and detection method |
| CN115678751B (en) * | 2022-12-30 | 2023-04-18 | 北京金诺美科技股份有限公司 | Sample tube |
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