CN115181652A - Detection device and detection method - Google Patents
Detection device and detection method Download PDFInfo
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- CN115181652A CN115181652A CN202110366006.0A CN202110366006A CN115181652A CN 115181652 A CN115181652 A CN 115181652A CN 202110366006 A CN202110366006 A CN 202110366006A CN 115181652 A CN115181652 A CN 115181652A
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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 provides a detection device and a detection method, wherein the detection device comprises a shell, a cracking cup, an amplification cup, a heating unit, a buffer cup, a liquid storage cup, a color development test paper and a puncture needle for sequentially puncturing the cracking cup, the amplification cup and the buffer cup; the detection method comprises the following steps: sampling, lysis, amplification, dilution and detection. The detection device and the detection method can realize integrated detection operation, the detection device has small volume, can carry out detection on site, does not need to be transferred to a laboratory, has short detection time, can obtain a detection result in time, and has high detection efficiency.
Description
Technical Field
The invention belongs to the technical field of material moving detection equipment, and particularly relates to a detection device and a detection method.
Background
When judging whether a human body is infected with virus, a nucleic acid detection method is generally adopted, and the current nucleic acid detection method is to transfer an acquired detection sample to a laboratory and analyze the sample through large-scale equipment, so that the nucleic acid detection time is long, a detection result cannot be obtained in time, the optimal time for isolation and treatment is delayed, and the risk of virus propagation is increased.
Disclosure of Invention
The invention aims to provide a detection device and a detection method, and aims to solve the problems that in the prior art, the detection time is long, and the detection result cannot be obtained in time.
In order to achieve the purpose, the invention provides a detection device which comprises a shell, a cracking cup for cracking a detection sample, an amplification cup for amplifying the detection sample to form amplification liquid, a heating unit for heating the amplification cup, a buffer cup for diluting the amplification liquid to reaction liquid, a liquid storage cup for containing the reaction liquid, a color development test paper for reacting with the reaction liquid to display a detection result, and a puncture needle for sequentially puncturing the cracking cup, the amplification cup and the buffer cup, wherein the cracking cup, the amplification cup, the buffer cup and the liquid storage cup are arranged in the shell and sequentially and correspondingly arranged from the top of the shell to the bottom of the shell, and a liquid suction end for developing color extends into the liquid storage cup.
The detection apparatus as described above, wherein the top and bottom of the lysis cup, the top and bottom of the amplification cup, and the top and bottom of the buffer cup are sealed by thin film layers.
The detection device as described above, wherein a window is provided on the housing, the window is opposite to the color development test paper, and the window is closed by a transparent plate.
The detection device as described above, wherein the heating unit is disposed in the casing, and the heating unit includes a heating sleeve sleeved on the amplification cup, a heating control board electrically connected to the heating sleeve, and a control switch, a power interface and an indicator light electrically connected to the heating control board respectively.
The detection device as described above, wherein the casing is provided with a first accommodating chamber and a second accommodating chamber which are not communicated with each other, the puncture needle is inserted into the first accommodating chamber, and the lysis cup, the amplification cup, the buffer cup and the liquid storage cup are fixed in the second accommodating chamber.
The detection device as described above, wherein the housing is formed by two housing bodies that are oppositely fastened, the two housing bodies are detachably connected, and the lysis cup, the amplification cup, the buffer cup and the liquid storage cup are fixedly clamped between the two housing bodies.
The detection apparatus as described above, wherein the lysis cup contains a lysis solution, and the lysis solution is a solution formed by diluting tris (2-carboxyethyl) phosphine hydrochloride, diethylamine tetraacetic acid, sodium hydroxide, and protease in diethylpyrocarbonate water.
The detection device as described above, wherein the amplification cup contains amplification powder, the amplification powder comprises a primer mixture and a reaction mixture, the primer mixture comprises a front inner primer, a rear inner primer, a front outer primer, a rear outer primer, a front circular primer and a rear circular primer; the reaction mixture includes deoxyribonucleic acid polymerase, reverse transcriptase, tris (hydroxymethyl) aminomethane hydrochloride, ammonium sulfate, potassium chloride, magnesium sulfate, and tween 20.
The invention also provides a detection method which adopts the detection device and comprises the following steps: sampling: collecting a detection sample on a detected person by using a swab; cracking: inserting the swab into the lysis cup, wherein the lysis solution in the lysis cup lyses the detection sample; amplification: puncturing the cracking cup by using the puncture needle, so that the cracking cup is communicated with the amplification cup, and the lysate in the cracking cup flows into the amplification cup; opening the heating unit to heat the amplification cup, and amplifying the detection sample in the lysis solution after the amplification powder in the amplification cup is heated to form an amplification solution; diluting: continuously pressing the puncture needle to puncture the amplification cup to enable the amplification cup to be communicated with the buffer solution cup, enabling amplification solution in the amplification cup to flow into the buffer solution cup, and diluting the amplification solution by using buffer solution in the buffer solution cup to form reaction solution; and (3) detection: and continuously pressing the puncture needle to puncture the buffer cup to enable the buffer cup to be communicated with the liquid storage cup, wherein the reaction liquid in the buffer cup flows into the liquid storage cup and reacts with the color test paper in the liquid storage cup, and the color test paper displays a detection result.
The detection method as described above, wherein a pH of a solution obtained by mixing the lysate with the amplification powder after the lysate flows into the amplification cup is 8.5 to 9.
The detection device and the detection method of the invention have the characteristics and advantages that:
1. according to the invention, the puncture needle, and the lysis cup, the amplification cup, the buffer cup and the liquid storage cup which correspond in sequence are arranged, so that during detection, a swab is adopted to collect a detection sample from a detection person, and the swab with the detection sample can be cracked by a lysate in the lysis cup; after the puncture needle pierces the lysis cup to communicate with the amplification cup, the lysis solution can flow into the amplification cup, and the amplification cup amplifies the detection sample in the lysis solution to form amplification solution under the heating of the heating unit; then the amplification cup is pierced by the puncture needle to be communicated with the buffer cup, and then the amplification liquid flows into the buffer cup and is diluted into reaction liquid; and after the puncture needle pierces the buffer liquid cup to be communicated with the liquid storage cup, the reaction liquid flows into the liquid storage cup and reacts with the color test paper to display the detection result. Therefore, the detection device can realize integrated detection operation, has small volume, can perform on-site detection, does not need to be transferred to a laboratory, has short detection time, can obtain a detection result in time and has high detection efficiency;
2. the window and the transparent plate are arranged, so that the detection result and the state of each component in the shell can be observed conveniently;
3. according to the invention, the cracking cup, the amplification cup, the buffer solution cup and the liquid storage cup are clamped by the two detachably connected shell bodies, so that the cracking cup, the amplification cup, the buffer solution cup and the liquid storage cup can be conveniently installed, maintained and replaced.
Drawings
The drawings are only for purposes of illustrating and explaining the present invention and are not to be construed as limiting the scope of the present invention. Wherein:
FIG. 1 is a schematic diagram of an external structure of a detecting device according to an embodiment of the present invention;
FIG. 2 is a schematic plan view of the internal structure of a detection apparatus according to an embodiment of the present invention;
FIG. 3 is a schematic perspective view of an internal structure of a detecting device according to an embodiment of the present invention;
FIG. 4 is a schematic structural view of the housing body of the present invention;
FIG. 5 is a flow chart of the detection method of the present invention.
Main element number description:
1. a housing; 10. a window; 11. a transparent plate; 12. a first accommodating cavity; 13. a second accommodating cavity;
14. a first opening; 15. a second opening; 150. a first annular groove; 16. a third opening;
160. a second annular groove; 17. a first sealing rubber plug; 18. a second sealing rubber plug; 181. a fourth ring-shaped flanging;
19. a third sealing rubber plug; 190. positioning holes; 191. a fifth annular flange; 100. a housing body;
101. a first card slot; 102. a second card slot; 103. a first positioning groove; 104. a second positioning groove;
105. a third positioning groove; 106. a first step portion;
2. puncturing needle; 21. positioning seats; 22. a second stepped portion;
3. a lysis cup; 31. a first annular flanging;
4. an amplification cup; 41. a second annular flanging;
5. a heating unit; 51. a heat-generating sleeve; 52. heating the control plate; 53. a control switch;
54. a power interface; 55. an indicator light;
6. a buffer cup; 61. a third annular flanging;
7. a liquid storage cup; 8. developing test paper; 9. a swab.
Detailed Description
In order to more clearly understand the technical features, objects, and effects of the present invention, embodiments of the present invention will now be described with reference to the accompanying drawings. Where adjective or adverbial modifiers "upper" and "lower", "top" and "bottom", "inner" and "outer" are used merely to facilitate relative reference between groups of terms, and do not describe any particular directional limitation on the modified terms. Furthermore, the terms "first", "second", etc. are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated, whereby a feature defined as "first", "second", etc. may explicitly or implicitly include one or more of such features. In the description of the present invention, "a plurality" means two or more unless otherwise specified. In the description of the present invention, unless otherwise indicated, the term "connected" should be understood broadly, for example, it may be a fixed connection, a detachable connection, a direct connection, or an indirect connection through an intermediate medium, and it is obvious to those skilled in the art that the above-mentioned terms are used in the patent in a specific sense.
Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present application. Thus, the appearances of the phrases "in one embodiment" or "in some embodiments" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.
Implementation mode one
As shown in fig. 1, 2, and 3, the present invention provides a detection device, which includes a housing 1, a lysis cup 3 for lysing a detection sample, an amplification cup 4 for amplifying the detection sample to form an amplification solution, a heating unit 5 for heating the amplification cup 4, a buffer cup 6 for diluting the amplification solution to a reaction solution, a storage cup 7 for containing the reaction solution, a color development test paper 8 for reacting with the reaction solution to display a detection result, and a puncture needle 2 for sequentially puncturing the lysis cup 3, the amplification cup 4, and the buffer cup 6, wherein the lysis cup 3, the amplification cup 4, the buffer cup 6, and the storage cup 7 are disposed in the housing 1 and sequentially disposed from the top of the housing 1 toward the bottom of the housing 1, the color development test paper 8 is disposed in the housing 1, and a liquid suction end of the color development test paper 8 extends into the storage cup 7.
When the detection device is used, the operation steps are as follows:
sampling: collecting a detection sample on a detected person by using a swab 9;
cracking: inserting the swab 9 into the lysis cup 3, and lysing the detection sample by using a lysate in the lysis cup 3;
amplification: puncturing the cracking cup 3 by using the puncture needle 2 to ensure that the cracking cup 3 is communicated with the amplification cup 4, and the lysate in the cracking cup 3 flows into the amplification cup 4; the heating unit 5 is opened to heat the amplification cup 4, and the amplification powder in the amplification cup 4 is heated to amplify the detection sample in the cracking liquid to form amplification liquid;
diluting: continuously pressing the puncture needle 2 to puncture the amplification cup 4, so that the amplification cup 4 is communicated with the buffer solution cup 6, the amplification solution in the amplification cup 4 flows into the buffer solution cup 6, and the buffer solution in the buffer solution cup 6 is used for diluting the amplification solution to form a reaction solution;
and (3) detection: and continuously pressing the puncture needle 2 to puncture the buffer cup 6, so that the buffer cup 6 is communicated with the liquid storage cup 7, the reaction liquid in the buffer cup 6 flows into the liquid storage cup 7 and reacts with the color test paper 8 in the liquid storage cup 7, and the color test paper 8 displays the detection result.
The detection device can realize integrated detection operation, has small volume, can carry out on-site detection, does not need to be transferred to a laboratory, has short detection time, can obtain a detection result in time and has high detection efficiency.
In one embodiment, the top and bottom of lysis cup 3, the top and bottom of amplification cup 4, and the top and bottom of buffer cup 6 are sealed by thin film layers that both seal and are easily punctured by puncture needle 2. The film layer is, for example, a plastic film.
During detection, in the amplification step, a puncture needle 2 is used for sequentially puncturing a top film layer, a bottom film layer and a top film layer of a cracking cup 3 and an amplification cup 4 so as to communicate the cracking cup 3 with the amplification cup 4; in the dilution step, the puncture needle 2 is continuously pressed to sequentially puncture the bottom film layer of the amplification cup 4 and the top film layer of the buffer cup 6 so as to communicate the amplification cup 4 with the buffer cup 6; in the detection step: and the puncture needle 2 is continuously pressed to puncture the bottom film layer of the buffer cup 6, so that the buffer cup 6 is communicated with the liquid storage cup 7, and the operation is simple and convenient.
In an embodiment, as shown in fig. 1 and 3, a window 10 is formed on the housing 1, the window 10 faces the color development test paper 8, the window 10 is sealed by a transparent plate 11, and an operator can visually see a detection result displayed on the color development test paper 8 through the transparent plate 11. The transparent plate 11 in this embodiment is convenient for observing the detection result and the state of each component in the housing 1, and can cover and seal the window 10, thereby preventing external dust and the like from entering the housing 1 to cause adverse effects on the detection result.
In one embodiment, as shown in fig. 2 and 3, the heating unit 5 is disposed in the housing 1, the heating unit 5 includes a heating sleeve 51 sleeved on the amplification cup 4, a heating control board 52 electrically connected to the heating sleeve 51, and a control switch 53, a power interface 54 and an indicator lamp 55 electrically connected to the heating control board 52, the heating sleeve 51 is controlled by the heating control board 52 to heat the amplification cup 4, the power interface 54 is used to energize the detection device with an external circuit, the control switch 53 is used to control the heating sleeve 51 to be manually turned on and off, so as to save energy and reduce cost, and the indicator lamp 55 is used to indicate the working state of the detection device and the temperature of the heating unit 5.
In this embodiment, the heating sleeve 51 is fitted over the amplification cup 4, so that the contact area between the heating sleeve 51 and the amplification cup 4 can be increased, and the heating efficiency of the amplification cup 4 can be improved. However, the present invention is not limited thereto, and the heating sleeve 51 may be replaced by an electric heating coil wound around the amplification cup 4, or may be replaced by an electric heating tube disposed around the amplification cup 4, wherein the electric heating tube has a vacuum structure inside, and a liquid medium is contained in the electric heating tube, and heat transfer is achieved by heating the liquid medium. Of course, in other embodiments, the heating sleeve 51 can be replaced by other structures capable of heating the amplification cup 4, and is not limited herein.
Further, as shown in fig. 1, a control switch 53 and a power interface 54 are installed on the outer wall of the housing 1 for operation, and an indicator lamp 55 is installed on the outer wall of the housing 1 for observation.
Further, a plurality of indicator lights 55, for example, three indicator lights 55, as shown in fig. 1 and 3, when the control switch 53 controls the heating unit 5 to be turned on, one indicator light 55 is on with the first color light, and the second indicator light 55 flashes with the second color light; when the heating unit 5 is heated to a constant temperature, the second indicator light 55 is turned off, and the third indicator light 55 is lit with a third color light; after the constant temperature is finished, the third indicator light 55 is turned off, and the amplification is finished; when the control switch 53 is turned off, all the indicator lamps 55 are turned off.
In an embodiment, as shown in fig. 4, a first accommodating chamber 12 and a second accommodating chamber 13 which are not communicated with each other are provided in the housing 1, please refer to fig. 3 together, the puncture needle 2 is inserted into the first accommodating chamber 12, when the puncture needle 2 needs to be used for puncturing, the puncture needle 2 is pulled out from the first accommodating chamber 12, the lysis cup 3, the amplification cup 4, the buffer cup 6 and the liquid storage cup 7 are fixed in the second accommodating chamber 13, because the first accommodating chamber 12 and the second accommodating chamber 13 are not communicated with each other, the puncture needle 2 is separated from other component cups when not in use, and misoperation is avoided, for example, the first accommodating chamber 12 and the second accommodating chamber 13 are separated by a partition plate.
Further, as shown in fig. 4, the top of the housing 1 is provided with a first opening 14 communicated with the first accommodating chamber 12, and a second opening 15 and a third opening 16 respectively communicated with the second accommodating chamber 13, the second opening 15 and the third opening 16 are respectively arranged opposite to the lysis cup 3, the second opening 15 is used for inserting the swab 9, and the third opening 16 is used for inserting the puncture needle 2; referring to fig. 3, the detecting device further includes a first sealing plug 17 installed in the first opening 14, a second sealing plug 18 installed in the second opening 15, and a third sealing plug 19 installed in the third opening 16.
As shown in fig. 3, the first sealing rubber plug 17 can seal the first opening 14, so as to seal and limit the puncture needle 2 in the first accommodating chamber 12, and prevent the puncture needle 2 from falling out of the first accommodating chamber 12. Similarly, the second hole 15 can be sealed by the second sealing rubber plug 18, and when the swab 9 needs to be inserted, the second sealing rubber plug 18 is opened, and the swab 9 is inserted into the lysis cup 3 through the second hole 15; the third sealing rubber plug 19 can seal the third opening 16, the third sealing rubber plug 19 is provided with a positioning hole 190 communicated with the second accommodating cavity 13, the puncture needle 2 can be guided to be inserted through the positioning hole 190, the third sealing rubber plug 19 does not need to be opened, and the operation is simpler and more convenient.
In an embodiment, as shown in fig. 3 and 4, the housing 1 is formed by two housing bodies 100 which are oppositely buckled, the two housing bodies 100 are detachably connected, and the lysis cup 3, the amplification cup 4, the buffer cup 6 and the liquid storage cup 7 are fixedly clamped between the two housing bodies 100, so that the lysis cup 3, the amplification cup 4, the buffer cup 6 and the liquid storage cup 7 can be conveniently installed, maintained and replaced.
For example, the two case bodies 100 are detachably connected by a snap, or detachably connected by a lock screw.
Further, as shown in fig. 3 and 4, two inner side walls of the two shell bodies 100 opposite to each other are correspondingly provided with first clamping grooves 101, and the two first clamping grooves 101 can allow corresponding ends of the color development test paper 8 to extend into the first clamping grooves 101. Through the cooperation of two first draw-in grooves 101, can realize fixed to the centre gripping of color development test paper 8 to improve the steadiness of color development test paper 8 installation.
Further, as shown in fig. 3 and 4, two opposite inner sidewalls of the two casing bodies 100 are correspondingly provided with second card slots 102, and each second card slot 102 is for a corresponding end of the transparent plate 11 to extend into. Through the cooperation of two second draw-in grooves 102, can realize fixed to the centre gripping of transparent plate 11 to improve the steadiness of transparent plate 11 installation.
Further, as shown in fig. 2 and fig. 4, the top peripheral surface of the cracking cup 3 is provided with a first annular flange 31 protruding outwards, two opposite inner side walls of the two shell bodies 100 are correspondingly provided with first positioning grooves 103, and the two first positioning grooves 103 can be used for the first annular flange 31 to be embedded. Through the cooperation of two first constant head tanks 103, can realize fixed to the centre gripping of schizolysis cup 3 to improve the steadiness of the installation of schizolysis cup 3.
Further, as shown in fig. 2 and fig. 4, the top peripheral surface of the amplification cup 4 is provided with a second annular flange 41 protruding outwards, two opposite inner side walls of the two shell bodies 100 are correspondingly provided with second positioning grooves 104, and the two second positioning grooves 104 are used for the second annular flange 41 to extend into. Through the cooperation of the two second positioning grooves 104, the amplification cup 4 can be clamped and fixed, so that the installation stability of the amplification cup 4 is improved.
Further, as shown in fig. 2 and 4, a third annular flange 61 is outwardly protruded from the outer peripheral surface of the top of the buffer cup 6, third positioning grooves 105 are correspondingly formed on two opposite inner sidewalls of the two casing bodies 100, and the third annular flange 61 can be inserted into the two third positioning grooves 105. Through the cooperation of two third constant head tank 105, can realize fixed to the centre gripping of buffer cup 6 to improve the steadiness of buffer cup 6 installation.
In an embodiment, as shown in fig. 4, the first accommodating cavity 12 is in a cylindrical configuration, the first accommodating cavity 12 and the first opening 14 are coaxially arranged, a diameter of the first opening 14 is greater than a diameter of the first accommodating cavity 12, and a transition joint of the first opening 14 and the first accommodating cavity 12 forms a first step portion 106; referring to fig. 2, the top of the puncture needle 2 is provided with a positioning seat 21, the positioning seat 21 is in a cylindrical configuration adapted to the first opening 14, the diameter of the positioning seat 21 is larger than that of the puncture needle 2, a second stepped portion 22 is formed at the joint of the positioning seat 21 and the puncture needle 2, and the second stepped portion 22 can be seated on the first stepped portion 106. The puncture needle 2 can be supported and resisted through the matching of the first step part 106 and the second step part 22.
In an embodiment, as shown in fig. 4, the inner circumferential surface of the second opening 15 is provided with a first annular groove 150, please refer to fig. 2 together, the outer circumferential surface of the second sealing rubber plug 18 is provided with a fourth annular flange 181 protruding outwards, the first annular groove 150 can clamp and fix the second sealing rubber plug 18, so as to improve the installation stability of the second sealing rubber plug 18.
In an embodiment, as shown in fig. 4, the inner circumferential surface of the third opening 16 is provided with a second annular groove 160, please refer to fig. 3 together, the outer circumferential surface of the third sealing rubber plug 19 is provided with a fifth annular flange 191 protruding outward, the second annular groove 160 can clamp and fix the third sealing rubber plug 19, so as to improve the installation stability of the third sealing rubber plug 19.
In one embodiment, the lysis cup 3 contains a lysis solution, and the lysis solution is a solution formed by diluting tris (2-carboxyethyl) phosphine hydrochloride, diethylamine tetraacetic acid, sodium hydroxide and protease in diethyl pyrocarbonate water. The lysis reaction is carried out on the detection sample by the lysis solution, and the nucleic acid substance in the detection sample can be extracted and used as a template for subsequent amplification.
Specifically, the components in the lysis solution comprise: the content of tris (2-carboxyethyl) phosphine hydrochloride may be 5 mmol; the content of ethylenediaminetetraacetic acid may be 2 millimolar; the sodium hydroxide content may be 29 millimoles; the protease may be proteinase K, and the concentration may be 100. Mu.g/mL.
In one embodiment, the amplification cup 4 contains amplification powder, the amplification powder comprises a primer mixture and a reaction mixture, the primer mixture comprises a front inner primer, a rear inner primer, a front outer primer, a rear outer primer, a front ring primer and a rear ring primer; the reaction mixture includes deoxyribonucleic acid polymerase, reverse transcriptase, tris (hydroxymethyl) aminomethane hydrochloride, ammonium sulfate, potassium chloride, magnesium sulfate, and tween 20. The reaction with the lysis solution can be realized by the amplification powder, and reverse transcription and isothermal amplification can be realized under the coordination of the heating unit 5.
Specifically, the primer mixture comprises the following components in percentage by weight: the content of the pre-inner primer can be 1.6 micromoles; the content of the rear inner primer can be 1.6 micromolar; the content of the front outer primer can be 0.2 micromolar; the content of the rear outer primer can be 0.2 micromolar; the content of the pre-loop primer may be 0.4 micromolar; the content of the latter loop primer may be 0.4 micromolar. The reaction mixture contained the following components: the DNA polymerase may be present in an amount ofEight units; the content of reverse transcriptase can be four units; the tris (hydroxymethyl) aminomethane hydrochloride content can be 20 millimoles; the ammonium sulfate content may be 10 millimoles; the potassium chloride content may be 50 millimoles; the content of magnesium sulfate may be 2 mmol; the mass fraction of tween 20 may be 0.1%. Wherein the primer mixture can be 5' -labeled with digoxin/Texas red/bright/dark
Biotin primer sequence.
In one embodiment, the sample pad, conjugate pad or absorption pad of the color test paper 8 is provided with nitrocellulose so that the reaction solution can be absorbed. Gold nanoparticles are concentrated on the conjugate pad and marked by avidin. Through the cellulose nitrate and the gold nanoparticles, lateral flow analysis can be realized so as to display the detection color on the color development test paper 8, and the detection result can be judged through the color.
Second embodiment
As shown in fig. 5, the present invention also provides a method for detecting viral nucleic acid in a human body by using the detection apparatus of the first embodiment, referring to fig. 1 to 3, the method comprising the steps of:
step S1: sampling: collecting a detection sample on a detected person by using a swab; specifically, the swab 9 is inserted into the nostril of the test person to collect the test sample.
Step S2: cracking: the swab is inserted into the lysis cup 3, and the detection sample is lysed by the lysate in the lysis cup 3. Specifically, the swab 9 for collecting the detection sample is broken, the second sealing rubber plug 18 is pulled out, the cotton head is placed into the cracking cup 3, then the second sealing rubber plug 18 is inserted into the second opening 15 for sealing, and the cracking reaction of the detection sample in the cracking cup 3 is realized, and can be completed within 5 minutes.
And step S3: amplification: puncturing the cracking cup 3 by using a puncture needle to communicate the cracking cup 3 with the amplification cup 4, and enabling the lysate in the cracking cup 3 to flow into the amplification cup 4; and (3) opening the heating unit 5 to heat the amplification cup 4, and amplifying the detection sample in the cracking liquid after the amplification powder in the amplification cup 4 is heated to form amplification liquid. Specifically, the first sealing rubber plug 17 is pulled out, and the puncture needle 2 is pulled out from the first accommodating cavity 12; then, the puncture needle 2 is aligned with the positioning hole 190 and inserted into the third sealing rubber plug 19; then the puncture needle 2 sequentially pierces the top thin film layer, the bottom thin film layer and the top thin film layer of the amplification cup 4 of the cracking cup 3, and the cracking solution flows into the amplification cup 4; finally, the control switch 53 is turned on, the heating unit 5 is turned on, and the heating sleeve 51 heats the amplification cup 4. Wherein the isothermal amplification temperature of the amplification cup 4 can be maintained at 65 ℃ and the amplification reaction can be completed within 30 minutes.
And step S4: diluting: and continuously pressing the puncture needle 2 to puncture the amplification cup 4, so that the amplification cup 4 is communicated with the buffer solution cup 6, the amplification solution in the amplification cup 4 flows into the buffer solution cup 6, and the buffer solution in the buffer solution cup 6 dilutes the amplification solution to form a reaction solution. Specifically, when the puncture needle 2 is pressed, the puncture needle 2 may puncture the bottom film layer of the amplification cup 4 and the top film layer of the buffer cup 6, and the amplification solution flows into the buffer cup 6. The buffer solution contained in the buffer solution cup 6 can dilute the amplification solution.
Step S5: and (3) detection: and continuously pressing the puncture needle 2 to puncture the buffer cup 6, so that the buffer cup 6 is communicated with the liquid storage cup 7, the reaction liquid in the buffer cup 6 flows into the liquid storage cup 7 and reacts with the color development test paper 8 in the liquid storage cup 7, and the color development test paper 8 displays the detection result. Specifically, when the puncture needle 2 is continuously pressed, the puncture needle 2 can puncture the bottom film layer of the buffer liquid cup 6, and the reaction liquid can flow into the liquid storage cup 7, so that the lateral flow analysis reaction with the color test paper 8 is realized. After the reaction is finished, the corresponding color is displayed on the color test paper 8, and an operator can visually observe the detection result through the window 10.
In one embodiment, the pH of the solution formed by mixing the lysate and the amplification powder after the lysate flows into the amplification cup 4 is 8.5 to 9, at which the reverse transcription and isothermal amplification effect is good, thereby improving the accuracy of the detection result. Specifically, the pH of the solution after mixing the amplification powder with the lysis solution may be 8.8.
The above description is only an exemplary embodiment of the present invention, and is not intended to limit the scope of the present invention. Any equivalent changes and modifications that can be made by one skilled in the art without departing from the spirit and principles of the invention should be considered within the scope of the invention. It should be noted that the components of the present invention are not limited to the above-mentioned whole application, and various technical features described in the present specification can be selected to be used alone or in combination according to actual needs, so that the present invention naturally covers other combinations and specific applications related to the invention.
Claims (10)
1. The detection device is characterized by comprising a shell, a cracking cup for cracking a detection sample, an amplification cup for amplifying the detection sample to form amplification liquid, a heating unit for heating the amplification cup, a buffer cup for diluting the amplification liquid into reaction liquid, a liquid storage cup for containing the reaction liquid, a color development test paper for reacting with the reaction liquid to display a detection result, and a puncture needle for sequentially puncturing the cracking cup, the amplification cup and the buffer cup, wherein the cracking cup, the amplification cup, the buffer cup and the liquid storage cup are arranged in the shell and sequentially and correspondingly arranged from the top of the shell to the bottom of the shell, and a liquid suction end of the color development test paper extends into the liquid storage cup.
2. The test device of claim 1, wherein the top and bottom of the lysis cup, the top and bottom of the amplification cup, and the top and bottom of the buffer cup are sealed by thin film layers.
3. The device as claimed in claim 1, wherein said housing has a window, said window is opposite to said color test paper, and said window is closed by a transparent plate.
4. The detection device according to claim 1, wherein the heating unit is disposed in the housing, and the heating unit comprises a heating sleeve sleeved on the amplification cup, a heating control board electrically connected to the heating sleeve, and a control switch, a power interface and an indicator light electrically connected to the heating control board respectively.
5. The detection device according to claim 1, wherein the housing has a first receiving chamber and a second receiving chamber that are not connected to each other, the puncture needle is inserted into the first receiving chamber, and the lysis cup, the amplification cup, the buffer cup and the liquid storage cup are fixed in the second receiving chamber.
6. The detection device as claimed in claim 1, wherein the housing is formed by two housing bodies which are fastened to each other, the two housing bodies are detachably connected, and the lysis cup, the amplification cup, the buffer cup and the reservoir cup are fixedly clamped between the two housing bodies.
7. The detecting device according to any one of claims 1 to 6, wherein a lysis solution is contained in the lysis cup, and the lysis solution is a solution formed by diluting tris (2-carboxyethyl) phosphine hydrochloride, diethylamine tetraacetic acid, sodium hydroxide and protease in diethylpyrocarbonate water.
8. The test device of any one of claims 1 to 6, wherein amplification powders are contained in the amplification cup, the amplification powders comprise a primer mixture and a reaction mixture, the primer mixture comprises a front inner primer, a rear inner primer, a front outer primer, a rear outer primer, a front loop primer and a rear loop primer; the reaction mixture includes deoxyribonucleic acid polymerase, reverse transcriptase, tris (hydroxymethyl) aminomethane hydrochloride, ammonium sulfate, potassium chloride, magnesium sulfate, and tween 20.
9. A detection method using the detection apparatus according to any one of claims 1 to 8, the detection method comprising the steps of:
sampling: collecting a detection sample on a detected person by using a swab;
cracking: inserting the swab into the lysis cup, wherein the lysis solution in the lysis cup lyses the detection sample;
amplification: puncturing the cracking cup by using the puncture needle, so that the cracking cup is communicated with the amplification cup, and the lysate in the cracking cup flows into the amplification cup; opening the heating unit to heat the amplification cup, and amplifying the detection sample in the lysis solution after the amplification powder in the amplification cup is heated to form an amplification solution;
diluting: continuously pressing the puncture needle to puncture the amplification cup to enable the amplification cup to be communicated with the buffer solution cup, enabling amplification solution in the amplification cup to flow into the buffer solution cup, and diluting the amplification solution by using buffer solution in the buffer solution cup to form reaction solution;
and (3) detection: and continuously pressing the puncture needle to puncture the buffer cup, so that the buffer cup is communicated with the liquid storage cup, the reaction liquid in the buffer cup flows into the liquid storage cup and reacts with the color development test paper in the liquid storage cup, and the color development test paper displays a detection result.
10. The method of claim 9, wherein the solution formed by mixing the lysate and the amplification powder after the lysate flows into the amplification cup has a PH of 8.5 to 9.
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|---|---|---|---|
| CN202110366006.0A CN115181652A (en) | 2021-04-06 | 2021-04-06 | Detection device and detection method |
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| CN202110366006.0A CN115181652A (en) | 2021-04-06 | 2021-04-06 | Detection device and detection method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117025379A (en) * | 2023-10-09 | 2023-11-10 | 迪飞医学科技(南京)有限公司 | RAPID isothermal amplification nucleic acid detection device and detection method |
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2021
- 2021-04-06 CN CN202110366006.0A patent/CN115181652A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117025379A (en) * | 2023-10-09 | 2023-11-10 | 迪飞医学科技(南京)有限公司 | RAPID isothermal amplification nucleic acid detection device and detection method |
| CN117025379B (en) * | 2023-10-09 | 2023-12-15 | 迪飞医学科技(南京)有限公司 | RAPID isothermal amplification nucleic acid detection device and detection method |
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