CN115322889A - Multi-section solvent reaction tube for biological nucleic acid detection and detection method thereof - Google Patents
Multi-section solvent reaction tube for biological nucleic acid detection and detection method thereof Download PDFInfo
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- CN115322889A CN115322889A CN202211265132.8A CN202211265132A CN115322889A CN 115322889 A CN115322889 A CN 115322889A CN 202211265132 A CN202211265132 A CN 202211265132A CN 115322889 A CN115322889 A CN 115322889A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
- C12Q1/686—Polymerase chain reaction [PCR]
Abstract
The invention relates to the technical field of nucleic acid detection devices, in particular to a multi-section solvent reaction tube for biological nucleic acid detection, which comprises a reaction tube body, wherein the reaction tube body comprises: the sampling tube is provided with caps at two ends; the top of the dilution pipe is provided with a top ring; the heating pipe is composed of a first inner pipe and a second inner pipe; the bottom of the pushing pipe is provided with a pressing ring, and a test paper clip arranged on the pressing ring is further arranged in the pushing pipe; the sampling pipe, the dilution pipe, the heating pipe and the pushing pipe are connected in sequence. The multi-section solvent reaction tube for detecting the biological nucleic acid can be subjected to whole-process closed operation in a common environment, is separated by the solution sealing film, and has no exposure links in the steps of extraction, dilution, amplification and detection. The method is simple to operate, adopts the operation steps of firstly loading and then unloading, punctures the solution sealing film to realize the butt joint of the pipe bodies, can accurately sample during detection, and has strong detection flexibility, high sample processing efficiency and reliable detection result.
Description
Technical Field
The invention relates to the technical field of nucleic acid detection devices, in particular to a multi-section solvent reaction tube for biological nucleic acid detection and a detection method thereof.
Background
The current nucleic acid detection process generally comprises sampling by front-end workers, freezing and storing samples, cold-chain transporting to a rear-end detection position, and detecting by rear-end professional operators. During detection, operation steps of uncovering, pipetting, extracting, amplifying and the like exist, the steps have high requirements on the environment, and detection is often required in a specific isolation laboratory in order to avoid the amplified PCR product polluting the environment and causing a false positive detection result. The existing nucleic acid rear-end detection steps are complex, detection needs to be carried out in a specific isolation laboratory, the sample treatment efficiency is low, the time consumption is long, and the detection error is easy to occur.
Disclosure of Invention
The present invention is directed to a multi-stage solvent reaction tube for detecting biological nucleic acid, which solves the above problems of the prior art.
The invention is realized by the following technical scheme:
a multi-section solvent reaction tube for biological nucleic acid detection comprises a reaction tube body, wherein the reaction tube body comprises:
the sampling tube is used for collecting samples of the swab head, and caps are arranged at two ends of the sampling tube, wherein the one-way cap arranged at the opening end is detachable, and the cap arranged at the tail end is a two-way cap;
the top of the dilution pipe is provided with a top ring, the bottom of the dilution pipe is provided with a needle body which penetrates through the bottom surface of the dilution pipe and extends into the heating pipe, the middle part of the needle body is provided with a through hole, and the aperture of the through hole is 0.25mm;
the heating pipe is used for PCR amplification and comprises an inner pipe I and an inner pipe II, the pipe diameter of the inner pipe I is smaller than that of the inner pipe II, freeze-dried microspheres and grinding ball steel balls are filled in the inner pipe I, and 2.5mL of diluent is arranged in the inner pipe II;
the test paper test device comprises a push pipe, a test paper clamp and a test paper strip, wherein a press ring is arranged at the bottom of the push pipe, a filter membrane is arranged on the press ring, the test paper clamp is arranged on the press ring, and the test paper strip is arranged on the test paper clamp;
the bottom end of the sampling pipe, the two ends of the dilution pipe, the two ends of the heating pipe and the connecting surface of the first inner pipe and the second inner pipe are respectively provided with a corresponding solution sealing film;
the sampling pipe, dilute the pipe, the heating pipe with the propelling movement pipe is for connecting in order.
Preferably, the outer wall surface of the top of the dilution pipe is provided with threads, the inner wall surface of the bottom of the dilution pipe is provided with threads, the outer wall surface of the first inner pipe is provided with a thread component, and the dilution pipe is in threaded connection with the sampling pipe and the dilution pipe is in threaded connection with the heating pipe.
As a further scheme of the invention, the wall surface of the inner pipe II is provided with a groove, ribs are distributed on the inner wall surface of the pushing pipe, and the ribs and the grooves are in one-to-one correspondence and clamped connection.
As a further scheme of the invention, the upper surface of the top ring is uniformly provided with barbed convex blocks, and the barbed convex blocks are arranged below the solution sealing film at the top end of the dilution pipe.
Preferably, the periphery cover of sampling socle portion is equipped with the silica gel gasket, the silica gel gasket is used for the body sealed, that prevent to connect not hard up.
Preferably, the periphery of the heating pipe is sleeved with an annular silica gel ring for sealing, and the annular silica gel ring is respectively located at the joint of the first inner pipe and the dilution pipe and the joint of the heating pipe and the propulsion pipe.
The invention also provides a detection method of the multi-section solvent reaction tube for biological nucleic acid detection, which comprises the following steps:
extruding the swab head after sampling in the sampling tube, performing a first-stage biological reaction with a biological reagent in the sampling tube, breaking off a cotton swab stick, covering a cap, screwing the cap to seal the sampling tube, and obtaining a fully dissolved biological sample;
after the sampling pipe and the dilution pipe are assembled, the dilution pipe is clockwise screwed along threads, the solution sealing films positioned at the top end of the dilution pipe and the bottom end of the sampling pipe are sequentially cut by using a top ring, so that the sampling pipe is communicated with the dilution pipe, the sampling pipe and the dilution pipe are vertically inverted and uniformly mixed, and the biological sample in the first stage and the biological reagent in the dilution pipe are subjected to a second-stage biological reaction to obtain a biological sample in a second stage;
screwing the dilution tube anticlockwise to an upper limit position, enabling the biological sample at the second stage in the dilution tube to quantitatively flow into the heating tube through the pinhole, uniformly mixing the solid preparation passing through the first inner tube, centrifuging, heating at 42 ℃ for 15min, and carrying out biological PCR reaction at the third stage to obtain a biological sample at the third stage;
after heating, clockwise screwing the dilution tube to a lower limit position, rotating the needle body to cut a solution sealing film on the connecting surface of the first inner tube and the second inner tube, and turning upside down to mix uniformly, so that a biological sample in a third stage and a biological reagent in the second inner tube carry out biological reaction in a fourth stage, and finally obtaining a biological sample in the fourth stage;
and inverting the reaction tube body, pushing the test paper clamp by using the compression ring, and bursting the solution sealing film at the tail end of the heating tube by using the test paper clamp, so that the test paper arranged on the test paper clamp is immersed in the biological sample obtained at the fourth stage, and after developing for 5min, obtaining a detection result through the test paper developing condition.
As a further scheme of the invention, an elastic limiting block is arranged in the inner tube II and is used for avoiding the test paper from being excessively immersed.
Preferably, the immersion depth of the test paper is 3 to 4mm.
Compared with the prior art, the invention has the beneficial effects that:
the multi-section solvent reaction tube for detecting the biological nucleic acid can be subjected to whole-process closed operation in a common environment, is separated by the solution sealing film, and has no exposure links in the steps of extraction, dilution, amplification and detection. The method is simple to operate, adopts the operation steps of firstly loading and then unloading, punctures the solution sealing film to realize the butt joint of the pipe bodies, can accurately sample during detection, and has strong detection flexibility, high sample processing efficiency and reliable detection result.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic view of the overall assembly of the present invention;
FIG. 2 is a front cross-sectional view of the present invention;
FIG. 3 is a schematic view of the internal structure of the present invention;
FIG. 4 is a schematic view of the construction of the top ring of the present invention;
FIG. 5 is a schematic structural view of a heating tube according to the present invention;
FIG. 6 is a schematic view of the push tube of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1:
as shown in fig. 1-6, the present embodiment provides a multi-stage solvent reaction tube for detecting biological nucleic acid, comprising a reaction tube body, wherein the reaction tube body comprises:
the sampling device comprises a sampling tube 1, wherein 400 mu L of lysate used for collecting a sample of a swab head 11 is arranged in the sampling tube 1, caps 12 are arranged at two ends of the sampling tube, the one-way cap arranged at the opening end can be detached, and the cap arranged at the tail end is a two-way cap;
the dilution tube 2 is internally provided with 3.6mL of diluent for diluting a sample solution, the top of the dilution tube 2 is provided with a top ring 21, the bottom of the dilution tube 2 is provided with a needle body 22 which penetrates through the bottom surface of the dilution tube 2 and extends into the heating tube 3, the middle part of the needle body 22 is provided with a through hole, and the aperture of the through hole is 0.25mm;
the heating pipe 3 is used for PCR amplification and comprises a first inner pipe 31 and a second inner pipe 32, the pipe diameter of the first inner pipe 31 is smaller than that of the second inner pipe 32, freeze-dried microspheres and grinding ball steel balls are filled in the first inner pipe 31, and 2.5mL of diluent is arranged in the second inner pipe 32;
the test paper strip pushing device comprises a pushing tube 4, a pressing ring 41 is arranged at the bottom of the pushing tube 4, a filter membrane 42 is arranged on the pressing ring 41, a test paper clamp 43 arranged on the pressing ring 41 is further arranged in the pushing tube 4, and the test paper strip is arranged on the test paper clamp 43;
the bottom end of the sampling tube 1, the two ends of the dilution tube 2, the two ends of the heating tube 3 and the connecting surface of the first inner tube 31 and the second inner tube 32 are provided with corresponding solution sealing films 5;
the sampling pipe 1, the dilution pipe 2, the heating pipe 3 and the pushing pipe 4 are connected in sequence, wherein,
the dilution pipe 2 is in threaded connection with the sampling pipe 1, and the dilution pipe 2 is in threaded connection with the heating pipe 3, the outer wall surface of the first inner pipe 31 is provided with a threaded component 33, the outer wall surface of the top of the dilution pipe 2 is provided with threads matched with the sampling pipe, and the inner wall surface of the bottom of the dilution pipe 2 is provided with threads matched with the threaded component 33;
the heating pipe 3 and the pushing pipe 4 are clamped, a groove 34 is formed in the wall surface of the inner pipe II 32, ribs 44 are distributed on the inner wall surface of the pushing pipe 4, and the ribs 44 are in one-to-one correspondence with the grooves 34 and are clamped.
The invention also provides a detection method using the multi-section solvent reaction tube for detecting the biological nucleic acid, which comprises the following steps:
extruding the sampled swab head 11 in the sampling tube 1 at least twice, performing a first-stage biological reaction on the swab head 11 and a lysate in the sampling tube 1, breaking off a cotton swab stick, covering a cap 12, screwing the cap 12, and sealing the sampling tube 1 to obtain a fully dissolved biological sample;
screwing the dilution tube 2 to an upper limit position anticlockwise, enabling the biological sample at the second stage in the dilution tube 2 to quantitatively flow into the heating tube 3 through a through hole formed by the needle body 22, uniformly mixing the solid preparation through the inner tube I31, centrifuging, heating at 42 ℃ for 15min, and carrying out biological PCR reaction at the third stage, wherein the aperture of a pinhole is 0.25mm, so that 50 mu L of the biological sample at the third stage can be quantitatively obtained;
after heating, clockwise screwing the dilution tube 2 to the lower limit position, the needle body 22 rotates to lacerate the solution sealing film 5 on the connection surface of the first inner tube 31 and the second inner tube 32, and the biological sample and the diluent in the second inner tube 32 are mixed by turning upside down, so that the volume of the biological sample and the diluent in the third stage are 1: diluting at the ratio of 50 to finally obtain a biological sample of the fourth stage;
and (3) inverting the reaction tube body, pushing the test paper clamp 43 to burst the solution sealing film 5 at the tail end of the heating tube 3 by using the pressing ring 41, so that the test paper arranged on the test paper clamp 43 is immersed in the biological sample obtained at the fourth stage, and after the biological sample is developed for 5min, obtaining a detection result according to the color development condition of the test paper.
An elastic limiting block 35 is arranged in the second inner tube 32 and used for avoiding the test paper from being immersed excessively, and the appropriate immersion depth of the test paper is 3-4 mm.
Example 2:
as shown in fig. 1-6, the present embodiment provides a multi-stage solvent reaction tube for detecting biological nucleic acid, the difference between the present embodiment and embodiment 1 is that a top ring 21 is formed by connecting a plurality of concentric rings, barb bumps 23 are uniformly distributed on the upper surfaces of the concentric rings located in the inner ring of the top ring 21, the barb bumps 23 are disposed below a solution sealing film 5 on the top end of a dilution tube 2, the barb bumps 23 are used for improving the bursting efficiency of the solution sealing film 5, so that the sampling tube 1 is communicated with the dilution tube 2, and the rest are the same as those in embodiment 1.
Example 3:
as shown in fig. 1-6, the present embodiment provides a multi-stage solvent reaction tube for detecting biological nucleic acid, which is different from embodiment 2 in that a silica gel gasket 13 is sleeved on the periphery of the bottom of a sampling tube 1, and the silica gel gasket 13 is used for sealing and preventing the connected tube body from loosening; an annular silica gel ring 36 for sealing is sleeved on the periphery of the heating pipe 3, two layers of annular silica gel rings 36 are arranged at the joint of the first inner pipe 31 and the dilution pipe 2, the outer diameter of each layer of annular silica gel ring 36 is 8mm, the outer diameter of the annular silica gel ring 36 arranged at the joint of the heating pipe 3 and the propulsion pipe 4 is 15mm, and other steps are consistent with those in embodiment 2.
The multi-section solvent reaction tube for detecting the biological nucleic acid can carry out whole-process closed operation in a common environment, realizes partition by a solution sealing film, and has no exposed links in the steps of extraction, dilution, amplification and detection. The method is simple to operate, adopts the operation steps of firstly loading and then unloading, punctures the solution sealing film to realize the butt joint of the pipe bodies, can accurately sample during detection, and has strong detection flexibility, high sample processing efficiency and reliable detection result.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand the invention for and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (9)
1. The utility model provides a biological nucleic acid detects with multistage formula solvent reaction pipe, includes the reaction tube body, its characterized in that, the reaction tube body includes:
the sampling tube is provided with caps at two ends;
the top of the dilution pipe is provided with a top ring, the bottom of the dilution pipe is provided with a needle body, and the middle part of the needle body is provided with a through hole;
the heating pipe is composed of a first inner pipe and a second inner pipe, and the pipe diameter of the first inner pipe is smaller than that of the second inner pipe;
the test paper test device comprises a push pipe, a test paper clamp and a test paper strip, wherein a press ring is arranged at the bottom of the push pipe, a filter membrane is arranged on the press ring, the test paper clamp is arranged on the press ring, and the test paper strip is arranged on the test paper clamp;
the bottom end of the sampling pipe, the two ends of the dilution pipe, the two ends of the heating pipe and the connecting surface of the first inner pipe and the second inner pipe are respectively provided with a corresponding solution sealing film;
the sampling pipe, dilute the pipe, the heating pipe with the propelling movement pipe is for connecting in order.
2. The multi-stage solvent reaction tube for detecting biological nucleic acid according to claim 1, wherein the dilution tube and the sampling tube, and the dilution tube and the heating tube are all screwed together.
3. The multi-stage solvent reaction tube for detecting biological nucleic acid according to claim 1, wherein a groove is formed on the wall surface of the inner tube II, ribs are distributed on the inner wall surface of the pushing tube, and the ribs and the groove are in one-to-one correspondence and clamped connection.
4. The multi-stage solvent reaction tube for detecting biological nucleic acid according to claim 1, wherein the top ring has barb protrusions uniformly distributed on the upper surface thereof.
5. The multi-stage solvent reaction tube for detecting biological nucleic acid according to claim 1, wherein a silica gel gasket is sleeved on the outer periphery of the bottom of the sampling tube, and the silica gel gasket is used for sealing and preventing looseness.
6. The multi-stage solvent reaction tube for detecting biological nucleic acid according to claim 1, wherein an annular silicone ring is sleeved on an outer circumference of the heating tube, and the annular silicone ring is respectively located at a joint of the first inner tube and the dilution tube and a joint of the heating tube and the propulsion tube.
7. A method for testing the multi-stage solvent reaction tube for testing biological nucleic acid according to any one of claims 1 to 6, comprising the following steps:
extruding the swab head after sampling in the sampling tube, performing a first-stage biological reaction with a biological reagent in the sampling tube, breaking off a cotton swab stick, covering a cap, screwing the cap to seal the sampling tube, and obtaining a fully dissolved biological sample;
after the sampling pipe and the dilution pipe are assembled, the dilution pipe is screwed clockwise along the threads, the solution sealing films positioned at the top end of the dilution pipe and the bottom end of the sampling pipe are sequentially cut by utilizing the top ring, so that the sampling pipe is communicated with the dilution pipe, the sampling pipe and the dilution pipe are turned upside down and mixed uniformly, and a biological sample in the first stage and a biological reagent in the dilution pipe are subjected to biological reaction in the second stage to obtain a biological sample in the second stage;
screwing the dilution pipe to an upper limit position anticlockwise, enabling the biological sample at the second stage in the dilution pipe to quantitatively flow into the heating pipe through the through hole, uniformly mixing the solid preparation through the inner pipe I, centrifuging, heating, and performing biological reaction at the third stage to obtain the biological sample at the third stage;
after heating, clockwise screwing the dilution tube to a lower limit position, rotating the needle body to cut a solution sealing film on the connecting surface of the first inner tube and the second inner tube, and turning upside down to mix uniformly, so that a biological sample in a third stage and a biological reagent in the second inner tube carry out biological reaction in a fourth stage, and finally obtaining a biological sample in the fourth stage;
the reaction tube body is inverted, the test paper clamp is pushed by the compression ring and is pushed to burst a solution sealing film at the tail end of the heating tube, so that test paper arranged on the test paper clamp is immersed in a biological sample obtained at the fourth stage, and after color development, a detection result is obtained through the color development condition of the test paper.
8. The detection method according to claim 7, wherein an elastic limiting block is arranged in the inner tube II and used for avoiding the test paper from being excessively immersed.
9. The detection method according to claim 7, wherein the test paper has an immersion depth of 3 to 4mm.
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| CN202211265132.8A CN115322889B (en) | 2022-10-17 | 2022-10-17 | Multi-section solvent reaction tube for biological nucleic acid detection and detection method thereof |
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| CN202211265132.8A CN115322889B (en) | 2022-10-17 | 2022-10-17 | Multi-section solvent reaction tube for biological nucleic acid detection and detection method thereof |
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| CN117165416A (en) * | 2023-11-03 | 2023-12-05 | 苏州雅睿生物技术股份有限公司 | Dilution tube, nucleic acid detection device and detection method thereof |
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| CN115322889B (en) | 2022-12-20 |
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