CN113667597B - Sampling gun and method integrating liquid collecting, drip sampling, sample processing and detection - Google Patents
Sampling gun and method integrating liquid collecting, drip sampling, sample processing and detection Download PDFInfo
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- CN113667597B CN113667597B CN202110976166.7A CN202110976166A CN113667597B CN 113667597 B CN113667597 B CN 113667597B CN 202110976166 A CN202110976166 A CN 202110976166A CN 113667597 B CN113667597 B CN 113667597B
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- 238000005070 sampling Methods 0.000 title claims abstract description 135
- 238000001514 detection method Methods 0.000 title claims abstract description 81
- 239000007788 liquid Substances 0.000 title claims abstract description 76
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000012545 processing Methods 0.000 title claims abstract description 10
- 238000003745 diagnosis Methods 0.000 claims abstract description 3
- 150000007523 nucleic acids Chemical class 0.000 claims description 12
- 102000039446 nucleic acids Human genes 0.000 claims description 12
- 108020004707 nucleic acids Proteins 0.000 claims description 12
- 230000003321 amplification Effects 0.000 claims description 10
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 230000009471 action Effects 0.000 claims description 7
- 238000011534 incubation Methods 0.000 claims description 5
- 238000003825 pressing Methods 0.000 claims description 4
- 238000003780 insertion Methods 0.000 claims description 2
- 230000037431 insertion Effects 0.000 claims description 2
- 238000007789 sealing Methods 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- 239000000945 filler Substances 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 49
- 244000052616 bacterial pathogen Species 0.000 abstract description 9
- 239000012472 biological sample Substances 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 description 6
- 230000004907 flux Effects 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 230000001717 pathogenic effect Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 241000700605 Viruses Species 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007847 digital PCR Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Classifications
-
- 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
-
- 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
- C12Q1/6851—Quantitative amplification
Abstract
The invention provides a liquid drop sampling, sample processing and detecting integrated sampling gun and a method thereof, wherein the sampling gun comprises the following components: a sampling gun body; the sampling gun comprises a negative pressure generating structure, an inserted detection chip, a temperature control module and an oil phase storage cavity, wherein the negative pressure generating structure, the inserted detection chip, the temperature control module and the oil phase storage cavity are sequentially arranged in the sampling gun body from the rear end to the front end, and sample channels for conveying sample liquid drops are arranged in the temperature control module and the oil phase storage cavity; a liquid droplet sampling gun head operably connected to a front end of a sampling gun body, comprising: an axially extending outer tube; and an inner capillary tube coaxially extending with the outer tube and fixed in the outer tube; and the hydrophilic filter structure is fixed at the front end of the outer layer pipeline, and a certain gap is kept between the hydrophilic filter structure and the front end of the inner layer capillary. According to the invention, a simple, efficient, reliable, rapid, high-flux and high-sensitivity biological sample detection method is provided, and an effective technical means and tool are hopefully provided for rapid diagnosis of instant fixed-point pathogenic bacteria.
Description
Technical Field
The invention relates to the field of liquid sampling devices, in particular to a sampling gun integrating liquid drop sampling, sample processing and detection and a method thereof.
Background
The detection of pathogenic bacteria is of great importance for human health and life safety. The existing pathogen detection method has high requirements on detection environment, long time period, low flux and large equipment volume, so that a closed, rapid, high-flux, portable and miniaturized instant and fixed-point detection system is the development direction of pathogen detection technology.
The liquid drop digital LAMP is a brand new absolute nucleic acid quantitative technology without a standard curve, which is generated in the last two years, and can accurately detect the concentration of pathogenic bacteria in a low-cost liquid drop and constant-temperature environment, so that the LAMP has a wide application prospect in the aspect of clinical rapid detection of pathogenic bacteria.
The existing liquid drop digital LAMP detection technology realizes the digital absolute quantitative result by a liquid drop method, and improves the detection sensitivity. However, most of pathogenic bacteria biological samples are still manually and directly sampled by a pipette in the process of treatment and detection, and because the samples are not transported in a 'packed' way, the samples are leaked or polluted by exogenous substances due to contact with air in the transportation, and the safety of detection personnel can be threatened in the process of detecting certain viruses. Meanwhile, the pump valve and the detection equipment are huge in size and are difficult to apply to on-site instant fixed-point detection. In summary, there is no instrument available at the research institution or enterprise company that can achieve immediate point-of-care collection under totally enclosed conditions and integrate manipulation and detection of pathogenic samples.
Disclosure of Invention
The invention aims to provide a sampling gun integrating liquid drop sampling, sample processing and detection and a method thereof, thereby solving the problems that the prior pathogenic bacteria detection technology is difficult to ensure a sampling closed environment, slow in detection, low in flux and huge in detection instrument in actual detection.
In order to solve the technical problems, the invention adopts the following technical scheme:
according to a first aspect of the present invention, there is provided a sampling gun integrating droplet collection, sample processing and detection, comprising: a sampling gun body; the negative pressure generating structure, the plug-in detection chip, the temperature control module and the oil phase storage cavity are sequentially arranged in the sampling gun body from the rear end to the front end, and sample channels for conveying sample liquid drops are arranged in the temperature control module and the oil phase storage cavity; a liquid drop sampling gun head operatively connected to a front end of the sampling gun body, comprising: an axially extending outer tube; and an inner capillary tube fixed in the outer tube and extending coaxially with the outer tube; the hydrophilic filter structure is fixed at the front end of the outer layer pipeline, and a certain gap is kept between the hydrophilic filter structure and the front end of the inner layer capillary tube; when the liquid drop sampling gun head is connected to the front end of the sampling gun body, a sample channel in the sampling gun body is communicated with an inner capillary tube in the liquid drop sampling gun head, oil phase in the oil phase storage cavity enters between an outer layer pipeline of the liquid drop sampling gun head and the inner capillary tube, under the action of the negative pressure generating structure, a water phase sample is collected into the inner capillary tube in a liquid drop mode through oil phase interval, and further is conveyed from the front end to the rear end through the sample channel, so that digital isothermal nucleic acid amplification reaction under the temperature control module, tiling and detection on the inserted detection chip are realized.
The plug-in detection chip comprises a chip shell with an inner cavity, and a liquid inlet hole for sample liquid drops to enter is formed in the chip shell.
The plug-in detection chip can be inserted along the side face of the sampling gun body, a flange which is convenient to hold is arranged on the chip shell, and a jack for inserting the plug-in detection chip is arranged on the side face of the sampling gun body.
Preferably, the sample channel meanders along an S-shaped path in the temperature control module so as to meet the time requirement of incubating the sample liquid drop at the constant temperature of 63 ℃ in the temperature control module and complete the digital isothermal nucleic acid amplification reaction.
Preferably, the oil phase storage cavity comprises a first oil phase storage cavity and a second oil phase storage cavity which are sequentially connected from the rear end to the front end, the second oil phase storage cavity has a radial size equivalent to that of an outer layer pipeline of the liquid drop sampling gun head, and the first oil phase storage cavity has a radial size which is obviously increased relative to that of the second oil phase storage cavity.
Preferably, the side wall of the oil phase storage cavity is provided with an oil filling hole for supplementing the oil phase.
Preferably, the liquid drop sampling gun head is connected with the front end of the sampling gun body through friction force.
According to a second aspect of the present invention, there is provided a method for integrating droplet sampling, sample processing and detection, comprising the steps of: s1: providing a sampling gun as described above, wherein the front end of the sampling gun body is connected with a liquid drop sampling gun head; s2: manually pressing the negative pressure generating structure to extend the liquid drop sampling gun head into the liquid sample to be collected, and loosening the negative pressure generating structure to collect the water phase sample into the liquid drop sampling gun head in a liquid drop form; s3: under the action of negative pressure, the liquid drop sample is further conveyed into a temperature control module through a sample channel to be heated, and digital isothermal nucleic acid amplification reaction is carried out; s4: and the liquid drops after the reaction are spread and stored on the inserted detection chip.
The method also comprises the steps of: s5: and the inserted detection chip stored with the sample is taken down from the sampling gun body and transferred to detection equipment, so that digital absolute quantitative detection is realized.
When the sampling gun is used for collecting samples in the centrifuge tube sealed by the rubber plug, the tip of the liquid drop sampling gun head is used for piercing the rubber plug, and the rubber plug automatically seals the centrifuge tube after sampling is finished, so that the full sealing in the sampling process can be ensured.
According to the invention, the high-flux sampling gun head capable of directly generating liquid drops is combined with the sampling gun with the functions of integrating the negative pressure and the temperature control system to construct integrated liquid drop sampling, transporting and incubating equipment, so that the detection efficiency of biological samples is improved. According to the sampling gun and the sampling method for directly sampling, generating liquid drops, transporting and incubating, which are provided by the invention, the sampling is carried out on the pathogenic bacteria outbreak site, the closed sampling in the form of liquid drops can be realized, and the water phase sample information can be processed and detected more efficiently, pollution-free and sensitively. The whole set of droplet microfluidic processes such as droplet sampling, digital LAMP reaction incubation, transportation, detection and the like are arranged on the integrated sampling gun. The liquid drop sampling device has the advantages of integration, no pollution, high sensitivity and the like, and is favorable for being applied to on-site instant detection so as to solve the problems of large volume, complex detection process and sample pollution of the existing sampling device. In the future, if the parallel automation is realized, not only can the liquid drops be directly generated for reaction and detection after sampling be realized, but also the automation and high-flux sampling and detection can be realized through multi-channel parallel connection.
Compared with the existing sampling equipment, the invention has the following advantages: firstly, the process from the sample to the detection is totally enclosed, and the samples carried by each droplet are transported and detected independently, so that the samples are not polluted and interfered with each other in the transportation process, and a foundation is laid for multichannel parallel sampling; secondly, a small amount of sample is divided into liquid drops with the diameter of about 500 mu m and the volume of about 0.5 mu L by the liquid drops, a large amount of independent reaction can be carried out on a small amount of sample at the same time, target molecules contained in each liquid drop are respectively detected in a back-end detection device, the signals of each reaction unit are subjected to statistical analysis after detection, absolute quantitative detection can be realized through digital result discrimination, and the detection sensitivity is improved; thirdly, integrate, from the sample to the whole flow that incubates is realized in the sampling rifle, has reduced equipment volume, has simplified work flow.
In summary, according to the present invention, there is provided a sampling gun and a method thereof integrating liquid droplet sampling, sample processing and detection, in which a sample is to be collected in the form of droplets, and then the droplets are used as reaction microcavities, and effective distribution of nucleic acid to be measured is achieved by the structure of the sampling gun. Meanwhile, detection such as digital LAMP or digital PCR can be realized on a chip by combining a droplet control technology. Based on the invention, the key bottleneck problems that the prior pathogenic bacteria detection technology is difficult to ensure a sampling closed environment, slow in detection, low in flux and huge in detection instrument in practical application can be overcome, the simple, convenient, efficient, reliable and rapid detection with high flux and high sensitivity can be realized, and the invention is expected to provide an effective technical means and tool for rapid diagnosis of the pathogenic bacteria at a point of care (POCT).
Drawings
FIG. 1 is a schematic view of the overall structure of a sampling gun according to a preferred embodiment of the present invention;
fig. 2 shows a process of an oil phase entering between an outer layer pipeline and an inner layer capillary of a liquid drop sampling gun head from an oil phase storage cavity, wherein A shows a state that a sampling gun body is just connected with the liquid drop sampling gun head, B shows a state that the oil phase enters the liquid drop sampling gun head from the sampling gun body under a negative pressure state, and C shows a state that the oil phase and sample liquid drops enter the inner layer capillary;
FIG. 3 is a perspective view of a plug-in test chip;
FIG. 4 is a schematic top view of a digital quantitative detection of a mid-droplet of an inserted detection chip after incubation, according to one embodiment of the invention;
fig. 5 is a schematic view of a sample collection in a centrifuge tube using the sampling gun of the present invention.
Detailed Description
The invention will be further illustrated with reference to specific examples. It should be understood that the following examples are illustrative of the present invention and are not intended to limit the scope of the present invention.
In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention.
Furthermore, the terms "first," "second," and the like, 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. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present invention, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.
As shown in fig. 1, a sampling gun according to a preferred embodiment of the present invention mainly includes: the sampling gun body 1 is sequentially provided with a negative pressure generating structure 2, an inserted detection chip 3, a temperature control module 4, an oil phase storage cavity 5 and a liquid drop sampling gun head 6 which are operatively connected with the front end of the sampling gun body 1 from top to bottom.
The sampling gun body 1 is a substantially rectangular parallelepiped housing having an internal cavity, and is used for mounting other components. The negative pressure generating structure 2 is arranged at the rear end of the sampling gun body 1 and comprises a handle capable of being pressed manually, and negative pressure can be provided in the sampling gun body 1 through manual pressing and loosening.
As shown in fig. 1, 3 and 4, the plug-in detection chip 3 includes a chip housing 31 having an internal cavity, and the chip housing 31 has a liquid inlet 32 for sample droplets to enter, and a flange 33 for easy holding. According to the preferred embodiment, the plug-in detection chip 3 can be inserted along the left side of the sampling gun body 1, and accordingly, the side of the sampling gun body 1 has a receptacle into which the plug-in detection chip 3 is inserted.
As shown in fig. 1, a temperature control module 4 is arranged below the inserted detection chip 3, and the sample droplet is heated at constant temperature by the module to perform a digital isothermal nucleic acid amplification reaction. The temperature control module 4 has a screen on its surface for displaying the temperature, so as to display the temperature in the module at this time in real time, for example, 63 ℃.
As shown in fig. 1, the oil phase storage cavity 5 is disposed below the temperature control module 4, and includes a first oil phase storage cavity 51 and a second oil phase storage cavity 52 sequentially connected from top to bottom, where the second oil phase storage cavity 52 has a radial dimension corresponding to an outer diameter of the droplet sampling gun head 6, and the first oil phase storage cavity 51 has a radial dimension that is significantly increased relative to the second oil phase storage cavity 52.
It should be understood that the shape of the oil phase storage chamber 5 is not limited to that shown in fig. 1, but may be any other suitable shape, by way of example only and not limitation.
Preferably, the side wall of the oil phase storage chamber 5 is also provided with an oil filling hole for replenishing the oil phase therein.
According to the preferred embodiment, the temperature control module 4 and the oil phase storage cavity 5 are provided with a sample channel 7 for conveying sample liquid drops, the sample channel 7 meanders along an S-shaped path in the temperature control module 4 so as to prolong the incubation time of the sample liquid drops in the temperature control module 4 as much as possible and complete the digital isothermal nucleic acid amplification reaction.
As shown in fig. 1, the liquid droplet sampling gun head 6 includes: an axially extending outer layer tube 61; and an inner capillary 62 coaxially extending therewith and fixed within the outer tube 61; a hydrophilic filter structure 63 fixed to the front end of the outer tube 61, the hydrophilic filter structure 63 being spaced apart from the front end of the inner capillary tube 62.
Similar to a common pipette, the liquid drop sampling gun head 6 is connected with the front end of the sampling gun body 1 through friction force.
The working principle of the sampling gun provided by the invention is described below with reference to fig. 2: when the liquid drop sampling gun head 6 is connected to the front end of the sampling gun body 1, the sample channel 7 in the sampling gun body 1 is communicated with the inner capillary 62 in the liquid drop sampling gun head 6, and at this time, only the space inside the oil phase storage cavity 5 and outside the sample channel 7 is filled with the oil phase O (as shown in A in FIG. 2); next, the oil phase O in the oil phase storage chamber 5 enters between the outer layer pipe 61 and the inner layer capillary 62 of the droplet-sampling gun head 6 (as shown in B in fig. 2); under the action of the negative pressure generating structure 2, the sample droplets D separated by the oil phase O are collected in the inner capillary 62, and then are conveyed from the front end to the rear end (as shown by C in fig. 2) through the sample channel 7, so that the digital isothermal nucleic acid amplification reaction under the temperature control module 4 and tiling and detection on the inserted detection chip 3 are realized.
According to another preferred embodiment of the present invention, there is provided a digital quantitative detection method including the steps of:
s1: providing a sampling gun as shown in fig. 1, wherein the front end of a sampling gun body 1 is connected with a liquid drop sampling gun head 6;
s2: manually pressing the negative pressure generating structure 2 to extend the liquid drop sampling gun head 6 into the liquid sample to be collected, loosening the negative pressure generating structure 2, generating negative pressure in the sampling gun body 1 at the moment, sucking the continuous water phase sample into the inner capillary 62 under the action of the negative pressure, and shearing the continuous water phase sample into water phase drops at intervals by the oil phase entering from the outer pipeline 61;
s3: under the action of negative pressure, the liquid drop sample is further conveyed into a temperature control module 4 through a sample channel to be heated, and digital isothermal nucleic acid amplification reaction is carried out;
s4: the liquid drops after the reaction are further spread and stored on the inserted detection chip 3;
s5: the insertion type detection chip 3 stored with the sample is taken down from the sampling gun body 1 and transferred to another detection device, so that the digitized absolute quantitative detection can be realized.
Fig. 4 is a schematic top view of digital quantitative detection of droplets after incubation in the plug-in detection chip 3, wherein a is a positive droplet and b is a negative droplet. After the optical equipment photographs and detects the target molecules, the original concentration of the target molecules in the sample can be calculated by identifying and counting the yin-yang liquid drops in the image.
According to a preferred embodiment of the present invention, as shown in fig. 5, when the sampling gun is used to collect the sample in the centrifuge tube 8 sealed by the rubber stopper 9, the tip of the liquid drop sampling gun head 6 is used to pierce the rubber stopper 9, and after the sampling is completed, the rubber stopper 9 automatically seals the centrifuge tube 8, so that a fully-closed sampling environment can be ensured.
The foregoing description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and various modifications can be made to the above-described embodiment of the present invention. All simple, equivalent changes and modifications made in accordance with the claims and the specification of the present application fall within the scope of the patent claims. The present invention is not described in detail in the conventional art.
Claims (10)
1. A sampling gun integrating liquid droplet sampling, sample processing and detection, comprising:
a sampling gun body which is a cuboid-shaped shell with an internal cavity;
the negative pressure generating structure, the plug-in detection chip, the temperature control module and the oil phase storage cavity are sequentially arranged in the sampling gun body from the rear end to the front end, and sample channels for conveying sample liquid drops are arranged in the temperature control module and the oil phase storage cavity;
a liquid drop sampling gun head operatively connected to a front end of the sampling gun body, comprising: an axially extending outer tube; and an inner capillary tube coaxially extending within the outer tube; the hydrophilic filter structure is fixed at the front end of the outer layer pipeline, and a certain gap is kept between the hydrophilic filter structure and the front end of the inner layer capillary tube;
when the liquid drop sampling gun head is connected to the front end of the sampling gun body, a sample channel in the sampling gun body is communicated with an inner capillary tube in the liquid drop sampling gun head, oil phase in the oil phase storage cavity enters between an outer layer pipeline of the liquid drop sampling gun head and the inner capillary tube, under the action of the negative pressure generating structure, a water phase sample is collected into the inner capillary tube in a liquid drop mode through oil phase interval, and further is conveyed from the front end to the rear end through the sample channel, so that digital isothermal nucleic acid amplification reaction under the temperature control module, tiling and detection on the inserted detection chip are realized.
2. The sampling gun of claim 1, wherein the plug-in detection chip comprises a chip housing having an internal cavity, the chip housing having a liquid inlet for sample droplets from a front end of the sampling gun body into the internal cavity.
3. The sampling gun of claim 2, wherein the chip housing has a flange thereon for facilitating handling, the plug-in detection chip being insertable along a side of the sampling gun body, the side of the sampling gun body having a receptacle for insertion of the plug-in detection chip.
4. The sampling gun of claim 1, wherein the sample channel meanders along an S-shaped path within the temperature control module to meet the time requirement for incubation of sample droplets at a constant temperature of 63 ℃ within the temperature control module and to complete a digital isothermal nucleic acid amplification reaction.
5. The sampling gun of claim 1, wherein the oil phase storage chamber comprises a first oil phase storage chamber and a second oil phase storage chamber connected in sequence from a rear end to a front end, the second oil phase storage chamber having a radial dimension comparable to an outer layer conduit of a droplet sampling gun head, the first oil phase storage chamber having a substantially increased radial dimension relative to the second oil phase storage chamber.
6. The sampling gun of claim 1, wherein the sidewall of the oil phase storage chamber is provided with a filler neck for replenishing the oil phase therein.
7. The sampling gun of claim 1, wherein the droplet sampling gun head is frictionally coupled to the sampling gun body front end.
8. A method for integrating drip sampling, sample processing and detection for diagnosis and treatment of non-disease, comprising the steps of:
s1: providing a sampling gun according to any one of claims 1-7, wherein the front end of the sampling gun body is connected with a liquid drop sampling gun head;
s2: manually pressing the negative pressure generating structure to extend the liquid drop sampling gun head into the liquid sample to be collected, and loosening the negative pressure generating structure to collect the water phase sample into the liquid drop sampling gun head in a liquid drop form wrapped by the oil phase;
s3: under the action of negative pressure, the liquid drop sample wrapped by the oil phase is further conveyed into a temperature control module through a sample channel to be heated, and digital isothermal nucleic acid amplification reaction is carried out;
s4: and (5) enabling the liquid drops after the reaction to enter an inserted detection chip for tiling and storing.
9. The method of claim 8, further comprising the step of:
s5: and the inserted detection chip stored with the sample is taken down from the sampling gun body and transferred to detection equipment, so that digital absolute quantitative detection is realized.
10. The method of claim 8, wherein when the sampling gun is used to collect the sample in the centrifuge tube sealed by the rubber plug, the tip of the liquid drop sampling gun head is used to pierce the rubber plug, and the rubber plug automatically seals the centrifuge tube after sampling is completed, so that the complete sealing in the sampling process can be ensured.
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| CN202110976166.7A CN113667597B (en) | 2021-08-24 | 2021-08-24 | Sampling gun and method integrating liquid collecting, drip sampling, sample processing and detection |
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| CN114674992A (en) * | 2022-03-04 | 2022-06-28 | 泉州市佳源环保科技有限公司 | Negative pressure sampling device of water quality ammonia nitrogen online analyzer |
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| US11130120B2 (en) * | 2018-10-01 | 2021-09-28 | Lifeng XIAO | Micro-pipette tip for forming micro-droplets |
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