CN111363664A - LAMP detection chip based on three-layer microchip detection and control method thereof - Google Patents
LAMP detection chip based on three-layer microchip detection and control method thereof Download PDFInfo
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Abstract
The invention discloses a LAMP detection chip based on three layers of microchips, which comprises a main body, a first injection layer, a second loading layer and a third reaction layer, wherein the main body comprises the three layers of microchips; wherein, the second layer of load layer comprises a plurality of load areas, and the third layer of reaction layer comprises a plurality of reaction areas, which are used for forming a system and carrying out reaction. The invention discloses a control method of an LAMP detection chip based on a three-layer microchip.
Description
Technical Field
The invention relates to the technical field of gene detection, in particular to an LAMP detection chip based on three-layer microchip detection and a control method thereof.
Background
Loop-mediated isothermal amplification (LAMP) can amplify nucleic acid in a short time (usually within one hour) under the condition of isothermal temperature (60-65 ℃), and is a simple, convenient, rapid, accurate and low-price gene amplification method; compared with the conventional PCR, the method does not need the processes of thermal denaturation, temperature cycling, electrophoresis, ultraviolet observation and the like of the template. The loop-mediated isothermal amplification method is a brand-new nucleic acid amplification method and has the characteristics of simplicity, rapidness and strong specificity. The technology can be comparable to or even superior to the PCR technology in the indexes such as sensitivity, specificity, detection range and the like, does not depend on any special instrument and equipment to realize on-site high-flux rapid detection, and has detection cost far lower than that of fluorescent quantitative PCR.
The microfluidic chip technology integrates basic operation units of sample preparation, reaction, separation, detection and the like in biological, chemical and medical analysis processes into a micron-scale chip, and automatically completes the whole analysis process. Due to its great potential in the fields of biology, chemistry, medicine and the like, the method has been developed into a new research field crossing the disciplines of biology, chemistry, medicine, fluid, electronics, materials, machinery and the like.
However, there are still significant problems in the prior art, including: (1) the prior art can only carry out qualitative detection, can not carry out quantitative detection, and can not accurately determine the content of pathogens; (2) the reaction container in the prior method has insufficient tightness, and aerosol pollution and false positive are easily caused after the reaction container is opened; (3) in the existing method, a sample and a reagent need to be added respectively, and dosage errors are easily caused in the process of multiple times of sample addition; (4) the result judgment is complex, the reaction result is judged to be easy to have errors through the precipitation change, and electrophoresis detection is still needed in most cases.
Disclosure of Invention
Based on the technical problems, the invention designs and develops the LAMP detection chip based on three-layer microchip detection, and aims to maintain the air tightness of a detection system under the condition of not opening the detection chip and carry out rapid quantitative detection without multiple sample adding.
The invention designs and develops a control method of an LAMP detection chip based on three-layer microchip detection, and aims to carry out rapid quantitative detection without multiple sample adding under the condition that the air tightness of a detection system is kept through an oil seal sample.
The technical scheme provided by the invention is as follows:
a LAMP detection chip based on three-layer microchip detection comprises:
the first injection layer comprises a first sample adding hole, a first oil filling hole and a plurality of first waste liquid pool exhaust holes;
the second layer of load layer comprises a second sample adding hole communicated with the first sample adding hole, a second oil filling hole communicated with the first oil filling hole and a plurality of load areas arranged corresponding to the first waste liquid pool exhaust hole;
the load area comprises a first oil dividing hole communicated with the second oil filling hole and a second waste liquid pool exhaust hole communicated with the first waste liquid pool exhaust hole;
the third reaction layer comprises a third sample adding hole communicated with the second sample adding hole and a plurality of reaction zones arranged corresponding to the load zone;
wherein the reaction zone comprises:
a reagent microchamber, one end of which is communicated with the third sample adding hole;
a mixing chamber, one end of which is communicated with the other end of the reagent microchamber;
a second oil dividing hole communicating with one end of the mixing chamber and communicating with the first oil dividing hole;
one end of the LAMP reaction zone is communicated with the other end of the mixing chamber;
a waste liquid pool which is communicated with the other end of the LAMP reaction zone;
and the third waste liquid pool exhaust hole is formed in the waste liquid pool and communicated with the second waste liquid pool exhaust hole.
Preferably, the load zone further comprises:
and one end of each of the plurality of oil distributing channels is communicated with the second oil filling hole, and the other end of each of the plurality of oil distributing channels is communicated with the first oil distributing hole.
Preferably, the method further comprises the following steps:
a plurality of first gas storage chambers respectively and correspondingly arranged in the load area;
a plurality of second gas storage chambers which are respectively and correspondingly arranged in the reaction zones and are arranged on a channel communicated between the mixing chamber and the LAMP reaction zone;
wherein the first air storage chamber is communicated with the second air storage chamber.
Preferably, the mixing chamber is arranged in a spiral.
Preferably, any one of the LAMP reaction zones in the plurality of reaction zones comprises:
a plurality of reaction microchambers which are communicated and arranged in a snake shape.
Preferably, the reaction microchamber is a plurality of reaction spaces separated by the bulges of the cuboid structure, the cuboid structure has the length of 375-425 μm, the width of 100-150 μm and the height of 69-79 μm.
Preferably, the interval between the cuboid structures is 80-120 mu m.
A control method of LAMP detection chip based on three-layer microchip detection is characterized in that the LAMP detection chip based on three-layer microchip detection is used, and comprises the following steps:
step one, performing an oil inlet process, injecting oil into an oil phase through the first oil filling hole, entering the second layer load layer through the second oil filling hole, dividing the oil phase into a first oil dividing hole connected with a plurality of oil passages through the oil dividing channel, and entering the third layer reaction layer through the first oil dividing hole;
the second oil dividing hole is used for receiving the oil phase of the second layer of load layer, the oil phase enters the main channel through the second oil dividing hole, flows through the mixing chamber, enters the LAMP reaction zone and is filled with the whole reaction microchamber, the redundant oil phase flows into the waste liquid pool, and the first oil inlet hole is sealed after the oil phase is filled with the reaction microchamber to ensure the pressure;
step two, carrying out a sample introduction process, wherein a sample flows through the second sample introduction hole through the first sample introduction hole and directly enters the third sample introduction hole, the sample flows through the reagent micro chamber to flush away reagent dry powder in the reagent micro chamber and enters the mixing chamber for sufficient mixing, at the moment, the first sample introduction hole is in a sealed state, the sample continuously enters the LAMP reaction zone and replaces an oil phase in the reaction micro chamber, and an excess water phase flows into the waste liquid pool;
injecting an oil phase through the first oil filling hole, sequentially flowing through the second oil filling hole, the oil distribution channel, the first oil distribution hole, the oil inlet channel, the second oil distribution hole and the mixing chamber to reach an LAMP reaction region to cut a water phase, combining and sealing the cut water phase with an oil layer remained in the reaction microchamber, and wrapping a sealed reaction system;
and step four, performing LAMP reaction detection.
Compared with the prior art, the invention has the following beneficial effects:
reaction detection integration need not to open reaction vessel and can obtain the reaction result, avoids aerosol pollution problem, and the chip is placed in with the form of dry powder in to the reagent, and the operation is more simple and convenient, has greatly shortened check-out time, need not the application of sample many times, avoids dosage error.
Drawings
FIG. 1 is a schematic structural diagram of a LAMP detection chip for three-layer microchip detection according to the present invention.
FIG. 2 is a schematic structural diagram of the first injection layer of the LAMP detection chip with three layers of microchips for detection according to the present invention.
FIG. 3 is a front view of the first injection layer of the LAMP detection chip with three-layer microchip detection according to the present invention.
FIG. 4 is a cross-sectional view of the first injection layer of the LAMP detection chip for three-layer microchip detection according to the present invention.
FIG. 5 is a schematic structural diagram of the second layer of the loading layer of the LAMP detection chip for three-layer microchip detection according to the present invention.
FIG. 6 is a front view of the second support layer of the LAMP detection chip with three layers of microchips for detection according to the present invention.
FIG. 7 is a cross-sectional view of the second support layer of the LAMP detection chip for three-layer microchip detection according to the present invention.
FIG. 8 is a schematic structural diagram of the third reaction layer of the LAMP detection chip with three layers of microchips for detection according to the present invention.
Fig. 9 is a partially enlarged view of "a" in fig. 8.
FIG. 10 is a front view of the third reaction layer of the LAMP detection chip with three-layer microchip according to the present invention.
FIG. 11 is a schematic view showing the structure of the reaction region in the third reaction layer of the LAMP detection chip with three-layer microchip according to the present invention.
FIG. 12 is a sectional view of the third reaction layer of the LAMP detection chip for three-layer microchip detection according to the present invention.
Detailed Description
The present invention is further described in detail below with reference to the attached drawings so that those skilled in the art can implement the invention by referring to the description text.
As shown in FIG. 1, the invention provides a LAMP detection chip based on three-layer microchip detection, the main body of which comprises three layers of microchips, a first injection layer 100, a second loading layer 200 and a third reaction layer 300; wherein, the second layer of supporting layer 200 comprises a plurality of supporting zones, and the third layer of reaction layer 300 comprises a plurality of reaction zones, which are used for forming a system and carrying out a reaction.
As shown in fig. 2 to 4, the first injection layer 100 includes a first filling hole 110, a first filling hole 120, and a plurality of first waste liquid pool vent holes 130; the first sample adding hole 110 is simultaneously communicated with the second sample adding hole 210 of the second loading layer 200 and the third sample adding hole 310 of the third reaction layer 300, the first oil filling hole 120 is simultaneously communicated with the second oil filling hole 220 of the second loading layer 200, and the first sample adding hole 110 and the first oil filling hole 120 are respectively used for injecting water phase and oil phase;
as shown in fig. 5 to 7, the second loading layer 200 includes a second sample hole 210, a second filling hole 220 and a plurality of loading areas; wherein the plurality of load zones include an oil distribution passage 250 gradually branched into a plurality of, a plurality of first oil distribution holes 240, a plurality of first gas storage chambers 260, and a plurality of second waste liquid pool vent holes 230; the second layer loading layer 200 is gradually divided into a plurality of oil dividing channels 250 from a second oil filling hole 220, and the oil dividing channels are respectively connected to a plurality of first oil dividing holes 240 entering the third layer reaction layer 300, and a plurality of first gas storage chambers 260 are additionally designed for storing the waste gas of the third layer reaction layer 300; the sample injection device specifically comprises a second sample injection hole 210 communicated with the first sample injection hole 110, a second oil filling hole 220 communicated with the first oil filling hole 120, and a plurality of second waste liquid pool exhaust holes 230 correspondingly communicated with a plurality of first waste liquid pool exhaust holes 130 respectively;
as shown in fig. 8 to 12, the third reaction layer 300 is composed of a third sample adding hole 310 and a plurality of reaction zones, and the reaction zones are respectively arranged corresponding to the load zones one by one; each reaction zone consists of a reagent microchamber 341, a second oil dividing hole 320, an oil inlet channel 350, a spiral mixer 342, a second gas storage chamber 360, an LAMP reaction zone 343, a waste liquid pool 370 and a third waste liquid pool exhaust hole 330; the LAMP reaction zone 343 comprises a serpentine channel 343a and a plurality of reaction microchambers 343b, the reagent microchamber 341 stores the primers and the dry powder such as the fluorescence detection reagent required for the reaction, the second oil distribution hole 320 of the third reaction layer 300 is communicated with the first oil distribution hole 240 of the second load layer 200, the spiral mixer 342 is used for uniformly mixing the sample and the reaction reagent, the second air storage chamber 360 of the third reaction layer 300 is communicated with the first air storage chamber 260 of the second load layer 200 and is used for storing the residual gas before the sample is added after the oil phase is injected, the serpentine channel 343a is connected with the plurality of reaction microchambers 343b to uniformly divide the reaction system into a plurality of parts, the waste liquid pool 370 is used for storing the redundant waste liquid, the LAMP reaction zone 343 comprises a plurality of serpentine channels 343a in total, and each row of the plurality of reaction microchambers 343b divides the total reaction system into a plurality of parts to realize digitization.
In another embodiment, as shown in fig. 5 to 7, the second loading layer 200 includes 1 second sample adding hole 210, 1 second oil adding hole 220, an oil separating channel 250 gradually branched into 8, 8 first oil separating holes 240, 8 first gas storage chambers 260, and 8 second waste liquid pool vent holes 230, the second loading layer 200 is gradually branched into 8 oil separating channels 250 from one second oil adding hole 220, the second oil adding holes 220 are respectively connected to 8 first oil separating holes 240 entering the third reaction layer 300, and 8 first gas storage chambers 260 are additionally designed for storing the waste gas of the third reaction layer 300.
In another embodiment, as shown in fig. 8, the third reaction layer 300 is composed of 8 reaction zones, and is respectively disposed in one-to-one correspondence with the oil distribution channel 250, the first oil distribution hole 240, the first gas storage chamber 260, and the second waste liquid pool exhaust hole 230 of the second loading layer 200.
In another embodiment, as shown in fig. 11, the serpentine channel 343a connects 360 reaction microchambers 343b, divides the reaction system into 360 parts uniformly, the waste liquid pool 370 is used to store the excess waste liquid, the LAMP reaction zone 343 contains 12 rows of serpentine channels 343a, each row contains 30 reaction microchambers 343b, and the total number of 360 reaction microchambers 343b divides the total reaction system into 360 parts, and the detection reagent is stored in the reagent microchamber 341 in the form of dry powder, so as to realize detection digitization.
In another embodiment, the interval between each row of serpentine channels 343a is 100 to 150 μm; preferably, the spacing between each row of serpentine channels 343a is 125 μm.
In another embodiment, the reaction microchamber 343b is a plurality of reaction spaces separated by protrusions of a rectangular parallelepiped structure, the rectangular parallelepiped structure has a length of 375 to 425 μm, a width of 100 to 150 μm, and a height of 69 to 79 μm; preferably, the rectangular parallelepiped structure has a length of 400 μm, a width of 120 μm and a height of 74 μm.
In another embodiment, the interval between the cuboid structures is 80-120 μm; preferably, the spacing between the cuboid structures is 100 μm.
The invention provides a flow of an LAMP detection chip based on three-layer microchip detection, which comprises four steps, wherein an oil phase protective layer is formed in the first step, a reagent and a sample are fed in the second step, mixing and digital sample introduction are carried out, a reaction system is sealed and then fed in the third step, and LAMP reaction detection is carried out in the fourth step, and the flow specifically comprises the following steps:
step one, carrying out an oil inlet process: the oil phase is connected with an oil source through the first oil filling hole 120 of the first injection layer 100, is injected, enters the second load layer 200 through the second oil filling hole 220, is divided into 8 oil passages by the oil dividing passage 250 to be connected with 8 first oil dividing holes 240, and enters the third reaction layer 300 through the second oil dividing holes 320 by the first oil dividing holes 240;
the second oil dividing hole 320 in the third reaction layer 300 is configured to receive the oil phase in the second load layer 200, enter the oil inlet channel 350 through the second oil dividing hole 320, flow through the spiral mixer 342 into the LAMP reaction region 343, fill the entire reaction microchamber 343b through the serpentine channel 343a, allow the excess oil phase to flow into the waste liquid pool 370, and seal the first oil inlet 120 after the oil phase fills the reaction microchamber to ensure pressure;
step two, carrying out a sample injection process: the sample flows through the second sample adding hole 210 of the second loading layer 200 through the first sample adding hole 110 of the first injection layer 100, and directly enters the third sample adding hole 310 of the third reaction layer 300, the sample flows through the reagent micro chamber 341 to flush the reagent dry powder in the reagent micro chamber 341, and then enters the spiral mixer 342 to be fully mixed, at this time, in order to ensure the pressure, the first oil adding hole 120 is in a sealed state, so that partial waste gas exists in the passage from the third sample adding hole 310 to the oil inlet passage 350, in the process of flowing through the second gas storage chamber 360, the redundant waste gas is fully discharged and stored in the first gas storage chamber 260 of the second loading layer 200, the influence of the gas on the filling rate of the reaction micro chamber 343b is avoided, the sample continues to flow into the LAMP reaction region 343 after passing through the second gas storage chamber 360, and replaces the oil phase in the reaction micro chamber 343b, because PDMS is subjected to oleophylic treatment, the water phase cannot completely replace the existing oil phase, a layer of oil film is left to wrap and prevent the evaporation of water, and the redundant water phase flows into the waste liquid pool 370;
step three, carrying out the oil inlet process again: after the second step is completed, the first sample adding hole 110 is sealed, the oil phase is injected through the first oil adding hole 120 again, sequentially flows through the second oil adding hole 220, the oil distribution channel 250, the first oil distribution hole 240, the oil inlet channel 350 and the spiral mixer 342 to reach the LAMP reaction region 343, the water phase is cut, and is combined and sealed with the oil layer remained in the reaction microchamber 343b to form a complete oil film to wrap the sealed reaction system;
step four, performing LAMP reaction: the sample template, six specific primers, Mix reagent, fluorescence detection reagent and the like required by the reaction are completely contained in the reagent microchamber 341 of each reaction zone, the detection chip is reacted for 1 hour at the constant temperature of 65 ℃, and the reaction result of the microchamber can be judged through the fluorescence reaction; the fluorescence light spots of 360 microchambers are collected through a CCD, and the reaction results of different microchambers are analyzed through computer Image J software.
While embodiments of the invention have been described above, it is not limited to the applications set forth in the description and the embodiments, which are fully applicable in various fields of endeavor to which the invention pertains, and further modifications may readily be made by those skilled in the art, it being understood that the invention is not limited to the details shown and described herein without departing from the general concept defined by the appended claims and their equivalents.
Claims (8)
1. The LAMP detection chip based on three-layer microchip detection is characterized by comprising:
the first injection layer comprises a first sample adding hole, a first oil filling hole and a plurality of first waste liquid pool exhaust holes;
the second layer of load layer comprises a second sample adding hole communicated with the first sample adding hole, a second oil filling hole communicated with the first oil filling hole and a plurality of load areas arranged corresponding to the first waste liquid pool exhaust hole;
the load area comprises a first oil dividing hole communicated with the second oil filling hole and a second waste liquid pool exhaust hole communicated with the first waste liquid pool exhaust hole;
the third reaction layer comprises a third sample adding hole communicated with the second sample adding hole and a plurality of reaction zones arranged corresponding to the load zone;
wherein the reaction zone comprises:
a reagent microchamber, one end of which is communicated with the third sample adding hole;
a mixing chamber, one end of which is communicated with the other end of the reagent microchamber;
a second oil dividing hole communicating with one end of the mixing chamber and communicating with the first oil dividing hole;
one end of the LAMP reaction zone is communicated with the other end of the mixing chamber;
a waste liquid pool which is communicated with the other end of the LAMP reaction zone;
and the third waste liquid pool exhaust hole is formed in the waste liquid pool and communicated with the second waste liquid pool exhaust hole.
2. The LAMP detection chip based on three-layer microchip detection according to claim 1, characterized in that the loading zone further comprises:
and one end of each of the plurality of oil distributing channels is communicated with the second oil filling hole, and the other end of each of the plurality of oil distributing channels is communicated with the first oil distributing hole.
3. The LAMP detection chip based on three-layer microchip detection according to claim 2, characterized by further comprising:
a plurality of first gas storage chambers respectively and correspondingly arranged in the load area;
a plurality of second gas storage chambers which are respectively and correspondingly arranged in the reaction zones and are arranged on a channel communicated between the mixing chamber and the LAMP reaction zone;
wherein the first air storage chamber is communicated with the second air storage chamber.
4. The LAMP detection chip based on three-layer microchip detection according to claim 3, characterized in that the mixing chamber is arranged in a spiral.
5. The LAMP detection chip based on three-layer microchip detection according to claim 1, characterized in that any one LAMP reaction zone in the plurality of reaction zones comprises:
a plurality of reaction microchambers which are communicated and arranged in a snake shape.
6. The LAMP detection chip based on three-layer microchip detection as claimed in claim 5, characterized in that the reaction microchamber is a plurality of reaction spaces separated by the projections of a rectangular parallelepiped structure, the length of the rectangular parallelepiped structure is 375-425 μm, the width is 100-150 μm, and the height is 69-79 μm.
7. The LAMP detection chip based on three-layer microchip detection as claimed in claim 6, characterized in that the interval between the rectangular parallelepiped structures is 80-120 μm.
8. A control method of LAMP detection chip based on three-layer microchip detection, characterized in that the LAMP detection chip based on three-layer microchip detection according to any one of claims 1 to 7 is used, and comprises the following steps:
step one, performing an oil inlet process, injecting oil into an oil phase through the first oil filling hole, entering the second layer load layer through the second oil filling hole, dividing the oil phase into a first oil dividing hole connected with a plurality of oil passages through the oil dividing channel, and entering the third layer reaction layer through the first oil dividing hole;
the second oil dividing hole is used for receiving the oil phase of the second layer of load layer, the oil phase enters the main channel through the second oil dividing hole, flows through the mixing chamber, enters the LAMP reaction zone and is filled with the whole reaction microchamber, the redundant oil phase flows into the waste liquid pool, and the first oil filling hole is sealed after the oil phase is filled with the reaction microchamber to ensure the pressure;
step two, carrying out a sample introduction process, wherein a sample flows through the second sample introduction hole through the first sample introduction hole and directly enters the third sample introduction hole, the sample flows through the reagent micro chamber to flush away reagent dry powder in the reagent micro chamber and enters the mixing chamber for sufficient mixing, at the moment, the first sample introduction hole is in a sealed state, the sample continuously enters the LAMP reaction zone and replaces an oil phase in the reaction micro chamber, and an excess water phase flows into the waste liquid pool;
injecting an oil phase through the first oil filling hole, sequentially flowing through the second oil filling hole, the oil distribution channel, the first oil distribution hole, the oil inlet channel, the second oil distribution hole and the mixing chamber to reach an LAMP reaction region to cut a water phase, combining and sealing the cut water phase with an oil layer remained in the reaction microchamber, and wrapping a sealed reaction system;
and step four, performing LAMP reaction detection.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114632558A (en) * | 2021-12-17 | 2022-06-17 | 上海交通大学医学院附属仁济医院 | Microfluidic chip and preparation method and application thereof |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013101154A (en) * | 2013-02-26 | 2013-05-23 | Sony Corp | Microchip and method of manufacturing microchip |
| CN206334683U (en) * | 2016-07-28 | 2017-07-18 | 上海速创诊断产品有限公司 | A kind of CD plate-likes micro-fluidic chip |
| CN107398307A (en) * | 2016-05-18 | 2017-11-28 | 博奥生物集团有限公司 | A kind of integrated micro-flow control chip |
| CN107988046A (en) * | 2018-01-23 | 2018-05-04 | 吉林大学 | Self-absorption multichannel detection of pathogens micro-fluidic chip based on LAMP |
| CN109682878A (en) * | 2019-03-01 | 2019-04-26 | 南京岚煜生物科技有限公司 | A kind of multi-channel fluid hemostasis examination chip with five-layer structure |
| CN109929749A (en) * | 2019-03-27 | 2019-06-25 | 深圳市尚维高科有限公司 | Micro-fluid self-driven micro-fluidic chip and its application method |
| CN110193385A (en) * | 2019-04-26 | 2019-09-03 | 长春技特生物技术有限公司 | Disc type high throughput microflow controlled biochip, automation lotion generating means and lotion generation method |
| CN110616138A (en) * | 2019-10-09 | 2019-12-27 | 山东百骏生物科技有限公司 | Chambered multi-index nucleic acid amplification micro-fluidic chip |
-
2020
- 2020-04-24 CN CN202010330178.8A patent/CN111363664B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013101154A (en) * | 2013-02-26 | 2013-05-23 | Sony Corp | Microchip and method of manufacturing microchip |
| CN107398307A (en) * | 2016-05-18 | 2017-11-28 | 博奥生物集团有限公司 | A kind of integrated micro-flow control chip |
| US20190143324A1 (en) * | 2016-05-18 | 2019-05-16 | Capitalbio Corporation | An integrated microfluidic chip and methods of use |
| CN206334683U (en) * | 2016-07-28 | 2017-07-18 | 上海速创诊断产品有限公司 | A kind of CD plate-likes micro-fluidic chip |
| CN107988046A (en) * | 2018-01-23 | 2018-05-04 | 吉林大学 | Self-absorption multichannel detection of pathogens micro-fluidic chip based on LAMP |
| CN109682878A (en) * | 2019-03-01 | 2019-04-26 | 南京岚煜生物科技有限公司 | A kind of multi-channel fluid hemostasis examination chip with five-layer structure |
| CN109929749A (en) * | 2019-03-27 | 2019-06-25 | 深圳市尚维高科有限公司 | Micro-fluid self-driven micro-fluidic chip and its application method |
| CN110193385A (en) * | 2019-04-26 | 2019-09-03 | 长春技特生物技术有限公司 | Disc type high throughput microflow controlled biochip, automation lotion generating means and lotion generation method |
| CN110616138A (en) * | 2019-10-09 | 2019-12-27 | 山东百骏生物科技有限公司 | Chambered multi-index nucleic acid amplification micro-fluidic chip |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114632558A (en) * | 2021-12-17 | 2022-06-17 | 上海交通大学医学院附属仁济医院 | Microfluidic chip and preparation method and application thereof |
| CN114632558B (en) * | 2021-12-17 | 2023-08-18 | 上海交通大学医学院附属仁济医院 | Microfluidic chip and preparation method and application thereof |
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