CN115521862A - Nucleic acid detection chip general structure combined with general test strip and use method - Google Patents
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- CN115521862A CN115521862A CN202211112772.5A CN202211112772A CN115521862A CN 115521862 A CN115521862 A CN 115521862A CN 202211112772 A CN202211112772 A CN 202211112772A CN 115521862 A CN115521862 A CN 115521862A
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- 102000039446 nucleic acids Human genes 0.000 title claims abstract description 31
- 108020004707 nucleic acids Proteins 0.000 title claims abstract description 31
- 150000007523 nucleic acids Chemical class 0.000 title claims abstract description 31
- 238000001514 detection method Methods 0.000 title claims abstract description 28
- 238000012812 general test Methods 0.000 title claims abstract description 9
- 238000000034 method Methods 0.000 title claims abstract description 7
- 238000006243 chemical reaction Methods 0.000 claims abstract description 94
- 238000012360 testing method Methods 0.000 claims abstract description 37
- 239000004205 dimethyl polysiloxane Substances 0.000 claims abstract description 27
- 235000013870 dimethyl polysiloxane Nutrition 0.000 claims abstract description 27
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 claims abstract description 27
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims abstract description 27
- 239000011229 interlayer Substances 0.000 claims abstract description 13
- 239000007795 chemical reaction product Substances 0.000 claims abstract description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 16
- 229910052782 aluminium Inorganic materials 0.000 claims description 10
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims description 10
- 239000011888 foil Substances 0.000 claims description 10
- 108090000623 proteins and genes Proteins 0.000 claims description 10
- 230000003321 amplification Effects 0.000 claims description 8
- 239000010410 layer Substances 0.000 claims description 8
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 8
- 239000000758 substrate Substances 0.000 claims description 7
- 108091005461 Nucleic proteins Proteins 0.000 claims description 6
- 230000001360 synchronised effect Effects 0.000 claims description 3
- 230000004888 barrier function Effects 0.000 claims 1
- 238000005842 biochemical reaction Methods 0.000 abstract description 9
- 239000000463 material Substances 0.000 abstract description 8
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 230000005484 gravity Effects 0.000 abstract description 4
- 238000002331 protein detection Methods 0.000 abstract description 4
- 230000036760 body temperature Effects 0.000 abstract description 3
- 238000010438 heat treatment Methods 0.000 abstract 1
- 239000002390 adhesive tape Substances 0.000 description 13
- 239000012295 chemical reaction liquid Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000011901 isothermal amplification Methods 0.000 description 3
- 230000002146 bilateral effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 238000010146 3D printing Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000018120 Recombinases Human genes 0.000 description 1
- 108010091086 Recombinases Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- 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
- C12Q1/6813—Hybridisation assays
- C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
- C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
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- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
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- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
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Abstract
The invention relates to a general structure of a nucleic acid detection chip combined with a general test strip and a use method thereof, belonging to the technical field of disease detection chips. According to the structure, a PDMS material is used for preparing a main body reaction chip with a specific shape, and nucleic acid or protein detection is realized by combining a detection test strip commonly used in the market. The main reaction chip comprises a sample reaction chamber and a sticker switch. Heating at the same temperature in the sample reaction cavity to carry out biochemical reaction, withdrawing the sticker switch after the reaction is finished, and breaking the PDMS interlayer, wherein the reaction product in the sample reaction cavity flows into the test strip due to gravity and the structural characteristics of the chip, and the test result is presented by the macroscopic strip color change of the test strip. The invention solves the problems that the reaction and the detection are carried out step by step and the reaction is carried out depending on laboratory equipment, can be matched with the existing test paper strips on the market for use, has small chip scale, and can be placed close to the skin to carry out the reaction by utilizing the body temperature so as to achieve the purpose of detecting the disease markers.
Description
Technical Field
The invention belongs to the technical field of disease detection chips, and relates to a nucleic acid detection chip general structure combined with a general test strip and a use method.
Background
Generally, in a laboratory, a nucleic acid amplification reaction or a protein biochemical reaction (such as an antigen-antibody reaction, an enzyme catalytic reaction, etc.) is performed by using Polymerase Chain Reaction (PCR) and other technologies, and then the nucleic acid or the protein is loaded on a test strip for detection. Due to different temperature cycle changes of nucleic acid amplification, complex control of protein biochemical reaction concentration, accurate quantitative reagent addition and other requirements, detection can be performed by professional equipment and professionals trained, and the requirement of convenient detection outside a laboratory is difficult to meet. By using techniques such as recombinase polymerase amplification, convenient isothermal amplification reaction of nucleic acid can be carried out at a single temperature. If a special universal structure is designed, and the isothermal amplification of nucleic acid or the biochemical reaction of protein is combined with the test paper strips commonly used in the market, the detection of nucleic acid or protein outside a laboratory can be realized by using the same device without professional equipment and technicians. Meanwhile, the chip can be detected by using the existing test paper strips in the market.
Disclosure of Invention
The invention combines nucleic acid constant temperature amplification reaction or protein biochemical reaction with a test strip which is used in the market, provides a general structure of a disease marking nucleic acid or protein detection test strip under constant temperature reaction, aims to solve the problem that reaction is needed in a laboratory in the prior art, and is convenient for detecting nucleic acid or protein outside the laboratory.
The technical scheme of the invention is as follows:
a nucleic acid detection chip general structure combined with a general test strip comprises a main reaction chip and a cover.
The main body reaction chip takes a PDMS substrate 1-4 as a main body, and a sample reaction cavity 1-1 and an internal horizontal channel for placing a sticker switch 1-2 are arranged on the PDMS substrate 1-4; the sample reaction cavity 1-1 is a funnel-shaped triangular frustum chamber, the bottom of the sample reaction cavity 1-1 is connected with a horizontal channel, and the horizontal channel extends to the edge of one side of the main body reaction chip from the position right below the sample reaction cavity 1-1; the sticker switch 1-2 is a bilaterally symmetrical integrated structure and comprises a tip 1-5 and a pull structure 1-6, wherein the tip 1-5 is consistent with the bottom of the sample reaction chamber 1-1 in shape and size, the tip 1-5 is positioned at the bottom of the sample reaction chamber 1-1, and the pull structure 1-6 extends from a horizontal channel to the outside of the main reaction chip; pouring a layer of PDMS between the upper surface of the tip 1-5 and the bottom of the sample reaction cavity 1-1 to form a PDMS interlayer 1-3;
the main structure of the cover is called as a cover body 2-7, the cover body 2-7 is provided with a bulge 2-8 with the same size as the sample reaction cavity 1-1, the cover is covered on the main reaction chip, and the bulge 2-8 is positioned in the sample reaction cavity 1-1;
the using method comprises the following steps: when the nucleic acid amplification or protein reaction is finished in the sample reaction cavity 1-1, and the pull structure 1-6 is pulled outwards, the PDMS interlayer 1-3 can be damaged by the tip 1-5, so that the reaction product in the sample reaction cavity 1-1 flows into a test strip sample pad below the main reaction chip, and the test strip generates a corresponding color reaction, thereby achieving the purpose of detection.
The cover is covered on the main reaction chip and can be sealed by an aluminum foil adhesive tape.
The main reaction chips can be used in parallel, and each main reaction chip is connected with a corresponding test strip to realize the synchronous detection of multiple target nucleic acids or proteins of multiple samples.
The main advantages of the invention are: the invention designs a special universal structure, which combines biochemical reaction before sample detection with a common test strip technology in the market, and the structure has small volume, the design width of the lower layer can be consistent with the width of a common test strip sample pad in the market, and the combination with the common test strip sample pad is convenient to realize. The invention can realize the detection of disease nucleic acid or protein markers outside the laboratory by simple operation by using body temperature without professional equipment and technicians. The invention overcomes the defect that the nucleic acid or protein detection test strip needs professional equipment and technicians to operate, and can increase the number of sample reaction tanks by connecting a plurality of main reaction chips in parallel, realize multi-index synchronous detection, improve the detection efficiency and effectively reduce the cost.
Drawings
FIGS. 1 (a) to 1 (c) are a top view, a front view and a side view of a main reaction chip, respectively;
FIGS. 2 (a) -2 (c) are top, front and side views, respectively, of a lid;
FIG. 3 is a schematic diagram of the assembly of the reaction chip and the test strip;
in the figure: 1-1 sample reaction chamber; 1-2 paster switches; 1-3PDMS spacers; 1-4PDMS substrate; 1-5 pointed ends; 1-6 drawing structure: 2-7 cover bodies; 2-8 bulges.
Detailed Description
The following further describes a specific embodiment of the present invention with reference to the drawings and technical solutions.
A nucleic acid detection chip general structure combined with a general test strip is a bilateral symmetry structure and comprises a main reaction chip and a cover. The main body reaction chip is manufactured by one-time pouring of PDMS materials through corresponding moulds in figures 1 (a) to 1 (c), a PDMS substrate 1-4 is used as a main body, and a sample reaction cavity 1-1 and an internal horizontal channel for placing a sticker switch 1-2 are arranged on the PDMS substrate 1-4. The main reaction chip performs nucleic acid amplification or protein reaction at the same temperature and provides a place for test strip detection. The overall height of the subject reaction chip in this example was 8mm, length 14mm, and width 17mm.
The sample reaction cavity 1-1 is designed into a funnel-shaped triangular frustum chamber, the sample reaction cavity 1-1 is a regular triangular frustum chamber, the lower surface of the sample reaction cavity is an isosceles triangle with a bottom edge of 4mm and a height of 4.8mm, and the upper surface of the sample reaction cavity is an isosceles triangle with a bottom edge of 10mm and a height of 12 mm. The funnel-shaped chamber has the following advantages: (1) the lower layer contact surface is easy to contact with the test strip sample pad; (2) the contact surface of the lower layer is small and easy to damage; (3) The reaction liquid is more easily concentrated on the lower side for outflow due to the chamber configuration and gravity.
A matched cover is made of hard material (such as 3D printing material) on the top of the sample reaction chamber 1-1 to prevent liquid in the chamber from splashing. The cover is of a left-right symmetrical structure, and is divided into a cover body 2-7 part (with the size of 14mm in length, 16mm in width and 2mm in height) and a downward bulge 2-8 part in the figures 2 (a) to 2 (c). The horizontal section size of part of the bulge 2-8 is the same as that of the sample reaction cavity 1-1, the height is 2mm, the lower surface is an isosceles triangle with the bottom edge being 10mm and the height being 12mm, and the bulge can be embedded into the sample reaction cavity 1-1. The sample reaction chamber 1-1 and the lid are sealed with an aluminum foil tape to prevent the reaction solution from splashing and to fix the lower chip.
The sticker switch 1-2 is of a bilateral symmetry structure and is 0.1mm thick. The adhesive tape is divided into two parts, namely a tip 1-5 and a drawing structure 1-6, wherein the bottom edge of the tip 1-5 is 1mm, the height of the tip is 1.2mm, the material is a PVC film, the material can also be made of softer materials such as an aluminum foil adhesive tape, and the PDMS2 layer below the sticker switch 1-2 needs to be correspondingly reduced, but the operation is difficult during final pouring; the tip 1-5 can also be made of metal sheets or wooden materials, and the finally formed PDMS interlayer 1-3 can be correspondingly enlarged, so that the integral manufacture is convenient. The switch tip 1-5 is completely consistent with the lower surface of the sample reaction cavity 1-1, and has the advantages of protecting the reaction liquid from leaking before detection, being easier to operate during filling and being easy to damage the PDMS interlayer 1-3 during extraction. During manufacturing, the upper surface and the lower surface of the tip 1-5 are coated with double-sided adhesive tapes, the lower surface is combined with the bulge of the film, the upper surface is combined with PDMS, after solidification, a small amount of PDMS solution forms a PDMS interlayer 1-3, the PDMS interlayer 1-3 on the bottom surface of the sample reaction cavity 1-1 can be damaged by drawing out the sticker switch 1-2, and reaction liquid can flow into a sample pad of a paper chip from a horizontal channel to develop color smoothly. The total length of the exposed parts of the drawing structures 1-6 is 10mm, and the drawing structures can be bent to save space and labor.
The thickness of the PDMS interlayer 1-3 is 0.1mm, and the sticker switch 1-2 is separated from the sample reaction chamber 1-1. After nucleic acid amplification or protein biochemical reaction is finished in the sample reaction cavity 1-1, a sample pad of a common test strip in the market is placed below the main reaction chip, the sticker switch 1-2 is pulled out, the PDMS interlayer 1-3 is damaged, and a reaction product in the sample reaction cavity 1-1 flows onto the sample pad of the paper chip through a channel structure and gravity and is absorbed by the test strip; the thickness of the PDMS layer below the sticker switch 1-2 is 0.8mm, the sticker switch 1-2 is wrapped by the PDMS interlayer 1-3, and the PDMS interlayer 1-3 is guaranteed to be damaged when the sticker switch is drawn out.
The test paper strip is a test paper strip commonly used in the market, a sample pad is exposed when the test paper strip is required to be used, after reaction, the lower surface of the main reaction chip is in contact with the sample pad, and an aluminum foil adhesive tape is used for wrapping the sample pad.
The aluminum foil adhesive tape is cut into segments with the length of 40mm and the width of 17mm, the lines are drawn at the positions 13mm away from the left side and the right side, the lower part of the central part of the aluminum foil adhesive tape is torn, the two sides of the cover and the chip are wrapped, the adhesive paper at the position of the ear of the aluminum foil adhesive tape is torn after the reaction, the adhesive tape is adhered to the lower surface of the test strip, and the test strip is fixed.
The specific use mode is (as shown in fig. 3): adding a nucleic acid isothermal amplification reagent or a protein biochemical reaction reagent and a sample to be detected into the sample reaction chamber 1-1 through an injector, covering a cover, and covering a layer of aluminum foil adhesive tape on the upper surface of the cover for sealing. Fixing the main reaction chip under the armpit by using an adhesive tape, taking out the main reaction chip after reacting for a period of time by using body temperature, placing a sample pad of a nucleic acid or protein detection test strip on the market on the lower surface of the main reaction chip, tearing off a sticker at the ear of an aluminum foil adhesive tape to adhere the adhesive tape on the lower surface of the test strip, drawing out a sticker switch 1-2, and destroying a PDMS interlayer 1-3. The liquid is absorbed on the test strip due to the self gravity, the chip structure and the water absorption of the test strip. The reaction product is subjected to color development reaction by a test strip, and then the detection result is judged by the color change of the strip. The body surface temperature is used as a reaction condition in the embodiment, the structure can also be carried out at other temperatures, and the biochemical reaction of nucleic acid or protein is combined with the detection of a test strip commonly used in the market.
Claims (4)
1. A nucleic acid detection chip general structure combined with a general test strip is characterized in that the nucleic acid detection chip general structure combined with the general test strip comprises a main reaction chip and a cover;
the main body reaction chip takes a PDMS substrate (1-4) as a main body, and a sample reaction cavity (1-1) and an internal horizontal channel for placing a sticker switch (1-2) are arranged on the PDMS substrate (1-4); the sample reaction cavity (1-1) is a funnel-shaped triangular frustum chamber, the bottom of the sample reaction cavity (1-1) is connected with a horizontal channel, and the horizontal channel extends to the edge of one side of the main body reaction chip from the position right below the sample reaction cavity (1-1); the sticker switch (1-2) is of a bilaterally symmetrical integrated structure and comprises a tip (1-5) and a drawing structure (1-6), wherein the shape and the size of the tip (1-5) are consistent with those of the bottom of the sample reaction cavity (1-1), the tip (1-5) is positioned at the bottom of the sample reaction cavity (1-1), and the drawing structure (1-6) extends to the outside of the main reaction chip from a horizontal channel; a layer of PDMS is poured between the upper surface of the tip (1-5) and the bottom of the sample reaction cavity (1-1) to form a PDMS interlayer (1-3);
the main structure of the cover is called as a cover body (2-7), the cover body (2-7) is provided with a bulge (2-8) with the same size as the sample reaction cavity (1-1), the cover is covered on the main reaction chip, and the bulge (2-8) is positioned in the sample reaction cavity (1-1).
2. The universal nucleic acid detecting chip structure combined with the universal test strip as claimed in claim 1, wherein the lid covers the main reaction chip and is sealed with aluminum foil tape.
3. The method of claim 1 or 2, wherein when the pull structure (1-6) is pulled outward after the nucleic acid amplification or protein reaction is completed in the sample reaction chamber (1-1), the tip (1-5) breaks the PDMS barrier (1-3) so that the reaction product in the sample reaction chamber (1-1) flows into the test strip sample pad under the main reaction chip through the horizontal channel, and the test strip generates a corresponding color reaction.
4. The general structure of a nucleic acid detection chip combined with a general test strip of claim 1 or 2, wherein a plurality of main reaction chips can be used in parallel, and each main reaction chip is connected with a corresponding test strip to realize synchronous detection of multiple target nucleic acids or proteins of multiple samples.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202211112772.5A CN115521862A (en) | 2022-09-14 | 2022-09-14 | Nucleic acid detection chip general structure combined with general test strip and use method |
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| CN202211112772.5A CN115521862A (en) | 2022-09-14 | 2022-09-14 | Nucleic acid detection chip general structure combined with general test strip and use method |
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
| CN211339511U (en) * | 2019-07-08 | 2020-08-25 | 中国人民解放军东部战区疾病预防控制中心 | Integrated test strip for LAMP or RT-LAMP |
| CN112899122A (en) * | 2021-03-02 | 2021-06-04 | 济南国益生物科技有限公司 | Closed lateral flow chromatography color development device |
| WO2021237396A1 (en) * | 2020-05-25 | 2021-12-02 | 杭州梓晶生物有限公司 | Integrated self-service nucleic acid detection device and use method thereor |
| CN114196519A (en) * | 2021-12-10 | 2022-03-18 | 中国检验认证集团辽宁有限公司 | Disease marker detection chip general structure utilizing body temperature reaction and use method thereof |
| WO2022151545A1 (en) * | 2021-01-14 | 2022-07-21 | 广东东阳光药业有限公司 | Method for detecting multi-nucleic acid amplification product and detection kit thereof |
-
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- 2022-09-14 CN CN202211112772.5A patent/CN115521862A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110218769A (en) * | 2019-07-08 | 2019-09-10 | 中国人民解放军东部战区疾病预防控制中心 | The multi-joint test strips of integration integral type for LAMP or RT-LAMP |
| CN211339511U (en) * | 2019-07-08 | 2020-08-25 | 中国人民解放军东部战区疾病预防控制中心 | Integrated test strip for LAMP or RT-LAMP |
| WO2021237396A1 (en) * | 2020-05-25 | 2021-12-02 | 杭州梓晶生物有限公司 | Integrated self-service nucleic acid detection device and use method thereor |
| WO2022151545A1 (en) * | 2021-01-14 | 2022-07-21 | 广东东阳光药业有限公司 | Method for detecting multi-nucleic acid amplification product and detection kit thereof |
| CN112899122A (en) * | 2021-03-02 | 2021-06-04 | 济南国益生物科技有限公司 | Closed lateral flow chromatography color development device |
| CN114196519A (en) * | 2021-12-10 | 2022-03-18 | 中国检验认证集团辽宁有限公司 | Disease marker detection chip general structure utilizing body temperature reaction and use method thereof |
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