CN214953571U - Chip for rapidly detecting dengue virus antibody marked by red fluorescent microspheres - Google Patents
Chip for rapidly detecting dengue virus antibody marked by red fluorescent microspheres Download PDFInfo
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- CN214953571U CN214953571U CN202120551409.8U CN202120551409U CN214953571U CN 214953571 U CN214953571 U CN 214953571U CN 202120551409 U CN202120551409 U CN 202120551409U CN 214953571 U CN214953571 U CN 214953571U
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Abstract
The utility model relates to an utilize dengue fever virus antibody short-term test chip of red fluorescence microballon mark, including the chip carrier, the chip front end is equipped with sample sampling hole, and the rear end is equipped with out the appearance hole, embedded microchannel of chip carrier, IgG reaction tank and IgM reaction tank, sample sampling hole is circular, sample sampling hole bottom sets up the through-hole, inside microchannel, IgG reaction tank and the IgM reaction tank of being equipped with of chip carrier, IgG reaction tank and IgM reaction tank have the adhesion after polylysine acid is handled, and reaction tank bottom average distribution IgG/IgM detects and uses the antibody, it passes through the microchannel with each reaction tank and is connected to go out the appearance hole. The invention has the advantages of simple and practical technology, high stability and good detection effect, and can well replace the prior ELISA detection technology in the aspect of detection.
Description
Technical Field
The utility model belongs to the technical field of the antibody detects, concretely relates to utilize dengue fever virus antibody short-term test chip of red fluorescence microballon mark.
Background
Aiming at the rapid detection of the dengue fever virus, the method has the disadvantage of inconvenient detection due to the reasons of longer process, complex operation and the like of the ELISA technology.
SUMMERY OF THE UTILITY MODEL
In order to solve the problem, the utility model provides an utilize dengue fever virus antibody short-term test chip of red fluorescence microballon mark, including the chip carrier, the chip front end is equipped with sample sampling hole, and the rear end is equipped with out the appearance hole, embedded microchannel of chip carrier, IgG reaction tank and IgM reaction tank, wherein, sample sampling hole is circular, sample sampling hole bottom sets up the through-hole, inside microchannel, IgG reaction tank and the IgM reaction tank of being equipped with of chip carrier, IgG reaction tank and IgM reaction tank have the adhesion after polylysine acid is handled, reaction tank bottom average distribution IgG IgM detects and uses the antibody, it passes through the microchannel with each reaction tank and is connected to go out the appearance hole.
Preferably, the chip carrier is a transparent visual structure.
In any of the above embodiments, preferably, the chip carrier is made of a polymethyl methacrylate material.
In any of the above embodiments, preferably, the IgG reaction tank is embedded with an anti-human IgG antibody, and the IgM reaction tank is embedded with an anti-human IgM antibody.
In any of the above schemes, preferably, the color fluorescent microspheres are prepared by copolymerizing rhodamine B fluorescent dye and modified styrene, the microspheres are pink (visible), and display red fluorescence under the irradiation of 532nm excitation wavelength, and the surface of the color fluorescent microsphere is grafted with dengue virus specific antigen through condensation reaction.
The utility model has the advantages that: the utility model provides an utilize dengue fever virus antibody short-term test chip of red fluorescence microballon mark, the invention adopts new red fluorescence mark point (diameter 300 nm's microballon) and antigen or antibody to carry out condensation reaction, form irreversible whole, realized the mark to target protein, and prepare out dengue fever virus antibody detection card, red fluorescence mark point comprises the diameter of the modified styrene that contains red fluorescent dye (rhodamine B) at 300 nm's microballon, the spheroid outside has hydroxyl functional group, can synthesize with target protein through condensation reaction and become integrative, finally form the protein of substitute red fluorescence mark.
Compared with the conventional detection methods such as ELISA and the like, the ELISA detection method has the advantages of simple and practical technology, high stability and good detection effect due to the reasons of longer process, complex operation and the like of the ELISA technology, and can well replace the existing ELISA detection technology in the aspect of detection.
Drawings
FIG. 1 is a schematic diagram of a preferred embodiment of a chip for rapid detection of dengue virus antibodies labeled with red fluorescent microspheres according to the present invention;
FIG. 2 is a side view of a preferred embodiment of the chip for rapid detection of dengue virus antibody labeled with red fluorescent microspheres according to the present invention.
The figures are labeled as follows: 1-a sampling hole; 2-a microchannel; a 3-IgG reaction tank; 4-IgM reaction tank; 5-sample outlet.
Detailed Description
In order to further understand the present invention, the present invention will be described in detail with reference to the following embodiments.
As shown in figures 1 and 2, the invention adopts a chip for rapidly detecting dengue viruses marked by red fluorescent microspheres, which comprises a chip carrier, wherein the front end of the chip carrier is provided with a sample sampling hole 1, the rear end of the chip carrier is provided with a sample outlet hole 5, the sampling hole 1 is in a round hole shape, the chip carrier is provided with a microchannel 2, the chip carrier is connected with an IgG reaction tank 3 and an IgM reaction tank 4, the IgG reaction tank 3 and the IgM reaction tank 4 are treated by poly-lysine and have adhesiveness, IgG/IgM detection antibodies are evenly distributed at the bottom of the reaction tanks, the sample outlet hole 5 is connected with each reaction tank through the microchannel 2, the IgG reaction tank 3 and the IgM reaction tank 4 are connected with the sample outlet hole 5 through the microchannel 3, when in use, the treated sample is guided into the reaction tank 3 and the IgM reaction tank 4 through the microchannel 2 IgG by an air compression pump according to a certain flow rate, the chip is horizontally placed on a table top, standing is carried out for five minutes at room temperature, a sample is led out of the chip from the sample outlet hole 5 through the micro channel 2 by using an air compression pump, a certain amount of PBS (containing 2% Tween 20) is led into the IgG reaction tank 3 and the IgM reaction tank 4 through the micro channel 2 at a certain flow rate by using the air compression pump, cleaning is carried out, the PBS buffer is led out of the chip from the sample outlet hole 5 through the micro channel 2 by using the air compression pump, repeating is carried out for 3 times, and then the red fluorescence value can be observed and quantitatively detected by using a related detection instrument.
Example two
Preparation of red fluorescent labeling ball:
1. preparing the red fluorescent microspheres into 5 mu M of solution by using saline;
2. the protein to be labeled is diluted to 5mg/ml by phosphate buffer (0.01M, pH 6.8);
3. adding a certain amount of microspheres into a reactor, starting stirring, sequentially adding the calculated amount of EDC aqueous solution (10mg/ml) and NHS aqueous solution (10mg/ml, adjusting the pH to about 7.0), and stirring for 0.5 h;
4. adding the protein to be marked (dengue specific antigen) according to the proportion, and stirring for 4 h;
5. after the reaction, the sample was filtered through a 0.22 μm needle filter and the concentration was measured. Storing at 4 deg.C; the method for detecting the sample to be processed comprises the following steps:
taking 100ul of blood sample (whole blood, serum or plasma) of a test patient, fully mixing the blood sample with 20ul of marked microsphere red fluorescent solution by using a micropipette, standing the mixture at room temperature for 30 minutes, and uniformly mixing the mixture once every 5 minutes by using the micropipette.
And (5) judging a result:
IgG quantitative detection: the IgG reaction tank 3 shows red color (red fluorescence under the irradiation of 532nm excitation wavelength), and the quantitative detection is carried out by a corresponding fluorescence detector; the serum to be detected (marked by red microspheres and combined with dengue virus recombinant antigen of red fluorescent microspheres into an antigen-antibody complex) is introduced into a micro-channel 2 through a sampling hole 1 by an air compression pump and then flows to an IgG reaction tank 3, firstly, on the antigen-antibody complex, the other binding site of the antibody to be detected is combined with an anti-human IgG antibody coated on the binding site to form a microsphere complex of a single antibody combined with an antibody and an antigen (double antibody sandwich), because the red fluorescent microspheres are deposited, the IgG reaction tank 3 is red, then PBS buffer solution (containing 2% Tween 20) is injected into the IgG reaction tank 3 by the air compression pump, cleaning is carried out, the operation is repeated for 3 times, and all waste liquid is led out of a chip from a sample outlet hole 5 through the micro-channel 2.
And (3) IgM quantitative detection: the IgM reaction tank 4 shows a red color (shows red fluorescence under the irradiation of 532nm excitation wavelength), and quantitative detection is carried out by a corresponding fluorescence detector; an air compression pump is utilized to introduce the serum to be detected (which is marked by the red microspheres and combined with the dengue virus recombinant antigen of the red fluorescent microspheres into an antigen-antibody complex) into the micro-channel 2 through the sampling hole 1, and then the serum flows to the IgM reaction tank 4. First, on the antigen-antibody complex, the other binding site of the antibody to be detected is bound to the anti-human IgM antibody coated thereon to form a microsphere complex in which a single antibody binds to one antibody and one antigen (double antibody sandwich), and the IgM reaction chamber 4 is colored red due to the deposition of red fluorescent microspheres thereon. Then injecting PBS buffer solution (containing 2% Tween 20) into the IgM reaction tank 4 by using an air compression pump, washing, repeating for 3 times, and leading all waste liquid out of the chip from the sample outlet hole 5 through the microchannel 2.
It will be understood by those skilled in the art that the chip for rapid detection of dengue virus antibody labeled with red fluorescent microspheres of the present invention includes any combination of the above-mentioned contents and embodiments of the present invention, and the portions shown in the drawings, which is limited to space and is not described in any combination for the sake of brevity. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. Utilize dengue fever virus antibody short-term test chip of red fluorescence microballon mark, including the chip carrier, chip carrier front end is equipped with sample sampling hole, and the rear end is equipped with out the appearance hole, embedded microchannel of chip carrier, IgG reaction tank and IgM reaction tank, its characterized in that, sample sampling hole is circular, sample sampling hole bottom sets up the through-hole, inside microchannel, IgG reaction tank and the IgM reaction tank of being equipped with of chip carrier, IgG reaction tank and IgM reaction tank have the adhesion after polylysine acid treatment, and reaction tank bottom evenly distributed IgG/IgM detects and uses the antibody, it passes through the microchannel with each reaction tank and is connected to go out the appearance hole.
2. The chip for rapidly detecting dengue virus antibody labeled with red fluorescent microspheres according to claim 1, wherein the chip carrier is a transparent visual structure.
3. The chip for rapidly detecting dengue virus antibody labeled with red fluorescent microspheres of claim 2, wherein the chip carrier is made of polymethyl methacrylate material.
4. The chip for rapidly detecting dengue virus antibody labeled with red fluorescent microspheres according to claim 1, wherein the IgG reaction tank is embedded with anti-human IgG antibody, and the IgM reaction tank is embedded with anti-human IgM antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN202120551409.8U CN214953571U (en) | 2021-03-17 | 2021-03-17 | Chip for rapidly detecting dengue virus antibody marked by red fluorescent microspheres |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202120551409.8U CN214953571U (en) | 2021-03-17 | 2021-03-17 | Chip for rapidly detecting dengue virus antibody marked by red fluorescent microspheres |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117849327A (en) * | 2024-03-06 | 2024-04-09 | 军科正源（北京）药物研究有限责任公司 | Methods for detecting a secukinumab-resistant antibody |
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2021
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117849327A (en) * | 2024-03-06 | 2024-04-09 | 军科正源（北京）药物研究有限责任公司 | Methods for detecting a secukinumab-resistant antibody |
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Effective date of registration: 20221028 Address after: Room 908-E, 9/F, Building D, Huoli Business Plaza, 185 Jumao Street, Yuanhe Street, Xiangcheng District, Suzhou City, Jiangsu Province, 215100 Patentee after: Suzhou Novi Bio Technology Co.,Ltd. Patentee after: JILIN SHUANGZHENG MEDICAL TECHNOLOGY CO.,LTD. Address before: Room 911, Kangyang building, 406 Chunfeng Road, Huangdai Town, Xiangcheng District, Suzhou City, Jiangsu Province Patentee before: Suzhou Novi Bio Technology Co.,Ltd. |
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