CN109239340B - Gel immunodiffusion tester - Google Patents
Gel immunodiffusion tester Download PDFInfo
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- CN109239340B CN109239340B CN201811245936.5A CN201811245936A CN109239340B CN 109239340 B CN109239340 B CN 109239340B CN 201811245936 A CN201811245936 A CN 201811245936A CN 109239340 B CN109239340 B CN 109239340B
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- 230000000951 immunodiffusion Effects 0.000 title claims abstract description 31
- 238000012360 testing method Methods 0.000 claims abstract description 77
- 239000000499 gel Substances 0.000 claims abstract description 43
- 238000011534 incubation Methods 0.000 claims abstract description 42
- 229920001817 Agar Polymers 0.000 claims abstract description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000008272 agar Substances 0.000 claims abstract description 27
- 229910001220 stainless steel Inorganic materials 0.000 claims abstract description 14
- 239000010935 stainless steel Substances 0.000 claims abstract description 14
- 239000011358 absorbing material Substances 0.000 claims abstract description 11
- 238000005485 electric heating Methods 0.000 claims abstract description 9
- 229920000742 Cotton Polymers 0.000 claims description 14
- 238000001556 precipitation Methods 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 210000003298 dental enamel Anatomy 0.000 claims description 6
- 239000011521 glass Substances 0.000 claims description 5
- 238000007789 sealing Methods 0.000 claims description 4
- 238000012546 transfer Methods 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 2
- 238000011160 research Methods 0.000 abstract description 5
- 239000000427 antigen Substances 0.000 description 20
- 102000036639 antigens Human genes 0.000 description 20
- 108091007433 antigens Proteins 0.000 description 20
- 230000000875 corresponding effect Effects 0.000 description 6
- 238000009792 diffusion process Methods 0.000 description 6
- 241000669298 Pseudaulacaspis pentagona Species 0.000 description 4
- 239000002775 capsule Substances 0.000 description 4
- 238000002768 Kirby-Bauer method Methods 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000004080 punching Methods 0.000 description 3
- 238000007711 solidification Methods 0.000 description 3
- 230000008023 solidification Effects 0.000 description 3
- 230000002745 absorbent Effects 0.000 description 2
- 239000002250 absorbent Substances 0.000 description 2
- 230000002457 bidirectional effect Effects 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 239000003779 heat-resistant material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 239000003973 paint Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
- G01N33/559—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody through a gel, e.g. Ouchterlony technique
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- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Dispersion Chemistry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a gel immunodiffusion tester, which is a special instrument for gel immunodiffusion test, and effectively improves test efficiency, and comprises: the lower part of the temperature control box is provided with an electric heating device, the outer surface of the lower box is provided with an electronic display screen, and a buzzer alarm and a timing key are also arranged for reminding the test time; the stainless steel shallow groove is arranged at the top of the electric heating device, and a water absorbing material piece is placed in the stainless steel shallow groove; the bottom of the incubation box is of a hollow shutter structure, and the incubation box is provided with a cover which is covered during the test; and a plurality of small boxes for making plates, which are arranged in the incubation box, wherein the small boxes for making plates are black opaque heat-resistant box bodies, the bottoms of the small boxes for making plates are provided with scales, and the small boxes for making plates are applicable to various hole patterns of double-expansion tests or single-expansion tests. The tester ensures that the test is carried out smoothly, avoids pollution caused by transferring the agar plate for a plurality of times in the test, improves the accuracy of test results, and brings convenience for scientific research and teaching.
Description
Technical Field
The invention relates to the field of immunological detection, in particular to a gel immunodiffusion tester.
Background
The soluble antigen and the corresponding antibody diffuse in the gel to form a concentration gradient, and macroscopic precipitation lines or precipitation rings are formed at the positions where the antigen and the antibody meet and the concentration ratio is proper. Common gels are agar, dextran or polyacrylamide gels. Gel immunodiffusion assays are classified into one-way diffusion assays and two-way diffusion assays.
Taking agarose gel as an example, a one-way diffusion test method is to punch holes in an agar plate containing specific antibodies, and add quantitative antigens into the holes, and when the antigens are diffused to the periphery, the antigens are combined with the antibodies in the agar, so that white precipitation rings are formed, and the diameter or the area of the white precipitation rings is positively correlated with the concentration of the antigens. And simultaneously, standard antigen or international reference protein is used for preparing a standard curve, so that the method can be used for quantitatively detecting the antigen concentration (mg/ml or U/ml) of an unknown specimen.
The two-way diffusion test method is to add antigen and antibody into different corresponding holes of agar plate, and the two are diffused freely in gel to form white precipitate line in proper proportion. The double-amplification can be used for estimating the antibody titer, the existence or non-existence of antigen or antibody in the specimen, the relative content of antigen and antibody, the molecular weight of the antigen and the antibody, and the like.
Gel immunodiffusion tests are carried out in the prior teaching and research, an agar plate is prepared on a plate or a glass slide, and a pipette is not easy to control when melted agar is poured into the agar plate to cause plate making failure; when the agar plate is perforated, in order to control the pitch of holes, a pen is needed to mark the bottom of a plate or a glass slide in advance, so that the agar plate is not accurate and is very complicated; the antigen-antibody reaction is carried out in a 37 ℃ moisturizing environment, and the method generally adopted at present is to put an agar plate in an enamel box with moist absorbent paper at the bottom, the enamel box is put in a 37 ℃ incubator for 24 or 48 hours for incubation, no specific incubation device is used for gel immunodiffusion test, the test process is very complicated, and the agar plate is easy to transfer for many times in operation and is easy to cause pollution.
At present, gel immunodiffusion tests are carried out in teaching and research, no special test instrument exists, the tests are carried out from plate making, punching, incubation to detection and other links, various test equipment is needed to be prepared, agar plates are transferred for multiple times, the process is tedious, and pollution is easy to cause.
Disclosure of Invention
The invention aims to provide a gel immunodiffusion tester, which provides a required test environment for gel immunodiffusion test, ensures smooth test, improves the accuracy of test results and brings convenience to scientific research and teaching.
Therefore, the invention provides a gel immunodiffusion tester, which is a special instrument for gel immunodiffusion test to reduce the transfer times of gel plates in the test, comprising: the temperature control box is provided with an electric heating device at the lower part, an electronic display screen for displaying the temperature and time in the box, a buzzer alarm and a timing key for reminding the test time; the stainless steel shallow groove is arranged at the top of the electric heating device, is provided with a water absorbing material piece, and is added with water during the test to ensure the humidity inside the test and prevent the agar plate from being dried in the test process; the incubation box is used for completing a reaction system in the incubation box during a test, the bottom of the incubation box is of a hollow shutter structure, and the incubation box is provided with a cover which is covered during the test to prevent the water of the agar plate from volatilizing; and a plurality of small plate boxes arranged in the incubation box and used for directly preparing the gel plate, wherein the bottom of the incubation box is provided with scales and various hole patterns suitable for double-expansion or single-expansion experiments, the incubation box is directly placed on the water absorbing material, and the shutter structure of the incubation box separates the small plate boxes from the water absorbing material.
Further, the temperature control box is provided with a transparent glass box door and is used for observing the internal condition of the instrument at any time, and the temperature control box is also provided with a door magnetic sealing device.
Further, the top or the side of the temperature control box is provided with an LED lamp, and the LED lamp is lightened when the box door is opened, so as to observe the internal condition of the temperature control box.
Further, the above-mentioned incubation box is a white stainless steel box or an enamel box, comprising a box body and a box cover.
Further, plugs and switches are arranged at the rear and the side of the temperature control box.
Further, the gel immunodiffusion tester is a special gel immunodiffusion tester.
Further, the electronic display screen is positioned on the outer surface of the lower box body of the temperature control box, and a temperature control key and a timing key are arranged beside the electronic display screen.
Further, the water absorbing material piece is a water absorbing cotton block.
Further, the small box for making the plate is a black opaque heat-resistant box body, the bottom is provided with white marked scales and various hole patterns suitable for double-expansion or single-expansion tests, when the gel plate is directly prepared in the small box for making the plate, the white hole patterns at the bottom are used for indicating a puncher to directly punch holes, a black background is convenient for observing a precipitation line formed by the test, and the scales are convenient for measuring the diameter of the precipitation line of the single-expansion test.
The invention provides a special instrument for gel immunodiffusion test, which provides all requirements for the gel immunodiffusion test, ensures that the test is carried out smoothly, avoids pollution caused by transferring the agar plate for many times in the test, improves the accuracy and the test efficiency of test results, and brings convenience to scientific research and teaching.
In addition to the objects, features and advantages described above, the present invention has other objects, features and advantages. The present invention will be described in further detail with reference to the drawings.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this specification, illustrate embodiments of the application and together with the description serve to explain the application. In the drawings:
FIG. 1 is a schematic structural view of a gel immunodiffusion tester according to the present invention;
FIG. 2 is a schematic structural view of a gel immunodiffusion tester according to the present invention;
FIG. 3 is a schematic view of the structure of an incubator's lid according to the present invention;
FIG. 4 is a schematic structural view of an incubation cassette according to the present invention;
FIG. 5 is a schematic view of the structure of the plate-making capsule according to the present invention;
FIG. 6 is a schematic view of the bottom of a set of plate cassettes according to the invention; and
FIG. 7 shows a schematic of an incubator according to the present invention equipped with a set of plate cassettes.
Detailed Description
It should be noted that, without conflict, the embodiments of the present application and features of the embodiments may be combined with each other. The application will be described in detail below with reference to the drawings in connection with embodiments.
Fig. 1-7 illustrate some embodiments according to the invention.
In one embodiment, as shown in fig. 1 and 2, the present test meter is designed as a box (or cabinet type), i.e., a temperature-controlled box 10 (or temperature-controlled cabinet), including a box body 11 and a box door 12.
Wherein, the chamber door 11 is made of transparent glass, which is convenient for observing the conditions in the chamber. The door edge is provided with a magnetic strip edge (door magnetic sealing structure) 13, and when in operation, the door is closed, so that the sealing of the tester is ensured, and the volatilization of the moisture in the box is prevented.
The lower part of the tester is provided with an electric heating device 40 for controlling the temperature in the box, the outer surface of the lower box is provided with an electronic display screen 17 for displaying the temperature and time in the box, a temperature control key 18 and a timing key 19 are arranged beside the electronic display screen, and a buzzer alarm (not shown in the figure) is arranged beside the electronic display screen. The rear and side of the tester are provided with plugs and switches 15.
Inside the tester, the top of the electric heating device is designed as a stainless steel shallow groove 16 (groove), and a small amount of water is added according to the test requirement. The test meter is equipped with a number of absorbent materials such as absorbent cotton block 30 which is sized to fit into the stainless steel recess and is placed in the recess for Cheng Shi of the water during the test.
The space above the stainless steel well is used for placing in an antigen-antibody reaction incubation device, i.e., an incubation box 20, matched with the tester.
The LED lamp 14 is equipped with at the top or side of tester, and when opening the tester door, the LED lamp lights, is convenient for observe the inside condition of tester.
As shown in fig. 3 and 4, the incubation cassette 20 is a white stainless steel or enamel cassette comprising a cassette body 20a and a cassette cover 20b.
As shown in fig. 4, the bottom of the box 20a is designed as a hollow shutter structure 21, and the shutter is in a vertical state with respect to the horizontal plane, for example, when the box is placed on the absorbent cotton, the bottom is not completely attached to the absorbent cotton, but stands on the absorbent cotton at a certain angle, so as to avoid water from adhering to other devices in the box.
As shown in fig. 3, the lid 20b is not connected to the case, and in use, is covered on the case, and a handle 22 is provided on the lid to facilitate opening.
As shown in fig. 5, the plate capsule 50 is entirely made of a black opaque heat resistant material, and the black plate capsule background facilitates the observation of the white precipitate line formed after the antigen-antibody incubation is completed.
In the invention, agar gel is taken as an example, a small plate box is selected according to test requirements and placed in a large box during gel immunodiffusion test, a tester is provided with small plate boxes with different specifications, white paint drawing scales 51a and hole patterns which are suitable for test requirements are designed at the bottoms 51 of the small plate boxes according to the requirements of agar unidirectional diffusion test and agar bidirectional diffusion, fig. 6 is the bottom surface of the small plate box with several specifications, only white 5cm scales 51a are drawn at the bottom (A) of the small box which is suitable for single expansion test, and several hole patterns such as white scales and triangular holes 51B, plum blossom holes (C) 51C, double row holes (D) 51D and the like are designed at the bottom of the small box (B) which is suitable for double expansion.
Fig. 7 shows the placement of selected cartridges in a large cartridge during the test. The tester is provided with a corresponding agar puncher.
Test example one: one-way agar diffusion test
Test principle: the antibody and the antigen to be detected are combined at a proper proportion to form a precipitation ring. The size of the loop is positively correlated with the concentration of antigen.
In the unidirectional agar gel immunodiffusion test, the gel immunodiffusion instrument is opened in advance, the temperature in the box is set to 37 ℃, a proper amount of water is added into a stainless steel shallow groove, and water absorbent cotton is put in to keep the water absorbent cotton in a wet state for 24-48 hours.
Selecting a small box with white scale marks at the bottom, mixing an antibody into 0.9% agarose (50 ℃), transferring a proper amount of agar, pouring the agar into the small box before solidification, solidifying the agar into a flat plate with the thickness of about 2-3mm at the bottom of the small box, punching (aperture of 3-5 mm) the white scale marks at the bottom of the small box by using a puncher according to test requirements, adding antigen solution and standard substances with different concentrations into the holes after solidification, covering a box cover, placing an incubation box in a preheated tester, and placing the incubation box on absorbent cotton, wherein the small box does not contact the absorbent cotton because a shutter at the bottom of the incubation large box has a certain angle with a horizontal plane.
The antigen-antibody reaction in the incubator was incubated at 37℃for 24 to 48 hours in a moist environment (preventing agar from drying). See if a white settling ring appears around the hole and measure the diameter of the ring according to the graduation marks at the bottom of the capsule. And the concentration of antigen was calculated.
Test example two: bidirectional agar diffusion test
Principle of: the antigen and the antibody are respectively added into different corresponding holes of the agar gel, and the two are freely diffused in the agar gel to form white precipitation lines at proper proportion.
In the two-way agar diffusion test, the gel immunodiffusion instrument is opened in advance, the temperature in the box is set to 37 ℃, a proper amount of water is added into a stainless steel shallow groove, and water absorbent cotton is put in to keep the water absorbent cotton in a moist state for 18-24 hours.
According to the test requirement, selecting the small box with corresponding specification.
Taking the detection of antibody titer as an example, selecting a plum blossom hole drawn at the bottom as an example, selecting a small box with white scale marks at the bottom, pouring a uniform agar gel thin layer, punching after solidification, wherein the agar thickness is generally 2-3 mm, the aperture is 3mm, the hole spacing is 3-5 mm, adding antigen or antibody into the corresponding hole, covering a box cover, and placing on absorbent cotton, wherein the small box is not contacted with the absorbent cotton due to the fact that the shutter at the bottom of the large box is at a certain angle with the horizontal plane.
The incubator is placed in a preheated tester, and after incubation is carried out for 18-24 hours at 37 ℃, the antigen and antibody diffused in the gel can form a visible white precipitation line at a proper concentration ratio. And judging the titer of the antibody according to the occurrence position of the precipitation line.
Compared with the prior art, the invention has the following characteristics/effects:
(1) A special tester special for gel immunodiffusion test is designed, so that pollution caused by transferring agar plates for multiple times in the test is avoided.
(2) The tester comprises an electric heating device, and can adjust the temperature to ensure the temperature required by the gel immunodiffusion test antigen antibody during incubation.
(3) The tester is provided with an electronic display screen for displaying the temperature and incubation time in the test process, and a buzzer alarm for reminding the test time.
(4) The stainless steel shallow groove and the water absorbing material are designed in the tester, a small amount of water is added during the test, and the humidity in the test is ensured to prevent the gel from being dried in the test process.
(5) The test apparatus is equipped with an incubation device, i.e., an incubation box, in which the reaction system is completed upon testing. The incubation device is made of enamel or stainless steel, the bottom of the incubation device is a hollow shutter structure, and the shutter structure is erected, so that the incubation device can be directly placed on a water absorbing material, but cannot contact a small box in the incubation box. The incubation box is provided with a cover, and the cover is covered during the test to prevent the water evaporation of the gel plate.
(6) The incubation box is internally provided with a plurality of small boxes which are made of black opaque heat-resistant materials, and the bottom of the small boxes is provided with white marked scales or is applicable to various holes of double expansion. Gel plates can be directly prepared in the small boxes, and the bottom white hole type is convenient for the puncher to directly punch holes. The black background is convenient for observing the precipitation line formed by the test, and the scale is convenient for measuring the diameter of the precipitation line of the single-expansion test.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. A gel immunodiffusion tester, which is a special instrument for gel immunodiffusion test to reduce the number of gel plate transfers in the test, comprising:
The temperature control box is provided with an electric heating device at the lower part, an electronic display screen for displaying the temperature and time in the box, a buzzer alarm and a timing key for reminding the test time;
The stainless steel shallow groove is arranged at the top of the electric heating device, is provided with a water absorbing material piece, and is added with water during the test to ensure the humidity inside the test and prevent the agar plate from being dried in the test process;
the incubation box is used for completing a reaction system in the incubation box during a test, the bottom of the incubation box is of a hollow shutter structure, and the incubation box is provided with a cover which is covered during the test to prevent the water of the agar plate from volatilizing; and
A plurality of small plate boxes which are arranged in the incubation box and are used for directly preparing gel plates, the bottoms of the small plate boxes are provided with scales and various hole patterns suitable for double-expansion or single-expansion experiments,
Wherein the incubation box is directly placed on the water absorbing material, the shutter structure of the incubation box separates the small plate-making box from the water absorbing material,
The temperature control box is provided with a transparent glass box door for observing the internal condition of the instrument at any time, and is also provided with a door magnetic sealing device,
The top or the side of the temperature control box is provided with an LED lamp which is lightened when the box door is opened for observing the internal condition of the temperature control box,
The small box for making the plate is a black opaque heat-resistant box body, the bottom is provided with white marked scales and various hole patterns suitable for double-expansion or single-expansion tests, when the gel plate is directly prepared in the small box for making the plate, the white hole patterns at the bottom are used for indicating a puncher to directly punch holes, a black background is convenient for observing a precipitation line formed by the test, and the scales are convenient for measuring the diameter of the precipitation line of the single-expansion test.
2. The gel immunodiffusion tester according to claim 1, wherein the incubation box is a white stainless steel box or an enamel box, comprising a box body and a box cover.
3. The gel immunodiffusion tester according to claim 1, wherein plugs and switches are mounted behind and to the sides of the temperature control box.
4. The gel immunodiffusion tester according to claim 1 wherein said piece of water absorbing material is a piece of water absorbing cotton.
5. The gel immunodiffusion tester according to claim 1, wherein the electronic display screen is positioned on the outer surface of the lower box body of the temperature control box, and a temperature control key is arranged beside the electronic display screen.
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| CN201811245936.5A CN109239340B (en) | 2018-10-24 | 2018-10-24 | Gel immunodiffusion tester |
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| CN201811245936.5A CN109239340B (en) | 2018-10-24 | 2018-10-24 | Gel immunodiffusion tester |
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| CN206223600U (en) * | 2016-12-08 | 2017-06-06 | 山东省滨州畜牧兽医研究院 | A kind of controllable warm type agar gel diffusion test moisture preservation box |
| CN106950385A (en) * | 2017-05-23 | 2017-07-14 | 北京乐普医疗科技有限责任公司 | A kind of platelet antibody detection kit and its application method |
| CN207457261U (en) * | 2017-11-15 | 2018-06-05 | 北京乐普医疗科技有限责任公司 | A kind of full-automatic fluorescence immune chromatography analytical equipment |
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