CN212514621U

CN212514621U - P16Ki67 and MCM2 immunocytochemistry detection kit

Info

Publication number: CN212514621U
Application number: CN202021110840.0U
Authority: CN
Inventors: 姚远颋; 许舟; 顾重建; 李克强
Original assignee: Chongqing Wokang Biological Technology Co ltd
Current assignee: Chongqing Wokang Biological Technology Co ltd
Priority date: 2020-06-16
Filing date: 2020-06-16
Publication date: 2021-02-09
Anticipated expiration: 2030-06-16

Abstract

The utility model discloses a P16Ki67, MCM2 immunocytochemistry detect reagent box, including box body and aseptic device hole, the inside of box body is provided with the liner, the aseptic device hole is located the container hole and keeps away from the one end at box body edge. This P16Ki67, MCM2 immunocytochemistry detect reagent box, compare with current ordinary detect reagent box, this equipment inner structure is compact, high durability and convenient carrying, good reproducibility, labour saving and time saving, it is high-efficient to detect, it is stable, can be fine satisfy the science, the needs of work, in the dyeing process, the sample is hatched after mixing the primary antibody of two kinds of albumen, detect the expression of two kinds of albumen on a sample film-making simultaneously, the detection speed of one time has almost accelerated, the pathology of experimenter is read the accuracy of piece, be convenient for observe, the detection accuracy improves, the possibility of false positive and false negative has been reduced, the possibility of erroneous judgement has been reduced, people's user demand has effectively been satisfied.

Description

P16Ki67 and MCM2 immunocytochemistry detection kit

Technical Field

The utility model relates to a cytochemistry check out test set technical field specifically is P16Ki67, MCM2 immunocytochemistry detect reagent box.

Background

The p16 gene is also named as multiple tumor suppressor gene (MTS), the expressed cancer suppressor protein p16 is a negative regulator protein of cell cycle progression, when human papilloma virus is integrated to infect cervical epithelial cells, the oncogene E7 is introduced and activated to promote the expression of the oncogenic protein E7, pRB is inhibited to combine with E2F protein to promote the proliferation of the cervical epithelial cells, when the cell proliferation is excessive, a negative feedback regulation mechanism of cell proliferation is activated, namely, the cancer suppressor gene p16 is activated to promote the expression of the cancer suppressor protein p16 and pRB is combined with E2F protein to inhibit the excessive proliferation of the cervical epithelial cells, the genes Ki-67 and MCM2 are genes related to the cell proliferation, the expression of the genes is different according to the cell cycle, the genes begin to be expressed in the G1 stage of the cell cycle, the expression is increased in the S stage and the G2 stage, the peak is reached to the M stage, and the silk mitosis is rapidly degraded or the later stage of losing antigen cluster, the gene is not expressed in the G0 stage, is not easy to be induced by growth factors due to short half-life period, and can be used as an index for evaluating the cell growth fraction.

In cervical cancer screening, the existing detection kit only detects P16 gene or only detects Ki-67 and MCM2 gene by a single immunocytochemistry method, the detection is time-consuming and labor-consuming, two samples need to be processed simultaneously, then the detection is carried out to obtain a result, the detection is complex, the staining solution is easy to become old, the staining effect is unstable, the accuracy of pathological interpretation of experimenters is influenced, the observation is inconvenient, the detection accuracy is low, the three times of single staining is time-consuming and labor-consuming, but if the three stains are combined, a large amount of reagents are required to be assembled in the same kit, the kit structure needs to be more perfect, after the three stains, the single staining is carried out simultaneously, the kit structure needs to be more perfect, the same three-staining kit has the advantages that the antibody concentrated solution is small in loading amount, the antibody concentrated solution can be loaded in smaller tubes, and if the antibody concentrated solution is loaded in large tubes, the reagent loss can be caused by overturning, the kit can not well meet the use requirements of people, and technical innovation is carried out on the basis of the existing detection kit aiming at the situation.

SUMMERY OF THE UTILITY MODEL

The utility model aims to provide a P16Ki67, MCM2 immunocytochemistry detect reagent box, in order to solve the above-mentioned background art and propose general detect reagent box in cervical carcinoma screening, the method of single through immunocytochemistry only detects P16 gene or only detects Ki-67, MCM2 gene, it wastes time and energy to detect, need to handle two samples simultaneously, then detect and obtain the result, comparatively complicated, the staining solution is easy old, and the dyeing effect is unstable, influence the accuracy of experimenter's pathology interpretation, be not convenient for observe, the rate of accuracy of detection is low, it wastes time and energy to singly dye for three times, but if the trichrome combines, just need a large amount of reagent to assemble in same reagent box, the reagent box structure just needs to be perfect more, after the trichrome, and singly dye simultaneously, the reagent box structure just needs to be perfect, same trichrome reagent box, the antibody concentrated solution can be subpackaged in smaller pipes due to smaller filling amount, and when the antibody concentrated solution is filled in a large pipe, reagent loss can be caused by overturning and reversing in the transportation process, so that the problem of use requirements of people can not be well met.

In order to achieve the above object, the utility model provides a following technical scheme: p16Ki67, MCM2 immunocytochemistry detect reagent box, including box body and aseptic device hole, the inside of box body is provided with the liner, and the container hole is installed to the one end of liner, aseptic device hole site is in the container hole and keeps away from the one end at box body edge, and the one end in aseptic device hole is provided with aseptic device.

Preferably, the container holes comprise p16 primary anti-concentration liquid, Ki-67 primary anti-concentration liquid, MCM2 primary anti-concentration liquid and antibody diluent, the container holes are four in the box body, and the p16 primary anti-concentration liquid, the Ki-67 primary anti-concentration liquid, the MCM2 primary anti-concentration liquid and the antibody diluent are respectively arranged in the container holes.

Preferably, the aseptic device hole includes that the work fluid is two to anti mouse of sheep, the work fluid is two to anti rabbit of sheep and DAB dyes substrate and reagent bottle, the aseptic device hole is provided with four in the inside of box body, and the inside of aseptic device hole is splendid attire respectively and is two work fluid, DAB dyes substrate and reagent bottle are two to anti mouse of sheep, anti rabbit of sheep.

Preferably, three sterile devices are arranged inside the box body, the shapes and the sizes of the sterile devices are the same, and the sterile devices are distributed in parallel.

Preferably, the container hole and the sterile device hole penetrate through the inside of the gasket, and the diameter of the outer surface of the reagent bottle is the same as the diameter of the inner surface of the sterile device hole.

Compared with the prior art, the beneficial effects of the utility model are as follows:

1. the utility model discloses a setting of aseptic device, aseptic device comprises aseptic plastic bag, independent lid, cell culture board, the buffer solution volume of laying in the container hole is less, in order to prevent toppling, adopt the bed hedgehopping design, different with several other pipe devices, and lay the buffer solution that the volume is great in the inside of aseptic device, so transversely lay, this equipment internal structure is compact, convenient to carry, good reproducibility, labour saving and time saving, P16/Ki-67, MCM 2's immunocytochemistry three-dyeing detects high-efficiently, and is stable, can be fine satisfy the needs of science, work;

2. the utility model discloses a setting of container hole etc, P16 Ki-67, MCM 2's three dyeings of immunocytochemistry, in dyeing process, with the sample of hatching behind the anti mixture of two kinds of proteins, detect the expression of two kinds of proteins on a sample film-making simultaneously, under the early prerequisite of accurate screening cervical carcinoma, can also accomplish cheap swiftly, almost accelerated the detection speed of one time, P16 Ki-67, the result of MCM 2's three dyeing methods of immunocytochemistry shows on same sample film-making, show the result on same sample film-making promptly, the accuracy of experimenter's pathology reading piece has been improved, be convenient for observe, the rate of accuracy that detects improves, the possibility of false positive and false negative has been reduced, the possibility of erroneous judgement has been reduced.

Drawings

FIG. 1 is a schematic perspective view of the present invention;

FIG. 2 is a schematic left side view, cross-sectional side view of the container port and sterile device port of the present invention;

fig. 3 is a schematic view of the cross-sectional structure of the aseptic device of the present invention.

In the figure: 1. a box body; 2. a liner; 3. a container aperture; 301. p16 primary antibody concentrate; 302. ki-67 primary antibody concentrate; 303. MCM2 primary antibody concentrate; 304. antibody diluent; 4. a sterile device aperture; 401. a goat anti-mouse secondary antibody working solution; 402. goat anti-rabbit secondary antibody working solution; 403. DAB staining substrate; 404. a reagent bottle; 5. and (4) a sterile device.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative work belong to the protection scope of the present invention.

Referring to fig. 1-3, the present invention provides a technical solution of P16Ki67 and MCM2 immunocytochemistry detection kit: the P16Ki67 and MCM2 immunocytochemistry detection kit comprises a box body 1 and an aseptic device hole 4, wherein a gasket 2 is arranged in the box body 1, a container hole 3 is arranged at one end of the gasket 2, the container hole 3 comprises P16-anti-concentrated solution 301, Ki-67-anti-concentrated solution 302, MCM 2-anti-concentrated solution 303 and antibody diluent 304, four container holes 3 are arranged in the box body 1, the P16-anti-concentrated solution 301, Ki-67-anti-concentrated solution 302, MCM 2-anti-concentrated solution 303 and the antibody diluent 304 are respectively arranged in the container hole 3, the aseptic device hole 4 is positioned at one end of the container hole 3, which is far away from the edge of the box body 1, an aseptic device 5 is arranged at one end of the aseptic device hole 4, the aseptic device 5 consists of an aseptic plastic bag, an independent box cover and a cell culture plate through the arrangement of the aseptic device 5, an independent packaging structure is adopted, and the situation that substances to be detected are mixed in the working process is avoided, the detection effect is influenced, the high efficiency and stability of immunocytochemistry double staining and Papanicolaou staining detection of P16/Ki-67 are ensured, and the requirements of science and work can be well met;

the sterile device hole 4 comprises a goat anti-mouse secondary working solution 401, a goat anti-rabbit secondary working solution 402, a DAB staining substrate 403 and reagent bottles 404, four sterile device holes 4 are arranged inside the box body 1, the goat anti-mouse secondary working solution 401, the goat anti-rabbit secondary working solution 402, the DAB staining substrate 403 and the reagent bottles 404 are respectively contained inside the sterile device hole 4, the container hole 3 and the sterile device hole 4 penetrate through the liner 2, the diameter of the outer surface of each reagent bottle 404 is the same as that of the inner surface of the sterile device hole 4, three sterile devices 5 are arranged inside the box body 1, the shapes and the sizes of the sterile devices 5 are the same, the sterile devices 5 are in a parallel distribution state, and the inner space of the equipment is fully utilized through the arrangement of the container hole 3, the sterile device hole 4 and the sterile device 5, so that the inner structure of the equipment is compact, and the waste of the space is reduced, the kit has the advantages of compact structure, convenient carrying, good repeatability, time and labor saving, and the aseptic device 5 adopts a horizontal lying design, so that the loose-leaf structure 6 needs to be lifted and then the reagent needs to be taken out for use when the reagent is taken out, thereby saving the space.

The working principle is as follows: when the P16Ki67 and MCM2 immunocytochemistry detection kit is used, firstly, equipment is unsealed, 1.85ml of antibody diluent 304 and 100 μ lp16 primary anti-concentration solution 301 are mixed and added into 100 μ lKi-67 and MCM2 primary anti-concentration solution 303 to prepare P16/lKi-67 and MCM2 primary anti-work mixed solution, a new label is pasted and stored in a dark place at 2-8 ℃ for later use, 50 μ l/piece of primary anti-work mixed solution is dripped, a 37 ℃ wet box is incubated for 30min, PBST is rinsed for 3min x 3 times and appropriately dried, 1ml of goat anti-mouse secondary anti-work mixed solution 401 is added into 1ml of goat anti-rabbit secondary antibody work solution 402 to prepare goat anti-mouse/rabbit secondary antibody work mixed solution, a new label is pasted and stored in a dark place at 2-8 ℃, 50 μ l/piece of secondary antibody is dripped for later use, the wet box is rinsed for 15min at 37 ℃, PBST 3min x 3 times, a buffer solution is rinsed and appropriately dried, preparing DAB working mixed liquid (according to the sample amount, taking a proper amount of DAB dyeing substrate 403 and DAB dyeing agent buffer solution to mix uniformly), preparing and using immediately, using up within 4h, storing in the dark at room temperature (20-25 ℃), dripping 50 mul/piece of DAB working mixed liquid, incubating in a wet box at room temperature (20-25 ℃) for 5-10 min, washing with running water for 3min, spin-drying moderately (non-dryable piece), preparing red working mixed liquid, using immediately, using up within 30min, storing in the dark at room temperature (20-25 ℃), dripping 50 mul/piece of red working mixed liquid, incubating in a wet box at room temperature (20-25 ℃) for 10-30 min, washing with running water for 3min, spin-drying moderately, placing cell slices into hematoxylin dye for dip dyeing, storing at room temperature (20-25 ℃) for 1s, re-dyeing, washing with running water for 3min, blow-drying, dripping 1 drop of neutral quick-sealing tablets, covering with a cover glass, evaluating, the working principle of the kit for detecting the immunocytochemistry of the P16Ki67 and MCM2 is the working principle.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

The P16Ki67 and MCM2 immunocytochemistry detection kit comprises a kit body (1) and a sterile device well (4), and is characterized in that: the inside of box body (1) is provided with liner (2), and container hole (3) are installed to the one end of liner (2), aseptic device hole (4) are located container hole (3) and keep away from the one end at box body (1) edge, and the one end in aseptic device hole (4) is provided with aseptic device (5).
2. The P16Ki67, MCM2 immunocytochemistry assay kit of claim 1, wherein: the container hole (3) comprises p16 primary anti-concentration liquid (301), Ki-67 primary anti-concentration liquid (302), MCM2 primary anti-concentration liquid (303) and antibody dilution liquid (304), the container hole (3) is provided with four in the box body (1), and the inside of the container hole (3) is respectively provided with p16 primary anti-concentration liquid (301), Ki-67 primary anti-concentration liquid (302), MCM2 primary anti-concentration liquid (303) and antibody dilution liquid (304).
3. The P16Ki67, MCM2 immunocytochemistry assay kit of claim 1, wherein: aseptic device hole (4) are including the anti mouse of sheep work fluid (401), the anti rabbit of sheep is two to work fluid (402) and DAB staining substrate (403) and reagent bottle (404), aseptic device hole (4) are provided with four in the inside of box body (1), and the inside of aseptic device hole (4) is splendid attire respectively to be equipped with the anti mouse of sheep and is two work fluid (401), the anti rabbit of sheep is two to work fluid (402), DAB staining substrate (403) and reagent bottle (404).
4. The P16Ki67, MCM2 immunocytochemistry assay kit of claim 1, wherein: the number of the aseptic devices (5) is three, the form and the size of the aseptic devices (5) are the same, and the aseptic devices (5) are distributed in parallel.
5. The P16Ki67, MCM2 immunocytochemistry assay kit of claim 1, wherein: the container hole (3) and the sterile device hole (4) penetrate through the inner part of the liner (2), and the diameter of the outer surface of the reagent bottle (404) is the same as that of the inner surface of the sterile device hole (4).