A rapid separation kit for mixed plaques without purification

Abstract

This application discloses a rapid separation kit for mixed plaques without purification, comprising a basket tube, a cap, and multiple centrifuge tubes. A limiting structure is installed on the inner wall of the centrifuge tubes and the outer wall of the basket tube. A sperm enrichment column is provided inside the basket tube, with an opening at its lower end. The basket tube is placed inside the centrifuge tubes and can be suspended within the centrifuge tubes by the limiting structure. The cap can be placed on the upper end of the basket tube. In use, the rapid separation kit for mixed plaques without purification, through the design of the centrifuge tube and basket tube, allows for the separation of female epithelial cell DNA and male sperm cell DNA from the mixed plaque sample simply by centrifugation in the basket tube. Furthermore, the female epithelial cell DNA and male sperm cell DNA can be used for STR-PCR detection without purification. Male sperm DNA can be extracted through multiple centrifugations and replacement of centrifuge tubes. Compared with the traditional two-step method, it eliminates repeated washing and liquid transfer steps, reduces the risk of contamination, and simplifies the operation.

Description

Technical Field

[0001] This application relates to the field of tube or bottle-type technology for enzymological or microbiological devices, specifically to one type. Background Technology

[0002] Mixed samples are commonly found in sex crime cases. These samples include vaginal swabs, underwear, toilet paper, bed sheets, and swabs from the suspect's external genitalia. These samples generally contain sperm, vaginal epithelial cells, and secretions, and are a mixture of male and female components.

[0003] The current mainstream method for separating mixed staining in laboratories is differential lysis. The first step in differential lysis is to lyse female component cells using proteases; the second step is centrifugation and washing; the third step involves adding DTT, proteases, SDS, etc., to lyse sperm cells, followed by purification to obtain single sperm DNA. However, differential lysis requires multiple pipetting operations and sperm DNA purification, which may result in sperm cell loss. This is detrimental to rapid detection in urgent cases and DNA extraction from samples with few sperm cells. Summary of the Invention

[0004] This invention aims to solve at least one of the technical problems existing in the prior art. To this end, this invention proposes a rapid separation kit for mixed plaques without purification, and the technical solution adopted includes:

[0005] A rapid separation kit for mixed plaques without purification includes a basket tube, a cap, and multiple centrifuge tubes. The inner wall of the centrifuge tubes and the outer wall of the basket tube are fitted with a limiting structure. The basket tube contains a sperm enrichment column with an opening at its lower end. The basket tube is placed inside the centrifuge tubes and can be suspended inside the centrifuge tubes by the limiting structure. The cap can be placed on the upper end of the basket tube.

[0006] The technical solution adopted by one embodiment of this utility model to solve its technical problem is as follows: the limiting structure includes an annular protrusion on the outer wall of the basket tube, the basket tube can be inserted into the centrifuge tube and the annular protrusion is placed at the upper end of the centrifuge tube to suspend the basket tube in the centrifuge tube.

[0007] The technical solution adopted by one embodiment of this utility model to solve its technical problem is: multiple tube caps are provided, and each tube cap is connected to a centrifuge tube in a one-to-one correspondence.

[0008] The technical solution adopted by one embodiment of this utility model to solve its technical problem is as follows: the tube cover includes a cover plate and a cylinder, the cylinder is installed on one side of the cover plate, and the outer diameter of the cylinder matches the inner diameter of the upper end of the basket tube and the centrifuge tube.

[0009] The technical solution adopted by one embodiment of this utility model to solve its technical problem is: the tube cap and the centrifuge tube are integrally formed.

[0010] The beneficial effects of this invention are as follows: When using the rapid separation kit for mixed plaques without purification, the design of the centrifuge tube and basket tube allows the mixed plaque sample to be separated from female epithelial cell DNA and male sperm cell DNA simply by centrifugation in the basket tube. Moreover, the female epithelial cell DNA and male sperm cell DNA can be used for STR-PCR detection without purification.

[0011] In the process of extracting male sperm DNA, the extraction of male sperm DNA is achieved by multiple centrifugations and replacement of centrifuge tubes. Compared with the traditional two-step method, it eliminates the need for repeated washing and liquid transfer steps, reduces the risk of contamination, and makes the operation simpler. Attached Figure Description

[0012] The above and / or additional aspects and advantages of this utility model will become apparent and readily understood from the description of the embodiments taken in conjunction with the following drawings, in which:

[0013] Figure 1 This is a schematic diagram of the structure of a rapid separation kit for mixed plaques without purification.

[0014] Figure 2 Exploded view of the structure of a rapid separation kit for mixed plaques without purification;

[0015] Figure 3 A cross-sectional view of a rapid separation kit for mixed plaques that does not require purification. Detailed Implementation

[0016] This section will describe in detail the specific embodiments of the present utility model. The preferred embodiments of the present utility model are shown in the accompanying drawings. The purpose of the drawings is to supplement the textual description with graphics, so that people can intuitively and vividly understand each technical feature and the overall technical solution of the present utility model, but they should not be construed as limiting the scope of protection of the present utility model.

[0017] In the description of this utility model, "multiple" means two or more; "greater than," "less than," and "exceeding" are understood to exclude the stated number; "above," "below," and "within" are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly specifying the number of indicated technical features or their sequential relationship.

[0018] In the description of this utility model, it should be understood that the directional descriptions, such as up, down, front, back, left, right, etc., indicate the directional or positional relationship based on the directional or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0019] In this utility model, unless otherwise explicitly defined, the terms "setting," "installing," and "connecting" should be interpreted broadly. For example, they can refer to a direct connection or an indirect connection through an intermediate medium; they can refer to a fixed connection, a detachable connection, or an integral molding; they can refer to a mechanical connection; they can refer to the internal connection of two components or the interaction between two components. Those skilled in the art can reasonably determine the specific meaning of the above terms in this utility model in conjunction with the specific content of the technical solution.

[0020] Reference Figure 1-3 As shown, an embodiment of this application is proposed. The rapid separation kit for mixed plaques without purification described in this embodiment includes a centrifuge tube 30, a basket tube 10, and a tube cap 20. A limiting structure 40 is installed on the inner wall of the centrifuge tube 30 and the outer wall of the basket tube 10. A sperm enrichment column 50 is provided inside the basket tube 10, and its lower end is provided with an opening. The basket tube 10 is used to be placed inside the centrifuge tube 30 and can be suspended inside the centrifuge tube 30 under the action of the limiting structure 40. The tube cap 20 can be placed on the upper end of the basket tube 10.

[0021] The method of using the rapid separation kit for mixed plaques without purification described in this embodiment includes:

[0022] Step 1: Place the mixed plaque sample into the basket tube 10, place the basket tube 10 into the centrifuge tube 30 and suspend it in the centrifuge tube 30, add 500 μL of epithelial cell lysis buffer to the basket tube 10, put the tube cap 20 on the top of the basket tube 10, and incubate the kit at 60°C for 40 min.

[0023] Step 2: Place the kit in a centrifuge and centrifuge at 12,000 rpm for three minutes. Sperm cells will bind to the sperm enrichment column 50. Epithelial cell lysis and female epithelial cells will pass through the sperm enrichment column 50 and flow into the centrifuge tube 30 through the opening at the lower end of the basket tube 10.

[0024] Step 3: Open the tube cap 20, take out the basket tube 10, collect the female DNA component in the centrifuge tube 30, and treat it at 95℃ for 12 minutes before using it for STR-PCR amplification.

[0025] Step 4: Place the basket tube 10 taken out in step 3 into a new secondary centrifuge tube 30, add 500 μL of epithelial cell DNA lysis remover to the basket tube 10, cover the basket tube 10 with the cap 20, and incubate at 60°C for 40 min.

[0026] Step 5: Place the reagent kit into a centrifuge and centrifuge at 12,000 rpm for three minutes;

[0027] Step 6: Open the tube cap 20, take out the basket tube 10, and discard the epithelial cell DNA lysis remover inside the secondary centrifuge tube 30.

[0028] Step 7: Place the basket tube 10 taken out in step 3 into a new triple centrifuge tube 30, add 500 microliters of cleaning buffer to the basket tube 10, and put the tube cap 20 on the basket tube 10.

[0029] Step 8: Place the reagent kit into a centrifuge and centrifuge at 12,000 rpm for three minutes;

[0030] Step 9: Open the tube cap 20, take out the basket tube 10, and discard the cleaning buffer in the three centrifuge tubes 30.

[0031] Step 10: Place the basket tube 10 taken out in step 9 into a new four-times centrifuge tube 30, add 100-200 μL of sperm lysis buffer to the basket tube 10, cover the basket tube 10 with the tube cap 20, and lyse at 95°C for 15 min.

[0032] Step 11: Place the reagent kit into a centrifuge and centrifuge at 12,000 rpm for three minutes;

[0033] Step 12: Open the tube cap 20, remove and discard the basket tube 10, and the male sperm DNA will be left in the four centrifuge tubes 30, which can be directly used for STR-PCR amplification.

[0034] The rapid separation kit for mixed plaques without purification described in this embodiment, through the design of centrifuge tube 30 and basket tube 10, allows the mixed plaque sample to be separated from female epithelial cell DNA and male sperm cell DNA simply by centrifugation in basket tube 10. Moreover, the female epithelial cell DNA and male sperm cell DNA can be used for STR-PCR detection without purification.

[0035] In the process of extracting male sperm DNA, the extraction of male sperm DNA is achieved by multiple centrifugations and replacement of centrifuge tubes. The extraction of epithelial cell DNA and male sperm cell DNA can be completed within 2 hours. Compared with the traditional two-step method, it eliminates the need for repeated washing and liquid transfer steps, reduces the risk of contamination, and makes the operation simpler.

[0036] In this embodiment, the limiting structure 40 includes an annular protrusion on the outer wall of the basket tube 10. The basket tube 10 can be inserted into the centrifuge tube 30 and the annular protrusion is placed at the upper end of the centrifuge tube 30 to suspend the basket tube 10 inside the centrifuge tube 30.

[0037] The basket tube 10 is placed on the upper end of the centrifuge tube 30 via an annular boss, so that the upper end of the basket tube 10 protrudes from the upper end of the centrifuge tube 30, making it easier to insert the basket tube 10 into or remove it from the centrifuge tube 30, thus making the operation more convenient.

[0038] In this embodiment, multiple tube caps 20 are provided, and each tube cap 20 is connected to a centrifuge tube 30 in a one-to-one correspondence. When replacing the centrifuge tube 30, the tube cap 20 is replaced at the same time, avoiding cross-contamination between multiple centrifugation steps.

[0039] Furthermore, as a preferred embodiment, the tube cover 20 includes a cover plate 21 and a cylinder 22, the cylinder 22 being installed on one side of the cover plate 21, and the outer diameter of the cylinder 22 matching the inner diameter of the upper end of the basket tube 10 and the centrifuge tube 30.

[0040] The tube cap 20 includes a cover plate 21 and a cylinder 22. The cylinder 22 is installed on one side of the cover plate 21 and is used to insert into and seal the upper end of the basket tube 10 or centrifuge tube 30. The inner diameter of the upper end of the tube cap 20 of the basket tube 10 matches the inner diameter of the upper end of the centrifuge tube 30.

[0041] The outer diameter of the cylinder 22 matches the inner diameter of the upper end of the basket tube 10 and the centrifuge tube 30, so that the cylinder 22 can be inserted into and sealed inside the upper end of the basket tube 10 and the centrifuge tube 30. In steps 3 and 12, the tube cap 20 is placed on the upper end of the corresponding centrifuge tube 30 to seal and preserve the female epithelial cell DNA and male sperm DNA in the centrifuge tube 30 respectively.

[0042] In other steps, the tube cap 20 is placed on the upper end of the basket tube 10 to facilitate incubation or centrifugation.

[0043] Preferably, the tube cap 20 is integrally formed with the centrifuge tube 30.

[0044] Of course, this utility model is not limited to the above-described embodiments. Those skilled in the art can make equivalent modifications or substitutions without departing from the spirit of this utility model. All such equivalent modifications and substitutions are included within the scope defined by the claims of this application.

Claims

1. A rapid separation kit for mixed plaques without purification, characterized in that, The device includes a basket tube (10), a tube cap (20), and multiple centrifuge tubes (30). The inner wall of the centrifuge tube (30) and the outer wall of the basket tube (10) are fitted with a limiting structure (40). The basket tube (10) is provided with a sperm enrichment column (50) and has an opening at its lower end. The basket tube (10) is used to be placed inside the centrifuge tube (30) and can be suspended inside the centrifuge tube (30) under the action of the limiting structure (40). The tube cap (20) can be placed on the upper end of the basket tube (10).

2. The rapid separation kit for purification-free mixed plaques according to claim 1, characterized in that, The limiting structure (40) includes an annular protrusion on the outer wall of the basket tube (10), the basket tube (10) can be inserted into the centrifuge tube (30) and the annular protrusion is placed at the upper end of the centrifuge tube (30) to suspend the basket tube (10) inside the centrifuge tube (30).

3. The rapid separation kit for purification-free mixed plaques according to claim 2, characterized in that, Multiple tube caps (20) are provided, and each tube cap (20) is connected to a centrifuge tube (30) in a corresponding manner.

4. The rapid separation kit for purification-free mixed plaques according to claim 3, characterized in that, The tube cap (20) includes a cover plate (21) and a cylinder (22). The cylinder (22) is installed on one side of the cover plate (21), and the outer diameter of the cylinder (22) matches the inner diameter of the upper end of the basket tube (10) and the centrifuge tube (30).

5. The rapid separation kit for purification-free mixed plaques according to claim 3, characterized in that, The cap (20) and the centrifuge tube (30) are integrally formed.

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