Self-mixing blood RNA preservation tube and use method

Abstract

The application discloses a self-mixing blood RNA storage tube and a use method, and belongs to the field of biomedical sample storage devices. The tube body is pre-stored with RNA stabilizing liquid, and the opening end is provided with a rotating sealing cover. The tube body is provided with a double-helix flow guide pipe, and the RNA stabilizing liquid is pre-installed at the bottom of the double-helix flow guide pipe. When blood flows into the tube body, the blood rotates and flows along the spiral flow guide pipe and is automatically mixed. The application sets the double-helix flow guide pipe in the tube body, and directly pre-installs the RNA stabilizing liquid at the bottom of the double-helix flow guide pipe, so that the blood rotates and flows along the spiral path after entering the tube body, the blood is fully contacted and uniformly mixed with the RNA stabilizing liquid in a short time, an additional mixing step is not needed, operation errors are reduced, the RNA storage conditions of all blood sampling samples are ensured to be consistent, and the stability of experimental data is improved. Through the cooperative design of the sealing rubber plug and the rotating sealing cover, the air tightness of the blood before entering the tube body is ensured, and external pollution is prevented.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical sample preservation devices, and specifically relates to a self-mixing blood RNA preservation tube and a method of use. It is widely applicable to fields requiring high-quality RNA sample preservation, such as clinical testing, molecular biology research, gene expression analysis, and early disease screening. Background Art

[0002] RNA (ribonucleic acid) is a key genetic information carrier for gene expression in cells and plays a vital role in the regulation of life activities, disease diagnosis, and biomedical research. Compared with DNA, the molecular structure of RNA is more fragile and is easily affected by the external environment, especially the degradation of RNase (ribonuclease). In addition, RNA is also affected by factors such as temperature changes, pH fluctuations, and oxidative stress during collection, transportation, and storage, making it very susceptible to degradation or inactivation. These factors not only affect the integrity of RNA, but may also affect the accuracy of gene expression analysis, thereby significantly affecting experimental results. Therefore, how to quickly and efficiently stabilize RNA after blood collection has become a major challenge in biomedical research and clinical applications.

[0003] At present, RNA preservation mainly relies on RNA preservation tubes. Common RNA preservation tubes mainly use the following two methods to preserve RNA: (1) Traditional blood collection tubes + RNA stabilizer: This method uses ordinary blood collection tubes to collect blood, and then manually adds RNA preservation solution to stabilize RNA molecules. However, this method has many drawbacks. First, the operation is complicated and requires additional manual addition steps, which increases the complexity of experimental operation and is prone to experimental errors, affecting the reproducibility of data; second, the risk of RNA degradation is high. Because the blood is exposed to non-ideal environments for a long time, the RNA may have been partially degraded before the RNA stabilizer is added, affecting the accuracy of subsequent analysis; in addition, the risk of contamination is high. This method requires open operation and is easily contaminated by the external environment, especially during RNA extraction and analysis, exogenous RNase may be introduced, further accelerating RNA degradation. (2) Blood collection tubes pre-filled with RNA preservation solution: In order to solve the problem caused by manually adding RNA stabilizer, some RNA preservation tubes are pre-filled with RNA stabilizing solution before blood collection, which can automatically mix after the blood enters the blood collection tube. However, this solution still has the following shortcomings: First, the preservation solution comes into direct contact with the human body. Since the RNA stabilization solution usually contains denaturants or other chemical inhibitors, it may have adverse effects on the blood collector, especially in the event of leakage during the blood collection process, which may lead to local irritation or toxicity risks; second, the mixing is uneven. After the blood enters the blood collection tube, the preservation solution is not fully mixed, which may cause some samples to fail to obtain effective RNA protection in time, affecting the subsequent extraction efficiency and RNA stability; in addition, the sealing performance is insufficient. Some existing RNA preservation tubes have poor sealing performance, and leakage or volatilization of the preservation solution may occur during transportation, thereby affecting the RNA preservation effect and reducing the stability and consistency of RNA extraction.

[0004] In summary, existing RNA storage tubes still present numerous challenges during use, including complex handling, high risk of RNA degradation, high risk of contamination, uneven mixing, and insufficient sealing, limiting their widespread application in clinical testing, RNA sequencing analysis, and biomedical research. Therefore, a more efficient, convenient, and safe RNA storage device is urgently needed to ensure RNA stability, improve experimental reproducibility, and meet the clinical and scientific demand for high-quality RNA samples. Summary of the Invention

[0005] The purpose of the present invention is to provide a self-mixing blood RNA storage tube and a method of use to solve the above-mentioned problems.

[0006] In order to solve the above problems, the present invention adopts the following technical solutions:

[0007] A self-mixing blood RNA storage tube comprises a tube body, one end of which is closed, the other end of which is open, and the interior of which is a hollow cavity;

[0008] A double-helix flow guide tube is disposed in the tube body;

[0009] RNA stabilizing liquid is distributed on the tube body and the bottom of the double-helix flow guide tube;

[0010] a rotary sealing cover, disposed at the open end of the tube body, for placing the cavity inside the tube body in a negative pressure state and sealing the open end;

[0011] When blood is collected, the rotary sealing cover is opened, and the blood flows into the tube body and rotates along the spiral guide tube, enters the RNA stabilizing liquid area from top to bottom, and automatically mixes with the RNA stabilizing liquid.

[0012] In a further embodiment, the double-helix flow guide tube includes two spiral flow guide tubes wound around each other, and the upper ends of the two spiral flow guide tubes are connected, and the lower ends are close to the bottom of the tube body and are arranged in opposite directions.

[0013] Through the above technical solution, the blood flow area is accelerated and the mixing uniformity is improved.

[0014] In a further embodiment, the spiral angle of the spiral flow guide tube is 20° to 30°.

[0015] Through the above technical solution, the blood flow rate is ensured to be moderate, blood is prevented from being retained in the tube body, and the mixing effect is enhanced.

[0016] In a further embodiment, the width of the double-helix flow guide tube is 5% to 10% of the inner diameter of the tube body.

[0017] Through the above technical solution, dead corners of blood flow are avoided and the mixing efficiency of blood and RNA stabilizing solution is improved.

[0018] In a further embodiment, a micro-bump spoiler zone is provided on the inner wall of the tube body, and the micro-bump spoiler zone includes a first micro-bump zone arranged at the center of the bottom of the tube body, a second micro-bump zone arranged in the transition area between the bottom of the tube body and the side wall, and a third micro-bump zone arranged at the edge of the bottom of the tube body.

[0019] Through the above technical solution, the smooth flow of blood is disrupted, turbulence is easily formed, and it is helpful to mix the RNA stabilizing solution and blood.

[0020] In a further embodiment, the sizes of the bumps in the second micro-bump area, the first micro-bump area and the third micro-bump area decrease successively.

[0021] Through the above technical solution, the micro-bumps are rationally partitioned and distributed, and the spacing and height are adjusted, which not only significantly improves the mixing effect of RNA stabilization liquid and blood, but also prevents the deposition of RNA stabilization liquid, ensuring the uniformity and stability of RNA samples.

[0022] In a further embodiment, a sealing rubber plug is embedded in the open end of the tube body, and a through hole is formed on the sealing rubber plug to match the connection point of the two spiral guide tubes;

[0023] Position marks are provided on the upper surface of the rotary sealing cover at positions corresponding to the through holes.

[0024] Through the above technical solution, the sealing rubber stopper and the rotating sealing cover work together to ensure airtightness after blood collection, improve the storage stability of the sample, and facilitate the insertion of the blood collection needle.

[0025] In a further embodiment, the tube body is made of a transparent material, and the outer surface is coated with a light-proof layer, and the inner wall is coated with a polytetrafluoroethylene coating.

[0026] The above technical solution can prevent ultraviolet radiation from degrading RNA and maintain the visibility of the sample for easy observation.

[0027] In a further embodiment, the rotary sealing cover is made of medical-grade polypropylene, and the tube body is injection-molded and adapted to fit the rotary sealing cover.

[0028] Through the above technical solution, the rotary sealing cap can withstand high temperature and high pressure sterilization, ensuring pollution-free during use, while reducing production costs and being suitable for disposable use.

[0029] The method for using the self-mixing blood RNA storage tube includes the following steps:

[0030] Step 1: Unscrew the rotary sealing cap and insert the blood collection needle into the marked position. The blood will flow from the top to the bottom along the double-helix guide tube and automatically mix with the RNA stabilizing solution.

[0031] Step 2: Close the rotary sealing cover.

[0032] Beneficial effects: By setting a double-helix flow guide tube in the tube body and pre-installing the RNA stabilizing liquid directly at the bottom of the double-helix flow guide tube, it is ensured that the blood flows along the spiral path after entering the tube body, so that the blood and the RNA stabilizing liquid are fully contacted and evenly mixed in a short time, without the need for additional mixing steps, reducing operational errors, ensuring that the RNA storage conditions of all blood samples are consistent, and improving the stability of experimental data; through the coordinated design of the sealing rubber stopper and the rotating sealing cover, the airtightness of the blood before entering the tube body is ensured to prevent external contamination; in addition, the tube body is made of transparent material and the outer surface is coated with a light-proof layer, and the inner wall is coated with a polytetrafluoroethylene coating to prevent RNA degradation and improve the quality of RNA preservation. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is a structural schematic diagram of the present invention;

[0034] Figure 2 Schematic diagram of the distribution of the micro-convex flow disturbance area at the bottom of the inner wall of the tube body in the present invention;

[0035] Figure 3 A top view of the rotary sealing cover in the present invention in a closed state;

[0036] Figure 4 This is a top view of the rotary sealing cover in the present invention in the open state.

[0037] Figure numerals: 1, tube body; 11, first micro-bump area; 12, second micro-bump area; 13, third micro-bump area; 2, double-helix flow guide tube; 3, RNA stabilizing liquid; 4, rotating sealing cover; 41, position mark; 5, sealing rubber stopper. DETAILED DESCRIPTION

[0038] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0039] This application proposes a self-mixing blood RNA storage tube, hereinafter referred to as the "tube." This tube utilizes a double-helix flow guide tube 2 and a rotating sealing cap 4, enabling automatic mixing of blood and RNA stabilization solution 3 after blood collection, ensuring RNA stability while preventing direct contact between the RNA stabilization solution 3 and the human body before blood collection, thereby improving sample safety and storage quality. This design not only optimizes the mixing efficiency of the RNA stabilization solution 3 and blood, but also enhances the sealing performance of the blood collection tube, effectively reducing the risk of sample contamination and stabilization solution volatilization, significantly improving the extraction quality and stability of RNA samples, and has broad application prospects in clinical testing and scientific research applications.

[0040] like Figures 1 to 3 As shown, the storage tube comprises a tube body 1, RNA stabilizing solution 3, a double-helix flow guide tube 2, and a rotating sealing cap 4. The tube body 1 has a closed end at one end and an open end at the other, with an interior cavity capable of accommodating an object. The double-helix flow guide tube 2 is disposed within the tube body 1 and comprises two intertwined spiral flow guide tubes. The RNA stabilizing solution 3 is pre-stored at the bottom of the tube body 1, and the rotating sealing cap 4 is mounted on the open end of the tube body 1. During blood collection, as blood flows into the tube body 1 through the lancet, it rotates along the spiral tube and enters the RNA stabilizing solution 3 region from top to bottom, initially mixing with the RNA stabilizing solution 3 as it rotates.

[0041] In this embodiment, the tube 1 is 120 mm long, with an outer diameter of 15 mm and an inner diameter of 12 mm. The upper portion of the tube 1 is the blood collection area with a blood collection capacity of 5 mL, and the bottom portion is filled with RNA stabilization solution 3 with a volume of 3 mL, suitable for preserving RNA from various types of whole blood.

[0042] Continue reading Figure 1 The upper ends of the two spiral guide tubes are connected, and the lower ends are located near the bottom of the tube body 1 and facing in opposite directions. This increases the blood flow area and accelerates the blood circulation speed, thereby improving the uniformity of mixing between the blood and the RNA stabilizing solution 3. The spiral angle of the spiral guide tube is 20° to 30°, which can increase the fluid speed and prevent blood retention. The width of the spiral guide tube is 5% to 10% of the inner diameter of the tube body 1. This design ensures that enough blood can pass through while avoiding dead corners for blood accumulation, thereby improving the mixing efficiency of the blood and the RNA stabilizing solution 3. Compared with traditional straight tubes, the spiral structure improves mixing uniformity by 30% and reduces blood deposition.

[0043] Continue reading Figure 2The RNA stabilizing liquid 3 is directly pre-installed at the bottom of the double-helix flow guide tube 2. The microfluidic principle is used to make the RNA stabilizing liquid 3 automatically diffuse and evenly mix after the blood enters. In order to make the RNA stabilizing liquid 3 fully merge with the blood at the bottom of the tube body 1, a micro-bump disturbance area is provided on the inner wall of the tube body 1. The micro-bump disturbance area includes a first micro-bump area 11, a second micro-bump area 12 and a third micro-bump area 13, wherein the first micro-bump area 11 is arranged at the central area of ​​the bottom of the tube body 11, which is the main accumulation area of ​​the RNA stabilizing liquid 3. The micro-bumps can promote the uniform dispersion of the fluid; the second micro-bump area 12 is arranged at the transition area between the bottom and the side wall of the tube body 1, so that when the blood rotates and flows in along the tube wall, the micro-bumps can break the smooth flow of the fluid and form turbulence, which is conducive to the mixing of the RNA stabilizing liquid 3 and the blood; the third micro-bump area 13 is arranged at the edge area of ​​the bottom of the tube body 1 to prevent the RNA stabilizing liquid 3 from being deposited on the tube wall without participating in the mixing. In summary, when blood is dissolved in the RNA stabilizing solution 3, the smooth flow of the blood is disrupted, turbulence is easily formed, and the mixing of the RNA stabilizing solution 3 and the blood is facilitated.

[0044] Furthermore, the sizes of the bumps in the second micro-bump area 12, the first micro-bump area 11 and the third micro-bump area 13 decrease successively. This design reasonably divides the micro-bumps into different areas and adjusts the spacing and height. It not only significantly improves the mixing effect of the RNA stabilizing liquid 3 and the blood, but also prevents the deposition of the RNA stabilizing liquid 3, thereby ensuring the uniformity and stability of the RNA sample.

[0045] Continue reading Figure 1 、 Figure 3 and Figure 4 A sealing rubber plug 5 is embedded in the open end of the tube body 1. The sealing rubber plug 5 is provided with a through hole that is compatible with the connection between the two spiral guide tubes. At the same time, the rotating sealing cover 4 is provided with a position mark 41 at the corresponding position of the through hole to facilitate the insertion of the blood collection needle; through the cooperation of the sealing rubber plug 5 and the rotating sealing cover 4, the airtightness inside the tube body 1 is ensured to prevent external contamination.

[0046] like Figure 3 As shown, before blood collection, the sealing cover 4 is rotated to seal the open end of the tube body 1 to prevent external contamination from entering the tube body 11. As shown in the figure, it is opened when blood is collected to facilitate the insertion of the blood collection needle; after blood collection is completed, it is closed to ensure airtightness.

[0047] The present application also provides a method for using a self-mixing blood RNA storage tube, comprising the following steps:

[0048] Step 1: Unscrew the rotary sealing cover 4, insert the blood collection needle into the position mark 41, and the blood will flow into the bottom of the tube body 1 along the double-helix guide tube 2 from top to bottom and automatically mix with the RNA stabilizing solution 3;

[0049] Step 2: Close the rotary sealing cover 4.

[0050] Specifically, the rotating sealing cover 4 cooperates with the sealing rubber plug 5 to ensure that the whole tube body 1 is sealed to prevent external pollution.

[0051] As a preferred solution, the tube body 1 is made of transparent material, and the outer surface is coated with a light-proof layer, and the inner wall is coated with a polytetrafluoroethylene coating. By adopting a double-layer transparent light-proof coating, the degradation and damage of RNA by ultraviolet light are prevented, and the visibility of the tube body 1 is maintained, facilitating observation of the sample.

[0052] As a preferred solution, the rotating sealing cover 4 is made of medical-grade polypropylene material, which can withstand high-temperature and high-pressure sterilization, ensuring no contamination during use, and the tube opening is designed with a medical-grade silicone sealing plug to ensure the airtightness of the tube body 11.

[0053] The storage tube is designed as a whole, manufactured by injection molding process, which simplifies the production process, reduces the manufacturing cost, and the product quality is stable, suitable for one-time use.

[0054] Compared with the prior art, the present application has the following advantages:

[0055] (1) Improve RNA mixing efficiency and prevent stratification. The design of the double-helix flow guide pipe 2 ensures that the blood rotates along the spiral path after entering the tube body 1, so that the blood and the RNA stable liquid 3 are fully contacted and uniformly mixed in a short time.

[0056] (2) Avoid RNA degradation and improve sample stability. Through the principle of spiral fluid dynamics, the blood is automatically dispersed during the flow into the tube body 1, so that the preservation liquid quickly covers the blood sample, reduces the time of RNA exposure in a non-ideal environment, and reduces the risk of degradation.

[0057] (3) Automatic mixing and improved accuracy. The automatic diffusion mixing mechanism allows the blood to naturally rotate along the spiral flow path after entering, and directly merges with the preservation liquid, without the need for additional mixing steps, reducing operational errors, ensuring that the RNA preservation conditions of all blood samples are consistent, and improving the stability of experimental data.

[0058] (4) Rotating sealing cover 4 design improves sealing performance. The rotating sealing cover 4 can completely seal the tube body 1 before blood collection to prevent external environmental pollution of the sample; after blood collection, the cover can be tightened to improve the sealing performance, avoid leakage during transportation, and ensure the integrity of the sample; in addition, the rotating sealing cover 4 combined with the sealing rubber plug 5 improves the negative pressure maintaining ability, ensuring that the blood does not leak after entering.

[0059] (5) The light-proof transparent tube wall is used to prevent RNA degradation. The light-proof layer design prevents RNA from being affected by ultraviolet rays, thereby improving the quality of RNA preservation. At the same time, the transparent outer wall facilitates experimental observation without the need for additional sample processing, thereby improving experimental operation efficiency.

[0060] (6) Low-cost manufacturing, suitable for disposable use. The tube body 1 adopts injection molding technology and structural integrated design to reduce production steps; the simplified production process ensures stable product quality, reduces medical costs, and improves market competitiveness.

[0061] (7) It is suitable for blood of different viscosities, improves versatility, optimizes the spiral tube diameter and inclination (20°-30°), ensures that blood of different viscosities can flow smoothly, and improves adaptability; at the same time, it reduces the blood residue on the tube wall, ensures that all blood samples can be fully mixed with RNA stabilizing solution 3, and improves sample integrity.

[0062] The above content is a detailed description of the present invention in conjunction with specific embodiments, and it cannot be considered that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art of the present invention, without departing from the concept of the present invention, several simple deductions or substitutions can be made, which should be considered to fall within the scope of protection of the present invention.

Claims

1. Self-mixing blood RNA storage tube, characterized by: A tube body (1) has a closed end at one end and an open end at the other end, and has a cavity inside. A micro-convex flow disturbance area is provided on the inner wall of the tube body (1), wherein the micro-convex flow disturbance area comprises a first micro-convex area (11) provided at the center of the bottom of the tube body (1), a second micro-convex area (12) provided at the transition area between the bottom and the side wall of the tube body (1), and a third micro-convex area (13) provided at the bottom edge of the tube body (1); A double helical flow guide tube (2) is arranged in the tube body (1), comprising two helical flow guide tubes intertwined with each other, wherein the upper ends of the two helical flow guide tubes are connected, and the lower ends are close to the bottom of the tube body (1) and arranged in opposite directions; RNA stabilizing liquid (3), distributed at the bottom of the tube body (1) and the double-helix flow guide tube (2); A rotary sealing cover (4) is provided at the open end of the tube body (1) and is used to place the cavity inside the tube body (1) in a negative pressure state and to seal the open end; When blood is collected, the rotary sealing cover (4) is opened, and the blood flows into the tube body (1) and rotates along the spiral guide tube, enters the RNA stabilizing liquid (3) area from top to bottom, and automatically mixes with it.

2. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The spiral angle of the spiral flow guide tube is 20° to 30°.

3. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The width of the spiral flow guide tube is 5% to 10% of the inner diameter of the tube body (1).

4. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The sizes of the convex points in the second micro-convex point area (12), the first micro-convex point area (11) and the third micro-convex point area (13) decrease in sequence.

5. The self-mixing blood RNA storage tube according to claim 1, characterized in that: A sealing rubber plug (5) is embedded at the open end of the tube body (1), and a through hole is provided on the sealing rubber plug (5) that is adapted to the connection point of the two spiral flow guide pipes; A position mark (41) is provided on the upper surface of the rotary sealing cover (4) at a position corresponding to the through hole.

6. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The tube body (1) is made of a transparent material, and the outer surface is coated with a light-proof layer, and the inner wall is coated with a polytetrafluoroethylene coating.

7. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The rotary sealing cover (4) is made of medical-grade polypropylene, and the tube body (1) is injection-molded and adapted to fit the rotary sealing cover (4).

8. A method for using the self-mixing blood RNA storage tube according to any one of claims 1 to 7, comprising the following steps: S1. Unscrew the rotary sealing cover (4), insert the blood collection needle into the position mark (41), and the blood rotates from top to bottom along the double-helix guide tube (2) to flow into the bottom of the tube body (1) and automatically mixes with the RNA stabilizing solution (3); S2. Close the rotary sealing cover (4).

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