Self-mixing blood RNA (Ribonucleic Acid) preservation tube and use method

Abstract

The invention discloses a self-mixing blood RNA preservation tube and a use method, and belongs to the field of biomedical sample preservation devices. Wherein RNA stabilizing liquid is pre-stored in the tube body, and a rotary sealing cover is arranged at the opening end of the tube body; a double-helix guide tube is arranged in the tube body, RNA stabilizing liquid is preloaded at the bottom of the double-helix guide tube, and when blood flows into the tube body, the blood rotationally flows along the spiral guide tube and is automatically mixed with the spiral guide tube. The double-helix guide tube is arranged in the tube body, and the RNA stabilizing solution is directly preloaded at the bottom of the double-helix guide tube, so that blood is ensured to rotationally flow along a spiral path after entering the tube body, the blood and the RNA stabilizing solution are fully contacted and uniformly mixed within a short time, an additional mixing step is not needed, the operation error is reduced, and the operation efficiency is improved. The consistency of RNA preservation conditions of all blood sampling samples is ensured, and the stability of experimental data is improved; through the collaborative design of the sealing rubber plug and the rotary sealing cover, the air tightness of blood before the blood enters 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, which 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, and is prone to degradation or inactivation. These factors not only affect the integrity of RNA, but may also affect the accuracy of gene expression analysis, thereby having a significant impact on experimental results. Therefore, how to quickly and efficiently stabilize RNA after blood collection has become an important 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 defects. First, the operation is complicated and requires additional manual addition steps, which increases the complexity of experimental operations and is prone to experimental errors, affecting the repeatability of data; second, the risk of RNA degradation is high. Since the blood is exposed to non-ideal environments for a long time, 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. In particular, exogenous RNase may be introduced during RNA extraction and analysis, further accelerating RNA degradation. (2) Blood collection tubes pre-loaded with RNA preservation solution: In order to solve the problem caused by manually adding RNA stabilizers, some RNA preservation tubes are pre-loaded with RNA stabilizing solution before blood collection, which can automatically mix the blood after it enters the blood collection tube. However, this solution still has the following shortcomings: first, the preservation solution is in 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 when there is leakage during the blood collection process, which may cause 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. The sealing performance of some existing RNA preservation tubes is poor, 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, the existing RNA storage tubes still have many problems during use, including complex operation, high risk of RNA degradation, high risk of contamination, uneven mixing, insufficient sealing performance, etc., which limit their wide 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 the stability of RNA, improve experimental reproducibility, and meet the needs of clinical and scientific research 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] 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 a closed end, the other end of which is an open end, 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 at the bottom of the tube body and the double-helix flow guide tube;

[0010] A rotating sealing cover is arranged at the open end of the tube body, and is used to put the cavity inside the tube body in a negative pressure state and seal 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 ports of the two spiral flow guide tubes are connected, and the lower ports are close to the bottom of the tube body and 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-protrusions are reasonably partitioned and distributed as well as the spacing and height are adjusted, which not only significantly improves the mixing effect of the RNA stabilizing solution and the blood, but also prevents the deposition of the RNA stabilizing solution, thereby ensuring the uniformity and stability of the RNA sample.

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

[0023] The upper surface of the rotary sealing cover is provided with a position mark at a position corresponding to the through hole.

[0024] Through the above technical solution, the sealing rubber stopper and the rotating sealing cover work together to ensure the 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-shielding layer, and the inner wall is coated with a polytetrafluoroethylene coating.

[0026] Through the above technical solution, the degradation and damage of RNA by ultraviolet rays can be prevented, and the visibility of the sample can be maintained 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 matched with the rotary sealing cover.

[0028] Through the above technical solution, the rotary sealing cover 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 comprises the following steps:

[0030] Step 1: Unscrew the rotary sealing cover, insert the blood collection needle into the position mark, and the blood will flow into the bottom of the tube from top to 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 in contact 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 RNA preservation quality. 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 when it is in a closed state;

[0036] Figure 4 It is a top view of the rotary sealing cover in the present invention after being opened.

[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 plug. DETAILED DESCRIPTION

[0038] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. 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 creative work are within the scope of protection of the present invention.

[0039] The present application proposes a self-mixing blood RNA preservation tube, hereinafter referred to as "the preservation tube". The preservation tube adopts a double-helix flow guide tube 2 structure and a rotating sealing cover 4, which can realize automatic mixing of blood and RNA stabilizing liquid 3 after blood collection, ensure the stability of RNA, and avoid direct contact of RNA stabilizing liquid 3 with the human body before blood collection, thereby improving the safety and preservation quality of the sample. This design not only optimizes the mixing efficiency of RNA stabilizing liquid 3 and blood, but also improves the sealing performance of the blood collection tube, effectively reduces the risk of sample contamination and volatilization of stabilizing liquid, greatly improves 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 includes a tube body 1, RNA stabilizing liquid 3, a double helix guide tube 2 and a rotating sealing cover 4. Among them, one end of the tube body 1 is a closed end, the other end is an open end, and the inside is a cavity that can accommodate objects; the double helix guide tube 2 is arranged in the tube body 1, and is composed of two spiral guide tubes intertwined with each other; the RNA stabilizing liquid 3 is pre-stored at the bottom of the tube body 1; the rotating sealing cover 4 is installed at the open end of the tube body 1; when blood is collected, when the blood flows into the tube body 1 through the blood collection needle, the blood rotates along the spiral tube and enters the RNA stabilizing liquid 3 area from top to bottom, and forms a preliminary mixture with the RNA stabilizing liquid 3 while rotating.

[0041] In this embodiment, the length of the tube body 1 is 120 mm, the outer diameter is 15 mm, and the inner diameter is 12 mm. The upper part of the tube body 1 is a blood collection area with a blood collection capacity of 5 mL, and the bottom is filled with RNA stabilizing liquid 3 with a volume of 3 mL, which is suitable for various types of whole blood RNA storage.

[0042] Continue reading Figure 1 The upper ports of the two spiral guide tubes are connected, and the lower ports are close to the bottom of the tube body 1 and arranged in opposite directions, which 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 enhance the fluid speed and prevent blood retention; the tube width of the spiral guide tube is 5% to 10% of the inner diameter of the tube body 1. This design can ensure that enough blood can pass through, and avoid the accumulation of blood in dead corners, thereby improving the mixing efficiency of the blood and the RNA stabilizing solution 3. Compared with the traditional straight tube, the spiral structure improves the mixing uniformity by 30% and reduces blood deposition.

[0043] Continue reading Figure 2, RNA stabilizing liquid 3 is directly pre-installed at the bottom of the double helix flow guide tube 2, and the RNA stabilizing liquid 3 is automatically diffused and evenly mixed after the blood enters by using the microfluidics principle. In order to make the RNA stabilizing liquid 3 fully merge with the blood at the bottom of the tube body 1, a micro-convex flow disturbance area is provided on the inner wall of the tube body 1. The micro-convex flow disturbance area includes a first micro-convex area 11, a second micro-convex area 12 and a third micro-convex area 13, wherein the first micro-convex 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, and the micro-convex points can promote the uniform dispersion of the fluid; the second micro-convex 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-convex points can break the smooth flow of the fluid, form turbulence, and help the mixing of the RNA stabilizing liquid 3 and the blood; the third micro-convex 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 blended into the RNA stabilizing solution 3, the smooth flow of blood can be disrupted, turbulence is easily formed, and it is helpful for mixing the RNA stabilizing solution 3 and blood.

[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 makes reasonable zoning distribution and spacing and height adjustment of the micro-bumps, which 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 at the open end of the tube body 1. The sealing rubber plug 5 is provided with a through hole that is compatible with the connecting part of the two spiral guide tubes. At the same time, a 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 during blood collection to facilitate the insertion of the blood collection needle; it is closed after blood collection is completed 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 rotates from top to bottom along the double helix guide tube 2 to flow into the bottom of the tube body 1 and automatically mix with the RNA stabilizing solution 3;

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

[0050] Specifically, the cooperation between the rotating sealing cover 4 and the sealing rubber plug 5 can ensure that the tube body 1 is fully sealed to prevent external contamination.

[0051] As a preferred solution, 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. The tube body 1 adopts a double-layer transparent light-proof coating to prevent ultraviolet rays from degrading RNA, while maintaining the visibility of the tube body 1 for easy observation of the sample.

[0052] As a preferred solution, the rotating sealing cover 4 is made of medical-grade polypropylene, which can withstand high temperature and high pressure sterilization to ensure pollution-free during use. At the same time, a medical-grade silicone sealing plug is designed at the tube mouth to ensure the airtightness of the tube body 11.

[0053] The storage tube is of integrated design and is manufactured by injection molding process, which simplifies the production process, reduces the manufacturing cost, and has stable product quality and is suitable for disposable use.

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

[0055] (1) Improving RNA mixing efficiency and preventing stratification. The design of the double-helix flow guide tube 2 ensures that the blood flows along the spiral path after entering the tube body 1, so that the blood and the RNA stabilizing solution 3 are fully contacted and evenly mixed in a short time.

[0056] (2) Avoid RNA degradation and improve sample stability. Through the principle of spiral fluid dynamics, the blood automatically disperses during the process of flowing into the tube body 1, so that the preservation solution quickly covers the blood sample, reducing the time RNA is exposed to non-ideal environment and reducing the risk of degradation.

[0057] (3) Automatic mixing to improve accuracy. The automatic diffusion mixing mechanism allows the blood to naturally rotate along the spiral flow path after entering the test tube and directly merge with the preservation solution. No additional mixing steps are required, which reduces operational errors, ensures that the RNA preservation conditions of all blood samples are consistent, and improves the stability of experimental data.

[0058] (4) The design of the rotating sealing cover 4 improves the sealing performance. The rotating sealing cover 4 can completely seal the tube body 1 before blood collection to prevent the external environment from contaminating 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 is combined with the sealing rubber plug 5 to improve the negative pressure holding ability to ensure that there will be no leakage after blood enters.

[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 the injection molding process and the structural integrated design to reduce the 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 the RNA stabilizing solution 3, and improves sample integrity.

[0062] The above content is a detailed description of the present invention in combination with specific embodiments, and it cannot be determined 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 regarded as falling within the scope of protection of the present invention.

Claims

1. A self-mixing blood RNA storage tube, characterized in that: A tube body (1) having a closed end at one end and an open end at the other end, and having a cavity inside; A double-helix flow guide tube (2) is arranged inside the tube body (1); RNA stabilizing liquid (3), distributed at the bottom of the tube body (1) and the double helix flow guide tube (2); A rotating sealing cover (4) is arranged 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 flow guide tube, enters the RNA stabilizing liquid (3) area from top to bottom, and automatically mixes with the RNA stabilizing liquid (3).

2. The self-mixing blood RNA storage tube according to claim 1, characterized in that: The double helical flow guide tube (2) comprises two helical flow guide tubes that are intertwined with each other, and the upper ports of the two helical flow guide tubes are connected, while the lower ports are close to the bottom of the tube body (1) and are arranged in opposite directions.

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

4. 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).

5. The self-mixing blood RNA storage tube according to claim 1, characterized in that: A micro-convex flow disturbance zone is provided on the inner wall of the tube body (1), and the micro-convex flow disturbance zone comprises a first micro-convex zone (11) arranged at the center of the bottom of the tube body (1), a second micro-convex zone (12) arranged in the transition area between the bottom and the side wall of the tube body (1), and a third micro-convex zone (13) arranged at the bottom edge of the tube body (1).

6. The self-mixing blood RNA storage tube according to claim 5, characterized in that: 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 in sequence.

7. The self-mixing blood RNA storage tube according to claim 2, characterized in that: A sealing rubber plug (5) is embedded at the open end of the tube body (1), and a through hole matching the connection between the two spiral flow guide pipes is provided on the sealing rubber plug (5); The upper surface of the rotary sealing cover (4) is provided with a position mark (41) at a position corresponding to the through hole.

8. 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.

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

10. The method for using the self-mixing blood RNA storage tube according to any one of claims 1 to 9, characterized in that it comprises the following steps: S1, unscrewing the rotary sealing cover (4), inserting the blood collection needle into the position mark (41), and the blood rotating along the double helix guide tube (2) from top to bottom flows 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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