CN213680675U - Virus transport storage tube - Google Patents

Abstract

The utility model discloses a virus conveying and storing tube, which comprises a tube body, wherein cell culture solution is stored in the tube body, and a host cell layer consisting of host cells is attached to the inner wall of the tube body; the cell culture solution is used for culturing host cells, and the host cell layer is used for culturing viruses; the utility model discloses a host cell cultivates the virus sample, realizes that the virus sample breeds transporting the in-process, reduces because the virus titer reduces the probability that leads to lou examining transporting the in-process and taking place to descend, has improved the positive rate of examining out the virus, need not low temperature moreover and preserves, the cost is reduced.

Description

Virus transport storage tube

All as the field of technology

The utility model relates to a virus detection technology field especially relates to a virus transports save pipe.

All the above-mentioned background techniques

Viruses are an acellular form of nucleic acid molecules (DNA or RNA) and proteins, living parasitically, and are an intermediate organic species between living and non-living organisms. It is composed of a protective shell enclosing a DNA or RNA, and is self-replicating by the host cell system via the mechanism of infection, but cannot grow and replicate independently.

In recent years, more and more new viruses such as SARS coronavirus, human highly pathogenic avian influenza virus, new influenza A H1N1 virus, new coronavirus COVID-19 and the like have been discovered. And in the face of infectious diseases caused by new viruses, whether accurate laboratory etiology diagnosis can be made in time is the key for effectively controlling epidemic situations. The etiological diagnosis of viruses involves the isolation and identification of viruses, the determination of infectious units, the detection of particles, serological detection, and the detection of nucleic acids; the storage and transportation of the virus sample is a very important link in the detection processes, and has a very important influence on the quality of the virus sample.

Currently, the virus detection method is to sample the virus with a sampling swab, and then put the virus into a virus transport preservation solution, such as a virus maintenance solution modified based on a transport medium, and a virus preservation solution modified based on a nucleic acid extraction lysate, for subsequent detection. Although existing methods of virus detection use viral-transport preservative solutions to provide nutrients or reduce factors that degrade viruses and nucleic acids. However, due to the limitations of the liquid composition, differences in individual samples, and differences in delivery conditions, viral titers and nucleic acid materials are still susceptible to various reductions during delivery. Meanwhile, the existing virus detection method needs to store the virus at low temperature, and the low-temperature storage and transportation cost is high.

All kinds of practical novel contents

The utility model aims at providing a virus transports save tube, reduces the probability that viral titer and nucleic acid material appear descending under the process that the sample transported.

In order to realize the utility model discloses a purpose, the utility model discloses a following technical scheme:

a virus conveying and preserving tube comprises a tube body, wherein cell culture solution is stored in the tube body, and a host cell layer which consists of host cells and is generated by a monolayer cell culture method is attached to the inner wall of the tube body; the cell culture fluid is used for culturing host cells, and the host cell layer is used for culturing viruses.

Further, the virus conveying and storing tube also comprises a screen, wherein the screen is assembled inside the tube body and is arranged at intervals with the inner wall of the tube body; the screen is used to contain a swab stained with a viral sample.

As a specific embodiment, the screen includes a screen mounting portion and a screen body; the outer diameter of the screen mounting part is larger than that of the screen main body, and the screen main body is clamped in the pipe body; a stepped hole for bearing the screen mounting part is formed in the inner wall of the pipe orifice of the pipe body; and sieve pores are formed in the side wall of the sieve mesh main body.

As a specific embodiment, the screen body comprises a first screen section and a second screen section; the first screen part is positioned between the screen mounting part and the second screen part; the screen cloth installation department is the ring-type, first screen cloth portion is cylindricly, second screen cloth portion is big-end-up's coniform.

As a specific embodiment, the stepped bore includes a first cylindrical bore and a second cylindrical bore; the first cylindrical hole is positioned above the second cylindrical hole, the diameter of the first cylindrical hole is larger than or equal to the outer diameter of the screen mounting part, and the diameter of the second cylindrical hole is smaller than the outer diameter of the screen mounting part.

In a specific embodiment, the diameter of the second cylindrical hole is smaller than the inner diameter of the pipe body at the lower side of the pipe orifice.

Further, the virus transportation and preservation tube also comprises a plurality of glass beads or ball bearings which are placed in the screen.

Further, the virus transportation and preservation tube also comprises a tube cover; the pipe cover is sleeved on the pipe orifice of the pipe body and detachably connected with the pipe body.

Further, the virus conveying and preserving tube also comprises a rubber plug; the top end face of the tube cover is provided with a rubber plug mounting hole, and the rubber plug is embedded into the rubber plug mounting hole from the inside of the tube cover.

As a specific embodiment, the longitudinal section of the rubber plug is in a shape of a Chinese character 'tu', and the rubber plug comprises a first rubber plug part and a second rubber plug part which are arranged from top to bottom; the diameter of the first rubber plug part is equal to the inner diameter of the rubber plug mounting hole; the diameter of the second rubber plug part is equal to the inner diameter of the annular side wall of the pipe cover.

The utility model has the advantages that:

the utility model discloses a virus transports save set passes through host cell culture virus sample, realizes that virus sample breeds transporting the in-process, reduces because the virus titer is transporting the probability that the in-process emergence decline leads to lou examining, has improved the positive rate of examining out the virus. Further, the utility model discloses a swab and body inner wall are kept apart to the screen cloth, avoid the virus to transport the preservation pipe and take place the oscillation transporting the in-process, lead to swab contact body inner wall, damage the host cell layer of establishing at the body inner wall, lead to the unable host cell that invades of virus to proliferate. Further, the utility model discloses a diameter that sets up the second cylinder hole is less than the inside lateral wall of body, when realizing that the screen cloth inserts the body inside, the first screen cloth portion and the second screen cloth portion of screen cloth are restricted in the within range of establishing of enclosing in the second cylinder hole, can not contact with the inside lateral wall of body, avoid screen cloth male in-process, contact the inside lateral wall of body, damage the host cell layer of establishing at the body inner wall. Furthermore, the utility model discloses a glass bead rubs the contact with the swab many times at the in-process of shaking, is convenient for elute the virus sample on the swab. Furthermore, the utility model discloses a plug that can puncture is embedded in the tube cap, realizes need not to unscrew the tube cap, alright pour into the material into in to the body. Further, the utility model discloses a second plug portion seals, and the guarantee virus transports the intraductal cell culture liquid of mixing with virus sample and host cell and can not take place to leak transporting the in-process, reaches the purpose of safe operation.

Description of the drawings

In order to more clearly illustrate the embodiments of the present invention, the drawings used in the embodiments will be briefly described below. The drawings in the following description are only examples of the present invention, and other drawings can be obtained by those skilled in the art without inventive efforts.

FIG. 1 is a schematic perspective view of a virus transportation and storage tube according to an embodiment of the present invention;

FIG. 2 is a cross-sectional view of a virus transport and storage tube according to an embodiment of the present invention;

FIG. 3 is an exploded view of a virus transport storage tube according to an embodiment of the present invention;

FIG. 4 is a cross-sectional view of the tube body of the virus transportation and storage tube according to the embodiment of the present invention;

FIG. 5 is a partial view of a host cell layer of a virus transport storage tube (in a resting state) according to an embodiment of the present invention;

FIG. 6 is a partial view of the host cell layer of a virus transport storage tube (after shaking) provided in accordance with an embodiment of the present invention with the screen removed;

FIG. 7 is a partial view of a host cell layer of a virus transport storage tube (after shaking) provided by an embodiment of the present invention;

fig. 8 is a schematic flow chart of a method for manufacturing a virus transportation storage tube according to an embodiment of the present invention;

fig. 9 is a schematic flow chart of a virus transportation and preservation method according to an embodiment of the present invention;

description of reference numerals: 100-a pipe body, 110-an internal thread, 120-a stepped hole, 121-a first cylindrical hole, 122-a second cylindrical hole, 200-a pipe cover, 210-an internal thread, 220-a rubber plug mounting hole, 300-a screen, 310-a screen mounting part; 320-a screen body; 321-a first screen part, 322-a second screen part, 400-a rubber plug, 410-a first rubber plug part, 420-a second rubber plug part.

(specific embodiments) in all cases

The present invention will be described in detail with reference to the accompanying drawings.

In order to make the objects, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

As shown in fig. 1 to 3, a virus transportation and preservation tube comprises a tube body 100, a tube cap 200, a screen 300 and a rubber plug 400; the pipe body 100 and the pipe cover 200 are both made of polypropylene materials through surface modification treatment; a host cell layer (not shown) for transporting viruses is attached to the inner wall (including the side inner wall and the bottom inner wall) of the tube 100, a cell culture solution (not shown) for culturing host cells is stored in the tube 100, and the cell culture solution is high enough to soak a swab placed in the tube 100; the host cell layer is a compact monolayer of host cells formed by adherent growth of the host cells under the culture of a monolayer cell culture method; the cell culture solution is used for providing nutrient substances required by the growth of the host cells; the screen 300 is assembled inside the pipe body 100 and spaced apart from the inner wall of the pipe body 100; a receiving space inside the mesh 300 for placing a swab stained with a virus sample; the pipe cover 200 is sleeved on the pipe orifice of the pipe body 100 and detachably connected with the pipe body 100; the top end face of the tube cover 200 is provided with a rubber plug mounting hole 220, and the rubber plug 400 is embedded into the rubber plug mounting hole 220 from the inside of the tube cover 200 and is used for adding lysis solution after being pierced.

In this embodiment, for different virus samples, different types of host cells, such as commercial cells, gene editing cells, or primary cells, can be selected in the tube 100 for culturing, so as to be used for transfection of the virus samples, thereby achieving propagation of the virus samples during transportation, reducing the probability of missed detection due to reduction of virus titer during transportation, and improving the positive rate of detected viruses (as shown in tables 4 to 6 below).

In this embodiment, the cell culture solution is prepared from basic powder, HEPES, BSA, phenol red, sucrose, sodium chloride, bacteriostatic agent, etc.

Table 1 shows the results of clinical tests for delivering influenza a specimens using a conventional viral delivery and storage tube (in which no host cell is used to culture influenza a specimens during the delivery of influenza a specimens) and using the viral delivery and storage tube provided in this example (in which host cells are used to culture influenza a specimens during the delivery of influenza a specimens).

As shown in table 1, when the influenza a virus specimen was transported by the ordinary virus transport storage tube, no influenza a positive test could be detected, whereas when the influenza a virus specimen was transported by the virus transport storage tube provided in this example, 4 influenza a positive tests were detected by the colloidal gold test, and the positive test rate was improved by 40%.

Table 2 shows the clinical test results of delivering the influenza a virus specimen using the common virus delivery/storage tube (in the process of delivering the influenza b virus specimen, the host cell is not used to culture the influenza b virus specimen) and the virus delivery/storage tube provided in this example (in the process of delivering the influenza b virus specimen, the host cell is used to culture the influenza b virus specimen).

As shown in table 2, when the common virus transport storage tube was used to transport the influenza b virus samples, 4 positive cases of influenza b virus were detected by the colloidal gold results, whereas when the virus transport storage tube provided in this example was used to transport the influenza b virus samples, 20 positive cases of influenza b virus were detected by the colloidal gold results, and the detection positive rate was improved by 72%.

Table 3 shows the results of clinical trials of adenovirus samples delivered by the conventional virus delivery storage tube (during the delivery of adenovirus samples, the adenovirus samples were not cultured by host cells) and the virus delivery storage tube provided in this example (during the delivery of adenovirus samples, the adenovirus samples were cultured by host cells).

As shown in table 3, 48 positive adenovirus samples were detected by the colloidal gold results when the adenovirus samples were transported by the common virus transport storage tube, whereas 22 positive adenovirus samples were detected by the colloidal gold results when the adenovirus samples were transported by the virus transport storage tube provided in this example, the detection positivity was improved by 45%.

As shown in tables 1 to 3, the virus transport culture tube provided in this embodiment can perform a sensitization effect on detection results of influenza a, influenza b, and adenovirus, such as qPCR, immunofluorescence, colloidal gold, and the like, thereby improving the detection positive rate, and further reducing the probability of missed detection due to the reduction of the virus titer in the transport process.

In this embodiment, since viruses are propagated in the transportation process, and normal temperature and high temperature are more favorable for infecting host cells by the viruses, the virus transportation culture tube provided by this embodiment can ensure that the titer of the viruses is not reduced without low-temperature cold chain transportation, and the cost is reduced. The cell-containing conveying pipe not only solves the problem that the original virus sample conveying method needs low-temperature cold chain conveying, but also changes the disadvantage of normal temperature or high temperature on sample storage into advantage, and the normal temperature or high temperature conveying is more beneficial to the infection of the virus in the sample on host cells, so that the effect of virus titer proliferation is achieved. Therefore, the titer of the virus sample can not be reduced and can be increased in the transportation process without cold chain conditions, the detection positive rate is finally improved, and the detection omission caused by the reduction of the sample quality is reduced.

In this example, the cell culture fluid transport and storage fluid for culturing the host cells is a serum-free culture fluid; the serum-free culture solution does not contain serum and contains various nutrient components (such as growth factors, tissue extracts and the like) which support cell proliferation and biological reaction, so that the virus can be better protected, the influence of the serum on virus infection is reduced, the virus can be favorably infected into host cells, the host cells can be directly inoculated without being washed, and the virus can be conveniently infected into the host cells for proliferation.

As shown in fig. 2 and 4, the outer side wall of the nozzle of the tube body 100 is provided with an external thread 110, the inner side wall of the ring-shaped tube cover 200 is provided with an internal thread 210, and the tube body 100 and the tube cover 200 are detachably connected through a threaded connection, so that the assembly and the disassembly are convenient.

As shown in fig. 2, 3 and 4, the screen 300 includes a screen mounting portion 310 and a screen main body 320 arranged in sequence from top to bottom; the screen body 320 includes a first screen section 321 and a second screen section 322; the mesh mounting part 310 is formed in a ring shape, and is used for clamping the mesh 300 in the pipe body 100; the first screen part 321 is cylindrical, and the second screen part 322 is conical with a large top and a small bottom; the side walls of the first screen part 321 and the second screen part 322 are both provided with screen holes 340 for communicating with the inside of the pipe body 100; the outer diameter of the screen mounting part 310 is larger than that of the first screen part 321, and the outer diameter of the first screen part 321 is equal to that of the top of the second screen part 322; the longitudinal section of the tube body 100 is approximately U-shaped, and the inner wall structure of the tube body 100 is smooth, so that the wall-attached growth of host cells is facilitated; the inner sidewall diameter of the tubular body 100 is greater than the outer diameter of the first screen section 321; a stepped hole 120 for receiving the screen mounting part 310 is formed in the inner side wall of the nozzle of the pipe body 100; the stepped hole 120 comprises a first cylindrical hole 121 and a second cylindrical hole 122, the first cylindrical hole 121 is positioned above the second cylindrical hole 122, the diameter of the first cylindrical hole 121 is larger than that of the screen mounting part 310, and the diameter of the screen mounting part 310 is larger than that of the second cylindrical hole 122; a stepped surface formed between the first cylindrical bore 121 and the second cylindrical bore 122 is adapted to receive the screen mounting portion 310.

In this embodiment, the virus transport and storage tube further comprises glass beads (not shown) for eluting the virus sample, the glass beads being disposed in the screen 300; after the swab is placed in the screen 300, the tube cover 200 is first tightened, and the virus carrying tube is shaken, and the glass beads are in multiple frictional contacts with the swab during shaking, so that the virus sample on the swab is eluted.

As shown in fig. 2 and 4, in the present embodiment, the diameter of the inner sidewall of the tube 100 is larger than the diameter of the second cylindrical hole 122, so that when the screen 300 is inserted into the tube 100, the first screen portion 321 and the second screen portion 322 of the screen 300 are limited within the range surrounded by the second cylindrical hole 122, and will not contact with the inner sidewall of the tube 100, thereby avoiding the damage to the host cell layer attached to the inner wall of the tube 100 due to the contact with the inner sidewall of the tube 100 during the insertion of the screen 300.

In this embodiment, after the swab stained with the virus sample is placed in the accommodating space inside the screen 300, the isolation effect of the screen 300 does not contact the inner wall of the tube 100, so as to prevent the virus from oscillating during transportation of the storage tube, and the swab shakes and contacts the inner wall of the tube 100, thereby damaging the host cell layer adhered to the inner wall of the tube 100, and causing the virus to be unable to invade the host cell for proliferation.

As shown in fig. 5, the swab is placed in a virus conveying and storing tube, and after standing for a period of time at the temperature of 2-8 ℃, the cells are seen to be dense and full in the visual field, the boundary is clear, and the adherence is good; as shown in fig. 6, the virus transporting and preserving tube without the screen and with the swab placed therein was subjected to low-speed shaking by using a shaker, and after 4 hours, it was seen that the cells were detached and suspended in the visual field, and partially broken, and a large amount of cell debris was present; as shown in FIG. 7, the virus-transporting and storing tube with the mesh and the swab placed therein was shaken at a low speed by a shaker, and after 4 hours, it was observed that the cells remained almost in the same state as in the standing state in the visual field, and a very small amount of the cells fell off.

As shown in fig. 3, the longitudinal section of the rubber plug 400 is in a shape of a Chinese character 'tu', and comprises a first rubber plug part 410 and a second rubber plug part 420 which are arranged from top to bottom; the diameter of the first rubber plug part 410 is equal to the inner diameter of the rubber plug mounting hole 220, and is used for being embedded into the rubber plug mounting hole 220, so that the rubber plug 400 is fixed on the tube cover 200, and the lysate can be conveniently added after being pierced; the diameter of the second packing part 420 is equal to the inner diameter of the annular sidewall of the cap 200 for sealing.

In this embodiment, due to the sealing effect of the second rubber plug part 420, the virus transporting and storing tube has good sealing performance, so that the cell culture fluid mixed with the virus sample and the host cells in the virus transporting tube is prevented from leaking during the transporting process, and the purpose of safe operation is achieved; when the detector needs to inject other substances, such as lysate, into the virus conveying and storing tube, the injection can be completed only by puncturing the rubber plug 400 with the syringe without opening the tube cap 200.

In this example, the lysate is prepared from a denaturant, a surfactant, a protease, a chelating agent, and the like.

As shown in fig. 8, a method for manufacturing a virus transportation storage tube includes the following steps:

unscrewing a tube cover, and adding cell culture solution and host cells into the tube body;

culturing cells by adopting a monolayer cell culture method to ensure that the host cells grow adherently to form a compact host cell layer;

the screen mesh is arranged in the pipe body, and the pipe cover is screwed tightly.

As shown in fig. 9, a virus transportation and preservation method includes the following steps:

unscrewing a pipe cover, putting the swab into a screen, and screwing the pipe cover;

oscillating the virus conveying and storing tube to elute the virus sample;

and transporting the virus conveying and storing tube to a detection site at normal temperature.

In this embodiment, the virus transportation and storage method further comprises attaching a label to the virus transportation and storage tube, and labeling the label with specimen information for easy management; the virus conveying and storing tube is transported to a detection place at normal temperature, such as a laboratory, and then can be directly detected or heated to 37 ℃ for 12-48 hours; if the cracking treatment is needed, the cracking solution can be injected into the tube body by puncturing the rubber plug.

It is only above the preferred embodiment of the utility model, the utility model discloses a scope of protection does not only confine above-mentioned embodiment, the all belongings to the utility model discloses a technical scheme under the thinking all belongs to the utility model discloses a scope of protection. It should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (10)

1. The utility model provides a virus transport preservation pipe, includes the body, its characterized in that: the cell culture solution is stored in the tube body, and a host cell layer consisting of host cells is attached to the inner wall of the tube body; the cell culture solution is used for culturing host cells, and the host cell layer is used for culturing viruses; the screen is assembled inside the pipe body and is arranged at intervals with the inner wall of the pipe body; the screen is used for accommodating a swab stained with a virus sample; the screen comprises a screen mounting part and a screen main body; the external diameter of screen cloth installation department is greater than the external diameter of screen cloth main part is used for with the screen cloth main part card is established inside the body.

2. The virus transport and storage tube of claim 1, wherein: a stepped hole for bearing the screen mounting part is formed in the inner wall of the pipe orifice of the pipe body; and sieve pores are formed in the side wall of the sieve mesh main body.

3. The virus transport and storage tube of claim 2, wherein: the screen body comprises a first screen section and a second screen section; the first screen part is positioned between the screen mounting part and the second screen part; the screen cloth installation department is the ring-type, first screen cloth portion is cylindricly, second screen cloth portion is big-end-up's coniform.

4. The virus transport and storage tube of claim 2, wherein: the stepped hole comprises a first cylindrical hole and a second cylindrical hole; the first cylindrical hole is positioned above the second cylindrical hole, the diameter of the first cylindrical hole is larger than or equal to the outer diameter of the screen mounting part, and the diameter of the second cylindrical hole is smaller than the outer diameter of the screen mounting part.

5. The virus transport and storage tube of claim 4, wherein: the diameter of the second cylindrical hole is smaller than the inner diameter of the pipe body at the lower side of the pipe orifice adjacent to the pipe orifice.

6. The virus transport and storage tube according to any one of claims 1 to 5, wherein: the device also comprises a plurality of balls; the balls are placed on the screen.

7. The virus transport and storage tube of claim 6, wherein: the ball adopts glass beads.

8. The virus transport and storage tube according to any one of claims 1 to 5, wherein: the device also comprises a tube cover; the pipe cover is sleeved on the pipe orifice of the pipe body and detachably connected with the pipe body.

9. The virus transport and storage tube of claim 8, wherein: also comprises a rubber plug; the top end face of the tube cover is provided with a rubber plug mounting hole, and the rubber plug is embedded into the rubber plug mounting hole from the inside of the tube cover.

10. The virus transport and storage tube of claim 9, wherein: the longitudinal section of the rubber plug is in a convex shape and comprises a first rubber plug part and a second rubber plug part which are arranged from top to bottom; the diameter of the first rubber plug part is equal to the inner diameter of the rubber plug mounting hole; the diameter of the second rubber plug part is equal to the inner diameter of the annular side wall of the pipe cover.

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