CN111944669B - Virus sampling and storing integrated device and use method thereof - Google Patents

Virus sampling and storing integrated device and use method thereof Download PDF

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CN111944669B
CN111944669B CN202010837277.5A CN202010837277A CN111944669B CN 111944669 B CN111944669 B CN 111944669B CN 202010837277 A CN202010837277 A CN 202010837277A CN 111944669 B CN111944669 B CN 111944669B
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sampling
pipe
virus
cover body
upper sleeve
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CN111944669A (en
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张玉基
葛行义
叶生宝
王鹏
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Nanjing Liding Medical Technology Co ltd
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Nanjing Liding Medical Technology Co ltd
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12M23/00—Constructional details, e.g. recesses, hinges
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/38—Caps; Covers; Plugs; Pouring means

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Abstract

The invention discloses a virus sampling and storing integrated device which comprises a rotary bottom tube, an upper sleeve, an upper cover, a lifting structure, a sampling structure and a virus liquid carrying tube. The upper sleeve is sleeved on the rotary bottom pipe, the sampling structure is arranged in the upper sleeve, and the sampling structure comprises a sampling base support, a sampling pipe and a sampling head; the sampling bottom support is provided with an opening at the bottom and is hollow inside; the sampling pipe is fixed on the top of the sampling bottom support, the sampling pipe is of a hollow steel pipe structure which is vertically communicated, and the lower end of the sampling pipe extends into an inner hole of the sampling bottom support; the sampling head is fixed at the top of the sampling pipe, is made of sponge and is communicated with the inside of the sampling pipe; the upper end of the virus liquid carrying pipe is sleeved in the inner hole of the sampling bottom support. The virus liquid carrying pipe is arranged in the rotary bottom pipe and the upper sleeve, the upper cover is opened, the sampling head collects virus samples, and virus liquid collected by the sampling head can directly enter the virus liquid carrying pipe through the sampling pipe, so that the virus can be rapidly and reasonably stored, and the activity and the integrity of virus storage are improved.

Description

Virus sampling and storing integrated device and use method thereof
Technical Field
The invention relates to the technical field of virus inspection and quarantine, in particular to a virus sampling and storing integrated device and a using method thereof.
Background
2019-nCoV is a novel coronavirus, and only research results at present show that the coronavirus is strong in infectivity, and the specific transmission infection characteristics, the retention state in the environment and the genetic variation rule of the coronavirus are not completely disclosed; meanwhile, the 2019-nCoV has strong pathogenicity, and no vaccine or specific clinical medicine for resisting 2019-nCoV infection exists. 2019-nCoV infection, the incubation period is 2-14 days, the initial infection is characterized by fever, dry cough, upper respiratory tract infection, low appetite, diarrhea and the like from asymptomatic to partial, partial cases gradually develop respiratory distress, pneumonia, pulmonary vitreous lesion, hemoptysis and the like, and severe cases are accompanied by lymphocyte reduction and increase of C-reactive protein and lactate dehydrogenase by 7-9. The detection of virus infection relies on methods such as virus isolation and identification, virus gene detection, virus-specific antigen detection, and the like, which must be established on the basis of the collection of high-quality specimens. The conventional virus sampling method is to use a swab to swab a site with a virus such as a pharynx. According to statistics, only 10% -20% of 2019-nCoV throat swab specimens can isolate influenza viruses, which greatly affects disease diagnosis, epidemiological research and research on vaccines and antiviral drugs, and is always a bottleneck of virology clinic and research work.
Patent CN204237790U discloses a sampling device that is used for collection, transportation and storage of viral particle, including the swab that is used for the sampling and the save pipe that contains virus sampling liquid, be provided with the lid on the save pipe, the inboard of lid is provided with the downward horn-shaped mouth of opening, the opening part on horn-shaped mouth upper portion is provided with the recess that is used for fixed swab, the swab includes head and pole portion, the inside a plurality of glass that is provided with of save pipe elutes the pearl. Patent CN206244786U discloses a virus sample thief, including tube cap, sealed, swab, sampling tube body, its characterized in that, the tube cap can be dismantled with the sampling tube body and be connected, the gasket is located the tube cap inboard, with sampling tube body opening looks adaptation, the swab is fixed in the tube cap, the sampling tube body divide into the sampling chamber that is located sampling tube body upper end, is located the sampling tube body middle part and is used for holding the solution chamber of sampling liquid, the barrier film between solution chamber and the sampling chamber to and be located the advancing device four bibliographic categories of sampling tube body bottom, the advancing device top is equipped with the puncture portion.
The problems with the above technique are as follows: after sampling, the swab needs to be soaked in buffer solution such as MEM and then sent to a laboratory for virus separation and detection of virus genes and antigens, but because a certain time interval exists between sample collection and laboratory detection, the samples need to be transported, stored at low temperature and the like, so that the virus activity is reduced, virus nucleic acids, antigens and the like are degraded, and the positive detection rate is far lower than that of clinical diagnosis.
Disclosure of Invention
The invention aims to provide a virus sampling and storing integrated device which can reasonably store viruses and improve the activity and the integrity of virus storage.
The invention solves the technical problems through the following technical means:
a virus sampling and storing integrated device comprises a rotary bottom tube, an upper sleeve, an upper cover, a sampling structure and a virus liquid carrying tube;
the upper sleeve is sleeved on the rotary bottom pipe, and the interior of the upper sleeve is communicated with the interior of the rotary bottom pipe; the upper cover is arranged at the top of the upper sleeve;
the sampling structure is arranged in the upper sleeve and comprises a sampling base support, a sampling pipe and a sampling head; the sampling bottom support is of a cylindrical structure with an opening at the bottom and a hollow interior; the sampling pipe is fixed on the top of the sampling bottom support, the sampling pipe is of a hollow steel pipe structure which is vertically communicated, and the lower end of the sampling pipe extends into an inner hole of the sampling bottom support; the sampling head is fixed at the top of the sampling pipe, is made of sponge and is communicated with the inside of the sampling pipe;
the virus liquid carrying pipe is fixed inside the rotary bottom pipe, the top of the virus liquid carrying pipe is open, the upper end of the virus liquid carrying pipe is sleeved in the inner hole of the sampling collet and is in sliding fit with the inner hole of the sampling collet in the length direction of the inner hole.
The virus liquid carrying pipe is arranged in the rotary bottom pipe and the upper sleeve, the upper cover is opened, the sampling head collects virus samples, and virus liquid collected by the sampling head can directly enter the virus liquid carrying pipe through the sampling pipe, so that the virus can be rapidly and reasonably stored, and the activity and the integrity of virus storage are improved.
Preferably, the lifting device further comprises a lifting structure, wherein two ends of the lifting structure are opened, and the lifting structure is hollow; the lifting structure is sleeved inside the rotating bottom pipe and sleeved outside the virus liquid carrying pipe, and the outer wall of the upper opening of the lifting structure is fixedly connected with the outer wall of the lower opening of the sampling pipe;
and the outer wall of the lifting structure is provided with an external thread.
Preferably, the outer wall of the rotary bottom pipe sequentially comprises a first area and a second area from top to bottom, and a step-shaped structure with the increased height is formed at the joint of the first area and the second area;
an annular bulge is fixedly arranged on the first area;
and the inner wall of the rotary bottom pipe is provided with internal threads.
Preferably, an annular groove matched with the annular bulge is formed in the inner wall of the lower end of the upper sleeve, and the lower end of the upper sleeve is sleeved on the first area of the rotary bottom pipe and is rotatably connected with the first area;
and a first magnet and a second magnet are respectively arranged on two sides of the upper opening wall of the upper sleeve.
Preferably, the upper cover comprises a first cover body and a second cover body, one end of the first cover body is opposite to one end of the second cover body, the other end of the first cover body is hinged with one end of the upper opening wall of the upper sleeve, and the other end of the second cover body is hinged with the other end of the upper opening wall of the upper sleeve; the first cover body and the second cover body form an integral cover structure;
a third magnet corresponding to the first magnet is fixedly arranged at the bottom of the first cover body; and a fourth magnet corresponding to the second magnet is fixedly arranged at the bottom of the second cover body.
Preferably, a first top rod which is vertically arranged is fixed at the bottom of the first cover body; a second ejector rod which is vertically arranged is fixed at the bottom of the second cover body; the first ejector rod and the second ejector rod respectively correspond to two sides of the top of the sampling bottom support;
the lifting structure ascends and moves to drive the sampling collet ascends and causes the top of the sampling collet to be in contact with the bottoms of the first ejector rod and the second ejector rod, and the sampling collet pushes the first ejector rod and the second ejector rod to move upwards and cause the first cover body and the second cover body to be opened towards two sides respectively.
Preferably, the bottom of the sampling tube is provided with a puncturing piece, and the sampling tube is of a hollow syringe needle structure which is communicated up and down.
Preferably, a blocking film is arranged at the top of the virus carrier liquid pipe; the lifting structure moves downwards to drive the sampling tube to move downwards, so that the puncture piece punctures the barrier membrane and extends into the virus carrier liquid tube.
Preferably, the sampling device further comprises a limiting structure arranged between the upper sleeve and the sampling bottom support, and the limiting structure comprises a limiting block and a connecting block;
the outer wall of the limiting block is fixedly connected with the inner wall of the upper sleeve, and a rectangular hole which is communicated up and down is formed in the limiting block;
the connecting block is of a rectangular block structure, is sleeved in the rectangular hole and is in sliding fit with the rectangular hole in the length direction of the rectangular hole; the inside mounting hole of having seted up of connecting block, sampling collet middle part cup joints in the mounting hole.
The invention also discloses a virus sampling and storing method using the virus sampling and storing integrated device, which comprises the following steps:
s1, opening an upper cover, enabling a lifting structure to perform ascending motion in a rotating bottom pipe and drive a sampling structure to perform ascending motion, and driving a sampling pipe and a sampling head to perform ascending motion while a sampling bottom support performs ascending motion until the sampling head extends out of the upper part of an upper sleeve;
s2, collecting virus liquid by a sampling head;
s3, after sampling is finished, the lifting structure performs descending motion in the rotary bottom pipe and drives the sampling structure to perform descending motion until the lower end of the sampling pipe extends into the virus carrier liquid pipe, and virus liquid collected by the sampling head enters the virus carrier liquid pipe through the sampling pipe;
and S4, closing the upper cover for sealed preservation.
Further, virus carrier liquid is arranged in the virus carrier liquid pipe, and the virus carrier liquid comprises the following raw materials in parts by weight: 5-10 parts of Hank's buffer solution, 1-10 parts of stabilizing agent, 0.1-1 part of antibiotic, 0.5-4 parts of protective agent, 0.001 part of indicator and the balance of purified water; the pH value of the virus carrier liquid is 6.9-7.4.
The invention has the advantages that:
1. the virus liquid carrying pipe is arranged in the rotary bottom pipe and the upper sleeve, the upper cover is opened, the sampling head collects virus samples, and virus liquid collected by the sampling head can directly enter the virus liquid carrying pipe through the sampling pipe, so that the virus can be rapidly and reasonably stored, and the activity and the integrity of virus storage are improved.
2. The sampling structure provided by the invention can move up and down through the lifting structure, so that the sampling structure can be rapidly extended and retracted, the sampling operation is safe, and the sampling risk is reduced.
3. The upper cover and the upper sleeve are respectively provided with the magnets, and the upper cover and the upper sleeve can be naturally covered due to the magnetic force between the magnets.
4. According to the sampling collet, the first ejector rod and the second ejector rod are arranged at the bottoms of the first cover body and the second cover body, and the sampling collet can push the first cover body and the second cover body to be opened towards two sides respectively through the first ejector rod and the second ejector rod in the ascending motion, so that time and labor are saved.
5. According to the invention, through the arrangement of the limiting structure, the sampling collet and the connecting block cannot rotate when moving up and down, and the connecting block can only slide up and down in the rectangular hole, so that when the sampling collet is used, the upper sleeve can be fixed firstly, and then the rotating bottom tube can be directly rotated.
Drawings
FIG. 1 is a schematic structural diagram of an integrated virus sampling and storing device according to the present invention;
FIG. 2 is a schematic structural diagram illustrating a usage status of the integrated virus sampling and storing device according to the present invention;
FIG. 3 is an enlarged schematic structural diagram of A according to an embodiment of the present invention;
FIG. 4 is an enlarged structural diagram of B in the embodiment of the present invention;
FIG. 5 is a schematic structural diagram of a limiting block according to an embodiment of the present invention;
FIG. 6 is a schematic structural diagram of a connecting block according to an embodiment of the present invention;
FIG. 7 is a schematic view of a connection structure of a connection block, a sampling bottom support and a limiting block according to an embodiment of the present invention;
FIG. 8 is a graph showing the results of preservation of virus activity and integrity with preservation solutions of different origins in accordance with examples of the present invention;
FIG. 9 is a graph showing the results of the virus activity according to the present invention in examples depending on the days of storage.
The reference numbers illustrate:
1. rotating the bottom tube; 11. a first region; 12. a second region; 13. an annular projection; 14. an internal thread; 2. sleeving a sleeve; 21. an annular groove; 22. a first magnet; 23. a second magnet; 3. an upper cover; 31. a first cover body; 32. a second cover body; 33. a third magnet; 34. a fourth magnet; 35. a first ejector rod; 36. a second ejector rod; 4. a lifting structure; 41. an external thread; 5. a sampling structure; 51. sampling a bottom support; 52. a sampling tube; 521. a piercing member; 53. a sampling head; 6. a virus-carrying tube; 61. a barrier film; 7. a limiting structure; 71. a limiting block; 711. a rectangular hole; 72. connecting blocks; 721. mounting holes; 8. a viral carrier fluid.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example one
As shown in fig. 1, the present embodiment discloses an integrated device for sampling and storing virus, which includes a rotary bottom tube 1, an upper sleeve 2, an upper cover 3, a lifting structure 4, a sampling structure 5, a virus-carrying tube 6 and a limiting structure 7.
As shown in fig. 1-3, the rotary bottom tube 1 of the present embodiment is a tubular structure with an open top end and a hollow interior, the outer wall of the rotary bottom tube 1 sequentially comprises a first region 11 and a second region 12 which are integrally formed from top to bottom, and a step-shaped structure with an increased height is formed at the joint of the first region 11 and the second region 12;
the first area 11 is provided with an annular bulge 13 which is integrally formed with the first area, and the annular bulge 13 can also be fixed on the first area 11 by adopting the existing welding mode;
as shown in fig. 3, the inner wall of the rotary bottom tube 1 is provided with an internal thread 14 integrally formed therewith.
As shown in fig. 1-3, the upper sleeve 2 of the present embodiment is a cylindrical structure with an upper opening and a lower opening and a hollow interior, an annular groove 21 is formed on an inner side wall of a lower end of the upper sleeve 2, the annular groove 21 is matched with the annular protrusion 13, the lower end of the upper sleeve 2 is sleeved on the first region 11 of the rotary bottom pipe 1, the lower end of the upper sleeve 2 is rotatably connected with the first region 11 of the rotary bottom pipe 1, and the interior of the upper sleeve 2 is communicated with the interior of the rotary bottom pipe 1;
as shown in fig. 1, and specifically referring to the orientation of fig. 1, the upper casing 2 of the present embodiment has a first magnet 22 and a second magnet 23 mounted on the left and right sides of the upper opening wall thereof, respectively.
As shown in fig. 1 and fig. 2, and specifically referring to the orientation of fig. 1, the upper cover 3 includes a first cover 31 and a second cover 32, the first cover 31 and the second cover 32 of this embodiment are both semi-circular disk structures, the right end of the first cover 31 is opposite to the left end of the second cover 32 and can be combined into a complete disk structure, and the left end of the first cover 31 is connected to the left end of the upper opening wall of the upper sleeve 2 through a hinge, and the right end of the second cover 32 is connected to the right end of the upper opening wall of the upper sleeve 2 through a hinge.
As shown in fig. 1-3, the lifting structure 4 is disposed inside the rotating bottom pipe 1, the lifting structure 4 of this embodiment has a hollow structure with openings at two ends, and an external thread 41 integrally formed with the lifting structure 4 is disposed on an outer wall of the lifting structure 4, the lifting structure 4 is sleeved inside the rotating bottom pipe 1, and the external thread 41 is in threaded fit with the internal thread 14 in a length direction thereof.
As shown in fig. 1 and 2, the sampling structure 5 includes a sampling shoe 51, a sampling tube 52 and a sampling head 53;
the sampling bottom support 51 is a cylindrical structure with an opening at the bottom and a hollow interior, and the lower opening wall of the sampling bottom support 51 is welded and fixed with the upper opening wall of the lifting structure 4, and can also be fixedly connected by adopting the existing bolt or screw connection mode;
the sampling pipe 52 is fixed on the top of the sampling support 51, the sampling pipe 52 is of a hollow steel pipe structure which is communicated up and down, the lower end of the sampling pipe 52 extends into an inner hole of the sampling support 51, and the outer wall of the sampling pipe 52 is welded and fixed with the sampling support 51;
the sampling head 53 is fixed on the top of the sampling pipe 52, the sampling head 53 is made of sponge, and the sampling head 53 is communicated with the inside of the sampling pipe 52;
the bottom of the sampling tube 52 is provided with a puncture piece 521 integrally formed with the sampling tube 52, and the sampling tube 52 is of a hollow syringe needle structure which is communicated up and down.
As shown in fig. 1 and fig. 2, the bottom of the virus liquid-carrying tube 6 is fixed to the bottom of the rotating bottom tube 1 by welding, the top of the virus liquid-carrying tube 6 is open, the lower end of the virus liquid-carrying tube 6 is sleeved in the inner hole of the lifting structure 4, the outer sidewall of the lower end of the virus liquid-carrying tube 6 is fixed to the inner hole wall of the lifting structure 4 by welding, and the upper end of the virus liquid-carrying tube 6 is sleeved in the inner hole of the sampling bottom support 51 and is in sliding fit with the inner hole of the sampling bottom support 51 in the length direction;
as shown in fig. 2 and 4, a blocking film 61 is disposed on the top of the virus-carrying tube 6, and the blocking film 61 is connected to the top port wall of the virus-carrying tube 6 by the existing adhesive bonding technique; the descending movement of the lifting structure 4 drives the sampling tube 52 to descend, so that the puncturing member 521 punctures the barrier film 61 and extends into the virus carrier tube 6.
As shown in fig. 1, fig. 2, and fig. 5-7, a limiting structure 7 is further disposed between the upper casing 2 and the sampling base 51, and the limiting structure 7 includes a limiting block 71 and a connecting block 72;
the outer wall of the limiting block 71 is welded and fixed with the inner wall of the upper sleeve 2, or can be fixedly connected by adopting the existing bolt or screw connection mode, and a rectangular hole 711 which is through up and down is formed in the limiting block 71;
the connecting block 72 is in a rectangular block structure, and the connecting block 72 is sleeved in the rectangular hole 711 and is in sliding fit with the rectangular hole 711 in the length direction of the connecting block; connecting block 72 is inside to be seted up mounting hole 721, and sampling collet 51 cup joints in mounting hole 721 in the middle part, and welded fastening between the outer sidewall middle part of sampling collet 51 and the inner wall of mounting hole 721 also can adopt current bolt or screwed connection's mode fixed connection.
The invention also discloses a virus sampling and storing method using the virus sampling and storing integrated device, which comprises the following steps:
s1, respectively opening a first cover body 31 and a second cover body 32, fixing an upper sleeve 2 and then rotating a rotary bottom tube 1, wherein due to the arrangement of a limiting structure 7, the sampling bottom support 51 and a connecting block 72 cannot rotate when moving up and down, and the connecting block 72 can only slide up and down in a rectangular hole 711, so that when the rotary bottom tube 1 is used, the upper sleeve 2 can be fixed firstly, and then the rotary bottom tube 1 can be directly rotated;
s2, the sampling head 53 collects virus liquid, and the sampling head 53 is made of sponge material and communicated with the sampling pipe 52, so that a collected virus liquid sample can be directly sucked into the sampling pipe 52 through the sampling head 53 and flows downwards along the inner hole of the sampling pipe 52;
s3, after sampling is finished, the lifting structure 4 descends in the rotary bottom tube 1 and drives the sampling structure 5 to descend, the puncture piece 521 at the bottom of the sampling tube 52 punctures the barrier film 61 at the top of the virus carrier tube 6 and extends into the virus carrier tube 6, and virus liquid collected by the sampling head 53 enters the virus carrier tube 6 through the sampling tube 52;
and S4, closing the upper cover 3 for sealed preservation.
Compared with the prior art, the invention has the following advantages: firstly, the sampling structure 5 of the invention realizes the up-and-down movement through the lifting structure 4, so that the sampling structure 5 can be quickly extended out and retracted, the sampling operation is safe, the sampling risk is reduced, and the sampling structure 5 completing the sampling retracts and extends into the virus carrying liquid pipe 6 by arranging the virus carrying liquid pipe 6 in the rotary bottom pipe 1, so that the reasonable storage of the virus in the virus carrying liquid pipe 6 is quickly realized, and the activity and the integrity of the virus storage are further improved. Secondly, according to the invention, through the arrangement of the limiting structure 7, the sampling bottom support 51 and the connecting block 72 cannot rotate when moving up and down, and the connecting block 72 can only slide up and down in the rectangular hole 711, so that the upper sleeve 2 can be fixed firstly when in use, and then the rotating bottom tube 1 can be directly rotated, and the sampling bottom support 51 connected with the lifting structure 4 cannot rotate because the rotating bottom tube 1 is in threaded fit with the lifting structure 5, so that the sampling bottom support 51 can only slide up and down.
Example two
The present embodiment differs from the above embodiments in that: as shown in fig. 1 and 2, a third magnet 33 corresponding to the first magnet 22 is mounted at the bottom of the first cover 31, and the first magnet 22 and the third magnet 33 are magnetically connected; a fourth magnet 34 corresponding to the second magnet 23 is mounted on the bottom of the second lid 32, and the second magnet 23 and the fourth magnet 34 are magnetically connected.
The upper cover 3 and the upper sleeve 2 are respectively provided with the magnets, and the upper cover 3 and the upper sleeve 2 can be naturally covered under the action of magnetic force between the magnets.
EXAMPLE III
The present embodiment differs from the above embodiments in that: as shown in fig. 1 and 2, a first push rod 35 vertically arranged is fixed at the bottom of the first cover 31, and the top end of the first push rod 35 is welded and fixed with the bottom of the first cover 31, and can also be fixedly connected by using the existing bolt or screw connection; a second top rod 36 which is vertically arranged is fixed at the bottom of the second cover body 32, and the top end of the second top rod 36 is welded and fixed with the bottom of the second cover body 32, and can also be fixedly connected in the existing bolt or screw connection mode; and the first and second push rods 35 and 36 correspond to both sides of the top of the sampling shoe 51, respectively.
In the embodiment, the first push rod 35 and the second push rod 36 are arranged at the bottoms of the first cover body 31 and the second cover body 32, the ascending motion of the lifting structure 4 drives the sampling bottom support 51 to ascend and causes the top of the sampling bottom support 51 to contact with the bottoms of the first push rod 35 and the second push rod 36, and the sampling bottom support 51 pushes the first push rod 35 and the second push rod 36 to move upwards and causes the first cover body 31 and the second cover body 32 to be opened towards two sides respectively.
Example four
The present embodiment differs from the above embodiments in that: the virus carrier liquid pipe 6 is internally provided with virus carrier liquid, and the virus carrier liquid 8 comprises the following raw materials in parts by weight: 5-10 parts of Hank's buffer solution, 1-10 parts of stabilizing agent, 0.1-1 part of antibiotic, 0.5-4 parts of protective agent, 0.001 part of indicator and the balance of purified water; the pH value of the virus carrier liquid is 6.9-7.4.
The stabilizer, the antibiotic, the protective agent and the purified water are all produced by Nanjing Litding medical technology Co., ltd, and the model of the stabilizer is LD-WDJ-01; the antibiotic is LD-KSS-02; the model of the purified water is LD-CS-11; the model of the protective agent is LD-BHJ-01.
EXAMPLE five
The present embodiment differs from the above embodiments in that: the virus carrier liquid 8 of the embodiment comprises the following raw materials in parts by weight: 5-8 parts of Hank's buffer solution, 5-10 parts of stabilizing agent, 0.5-1 part of antibiotic, 0.5-2 parts of protective agent, 0.001 part of indicator and the balance of purified water, wherein the pH value of the virus carrier liquid 8 is 6.9-7.4.
EXAMPLE six
The present embodiment is a virus carrier liquid of the present invention, where the carrier liquid includes a reagent a: 8-10 parts of Hank's buffer solution, 1-5 parts of stabilizing agent, 0.1-0.5 part of antibiotic, 2-4 parts of protective agent and 0.001 part of indicator, wherein the pH value of virus carrier liquid 8 is 6.9-7.4.
Comparative example 1
This comparative example was a virus preservation solution purchased from another company.
EXAMPLE seven
This example uses an agar plaque assay to test the activity and integrity of the viral carrier fluid 8 of examples 4-5 against virions.
Experimental materials:
1. 30ml of maintenance liquid; 1 6-well plate and 12-well plate (agar final concentration 0.8%)
(1)10mlddH 2 O +0.3g of low melting agar (0.03 g/ml);
(2) 19mlDMEM +10% FBS (3 ml) preheated at 37 ℃;
(3) Mixing 10mlddH 2 O +0.3g low melting point agar (0.03 g/ml) and 19mlDMEM +10% FBS (3 ml) to prepare maintenance liquid;
(4) 8ml Low melting agar +19mlDMEM +3mlFBS =30ml.
2. Fixing liquid: 75% ethanol +25% acetic acid.
3. 0.1% of Crystal Violet: dyeing process
Figure RE-GDA0002678443380000131
Storing at room temperature in dark.
4. Cell line: hela cells.
5. Virus: and (4) VSV.
The experimental steps are as follows:
1. hela cells were seeded in 6-well plates (3 plates, two time points) at 5X 10 5 2ml per well (density about 60% -70%); virus CVB3 (4 ℃) was preserved simultaneously at 7d/14d/21d using viral carrier fluid 8 of a different origin.
2. Cell supernatants were collected separately at 12 h/h of culture and cell lysates were collected separately at the corresponding time.
3. Progeny CVB3 virus PFU assay (virus PFU in each sample was measured separately by the plaque method):
1) Hela cells were seeded in 6-well plates (3 plates were spread), 10 5 2ml per well;
2) Removing the culture medium by suction, washing the cells for 1-2 times by PBS, and adding the VSV virus solution collected before for incubation for 1 hour;
3) And the venom is absorbed and discarded, and the washing is carried out for 1 to 2 times. Adding 2ml of maintenance liquid into each hole, standing and solidifying, and not moving the plate for 20min in the solidification process;
4) Culturing at 37 deg.C in 5% carbon dioxide incubator for 2d without shaking the plate;
5) After the formation of plaques was observed under a microscope, 1.5ml of the fixative was added to each well for 20min. Scraping off agar in the pore plate, adding 1ml of crystal violet solution into each pore, and dyeing for 3min at room temperature;
6) Absorbing the crystal violet solution, putting the pore plate into bleaching water for a few seconds, taking off the cover, taking out the pore plate after a few seconds, and slowly washing with water;
7) And a few plaques, and the protective effect of the preservation solution on the active organisms of the virus was evaluated.
The results are shown in fig. 8, and it can be seen from fig. 8 that the plaque experiments performed on the collected venom show that the virus preserved in the preservation fluid from different sources still has a relatively obvious difference in virus activity and integrity, wherein the cells in the virus carrier fluid No. 1 and the virus carrier fluid No. 2 are obviously completely lysed, which indicates that the virus activity and toxicity are very strong. FIG. 9 shows the course of viral activity over days of storage, with no significant change in activity observed with increasing days for carrier 1 and carrier 2. The IBL/TSZ increase in cells over the course of days and virus viability decreases.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims (10)

1. The utility model provides a virus sampling and save integrated device which characterized in that: comprises a rotary bottom tube, an upper sleeve, an upper cover, a sampling structure and a virus liquid carrying tube;
the upper sleeve is sleeved on the rotary bottom pipe, and the interior of the upper sleeve is communicated with the interior of the rotary bottom pipe; the upper cover is arranged at the top of the upper sleeve;
the sampling structure is arranged in the upper sleeve and comprises a sampling bottom support, a sampling pipe and a sampling head; the sampling bottom support is of a cylindrical structure with an opening at the bottom and a hollow interior; the sampling pipe is fixed on the top of the sampling bottom support, the sampling pipe is of a hollow steel pipe structure which is communicated up and down, and the lower end of the sampling pipe extends into an inner hole of the sampling bottom support; the sampling head is fixed at the top of the sampling pipe, is made of sponge and is communicated with the inside of the sampling pipe;
the virus liquid carrying pipe is fixed inside the rotary bottom pipe, the top of the virus liquid carrying pipe is provided with an opening, and the upper end of the virus liquid carrying pipe is sleeved in the inner hole of the sampling collet and is in sliding fit with the inner hole of the sampling collet in the length direction of the sampling collet;
the sampling device also comprises a limiting structure arranged between the upper sleeve and the sampling bottom support, wherein the limiting structure comprises a limiting block and a connecting block;
the outer wall of the limiting block is fixedly connected with the inner wall of the upper sleeve, and a rectangular hole which is communicated up and down is formed in the limiting block;
the connecting block is of a rectangular block structure, is sleeved in the rectangular hole and is in sliding fit with the rectangular hole in the length direction of the rectangular hole; the connecting block is internally provided with a mounting hole, and the middle part of the sampling collet is sleeved in the mounting hole.
2. The integrated virus sampling and storing device of claim 1, wherein: the lifting structure is provided with openings at two ends and is hollow inside; the lifting structure is sleeved inside the rotary bottom pipe and sleeved outside the virus liquid carrying pipe, and the outer wall of the upper opening of the lifting structure is fixedly connected with the outer wall of the lower opening of the sampling structure;
and the outer wall of the lifting structure is provided with an external thread.
3. The integrated virus sampling and storage device of claim 1, wherein: the outer wall of the rotary bottom pipe sequentially comprises a first area and a second area from top to bottom, and a step-shaped structure with the height increased is formed at the joint of the first area and the second area;
an annular bulge is fixedly arranged on the first area;
the inner wall of the rotary bottom pipe is provided with internal threads, and the internal threads on the inner wall of the rotary bottom pipe are in threaded fit with the external threads on the outer wall of the lifting structure.
4. The integrated virus sampling and storage device of claim 3, wherein: an annular groove matched with the annular bulge is formed in the inner wall of the lower end of the upper sleeve, and the lower end of the upper sleeve is sleeved on the first area of the rotary bottom pipe and is in rotary connection with the first area;
and a first magnet and a second magnet are respectively arranged on two sides of the upper opening wall of the upper sleeve.
5. The integrated virus sampling and storage device of claim 4, wherein: the upper cover comprises a first cover body and a second cover body, one end of the first cover body is opposite to one end of the second cover body, the other end of the first cover body is hinged with one end of the upper opening wall of the upper sleeve, and the other end of the second cover body is hinged with the other end of the upper opening wall of the upper sleeve; the first cover body and the second cover body form an integral cover structure;
a third magnet corresponding to the first magnet is fixedly arranged at the bottom of the first cover body; and a fourth magnet corresponding to the second magnet is fixedly arranged at the bottom of the second cover body.
6. The integrated virus sampling and storage device of claim 5, wherein: a first ejector rod which is vertically arranged is fixed at the bottom of the first cover body; a second ejector rod which is vertically arranged is fixed at the bottom of the second cover body; the first ejector rod and the second ejector rod respectively correspond to two sides of the top of the sampling bottom support;
the lifting structure ascends and moves to drive the sampling collet ascends and causes the top of the sampling collet to be in contact with the bottoms of the first ejector rod and the second ejector rod, and the sampling collet pushes the first ejector rod and the second ejector rod to move upwards and cause the first cover body and the second cover body to be opened towards two sides respectively.
7. The integrated virus sampling and storage device of claim 6, wherein: the sampling pipe bottom is provided with the piercing member, the sampling pipe is the hollow syringe needle structure that link up from top to bottom.
8. The integrated virus sampling and storage device of claim 7, wherein: a blocking film is arranged at the top of the virus liquid carrying pipe; the lifting structure moves downwards to drive the sampling tube to move downwards, so that the puncture piece punctures the barrier membrane and extends into the virus carrier liquid tube.
9. A method for sampling and storing a virus using the integrated device for sampling and storing a virus according to claim 1, comprising the steps of:
s1, opening an upper cover, enabling a lifting structure to perform lifting motion in a rotary bottom pipe and drive a sampling structure to perform lifting motion, and driving a sampling pipe and a sampling head to perform lifting motion while the sampling bottom support performs lifting motion until the sampling head extends out of the upper part of an upper sleeve;
s2, collecting virus liquid by a sampling head;
s3, after sampling is finished, the lifting structure performs descending motion in the rotary bottom pipe and drives the sampling structure to perform descending motion until the lower end of the sampling pipe extends into the virus carrier liquid pipe, and virus liquid collected by the sampling head enters the virus carrier liquid pipe through the sampling pipe;
and S4, closing the upper cover for sealing and storing.
10. The method for sampling and storing a virus according to claim 9, wherein: virus carrier liquid is arranged in the virus carrier liquid pipe and comprises the following raw materials in parts by weight: 5-10 parts of Hank's buffer solution, 1-10 parts of stabilizing agent, 0.1-1 part of antibiotic, 0.5-4 parts of protective agent, 0.001 part of indicator and the balance of purified water; the pH value of the virus carrier liquid is 6.9-7.4.
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