Rapid detection kit for micropterus salmoides cell swelling iridovirus

Abstract

The utility model relates to the field of rapid detection kits for cell swelling iridovirus of micropterus salmoides, in particular to a rapid detection kit for cell swelling iridovirus of micropterus salmoides. Comprising a plurality of reagent and raw material storage places, a plurality of reaction liquid preparation box chambers with stirring, mixing, heating and heat preservation functions, a detection mechanism utilizing colloidal gold test strips and the like. Compared with the prior art, the rapid detection kit for the cell swelling iridovirus of the micropterus salmoides has the advantages that the rapid detection kit for the cell swelling iridovirus of the micropterus salmoides is provided under the support of a recombinase polymerase amplification (RPA) and a lateral flow chromatography technology, and the cell swelling iridovirus can be conveniently and rapidly diagnosed by adopting the rapid detection kit for the cell swelling iridovirus of the micropterus salmoides.

Description

Technical Field

[0001] The utility model relates to the field of a rapid detection kit for California sea bass cell swelling iridescent virus, in particular to a rapid detection kit for California sea bass cell swelling iridescent virus. Background Art

[0002] Megalocytivirus, belonging to the family Iridovirudae, is an icosahedral, cytoplasmic DNA virus and one of the important viral pathogens of fish. The diseased fish of California bass infected with the disease mainly swim slowly and independently on the water surface, with white gills or bleeding spots, and enlarged liver, spleen and kidneys. The mortality rate after the disease ranges from 30% (adult fish stage) to 100% (seedling stage). Therefore, the development of a rapid and efficient method for detecting Megalocytivirus is crucial for the prevention and treatment of Megalocytivirus disease of California bass.

[0003] Existing detection methods mostly use PCR amplification, which has high hardware requirements, relies on professional scientific researchers, and is time-consuming. Therefore, it is necessary to design a rapid detection kit for California bass cell swelling iris virus to achieve rapid and economical detection of cell swelling iris virus. Utility Model Content

[0004] 1. Technical Problems Solved

[0005] The technical problem to be solved by the utility model is that the existing methods for detecting California sea bass cell swelling iris virus mostly adopt PCR amplification technology as the basis, which has high requirements on equipment and personnel during detection, thus causing inconvenience in detection and relatively high cost.

[0006] 2. Technical Solution

[0007] In order to solve the above technical problems, the technical solution provided by the utility model is: a rapid detection kit for California sea bass cell swelling iris virus, including a box body, a detection box is connected to the box body, a plurality of slots are connected to one side of the box body, and a colloidal gold test strip matching the detection box is connected to the box body in the slot.

[0008] A powder trough is connected to the top of the box body, a powder tank is rotatably connected to the box body in the powder trough, a discharge port is connected to the bottom of the powder tank, a feed pipe matching the discharge port is downwardly connected to the box body at the bottom of the powder trough, a plurality of reagent chambers are connected to the box body at the outside of the powder tank, a reaction box connected to the bottom of the feed pipe is located below the powder tank, a reagent conduit is commonly connected between the reagent chamber and the reaction box, a sample box is connected to the side of the top of the box body away from the detection box, a support shaft is rotatably connected to the sample box, a plurality of blades are connected to the support shaft, and a sample conduit is commonly connected between the sample box and the reaction box.

[0009] A collection funnel is connected to the top of the reaction box, a mixing chamber connected to the bottom of the collection funnel is connected to the bottom of the reaction box, a plurality of electric heating columns are connected to the reaction box located outside the mixing chamber and on both sides of the sample box, a connecting pipe connected to the mixing chamber is commonly connected between the reaction box and the detection box, a liquid storage tank is connected to the side of the box body close to the detection box, an infusion tube is commonly connected between the liquid storage tank and the detection box, a transmission shaft is rotatably connected to the reaction box located inside the mixing chamber and at the bottom of the detection box, a plurality of stirring blades are connected to the transmission shaft, a waste water tank is connected to the box body located below the sample box, a waste pipe is commonly connected between the sample box, the reaction box and the detection box and the waste water tank, and a sewage outlet matching the waste water tank is connected to one side of the bottom of the box body.

[0010] Furthermore, the reagent conduit is connected to a solenoid valve for allowing the reagent in the reagent chamber to flow into a reaction box below under the action of gravity.

[0011] Furthermore, the sample conduit, connecting tube, infusion tube, waste pipe and sewage outlet are all connected with a water pump for making the liquid flow in each box through the corresponding pipeline.

[0012] Furthermore, the powder tank, the transmission shaft and one end of the support shaft are all connected with a driving motor to provide power for the rotation of the corresponding matching parts.

[0013] Furthermore, the reagent chamber, powder tank and liquid storage box are all connected with feed ports at the top to facilitate replenishment to the corresponding storage positions of each raw material involved in the reaction detection. The top of the sample box is connected with a sliding cover for opening or closing the sample box.

[0014] 3. Beneficial Effects

[0015] The advantages of the utility model compared with the prior art are: the device adopts recombinase polymerase amplification (RPA) and lateral flow chromatography technology, and with the support of this technology, provides a California sea bass cell swelling iris virus rapid detection kit, using this kit, cell swelling iris virus disease can be conveniently and quickly diagnosed. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The utility model is a schematic diagram of the appearance structure of a rapid detection kit for California sea bass cell swelling iris virus.

[0017] Figure 2 The utility model is a schematic diagram of the appearance structure of a rapid detection kit for California sea bass cell swelling iris virus when in use.

[0018] Figure 3 This is a schematic diagram of the internal structure of a California sea bass cell swelling iris virus rapid detection kit of the utility model Figure 1 .

[0019] Figure 4 This is a schematic diagram of the internal structure of a California sea bass cell swelling iris virus rapid detection kit of the utility model Figure 2 .

[0020] Figure 5 This is a schematic diagram of the internal structure of a California sea bass cell swelling iris virus rapid detection kit of the utility model Figure 3 .

[0021] Figure 6 This is a schematic diagram of the internal structure of a California sea bass cell swelling iris virus rapid detection kit of the utility model Figure 4 .

[0022] Figure 7 It is a schematic diagram of the structure of part A.

[0023] As shown in the figure: 1. Box body, 2. Reagent chamber, 3. Powder tank, 4. Powder trough, 5. Driving motor, 6. Feeding port, 7. Discharging port, 8. Feed pipe, 9. Reagent conduit, 10. Solenoid valve, 11. Reaction box, 12. Aggregate funnel, 13. Mixing chamber, 14. Transmission shaft, 15. Stirring blade, 16. Water pump, 17. Electric heating column, 18. Connecting pipe, 19. Sample box, 20. Sliding cover, 21. Support shaft, 22. Blade, 23. Sample conduit, 24. Waste pipe, 25. Test box, 26. Liquid storage tank, 27. Infusion tube, 28. Wastewater tank, 29. Sewage outlet, 30. Slot, 31. Colloidal gold test strip. DETAILED DESCRIPTION

[0024] The utility model is further described in detail below in conjunction with the accompanying drawings.

[0025] Embodiment 1

[0026] Combined with Figure 1-3 In order to solve the above technical problems, the technical solution provided by the utility model is: a rapid detection kit for California sea bass cell swelling iris virus, including a box body 1, a detection box 25 is connected to the box body 1, a plurality of slots 30 are connected to one side of the box body 1, and a colloidal gold test strip 31 matching the detection box 25 is connected to the box body 1 located in the slot 30.

[0027] The colloidal gold test strip 31 is inserted into the slot 30 on the box body 1, and the bottom of the colloidal gold test strip 31 extends into the detection box 25 and contacts with the prepared detection liquid. Then, the detection result is obtained through the corresponding scale lines on the colloidal gold test strip 31, namely the quality control line, the detection line and the sample line.

[0028] Embodiment 2

[0029] Combined with Figure 1 , Attachment Figure 2 , Attachment Figure 4 , Attachment Figure 5 , Attachment Figure 7 The top of the box body 1 is connected with a powder trough 4, the box body 1 is rotatably connected with a powder tank 3 in the powder trough 4, the top of the powder tank 3 is connected with a feeding port 6 with a plugging cover, the box body 1 is located in the bottom of the powder trough 4 and is connected with a driving motor 5 that matches the powder tank 3, the bottom of the powder tank 3 is connected with a discharge port 7, the box body 1 is located at the bottom of the powder trough 4 and is downwardly connected with a feeding pipe 8 that matches the discharge port 7, the box body 1 is located outside the powder tank 3 and is connected with a plurality of reagent chambers 2, the top of the reagent chamber 2 is connected with a feeding port 6 with a plugging cover, the box body 1 is located below the powder tank 3 and is connected with a reaction box 11 that is connected to the bottom of the feeding pipe 8, the reagent A reagent conduit 9 is commonly connected between the chamber 2 and the reaction box 11, and a solenoid valve 10 is connected to the reagent conduit 9 for allowing the reagent in the reagent chamber 2 to flow into the reaction box 11 below under the action of gravity. A sample box 19 is connected to the top of the box body 1 away from the detection box 25, a sliding cover 20 is connected to the top of the sample box 19, a support shaft 21 is connected to the sample box 19 for upward rotation, a driving motor 5 matching the support shaft 21 is connected to the bottom of the sample box 19, a plurality of blades 22 are connected to the support shaft 21, a sample conduit 23 is commonly connected between the sample box 19 and the reaction box 11, and a water pump 16 is connected to the sample conduit 23.

[0030] A certain amount of test materials are stored in the multiple reagent chambers 2 and powder tanks 3 arranged in the box body 1. RPA freeze-dried enzyme powder is stored in the powder tank, and RPA reaction buffer, upper primer, lower primer, probe, ultrapure sterile water and control reagent are placed in the multiple reagent chambers 2 respectively. The control reagent is divided into negative control reagent and positive control reagent. The powder tank 3 rotates in the powder trough 4. When the discharge port 7 at the bottom and the feed pipe 8 are engaged with each other, the RPA freeze-dried enzyme powder in the powder tank 3 falls into the reaction box 11 below. The amount added to the reaction box 11 can be controlled by rotation. The reagent in the reagent chamber 2 is conveniently added to the reaction box 11 through the reagent conduit 9. A small amount of California bass liver or spleen debris is added to the sample box 19, which is cut and crushed by a rotating tool and mixed with PBS buffer to make a virus suspension. The template is obtained under the heat supply of the heating columns on both sides, and the template enters the reaction box 11 through the sample conduit 23.

[0031] Embodiment 3

[0032] Combined with Figure 1-7 A collection funnel 12 is connected to the top of the reaction box 11, and a mixing chamber 13 connected to the bottom end of the collection funnel 12 is connected to the bottom of the reaction box 11. A plurality of electric heating columns 17 are connected to the reaction box 11 located outside the mixing chamber 13 and on both sides of the sample box 19. A connecting pipe 18 connected to the mixing chamber 13 is commonly connected between the reaction box 11 and the detection box 25, and a water pump 16 is connected to the connecting pipe 18. A liquid storage tank 26 is connected to the side of the box body 1 close to the detection box 25, and a feeding port 6 with a plugging cover is connected to the top of the liquid storage tank 26. A liquid infusion pipe 27 is commonly connected between the liquid storage tank 26 and the detection box 25, and a liquid infusion pipe 27 is connected to the liquid infusion pipe 27. A water pump 16 is connected, the reaction box 11 is located in the mixing chamber 13 and in the bottom of the detection box 25, and is rotatably connected to a transmission shaft 14. The outsides of the reaction box 11 and the detection box 25 are connected to a driving motor 5 that matches the transmission shaft 14, and a plurality of stirring blades 15 are connected to the transmission shaft 14. The box body 1 is located below the sample box 19 and is connected to a wastewater tank 28. A waste pipe 24 is commonly connected between the sample box 19, the reaction box 11, the detection box 25 and the wastewater tank 28, and the waste pipe 24 is connected to the water pump 16. A sewage outlet 29 that matches the wastewater tank 28 is connected to one side of the bottom of the box body 1, and the sewage outlet 29 is connected to the water pump 16.

[0033] The mixed reaction liquid is obtained through the mixing chamber 13 with stirring and heating functions in the reaction box 11. The detection box 25 can obtain the reaction liquid through the connecting pipe 18 and mix it with the liquid in the liquid storage tank 26. The liquid storage tank 26 is filled with LFD buffer. The internal stirring mechanism can make the detection liquid suitable for detection operation. Through the waste pipe 24 connected to the operation box body, the excess waste liquid of the detection can be discharged centrally. The waste water tank 28 can be used to receive and store the waste liquid for a short time.

[0034] During the specific implementation of the utility model, when the detection kit device is used to detect cell swelling iris virus in California sea bass, a sufficient amount of RPA reaction buffer, upper primer, lower primer, probe and ultrapure sterile water are respectively placed in each reagent chamber 2, and a sufficient amount of RPA freeze-dried enzyme powder is placed in the powder tank, and a small amount of California sea bass liver or spleen debris is placed in the sample box 19 on one side. The support shaft 21 at the bottom drives the cutter to cut and crush, and then 1 mL of PBS buffer is added to the sample box 19 to mix with the crushed sample tissue and cells to form a virus suspension, and the heating function of the chamber is started, and the virus suspension is boiled at 100 degrees for 10 minutes to obtain a sample template, and 2 microliters of the template is input into the reaction chamber through the sample conduit 23. Prior to this, 2.5 microliters and 29.4 microliters of RPA freeze-dried enzyme powder and RPA reaction buffer have been added to the mixing chamber 13 of the reaction box 11, and the chamber is equipped with Under the action of the stirring mechanism, the mixture is mixed evenly. While adding the template to the reaction box 11, 2 microliters of the upper primer, 2 microliters of the lower primer, 0.6 microliters of the probe and 11.5 microliters of ultrapure sterile water are added from the corresponding reagent chamber 2 to the reaction box 11. The above stock solutions are collected from the collection funnel 12 and fall into the mixing chamber 13 together. After the addition is completed, the mixture is mixed evenly and heated to 37 degrees. It is left to stand for 10 minutes, and then the mixed reaction mixture is connected to the connecting tube 18. 10 microliters of the test liquid is transported to the detection box 25. At the same time, 190 microliters of LFD buffer solution taken from the liquid storage tank 26 is also in the detection box 25. The mixture is evenly mixed under the action of the stirring mechanism in the box. Then, the colloidal gold test strip 31 is taken out and inserted into the slot 30. The bottom of the colloidal gold test strip 31 contacts the evenly mixed test liquid in the detection box 25. The quality control line and the test line are observed within 5 minutes to determine the result. When preparing the reaction solution, the amount of each reagent added is as follows:

[0035]

[0036] After the sample is used for testing, it is necessary to set up a yin-yang control group, add a negative control reagent or a positive control reagent into the corresponding reagent chamber 2, and use the control reagent as a template in the absence of a sample to perform the above-mentioned detection reaction. Each time the reaction is completed, excess waste liquid in each chamber of the box body 1 can be discharged through the waste pipe 24 at the bottom of the corresponding chamber and temporarily stored in the waste water tank 28. When unified cleaning is required, it can be discharged as a whole through the sewage outlet 29 with a water pump 16.

[0037] The above description of the utility model and its implementation methods is not restrictive. The drawings show only one implementation method of the utility model, and the actual structure is not limited thereto. In short, if ordinary technicians in this field are inspired by it and design structural methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention of the utility model, they should all fall within the protection scope of the utility model.

Claims

1. A rapid detection kit for California sea bass cell swelling iris virus, comprising a box body (1), wherein a detection box (25) is connected to the box body (1), a plurality of slots (30) are connected to one side of the box body (1), and a colloidal gold test strip (31) matched with the detection box (25) is connected to the box body (1) in the slots (30), characterized in that: The top of the box body (1) is connected to a powder trough (4), the box body (1) is rotatably connected to a powder tank (3) in the powder trough (4), the bottom of the powder tank (3) is connected to a discharge port (7), the box body (1) is located at the bottom of the powder trough (4) and is downwardly connected to a feed pipe (8) that matches the discharge port (7), the box body (1) is located outside the powder tank (3) and is connected to a plurality of reagent chambers (2), the box body (1) is located below the powder tank (3) and is connected to a feed pipe (8) that matches the discharge port (7) The bottom end of the tube (8) is connected to a reaction box (11), a reagent conduit (9) is commonly connected between the reagent chamber (2) and the reaction box (11), a sample box (19) is commonly connected to the top of the box body (1) away from the detection box (25), a support shaft (21) is rotatably connected to the sample box (19), a plurality of blades (22) are commonly connected to the support shaft (21), and a sample conduit (23) is commonly connected between the sample box (19) and the reaction box (11); The top of the reaction box (11) is connected to a material collecting funnel (12), the bottom of the reaction box (11) is connected to a mixing chamber (13) connected to the bottom end of the material collecting funnel (12), the reaction box (11) is located outside the mixing chamber (13) and is connected to a plurality of electric heating columns (17) on both sides of the sample box (19), the reaction box (11) and the detection box (25) are connected to a connecting pipe (18) connected to the mixing chamber (13), the box body (11) is connected to a liquid storage tank (26) on the side close to the detection box (25), and the liquid storage tank (26) is connected to the detection box (25). ) are commonly connected with an infusion tube (27), the reaction box (11) is located in the mixing chamber (13) and is rotatably connected with a transmission shaft (14) at the bottom of the detection box (25), and a plurality of stirring blades (15) are connected to the transmission shaft (14), the box body (1) is located below the sample box (19) and is connected with a waste water tank (28), the sample box (19), the reaction box (11) and the detection box (25) are commonly connected with the waste water tank (28) with a waste discharge pipe (24), and one side of the bottom of the box body (1) is connected with a sewage outlet (29) that matches the waste water tank (28).

2. A rapid detection kit for California sea bass cell swelling iris virus according to claim 1, characterized in that: The reagent conduit (9) is connected to a solenoid valve (10).

3. A rapid detection kit for California sea bass cell swelling iris virus according to claim 1, characterized in that: The sample conduit (23), the connecting tube (18), the infusion tube (27), the waste discharge tube (24) and the sewage discharge port (29) are all connected to a water pump (16).

4. A rapid detection kit for California sea bass cell swelling iris virus according to claim 1, characterized in that: The powder tank (3), the transmission shaft (14) and one end of the support shaft (21) are all connected with a driving motor (5).

5. A rapid detection kit for California sea bass cell swelling iris virus according to claim 1, characterized in that: The reagent chamber (2), the powder tank (3) and the liquid storage box (26) are all connected to the top with a feeding port (6), and the sample box (19) is connected to the top with a sliding cover (20).

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