Device for extracting soil ralstonia solanacearum obligate bacteriophage lysate
By designing a device that includes centrifuge tubes and enrichment tubes, and utilizing filter membranes with different pore sizes to complete the enrichment and separation of phages in the same device, the problem that the enrichment and separation of phage lysate cannot be completed in the same device in the existing technology is solved, and a highly efficient phage separation effect is achieved.
Patent Information
- Application Number
- CN202422934824.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-28
- Publication Date
- 2025-11-07
- Estimated Expiration
- 2034-11-28
Smart Images
Figure CN223522513U_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The utility model relates to the enrichment separation technical field of bacteriophage, especially in kind be used for extracting the device of soil bacterial wilt specific bacteriophage lysate. BACKGROUND
[0002] Tomato bacterial wilt is one of the diseases that harm greatly in tomato production, caused by Ralstonia solnaceatμm with strong transmission and survival ability, and the disease is increasingly severe and continues to break out.
[0003] Since the 1990s, bacteriophage has been valued and concerned in the prevention and treatment of tomato bacterial wilt. The virulent bacteriophage can target host-specific receptors and lyse host bacteria. According to this characteristic, the virulent bacteriophage that can specifically infect Ralstonia solnaceatμm is screened as a biocontrol agent, which can greatly reduce the occurrence of crop bacterial wilt. However, the current enrichment and separation equipment of virulent bacteriophage of Ralstonia solnaceatμm has the defects of long use time and high material cost, and the enrichment and separation cannot be completed in the same equipment. UTILITY MODEL CONTENT
[0004] In view of the problem that the enrichment and separation of bacteriophage lysate cannot be completed in the same equipment in the prior art, the utility model provides a device for extracting soil bacterial wilt specific bacteriophage lysate, which can complete the enrichment and separation of bacteriophage in the same equipment.
[0005] To achieve the above purpose, the utility model provides the following technical scheme:
[0006] A device for extracting soil bacterial wilt specific bacteriophage lysate, comprising:
[0007] A centrifuge tube having a sample inlet and a sample outlet, the inside of the centrifuge tube having a containing cavity communicating with the sample inlet and the sample outlet;
[0008] An enrichment tube detachably installed in the containing cavity, the enrichment tube having a sample receiving port and a filter port, the sample receiving port communicating with the filter port, the sample receiving port being arranged towards the sample inlet, and the filter port being provided with a filter screen.
[0009] The device for extracting soil bacterial wilt specific bacteriophage lysate as described above further comprises a centrifuge tube cap, which is threadedly connected at the sample inlet.
[0010] The device for extracting soil bacterial wilt specific bacteriophage lysate as described above further comprises a first sealing cover, which is hingedly connected at the sample receiving port.
[0011] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0012] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0013] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0014] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0015] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0016] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port.
[0017] The device for extracting soil Ralstonia solanacearum specific phage lysate as described above, further comprises a second sealing cover, which is arranged at the sampling port. BRIEF DESCRIPTION OF DRAWINGS
[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings needed in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can be obtained by those skilled in the art without creative effort on the basis of these drawings.
[0019] Figure 1 The device structure schematic view in the embodiments of the present application;
[0020] Figure 2 The structure schematic view of the enrichment tube in the embodiments of the present application;
[0021] Figure 3The top view of the buckle in the embodiment of the utility model;
[0022] In the figure: 1, centrifugal tube cap; 2, internal thread; 3, first sealing cover; 4, external thread; 5, enrichment tube; 6, filter screen; 7, sampling port; 8, second sealing cover; 9, upper cavity; 10, centrifugal tube; 11, filter membrane; 12, lower cavity; 13, sample port; 14, buckle. DETAILED DESCRIPTION
[0023] The technical solutions in the embodiments of the utility model will be clearly and completely described below with reference to the drawings in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the present application, not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the scope of protection of the present application.
[0024] Embodiment:
[0025] It should be noted that the terms "first", "second" and the like in the description and claims of the utility model and the above drawings are used to distinguish similar objects, and do not have to be used to describe a particular order or sequence. It should be understood that the data used in this way can be interchanged under appropriate circumstances, so that the embodiments of the utility model described herein can be implemented in an order other than those illustrated or described herein. In addition, the terms "include" and "have" and any variations thereof in the embodiments of the utility model are intended to cover non-exclusive inclusion, for example, a process, method, system, product or device including a series of steps or units does not have to be limited to the clearly listed steps or units, but can include other steps or units not clearly listed or inherent to these processes, methods, products or devices.
[0026] It should be understood that the terms "center", "longitudinal", "transverse", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like indicate the orientation or positional relationship based on the orientation or positional relationship shown in the drawings, and are only for the convenience of describing the utility model and simplifying the description, and do not indicate or imply that the indicated device or element must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the utility model.
[0027] In the description of the utility model, the meaning of "multiple" is at least two, such as two, three, etc., unless there is a specific definition. In addition, unless there is a specific provision and limitation, the terms "installation", "connection", "connection" should be understood broadly, for example, it can be fixed connection, or detachable connection, or integrally connected;It can be mechanical connection, or electrical connection;It can be directly connected, or indirectly connected through an intermediate medium, or the communication between two elements. For ordinary skilled in the art, the specific meaning of the above terms in the utility model can be understood according to the specific circumstances.
[0028] In the utility model, unless there is a specific provision and limitation, the first feature is "on" or "under" the second feature. The first and second features can be in direct contact, or the first and second features can be indirectly contacted through an intermediate medium. Moreover, the first feature "above", "above" and "above" of the second feature can be directly above or obliquely above the first feature, or only indicate that the horizontal height of the first feature is higher than that of the second feature. The first feature "below", "below" and "below" of the second feature can be directly below or obliquely below the first feature, or only indicate that the horizontal height of the first feature is less than that of the second feature.
[0029] The utility model provides a technical scheme: a device for extracting soil pectobacterium specific bacteriophage lysis solution, it includes centrifugal tube 10 and enrichment tube 5, centrifugal tube 10 has sample inlet and sampling port 7, the inside of centrifugal tube 10 has the accommodation cavity that communicates with sample inlet and sampling port 7, enrichment tube 5 can be detachably installed in the accommodation cavity, enrichment tube 5 has sample inlet 13 and filter port, sample inlet 13 communicates with filter port, sample inlet 13 is set to sample inlet, and filter port is provided with filter screen 6.
[0030] Specifically, referring to Figure 1 , the device mainly includes centrifugal tube 10 and enrichment tube 5, the accommodation cavity of centrifugal tube 10 is divided into two parts of upper cavity and lower cavity, enrichment tube 5 is detachably installed in the upper cavity part, and the sample inlet of enrichment tube 5 is close to the sample inlet of centrifugal tube 10, in addition, in order to facilitate the sample to enter the enrichment tube 5 as much as possible, the size of the sample inlet can be designed to be slightly smaller than the size of the sample inlet, so that when the sample enters the centrifugal tube 10, it will first enter the enrichment tube 5 for enrichment, and then pass through the filter screen 6 at the filter port of the enrichment tube 5 and enter the lower cavity of the centrifugal tube 10, a sampling port 7 is arranged at the lower cavity part, and the bacteriophage lysis solution after enrichment and centrifugal separation is taken out through the sampling port 7. In addition, the detachable installation mode can include but is not limited to buckle connection, socket connection and the like. Therefore, the device can complete the enrichment and separation of bacteriophage in the same equipment.
[0031] As an optional implementation, in some embodiments, a centrifugal tube cap 1 is further included, which is screwed at the sample inlet. In order to avoid contamination of the sample, the centrifugal tube cap 1 is installed at the sample inlet, and the inner side wall of the centrifugal tube cap 1 is provided with an internal thread, and the outer side wall of the centrifugal tube 10 near the sample inlet is provided with an external thread, and the two are screwed together by the threads.
[0032] As an optional implementation, in some embodiments, a first sealing cover 3 is further included, which is hinged at the sample inlet 13. Referring to Figure 2 In this way, the sealing performance of the enrichment tube 5 can be improved.
[0033] As an optional implementation, in some embodiments, a second sealing cover 8 is further included, which is arranged at the sample outlet. In this way, the sealing performance of the centrifugal tube 10 can be improved, and the bacteriophage lysate can be prevented from leaking out of the sample outlet.
[0034] As an optional implementation, in some embodiments, a filter membrane 11 for filtering bacteria is further arranged in the accommodating cavity, the cross section of the filter membrane 11 is parallel to the cross section of the centrifugal tube 10, and the filter membrane 11 is arranged between the filter outlet and the sample outlet 7. In order to further improve the filtering effect, the filter membrane 11 is further arranged below the filter screen 6 of the enrichment tube 5 (i.e. between the upper and lower cavities of the centrifugal tube 10), so that the bacteria such as P. solanacearum is retained in the upper cavity through secondary filtration, and the bacteriophage lysate with higher purity is obtained.
[0035] In the above embodiments, further, the pore size of the filter membrane 11 is 0.20-0.25 μm. The pore size of the filter membrane 11 can be adjusted according to actual use, so as to retain the bacteria such as P. solanacearum in the upper cavity. Preferably, the pore size of the filter membrane 11 is 0.22 μm, and the filtering effect of this pore size is better.
[0036] As an optional implementation, in some embodiments, the inner side wall of the centrifugal tube 10 is provided with a buckle 14 for fixing the enrichment tube 5. Referring to Figure 3 The device uses the buckle connection mode to fix the enrichment tube 5 in the centrifugal tube 10, and this fixing mode is simple and has good stability.
[0037] As an optional implementation, in some embodiments, the sample outlet 7 is arranged on the side near the bottom of the centrifugal tube 10, and the sample outlet 7 is arranged in an upward inclined manner. In this way, when the second sealing cover at the sample outlet 7 is not well shielded and the bacteriophage lysate in the centrifugal tube 10 is too much, the bacteriophage lysate can be prevented from directly leaking out of the sample outlet 7.
[0038] As an optional implementation, in some embodiments, the pore size of the filter screen 6 is 30-50 μm. The pore size of the filter screen 6 can be adjusted according to actual use, so as to retain the sample soil in the enrichment tube 5. Preferably, the pore size of the filter screen 6 is 40 μm, which has better filtering effect.
[0039] Method for use: when used, the enrichment tube 5 is placed in the buckle 14 of the centrifugal tube 10, and the soil sample sieved by a 2 mm sieve, the bacterial liquid OD 600 = 0.5 and 3 times of the CPG liquid medium (formula: acid hydrolysis casein peptone 3 g / L, anhydrous glucose 15 g / L, bacteriological peptone 30 g / L, preparation method: take the above medium, add 1000 mL of deionized water, stir until completely dissolved, adjust the pH of the solution to 7.0 with 1 mol / L sodium hydroxide, and divide into triangular flasks, sterilize at 121℃ for 15 min, and cool) are added into the enrichment tube 5 in a ratio of 1:1:1, and the first sealing cover 3 and the centrifugal tube cap 1 are covered, and the device is placed in a shaking bed at room temperature for 15 min. Then the whole centrifugal tube 10 is placed in a shaking bed at 30℃, 180 r / min for 12 h, and after being taken out, it is static for 2 hours, and then the device is placed in a centrifuge for centrifugation at 10000 r / min for 10 min. The filter screen 6 of the enrichment tube 5 retains the soil in the enrichment tube 5, and the filter membrane 11 between the upper cavity and the lower cavity retains the bacteria such as Ralstonia solanacearum in the upper cavity, and the bacteriophage lysis liquid is filtered into the lower cavity, and finally the bacteriophage lysis liquid is taken out from the sampling port 7 using a pipette, so as to complete the separation of the bacteriophage.
[0040] Therefore, the device can complete the enrichment and separation of the bacteriophage in the same device, and the separation efficiency of the bacteriophage is improved, and the separation effect is good.
[0041] In the description of the present specification, the description of the terms "one embodiment", "some embodiments", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In the present specification, the illustrative description of the above terms is not necessarily for the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, the skilled in the art can combine and combine the different embodiments or examples described in the present specification and the features of the different embodiments or examples without contradiction.
[0042] The above examples are only for illustrating the technical concept and characteristics of the present application, and the purpose is to enable those skilled in the art to understand the content of the present application and to implement it, and cannot limit the protection scope of the present application. Any equivalent changes or modifications made according to the essence of the present application shall be covered within the protection scope of the present application.
Claims
1. A device for extracting a Pseudomonas soli specific bacteriophage lysate, characterized by, The application relates to a centrifugal tube for enriching bacteria, comprising: a centrifugal tube with a sample inlet and a sample outlet, the inside of the centrifugal tube having a containing cavity in communication with the sample inlet and the sample outlet; an enrichment tube detachably installed in the containing cavity, the enrichment tube having a sample receiving port and a filtering port, the sample receiving port being in communication with the filtering port, the sample receiving port being arranged towards the sample inlet, and the filtering port being provided with a filter screen.
2. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, A centrifugal tube cap is further provided and is screwed at the sample inlet.
3. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, A first sealing cover is further provided and is hinged at the sample receiving port.
4. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, A second sealing cover is further provided and covers the sample outlet.
5. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, A filter membrane for filtering bacteria is further provided in the containing cavity, the cross section of the filter membrane being parallel to the cross section of the centrifugal tube, and the filter membrane being arranged between the filtering port and the sample outlet.
6. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, The inner side wall of the centrifugal tube is provided with a buckle for fixing the enrichment tube.
7. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, The sample outlet is arranged on the side close to the bottom of the centrifugal tube and is arranged in an upwardly inclined manner.
8. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 1, characterized in that, The pore size of the filter screen is 30-50 mu m.
9. The device for extracting soil Pseudomonas soli specific bacteriophage lysate according to claim 5, characterized in that, The pore size of the filter membrane is 0.20-0.25 mu m.