Device and method for detecting antibiotic resistance genes in soil

Abstract

The present invention relates to a device and method for detecting antibiotic resistance genes in soil, and relates to the field of environmental testing technology. The device comprises a housing and a lid located on top of the housing and snapped into place. Adjustable, retractable support rods are provided at the four corners of the bottom lid to support the lid's elevation. The lid is integrated with an air curtain unit for forming a closed testing space above the housing. The air curtain unit includes at least one filter element located within a cavity within the lid, and an air pump located at the bottom of the lid and connected to the cavity containing the filter element. The top of the lid is provided with at least one air inlet for delivering air to the filter element. The detection device is easy to carry and can be quickly assembled and used on-site, facilitating on-site preparation of PCR reaction systems. During testing, an air barrier is formed around the lid, creating a closed testing space, enabling testing in a dust-free environment and significantly improving the accuracy of experimental results. The device is suitable for outdoor use.

Description

Technical Field

[0001] The present invention belongs to the technical field of environmental detection, and in particular relates to a device and method for detecting antibiotic resistance genes in soil. Background Art

[0002] Detection of antibiotic resistance genes is an important means of assessing the level of antibiotic resistance in soil environments. With the widespread use and even abuse of antibiotics in fields such as human medicine and animal husbandry, the emergence and spread of antibiotic resistance genes in bacteria is becoming increasingly serious. These resistance genes can spread not only between bacteria of the same species, but also between different species through horizontal gene transfer, posing a threat to public health and the ecological environment. Detection of antibiotic resistance genes is crucial for assessing the level of antibiotic resistance in the environment and developing effective prevention and control strategies. Regular monitoring of antibiotic resistance genes in the environment can help promptly detect and control the spread of resistance, safeguarding public health and ecological security.

[0003] Devices for detecting antibiotic resistance genes in soil are widely used in environmental monitoring, agricultural ecology, public health and other fields. By detecting resistance genes in the soil, the degree of antibiotic resistance contamination can be assessed, providing a scientific basis for formulating prevention and control strategies. However, existing detection devices generally test soil samples in the laboratory, which is not convenient to carry outdoors. When used outdoors, particulates in the air can easily contaminate the PCR reaction system, causing certain errors in the accuracy of the experimental results. Existing detection devices are not convenient for outdoor preparation, and manual pipetting of reagents is prone to splashing or cross-contamination, and the control of the pipette is not convenient.

[0004] To this end, we provide a device and method for detecting antibiotic resistance genes in soil to solve the above problems. Summary of the Invention

[0005] The purpose of the present invention is to provide a device and method for detecting antibiotic resistance genes in soil in response to the problems of the background technology.

[0006] The present invention achieves the above-mentioned purpose through the following technical solutions:

[0007] A device for detecting antibiotic resistance genes in soil comprises a housing and a lid positioned on top of the housing and snapped into place therewith. Adjustable, retractable support rods for supporting the lid at its four corners are provided. An air curtain unit is integrated into the lid to form a closed detection space above the housing. The air curtain unit comprises at least one filter element positioned within a cavity within the lid and an air pump positioned at the bottom of the lid and communicating with the cavity within which the filter element is positioned. At least one air inlet is provided at the top of the lid for conveying air into the filter element, and a plurality of evenly distributed air outlets are provided at the bottom edge of the lid. A PCR instrument, a batching tube for preparing a PCR reaction system, and reagent tubes for storing various reagents are provided in the interior cavity of the housing.

[0008] As a further optimization solution of the present invention, elastic clamping columns for locking the box cover are provided on both sides of the box body; and a handle is provided on the top of the box cover.

[0009] As a further optimization solution of the present invention, the air curtain unit also includes side panels symmetrically distributed on both sides of the box cover, one end of the side panel is hinged to the box cover and locked by fasteners; the side panel is also provided with a plurality of evenly distributed air outlet holes.

[0010] As a further optimization solution of the present invention, the box cover is further provided with a solar panel located on the top of the box cover and a rechargeable battery pack located at the bottom of the box cover for power supply.

[0011] As a further optimization scheme of the present invention, a first placement slot for placing a PCR instrument is provided at the top of the box near its back, and a configuration slot for placing a batching tube is provided at the top of the box near its front. Two rows of equally spaced second placement slots for placing reagent tubes are provided on both sides of the configuration slot. The multiple reagent tubes respectively store template DNA reagents, primers, dNTPs, heat-resistant DNA polymerase and PCR buffer; an auxiliary pipetting component is provided above the reagent tubes, which can assist in the rapid preparation of the PCR reaction system and compress the reagent tubes when the detection device is transported.

[0012] As a further optimization solution of the present invention, four evenly distributed telescopic suspension rods are provided between the PCR instrument and the box cover; and a rubber pressing block for pressing the PCR instrument is also provided at the bottom of the box cover.

[0013] As a further optimization scheme of the present invention, an oscillation component for mixing the material in the dispensing tube is provided in the configuration groove; the oscillation component includes a vibration motor and a cylinder fixed on the top of the vibration motor, and a plurality of springs evenly distributed along the circumference are provided between the cylinder and the configuration groove.

[0014] As a further optimization solution of the present invention, a tube plug is provided at the top end of the reagent tube, and a pipette is fixedly provided inside the tube plug.

[0015] As a further optimization scheme of the present invention, the auxiliary pipetting assembly includes a frame body and four rod bodies fixed at the four corners of the bottom of the frame body; the rod bodies are movably inserted into the box body, and the outer surface of the box body is provided with a top screw knob for locking the rod body, and a slot hole is provided in the frame body, and a telescopic rod is passed through the slot hole, and the top end of the telescopic rod is provided with a rubber anti-slip sleeve, and the bottom end of the telescopic rod is fixedly connected to the tube plug; the bottom of the frame body is provided with a rubber compression strip for compressing the reagent tube.

[0016] A method for detecting antibiotic resistance genes in soil comprises the following steps:

[0017] S1. Bring the detection device to the outdoor location where the antibiotic resistance gene in the soil needs to be detected. After placing it, pull the lid upward to unfold the detection device. As the lid is lifted, the PCR instrument is also lifted.

[0018] S2. Start the air curtain unit. External air enters the filter element through the air inlet. After being filtered by the filter element, the air enters the air pump and is finally ejected from multiple air outlets, forming an air barrier around the box cover.

[0019] S3. The tester's arm passes through the air barrier and adds each substance in the reagent tube into the ingredient tube one by one, mixes it into a PCR reaction system, and finally tests it through a PCR instrument to achieve on-site rapid detection of antibiotic resistance genes in the soil.

[0020] The beneficial effects of the present invention are:

[0021] 1. The present invention designs the detection device into a compact suitcase form by arranging a box body, a box cover and a PCR instrument, which is convenient for carrying and quick on-site assembly and use, and facilitates on-site preparation of the PCR reaction system.

[0022] 2. The present invention forms an air barrier around the box cover by arranging a box cover and an air curtain unit, forming a closed detection space, realizing detection in a dust-free environment, effectively preventing particulate matter in the air from contaminating the PCR reaction system, helping to maintain the cleanliness of the experimental environment, and greatly improving the accuracy of the experimental results.

[0023] 3. By providing an auxiliary pipetting component, the present invention can more accurately locate and move the reagents in the reagent tube, reduce the risk of reagent splashing or cross contamination, and can achieve rapid pipetting, thereby improving experimental efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 A three-dimensional schematic diagram of the overall structure of the present invention Figure 1 ;

[0025] Figure 2 A three-dimensional schematic diagram of the overall structure of the present invention Figure 2 ;

[0026] Figure 3 Schematic diagram of the internal structure of the box of the present invention;

[0027] Figure 4 This is a schematic diagram of the top surface structure of the box cover of the present invention;

[0028] Figure 5 This is a schematic diagram of the bottom structure of the box cover of the present invention;

[0029] Figure 6 It is a schematic structural diagram of the oscillation component of the present invention;

[0030] Figure 7 Schematic diagram of the structure of the auxiliary pipetting component of the present invention.

[0031] In the picture:

[0032] 1. Box body; 101. Elastic clamping column; 102. First placement slot; 103. Configuration slot; 104. Oscillation assembly; 104a. Vibration motor; 104b. Cylinder; 104c. Spring; 105. Second placement slot; 106. Reagent tube; 106a. Tube plug; 106b. Pipette; 2. Box cover; 201. Adjustable telescopic support rod; 202. Handle; 203. Filter element; 204. Air pump; 205. Air inlet; 206. Air outlet; 207. Side panel; 208. Solar panel; 209. Rechargeable battery pack; 210. Rubber pressure block; 3. PCR instrument; 301. Telescopic boom; 4. Auxiliary pipetting assembly; 401. Frame; 402. Slot; 403. Telescopic rod; 404. Rubber anti-slip cover; 405. Rod body; 406. Rubber pressure strip. DETAILED DESCRIPTION

[0033] The present application is described in further detail below in conjunction with the accompanying drawings. It is necessary to point out that the following specific implementation methods are only used to further illustrate the present application and cannot be understood as limiting the scope of protection of the present application. Technicians in this field can make some non-essential improvements and adjustments to the present application based on the above application content.

[0034] Example 1

[0035] In order to solve the problem that when existing detection devices are used outdoors, particles in the air can easily contaminate the PCR reaction system, causing certain errors in the accuracy of the experimental results, please refer to Figure 1-Figure 2 、 Figure 4The present invention provides a device for detecting antibiotic resistance genes in soil, comprising a housing 1 and a lid 2 located on top of the housing 1 and snapped into place therein. Elastic latches 101 are provided on both sides of the housing 1 for locking the lid 2. Adjustable telescopic support rods 201 are provided at the four corners of the bottom of the lid 2 for supporting the lid 2 in its elevation. A handle 202 is provided on the top of the lid 2. An air curtain unit is integrated into the lid 2 to form a closed detection space above the housing 1. The detection device is designed in the form of a compact suitcase, making it easy to carry and quickly assemble on site. The air curtain unit can form a closed detection space, enabling detection in a dust-free environment and effectively preventing airborne particulates from contaminating the PCR reaction system.

[0036] like Figure 4-Figure 5 As shown, the air curtain unit includes at least one filter element 203 located in the internal cavity of the box cover 2 and an air pump 204 located at the bottom of the box cover 2 and connected to the cavity where the filter element 203 is located. For example, four filter elements 203 may be provided, two for normal use and two for standby. The filter element 203 may be configured as an activated carbon filter or a HEPA filter, which can effectively remove dust, bacteria, and other pollutants from the air. It works in conjunction with other components of the air curtain unit to provide a purer testing environment. The filter element 203 is replaced regularly to ensure its filtering effect. The top of the box cover 2 is provided with at least one air inlet 205 for conveying air into the filter element 203, and the bottom edge of the box cover 2 is provided with multiple evenly distributed air outlets 206. An air barrier is formed around the box cover 2. When configuring the PCR reaction system, the risk of external pollutants entering the experimental area is reduced, which helps to maintain the cleanliness of the experimental environment and greatly improves the accuracy of the experimental results.

[0037] In order to form a more comprehensive barrier, avoid the gap under the arm when the inspector's arm passes through, such as Figure 4-Figure 5 As shown, the air curtain unit also includes side panels 207 symmetrically distributed on either side of the lid 2. One end of the side panels 207 is hinged to the lid 2 and secured with fasteners. The side panels 207 are also provided with multiple evenly distributed air outlets 206. Air curtains are ejected through the side panels 207, creating a double air barrier on both sides of the lid 2. When an inspector's arm passes through, the air ejected from the upper air outlets 206 is blocked, leaving an air gap below the arm. The air curtains ejected from the side panels 207 can then supplement the main air curtain, forming a more comprehensive barrier.

[0038] In order to meet the needs of testing devices to be tested outdoors without conventional power supply, such as Figure 4-Figure 5 As shown, the box cover 2 is also provided with a solar panel 208 located at the top of the box cover 2 for power supply and a rechargeable battery pack 209 located at the bottom of the box cover 2. Equipped with a portable power supply, it is ensured that it can work for a long time in an environment without power supply.

[0039] Example 2

[0040] On the basis of Example 1, in order to facilitate the on-site deployment of the PCR reaction system, as Figure 1-Figure 3 As shown, the internal cavity of the box 1 is equipped with a PCR instrument 3, a batching tube for preparing the PCR reaction system, and reagent tubes 106 for storing various reagents. A first placement slot 102 for placing the PCR instrument 3 is located near the back of the top of the box 1, and a configuration slot 103 for placing the batching tubes is located near the front of the top of the box 1. Two rows of equally spaced second placement slots 105 for placing reagent tubes 106 are located on both sides of the configuration slot 103. The multiple reagent tubes 106 store template DNA reagents, primers, dNTPs, heat-resistant DNA polymerase, PCR buffer, or other optional reagents such as enhancers. The various substances in the reagent tubes 106, such as primers, template DNA, and other reagents, are added to the batching tubes one by one and mixed thoroughly. The mixture is then tested using the PCR instrument 3, enabling on-site rapid detection of antibiotic resistance genes in soil.

[0041] Four evenly spaced telescopic booms 301 are positioned between the PCR instrument 3 and the lid 2. A rubber clamp 210 is located at the bottom of the lid 2 to hold the PCR instrument 3 in place. The PCR instrument 3 can be raised and lowered synchronously with the lid 2. When the lid 2 is raised, it pulls the PCR instrument 3 out of the first placement slot 102, facilitating subsequent use. When the entire testing device is not in use, the rubber clamp 210 holds the PCR instrument 3 in place, preventing damage to the PCR instrument 3 from shaking during transport.

[0042] In order to quickly achieve the mixing of various components in the batching pipe, such as Figure 6 As shown, an oscillating assembly 104 for mixing the materials in the dispensing tube is located within the dispensing tank 103. The oscillating assembly 104 comprises a vibration motor 104a and a cylinder 104b secured to the top of the vibration motor 104a. Multiple springs 104c are evenly distributed along the circumference of the cylinder 104b and the dispensing tank 103. Vibrating motor 104a drives cylinder 104b to vibrate, which in turn drives the dispensing tube within it to vibrate, rapidly mixing the various components. The presence of springs 104c further enhances the vibration effect.

[0043] like Figure 7 As shown, a pipe plug 106a is provided at the top of the reagent tube 106, and a pipette 106b is fixedly provided inside the pipe plug 106a. The reagent in the reagent tube 106 is moved into the dispensing tube by the pipette 106b.

[0044] Example 3

[0045] On the basis of the first and second embodiments, in order to more accurately locate and move the reagents in the reagent tube 106, reduce the risk of reagent splashing or cross contamination, and to quickly pipette and improve the experimental efficiency, as shown in FIG. Figure 1 、 Figure 7 As shown, an auxiliary pipetting assembly 4 is provided above the reagent tube 106 to assist in the rapid preparation of the PCR reaction system and to hold the reagent tube 106 in place during transport of the detection device. The auxiliary pipetting assembly 4 not only accurately positions and moves the reagents within the reagent tube 106, but also acts as a pressure plate. When the entire detection device is not in use, the auxiliary pipetting assembly 4 can hold the reagent tube 106 in place to prevent it from shaking and potentially being damaged during transport.

[0046] The auxiliary pipetting assembly 4 includes a frame 401 and four rods 405 fixed at the four corners of the bottom of the frame 401; the rods 405 are movably inserted into the box body 1, and a top screw knob for locking the rods 405 is provided on the outer surface of the box body 1. A slot 402 is provided in the frame 401, and a telescopic rod 403 is passed through the slot 402. The top end of the telescopic rod 403 is provided with a rubber anti-slip sleeve 404, and the bottom end of the telescopic rod 403 is fixedly connected to the tube plug 106a; a rubber compression strip 406 for compressing the reagent tube 106 is provided at the bottom of the frame 401. When in use, loosen the top screw knob on the outer side of the box body 1, lift the frame 401 upward to the required height, and then lock the top screw knob to unfold the auxiliary pipetting assembly 4. The frame 401 drives the telescopic rod 403 to extend during the rising process. The telescopic rod 403 is configured to be magnetic, that is, the rods are attracted by magnets. When the rods are close to each other, the magnetic fields generated by the magnets will attract each other, thereby achieving a stable connection. No additional fixing device or locking mechanism is required, and the operation is simple and quick. When pipetting, the reagent in the reagent tube 106 is absorbed through the pipette 106b, and then the tube plug 106a is loosened and the tube plug 106a is lifted upward until the telescopic rods 403 are attracted to each other under the magnetic force of the magnet inside them. Then, manually move the top end of the telescopic rod 403 to make it move along the slot 402 to the top of the dispensing tube, and then pull the telescopic rod 403 downward to drip the reagent in the pipette 106b into the dispensing tube.

[0047] The present invention also provides a method for detecting antibiotic resistance genes in soil, comprising the following steps:

[0048] S1. Bring the detection device to the outdoor location where the antibiotic resistance gene in the soil needs to be detected. After placing it, pull the box cover 2 upward to unfold the detection device. As the box cover 2 is lifted, the PCR instrument 3 is also lifted.

[0049] S2. Start the air curtain unit. External air enters the filter element 203 through the air inlet 205. After being filtered by the filter element 203, the air enters the air pump 204 and is finally ejected from the multiple air outlets 206, forming an air barrier around the box cover 2.

[0050] S3. The inspector's arm passes through the air barrier and adds the substances in the reagent tube 106 to the ingredient tube one by one to prepare a PCR reaction system. Finally, the system is tested by the PCR instrument 3 to achieve on-site rapid detection of antibiotic resistance genes in the soil.

[0051] The above embodiment merely represents one embodiment of the present invention. While the description is relatively specific and detailed, it should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art may make various modifications and improvements without departing from the scope of the present invention, and such modifications and improvements are all within the scope of protection of the present invention.

Claims

1. A device for detecting antibiotic resistance genes in soil, comprising a housing (1) and a housing cover (2) located on top of the housing (1) and inserted into the housing, characterized in that: Adjustable telescopic support rods (201) for supporting the box cover (2) to be raised are provided at the four corners of the bottom of the box cover (2); an air curtain unit for forming a closed detection space above the box body (1) is integrated on the box cover (2); the air curtain unit comprises at least one filter element (203) located in the internal cavity of the box cover (2) and an air pump (204) located at the bottom of the box cover (2) and connected to the cavity where the filter element (203) is located; at least one air inlet hole (205) for conveying air to the filter element (203) is provided at the top of the box cover (2); and a plurality of evenly distributed air outlet holes (206) are provided at the bottom edge of the box cover (2); The air curtain unit further comprises side panels (207) symmetrically distributed on both sides of the box cover (2), one end of the side panels (207) being hinged to the box cover (2) and locked by fasteners, and the side panels (207) are also provided with a plurality of evenly distributed air outlet holes (206); the internal cavity of the box body (1) is respectively provided with a PCR instrument (3), a material pipe for preparing a PCR reaction system, and a reagent tube (106) for storing various reagents; four evenly distributed telescopic suspension rods (301) are provided between the PCR instrument (3) and the box cover (2), and a rubber pressing block (210) for pressing the PCR instrument (3) is also provided at the bottom of the box cover (2); An auxiliary pipetting assembly (4) is provided above the reagent tube (106) and is capable of assisting in the rapid preparation of the PCR reaction system and pressing the reagent tube (106) when the detection device is transported. The auxiliary pipetting assembly (4) includes a frame (401) and four rods (405) fixed at the four corners of the bottom of the frame (401). A rubber pressing strip (406) for pressing the reagent tube (106) is provided at the bottom of the frame (401); The rod body (405) is movably inserted into the box body (1), and a top screw knob for locking the rod body (405) is provided on the outer surface of the box body (1). A slot hole (402) is provided in the frame body (401), and a telescopic rod (403) is provided through the slot hole (402).

2. The device for detecting antibiotic resistance genes in soil according to claim 1, characterized in that: Both sides of the box body (1) are provided with elastic clamping columns (101) for locking the box cover (2); and the top of the box cover (2) is provided with a handle (202).

3. The device for detecting antibiotic resistance genes in soil according to claim 1, characterized in that: The box cover (2) is also provided with a solar panel (208) located at the top of the box cover (2) for power supply and a rechargeable battery pack (209) located at the bottom of the box cover (2).

4. The device for detecting antibiotic resistance genes in soil according to claim 1, characterized in that: The top of the box (1) is provided with a first placement slot (102) for placing a PCR instrument (3) near its back, and the top of the box (1) is provided with a configuration slot (103) for placing a material tube near its front. Both sides of the configuration slot (103) are provided with two rows of second placement slots (105) for placing reagent tubes (106) at equal intervals. The multiple reagent tubes (106) respectively store template DNA reagents, primers, dNTPs, heat-resistant DNA polymerase and PCR buffer.

5. The device for detecting antibiotic resistance genes in soil according to claim 4, characterized in that: An oscillating assembly (104) for mixing materials in a dispensing pipe is provided in the dispensing tank (103); the oscillating assembly (104) comprises a vibration motor (104a) and a cylinder (104b) fixed on the top of the vibration motor (104a); a plurality of springs (104c) evenly distributed along the circumference are provided between the cylinder (104b) and the dispensing tank (103).

6. The device for detecting antibiotic resistance genes in soil according to claim 4, characterized in that: A tube plug (106a) is provided at the top end of the reagent tube (106), and a pipette (106b) is fixedly provided inside the tube plug (106a).

7. The device for detecting antibiotic resistance genes in soil according to claim 6, characterized in that: The top end of the telescopic rod (403) is covered with a rubber anti-slip sleeve (404), and the bottom end of the telescopic rod (403) is fixedly connected to the pipe plug (106a).

8. A method for detecting antibiotic resistance genes in soil, using a device for detecting antibiotic resistance genes in soil according to any one of claims 1 to 7, characterized in that: The following steps are involved: S1. The detection device is carried to the outdoor location where the antibiotic resistance gene in the soil is to be detected. After placement, the cover (2) is pulled upward to unfold the detection device. During the lifting process of the cover (2), the PCR instrument (3) is driven to rise. S2, start the air curtain unit, external air enters the filter element (203) through the air inlet (205), the air is filtered by the filter element (203) and enters the air pump (204), and finally is ejected from the multiple air outlets (206), forming an air barrier around the box cover (2); S3. The inspector's arm passes through the air barrier and adds the substances in the reagent tube (106) to the ingredient tube one by one to prepare a PCR reaction system. Finally, the system is tested using a PCR instrument (3) to achieve on-site rapid detection of antibiotic resistance genes in the soil.

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