Device and method for detecting antibiotic resistance genes in soil

Abstract

The invention relates to a device and method for detecting antibiotic resistance genes in soil, and relates to the technical field of environmental detection.The device comprises a box body and a box cover located at the top of the box body and clamped into the box body, and adjustable telescopic supporting rods used for supporting the box cover to rise are arranged at the four corners of the bottom of the box cover; an air curtain unit used for forming a closed detection space above the box body is integrated on the box cover, the air curtain unit comprises at least one filter element located in an inner cavity of the box cover and an air pump located at the bottom of the box cover and communicated with the cavity where the filter element is located, and at least one air inlet hole used for conveying air into the filter element is formed in the top of the box cover. The detection device is convenient to carry and rapidly assemble and use on site, a PCR reaction system is convenient to deploy on site, during detection, an air barrier can be formed around the box cover, a closed detection space is formed, detection in a dust-free environment is achieved, the accuracy of an experimental result is greatly improved, and the detection 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] The detection of antibiotic resistance genes is an important means to assess the level of antibiotic resistance in the soil environment. With the widespread use and even abuse of antibiotics in the fields of human medicine, animal husbandry, etc., the emergence and spread of antibiotic resistance genes in bacteria are becoming increasingly serious. These resistance genes can not only spread between bacteria of the same species, but also between different species of bacteria through horizontal gene transfer, thus posing a threat to public health and the ecological environment. The detection of antibiotic resistance genes is of great significance for assessing the level of antibiotic resistance in the environment and formulating effective prevention and control strategies. By regularly monitoring antibiotic resistance genes in the environment, the spread and spread of resistance can be discovered and controlled in a timely manner to ensure public health and ecological safety.

[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 pollution can be evaluated, 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 for use. 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 deployment, and completely manual pipetting of reagents is prone to splashing or cross-contamination, and the control of the pipette is not convenient enough.

[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 box body and a box cover located on the top of the box body and inserted into the inside thereof, wherein adjustable telescopic support rods for supporting the box cover to rise are arranged at the four corners of the bottom; an air curtain unit for forming a closed detection space above the box body is integrated on the box cover, wherein the air curtain unit comprises at least one filter element located in the inner cavity of the box cover and an air pump located at the bottom of the box cover and connected to the cavity where the filter element is located, at least one air inlet hole for conveying air to the filter element is arranged on the top of the box cover, and a plurality of evenly distributed air outlet holes are arranged at the bottom edge of the box cover; a PCR instrument, a batching pipe for preparing a PCR reaction system, and a reagent tube for storing various reagents are arranged in the inner cavity of the box body.

[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 scheme 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 outlets.

[0010] As a further optimization solution of the present invention, the box cover is also 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 body near its back, a configuration slot for placing a preparation tube is provided at the top of the box body near its front, and two rows of second placement slots for placing reagent tubes are provided at equal intervals on both sides of the configuration slot, and 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 materials in the batching 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 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 the frame body is provided with a slot hole, 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 box cover upward to unfold the detection device. During the lifting process of the box cover, the PCR instrument is driven to rise;

[0018] S2. Start the air curtain unit, and the external air enters the filter element through the air inlet. After being filtered by the filter element, the air enters the air pump and finally sprays out from multiple air outlets to form 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 providing a box body, a box cover and a PCR instrument, which is convenient for carrying and quick on-site assembly and use, and is convenient for on-site preparation of the PCR reaction system.

[0022] 2. The present invention forms an air barrier around the box cover and an air curtain unit by arranging the box cover, forming a closed detection space, realizing detection in a dust-free environment, effectively preventing particles 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 The overall structure of the present invention is shown in three dimensions. Figure 1 ;

[0025] Figure 2 The overall structure of the present invention is shown in three dimensions. Figure 2 ;

[0026] Figure 3 It is a schematic diagram of the internal structure of the box of the present invention;

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

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

[0029] Figure 6 It is a schematic diagram of the structure of the oscillating assembly of the present invention;

[0030] Figure 7 It is a schematic diagram of the structure of the auxiliary pipetting component of the present invention.

[0031] In the figure:

[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 compression block; 3. PCR instrument; 301. Telescopic suspension rod; 4. Auxiliary pipetting assembly; 401. Frame; 402. Slot; 403. Telescopic rod; 404. Rubber anti-slip sleeve; 405. Rod body; 406. Rubber compression strip. DETAILED DESCRIPTION

[0033] The present application is further described in detail below in conjunction with the accompanying drawings. It is necessary to point out here 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. Technical personnel in this field can make some non-essential improvements and adjustments to the present application based on the above application content.

[0034] Embodiment 1

[0035] In order to solve the problem that when the existing detection device is used outdoors, the particles in the air are easy to contaminate the PCR reaction system, causing a certain error in the accuracy of the experimental results, please refer to Figure 1-Figure 2 , Figure 4The present invention provides a soil antibiotic resistance gene detection device, comprising a box body 1 and a box cover 2 located on the top of the box body 1 and inserted into the inside thereof, both sides of the box body 1 are provided with elastic clamping columns 101 for locking the box cover 2, the bottom four corners of the box cover 2 are provided with adjustable telescopic support rods 201 for supporting the box cover 2 to rise, and the top of the box cover 2 is provided with a handle 202; the box cover 2 is integrated with an air curtain unit for forming a closed detection space above the box body 1. The detection device is designed in a compact suitcase form, which is convenient for carrying and quick on-site assembly and use, and can form a closed detection space through the air curtain unit, realize detection in a dust-free environment, and effectively prevent particulate matter in the air 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 inner 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 set as an activated carbon filter or a HEPA filter, which can effectively remove dust, bacteria and other pollutants in the air, and work 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 to the filter element 203, and the bottom edge of the box cover 2 is provided with a plurality of evenly distributed air outlets 206. An air barrier is formed around the box cover 2, which reduces the risk of external pollutants entering the experimental area when configuring the PCR reaction system, 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 passes through, such as Figure 4-Figure 5 As shown, the air curtain unit also includes side panels 207 symmetrically distributed on both sides of the box cover 2, one end of the side panels 207 is hinged to the box cover 2 and locked by fasteners; the side panels 207 are also provided with a plurality of evenly distributed air outlets 206. The air curtain is sprayed through the side panels 207, so that two layers of air barriers can be formed on both sides of the box cover 2. When the inspector's arm passes through, the gas sprayed from the upper air outlet 206 is blocked, and a gap is formed under the arm. At this time, the air curtain sprayed from the side panels 207 can supplement the deficiency of the main air curtain and form a more comprehensive barrier.

[0038] In order to meet the needs of testing devices in outdoor environments 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 can ensure long-term operation even in an environment without power supply.

[0039] Embodiment 2

[0040] On the basis of Example 1, in order to facilitate the on-site preparation of the PCR reaction system, Figure 1-Figure 3 As shown, the internal cavity of the box 1 is provided with a PCR instrument 3, a batching tube for preparing a PCR reaction system, and a reagent tube 106 for storing various reagents; a first placement slot 102 for placing the PCR instrument 3 is provided at the top of the box 1 near its back, and a configuration slot 103 for placing the batching tube is provided at the top of the box 1 near its front, and two rows of second placement slots 105 for placing the reagent tubes 106 are provided on both sides of the configuration slot 103 at equal intervals. Template DNA reagents, primers, dNTPs, heat-resistant DNA polymerases, and PCR buffers, or other optional reagents, such as enhancers, are stored in the multiple reagent tubes 106. 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, mixed, and then detected by the PCR instrument 3 to achieve on-site rapid detection of antibiotic resistance genes in soil.

[0041] Four evenly distributed telescopic suspension rods 301 are provided between the PCR instrument 3 and the box cover 2; a rubber pressing block 210 for pressing the PCR instrument 3 is also provided at the bottom of the box cover 2. The PCR instrument 3 can be raised and lowered synchronously with the box cover 2. When the box cover 2 is being raised, the PCR instrument 3 is driven to leave the first placement slot 102 to facilitate the subsequent use of the PCR instrument 3. When the entire detection device is not in use, the PCR instrument 3 can be pressed by the rubber pressing block 210 to prevent the PCR instrument 3 from shaking and being damaged during the transportation of the detection device.

[0042] In order to quickly achieve the mixing of the various components in the batching pipe, Figure 6 As shown, an oscillating assembly 104 for mixing the materials in the batching tube is provided in the configuration tank 103; the oscillating assembly 104 includes a vibration motor 104a and a cylinder 104b fixed on the top of the vibration motor 104a, and a plurality of springs 104c evenly distributed along the circumference are provided between the cylinder 104b and the configuration tank 103. The vibration motor 104a drives the cylinder 104b to vibrate, and the cylinder 104b drives the batching tube therein to vibrate, so that the components of the materials are quickly mixed, and the provision of the spring 104c can further enhance 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 to penetrate through the inside of the reagent tube 106. The reagent in the reagent tube 106 is moved into the batching tube by the pipette 106b.

[0044] Embodiment 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 transfer liquids and improve experimental efficiency, as shown in FIG. Figure 1 , Figure 7 As shown, an auxiliary pipetting assembly 4 is provided above the reagent tube 106, which can assist in the rapid preparation of the PCR reaction system and press the reagent tube 106 when the detection device is transported. The auxiliary pipetting assembly 4 can more accurately locate and move the reagent in the reagent tube 106 on one hand, and acts as a pressure plate on the other hand. When the entire detection device is not in use, the reagent tube 106 can be pressed by the auxiliary pipetting assembly 4 to prevent the reagent tube 106 from shaking and being damaged during the transportation of the detection device.

[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 passes through the slot 402. 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 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 component 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 adsorbed 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 by 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 adsorbed to each other under the magnetic force of the magnet inside them, and then the top end of the telescopic rod 403 is manually moved to make it move along the slot 402 to the top of the dispensing tube, and then the telescopic rod 403 is pulled 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, carry 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. During the lifting process of the box cover 2, the PCR instrument 3 is driven to rise;

[0049] S2, start the air curtain unit, the 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;

[0050] S3. The tester's arm passes through the air barrier and adds the substances in the reagent tube 106 into the mixing 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-mentioned embodiment only expresses one implementation mode of the present invention, and its description is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope 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 the top of the housing (1) and inserted into the interior thereof, 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); The box cover (2) is integrated with an air curtain unit for forming a closed detection space above the box body (1), the air curtain unit comprising at least one filter element (203) located in the inner 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, the top of the box cover (2) is provided with at least one air inlet hole (205) for conveying air to the filter element (203), and the bottom edge of the box cover (2) is provided with a plurality of evenly distributed air outlet holes (206); A PCR instrument (3), a mixing tube for mixing a PCR reaction system, and reagent tubes (106) for storing various reagents are respectively arranged in the internal cavity of the box (1).

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); A handle (202) is provided on the top of the box cover (2).

3. The device for detecting antibiotic resistance genes in soil according to claim 1, characterized in that: The air curtain unit further comprises side panels (207) symmetrically distributed on both sides of the box cover (2), one end of the side panel (207) being hinged to the box cover (2) and locked by a fastener; The side plate (207) is also provided with a plurality of evenly distributed air outlet holes (206).

4. 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) and used for power supply, and a rechargeable battery pack (209) located at the bottom of the box cover (2).

5. The device for detecting antibiotic resistance genes in soil according to claim 1, characterized in that: A first placement slot (102) for placing a PCR instrument (3) is provided at the top of the box (1) near its back, and a configuration slot (103) for placing a batching tube is provided at the top of the box (1) near its front, and two rows of second placement slots (105) for placing reagent tubes (106) are provided at equal intervals on both sides of the configuration slot (103), wherein the plurality of reagent tubes (106) respectively store template DNA reagents, primers, dNTPs, heat-resistant DNA polymerase and PCR buffer; An auxiliary liquid transfer component (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.

6. The device for detecting antibiotic resistance genes in soil according to claim 5, characterized in that: Four evenly distributed telescopic suspension rods (301) are provided between the PCR instrument (3) and the box cover (2); The bottom of the box cover (2) is also provided with a rubber pressing block (210) for pressing the PCR instrument (3).

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

8. The device for detecting antibiotic resistance genes in soil according to claim 5, 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).

9. The device for detecting antibiotic resistance genes in soil according to claim 8, characterized in that: The auxiliary pipetting assembly (4) comprises a frame (401) and four rods (405) fixed at the four corners of 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 passed through the slot hole (402). 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); The bottom of the frame (401) is provided with a rubber compression strip (406) for compressing the reagent tube (106).

10. A method for detecting antibiotic resistance genes in soil according to any one of claims 1 to 9, 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 needs to be detected. After the location is placed, the box cover (2) is pulled upward to unfold the detection device. During the lifting process of the box 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 then enters the air pump (204), and finally is ejected from the plurality of air outlets (206), forming an air barrier around the box cover (2); S3. The arm of the inspector passes through the air barrier and adds each substance in the reagent tube (106) into the ingredient tube one by one to prepare a PCR reaction system. Finally, the system is tested by a PCR instrument (3) to achieve on-site rapid detection of antibiotic resistance genes in the soil.

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