Intelligent extraction system for microbial nucleic acid in fecal and soil samples

Abstract

This invention provides an intelligent nucleic acid extraction system for microorganisms from fecal and soil samples. The system includes a nucleic acid extractor body, and further includes an injection device, a pipetting device, a positive pressure device, a negative pressure device, a sample tube rack, a processing tube, and a collection device disposed within the extractor body. In use, simply place the fecal or soil sample into the processing tube, tighten the cap, and place the processing tube into the sample tube rack. By sequentially activating the injection device, pipetting device, negative pressure device, and positive pressure device, the solution in the processing tube passes through a filter screen, removing impurities. The processed solution and microorganisms pass through the filter screen and enter through the water inlet, exiting from the bottom of the sealed tube to the collection device for subsequent nucleic acid extraction steps. No human intervention is required, avoiding errors caused by human operation.

Description

Technical Field

[0001] This invention relates to the field of nucleic acid extraction technology, and in particular to an intelligent system for extracting microbial nucleic acids from fecal and soil samples. Background Technology

[0002] Nucleic acid extractors are instruments that use matching nucleic acid extraction reagents to automatically complete the extraction of nucleic acids from samples. They are widely used in various fields such as disease control centers, clinical disease diagnosis, blood transfusion safety, forensic identification, environmental microbiology testing, food safety testing, animal husbandry, and molecular biology research. Nucleic acid extraction includes the following basic steps: 1. Lysis: Lysis buffer is added to the sample. Through mechanical movement and heating, the reaction solution is mixed and fully reacted, causing cell lysis and releasing nucleic acid; 2. Adsorption: Magnetic beads are added to the sample lysis buffer and mixed thoroughly. Utilizing the strong affinity of magnetic beads for nucleic acids at high salt and low pH, nucleic acids are adsorbed, and the magnetic beads are separated from the lysis buffer under the action of an external magnetic field; 3. Washing: The magnetic beads with adsorbed nucleic acid are transferred to a new washing buffer and mixed thoroughly to wash away impurities. Then, the magnetic beads are separated from the washing buffer under the action of an external magnetic field; 4. Elution: The magnetic beads are transferred to the elution buffer, the external magnetic field is removed, and the magnetic beads are mixed thoroughly with the elution buffer. The bound nucleic acid is then separated from the magnetic beads and mixed into the elution buffer, thus obtaining purified nucleic acid.

[0003] Currently, nucleic acid extraction from microorganisms in feces and soil generally requires manual operation by laboratory technicians. The feces or soil sample is placed in a sample tube, then a processing solution is added, followed by centrifugation to obtain the supernatant containing dissolved microorganisms. The supernatant is then transferred to a lysis tube using a pipette, and the lysis tube is placed into a nucleic acid extractor for nucleic acid extraction. The operation steps are complex, and manual operation is prone to errors in the extraction results. Furthermore, manual operation can be nauseating and uncomfortable for laboratory technicians when extracting feces and contaminated soil samples. Therefore, an intelligent nucleic acid extraction system for microorganisms in feces and soil samples is proposed. Summary of the Invention

[0004] The purpose of this invention is to provide an intelligent extraction system for microbial nucleic acids from fecal and soil samples, addressing the problems of errors in extraction results due to manual operation in existing technologies, and the discomfort and nausea experienced by lab technicians when manually extracting fecal and contaminated soil samples. The specific technical solution is as follows:

[0005] To achieve the above and other related objectives, this invention provides an intelligent extraction system for microbial nucleic acids from fecal and soil samples, comprising a nucleic acid extractor body, and further comprising an injection device, a pipetting device, a positive pressure device, a negative pressure device, a sample tube rack, a processing tube, and a collection device disposed within the nucleic acid extractor body; wherein,

[0006] The injection device is located above the sample tube rack and is used to insert into the processing tube. The injection device is connected to a pipette, a positive pressure device, and a negative pressure device. The pipette is used to draw up the processing liquid and inject it into the processing tube through the injection device. The positive pressure device injects compressed air into the processing tube through the injection device, and the negative pressure device draws out the air from the processing tube through the injection device.

[0007] The processing tube is placed in the sample tube rack. The processing tube includes a tube body, a tube cap, a sealing plug, a sealing ring, a sealing cylinder, a filter screen, and a limiting ring. The tube body is provided with a tube cap at the upper end, and a sealing plug is provided in the middle of the tube cap. The sealing plug is used for the insertion of the injection device. A sealing cylinder slides through the bottom of the tube body. The sealing cylinder is a hollow tubular structure with an open bottom. Multiple water inlet holes are provided on the side wall of the sealing cylinder. The sealing ring is fixedly connected to the inner wall of the tube body and is sleeved on the outer side of the sealing cylinder. The filter screen is provided on the inner wall of the tube body and is located above the sealing cylinder. The limiting ring is fixedly connected to the sealing cylinder and slidably connected to the inner wall of the tube body to limit the displacement distance of the sealing cylinder. Multiple locking rods are provided on the upper side of the sealing ring. The locking rods are used to lock onto the water inlet holes.

[0008] The liquid collection device is located below the processing tube and is used to receive the filtrate discharged from the processing tube.

[0009] Preferably, a mounting block is fixedly connected to the lower part of the sealing plug via multiple connecting rods, and a spring plate is fixedly connected to the lower side of the mounting block. The spring plate is used to lift the sample. An electromagnet is provided on the sample tube rack to attract the spring plate. A protrusion is provided on the outer wall of the tube cap, and a positioning groove corresponding to the protrusion is provided on the sample tube rack.

[0010] Preferably, the injection device includes a lifting mechanism, a four-way valve, and an injection needle. The lifting mechanism is located inside the nucleic acid extractor body and is equipped with a four-way valve. The lifting mechanism is used to control the up and down movement of the four-way valve. An injection needle is provided at the lower end of the four-way valve. The lifting mechanism controls the four-way valve to move down, so that the injection needle is inserted into the sealing plug. The three upper ends of the four-way valve are connected to a pipetting device, a positive pressure device, and a negative pressure device respectively through pipelines.

[0011] Preferably, the liquid collection device includes a translation mechanism, a heating mechanism, and a lysis tube. The translation mechanism is installed inside the nucleic acid extractor body, and a heating mechanism is provided on the translation mechanism. A blind hole for placing the lysis tube is opened above the heating mechanism, and the lysis tube is placed on the heating mechanism.

[0012] Preferably, the mounting block has a conical structure.

[0013] Preferably, the sample tube rack is installed in the nucleic acid extractor body via a ball bearing slide rail, and a fixing mechanism is provided between the sample tube rack and the nucleic acid extractor body to fix the position of the sample tube rack.

[0014] Preferably, the nucleic acid extractor body is provided with a waste liquid box at the bottom, which is located directly below the processing tube. The waste liquid box is used to collect residual liquid dripping from the lower end of the processing tube.

[0015] Preferably, the pipetting device uses a peristaltic pump of model SF-ANR.

[0016] Preferably, the negative pressure device is a vacuum pump, and the exhaust port of the vacuum pump is equipped with an activated carbon adsorption box, which is used to absorb odors.

[0017] Preferably, the positive pressure device is a pressure pump.

[0018] The present invention provides an intelligent extraction system for microbial nucleic acids from fecal and soil samples, which has the following beneficial effects:

[0019] This invention provides an intelligent nucleic acid extraction system for microorganisms from fecal and soil samples. The system incorporates an injection device, a pipetting device, a positive pressure device, a negative pressure device, and a collection device within the nucleic acid extractor. In use, the fecal or soil sample is simply placed into the processing tube, the cap is tightened, and the tube is placed in the sample holder. By sequentially activating the injection device, pipetting device, negative pressure device, and positive pressure device, the solution in the processing tube passes through a filter screen, removing impurities. The processed solution and microorganisms pass through the filter screen and enter through the inlet, exiting from the bottom of the sealed tube to the collection device for subsequent nucleic acid extraction. No human intervention is required, avoiding errors caused by manual operation. Attached Figure Description

[0020] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0021] Figure 1 This is a schematic diagram of the structure of an intelligent microbial nucleic acid extraction system for fecal and soil samples provided by the present invention.

[0022] Figure 2 This is a schematic diagram of the structure of the processing tube provided by the present invention.

[0023] Figure 3 yes Figure 2Enlarged view of a portion of point A in the middle.

[0024] Figure 4 This is a schematic diagram of the injection device provided by the present invention. Detailed Implementation

[0025] The following detailed description, in conjunction with the accompanying drawings and specific embodiments, provides a more detailed explanation of the intelligent microbial nucleic acid extraction system for fecal and soil samples proposed in this invention. The advantages and features of this invention will become clearer from the following description. It should be noted that the accompanying drawings are all in a very simplified form and use non-precise proportions, and are only used to facilitate and clarify the illustration of the embodiments of this invention.

[0026] In the description of this invention, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified.

[0027] It should be noted that the illustrations provided in the following embodiments are only schematic representations of the basic concept of the present invention. Therefore, the illustrations only show the components related to the present invention and are not drawn according to the actual number, shape and size of the components in the actual implementation. In the actual implementation, the shape, quantity and proportion of each component can be arbitrarily changed, and the layout of the components may also be more complex.

[0028] Therefore, this invention provides an intelligent extraction system for microbial nucleic acids from fecal and soil samples, such as... Figure 1-4 As shown, the instrument includes a nucleic acid extractor body 100, and further includes an injection device 200, a pipetting device 300, a positive pressure device 400, a negative pressure device 500, a sample tube rack 600, a processing tube 700, and a liquid collection device 800 disposed inside the nucleic acid extractor body 100; wherein,

[0029] The injection device 200 is positioned above the sample tube rack 600 and is used to insert into the processing tube 700. The injection device 200 is connected to a pipette 300, a positive pressure device 400, and a negative pressure device 500. The pipette 300 is used to draw up the processing liquid and inject it into the processing tube 700 through the injection device 200. The positive pressure device 400 injects compressed air into the processing tube 700 through the injection device 200. The negative pressure device 500 draws out the air from the processing tube 700 through the injection device 200.

[0030] The processing tube 700 is placed in the sample tube rack 600. The processing tube 700 includes a tube body 701, a tube cap 702, a sealing plug 703, a sealing ring 704, a sealing cylinder 705, a filter screen 706, and a limiting ring 707. The tube cap 702 is provided at the upper end of the tube body 701, and the sealing plug 703 is provided in the middle of the tube cap 702. The sealing plug 703 is used for the insertion of the injection device 200. The sealing cylinder 705 slides through the bottom of the tube body 701. The sealing cylinder 705 is a hollow tubular structure with an open bottom. Multiple water inlet holes 708 are provided on the side wall of the sealing cylinder 705. The sealing ring 704 is fixedly connected to the inner wall of the pipe body 701. The sealing ring 704 is sleeved on the outside of the sealing cylinder 705 and blocks the water inlet 708. The filter screen 706 is set on the inner wall of the pipe body 701 and is located above the sealing cylinder 705. The limiting ring 707 is fixedly connected to the sealing cylinder 705 and is slidably connected to the inner wall of the pipe body 701 to limit the displacement distance of the sealing cylinder 705. The upper side of the sealing ring 704 is provided with multiple locking rods 709, which are used to lock on the water inlet 708.

[0031] The liquid receiving device 800 is located below the processing tube 700 and is used to receive the filtrate discharged from the processing tube 700.

[0032] In use, first place the fecal or soil sample into the processing tube 700, tighten the tube cap 702, and place the processing tube 700 into the sample tube rack 600. At this time, the injection device 200 is activated, inserting and penetrating the sealing plug 703. Then, the pipette 300 is activated, injecting the processing solution into the processing tube 700 to dissolve the sample. After processing, the negative pressure device 500 is activated, extracting the air from the processing tube 700, causing the sealing cylinder 705 to slide upwards under atmospheric pressure. At this time, the water inlet 708 disengages from the sealing ring 704, and the locking rod 709... The device is engaged with the water inlet 708, preventing the sealing cylinder 705 from moving downwards. At this time, the positive pressure device 400 is activated, filling the processing tube 700 with compressed air. The solution in the processing tube 700 passes through the filter screen 706, filtering out impurities. The processing solution and microorganisms pass through the filter screen 706 and enter through the water inlet 708, exiting from the lower end of the sealing cylinder 705 to the liquid collection device 800 for subsequent nucleic acid extraction steps. The injection device 200, pipetting device 300, positive pressure device 400, and negative pressure device 500 can all be controlled by a microcontroller, achieving full automation without human intervention and avoiding errors caused by human operation.

[0033] In one embodiment of the present invention, a mounting block 710 is fixedly connected to the lower part of the sealing plug 703 via multiple connecting rods. A spring plate 711 is fixedly connected to the lower side of the mounting block 710. The spring plate 711 is used to lift the sample. An electromagnet 601 is provided on the sample tube rack 600. The electromagnet 601 is used to attract the spring plate 711, causing the spring plate 711 to vibrate, so that the sample can be fully mixed with the processing liquid. A protrusion 712 is provided on the outer wall of the tube cap 702. The sample tube rack 600 is provided with a positioning groove corresponding to the protrusion 712. When the processing tube 700 is inserted into the sample tube rack 600, the protrusion 712 slides into the positioning groove. At this time, the spring plate 711 is perpendicular to the electromagnet 601, so that the electromagnet 601 can attract the spring plate 711 to bend.

[0034] When the pipette 300 injects the processing solution into the processing tube 700, the electromagnet 601 works intermittently. When the electromagnet 601 is activated, the spring plate 711 is attracted and deformed. When the electromagnet 601 is turned off, the spring plate 711 vibrates, thereby quickly mixing the sample with the processing solution evenly, thus greatly reducing the extraction time and improving the extraction efficiency.

[0035] In one embodiment of the present invention, the injection device 200 includes a lifting mechanism 201, a four-way valve 202, and an injection needle 203. The lifting mechanism 201 is disposed inside the nucleic acid extractor body 100. The four-way valve 202 is disposed on the lifting mechanism 201. The lifting mechanism 201 is used to control the up and down movement of the four-way valve 202. The injection needle 203 is disposed at the lower end interface of the four-way valve 202. The lifting mechanism 201 controls the four-way valve 202 to move down, so that the injection needle 203 is inserted into the sealing plug 703. The three upper interfaces of the four-way valve 202 are respectively connected to the pipetting device 300, the positive pressure device 400, and the negative pressure device 500 through pipelines.

[0036] In one embodiment of the present invention, the liquid collection device 800 includes a translation mechanism 801, a heating mechanism 802, and a lysis tube 803. The translation mechanism 801 is installed inside the nucleic acid extractor body 100. The heating mechanism 802 is provided on the translation mechanism 801. A blind hole for placing the lysis tube 803 is opened above the heating mechanism 802. The lysis tube 803 is placed on the heating mechanism 802. Before the positive pressure device 400 is activated, the translation mechanism 801 drives the heating mechanism 802 to translate, so that the lysis tube 803 moves directly below the processing tube 700, thereby collecting the filtered filtrate. After collection, the translation mechanism 801 resets the heating mechanism 802, so that the lysis tube 803 returns to its initial position. At this time, the magnetic rod and magnetic rod sleeve in the nucleic acid extractor body 100 work together to extract nucleic acid, thereby realizing automatic extraction of sample nucleic acid from the processing tube 700.

[0037] In one embodiment of the present invention, the mounting block 710 has a conical structure. When the injection needle 203 is inserted into the sealing plug 703 and the treatment liquid is sprayed onto the mounting block 710, the conical mounting block 710 can guide the treatment liquid to the wall of the treatment tube 700, thereby flushing the sample attached to the tube wall to the bottom.

[0038] In one embodiment of the present invention, the sample tube rack 600 is installed inside the nucleic acid extractor body 100 via a ball bearing slide rail 602. By pulling the sample tube rack 600, the tubes 700 can be easily inserted and removed. A fixing mechanism is provided between the sample tube rack 600 and the nucleic acid extractor body 100. The fixing mechanism is used to fix the position of the sample tube rack 600. Specifically, the fixing mechanism can be fixed by magnetic attraction or snap-fit. A position sensor can also be provided to determine the position of the fixing mechanism.

[0039] In one embodiment of the present invention, a waste liquid box 804 is provided at the bottom of the nucleic acid extractor body 100. The waste liquid box 804 is located directly below the processing tube 700 and is used to collect residual liquid dripping from the lower end of the processing tube 700.

[0040] In one embodiment of the present invention, the pipetting device 300 uses a peristaltic pump of model SF31-ANR 20250010.

[0041] In one embodiment of the present invention, the negative pressure device 500 adopts a vacuum pump, and the exhaust port of the vacuum pump is equipped with an activated carbon adsorption box. The activated carbon adsorption box is used to absorb odors and protect the air quality of the laboratory environment.

[0042] In one embodiment of the present invention, the positive pressure device 400 is a pressure pump.

[0043] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A smart system for extracting microbial nucleic acids from fecal and soil samples, comprising a nucleic acid extractor body (100), characterized in that, It also includes an injection device (200), a pipette device (300), a positive pressure device (400), a negative pressure device (500), a sample tube rack (600), a processing tube (700), and a liquid collection device (800) disposed inside the nucleic acid extractor body (100); wherein, the injection device (200) is disposed above the sample tube rack (600) and is used to insert into the processing tube (700); the injection device (200) is connected to the pipette device (300), the positive pressure device (400), and the negative pressure device (500) respectively; the pipette device (300) is used to extract the processing liquid and, through... The injection device (200) injects the treatment liquid into the treatment tube (700). The positive pressure device (400) injects compressed air into the treatment tube (700) through the injection device (200), and the negative pressure device (500) extracts air from the treatment tube (700) through the injection device (200). The treatment tube (700) is placed in the sample tube rack (600). The treatment tube (700) includes a tube body (701), a tube cap (702), a sealing plug (703), a sealing ring (704), a sealing cylinder (705), a filter screen (706), and a limiting ring (707). The upper end of the tube body (701) is provided with a tube cap (702), and a sealing plug (703) is provided in the middle of the tube cap (702). The sealing plug (703) is used for the insertion of the injection device (200). A sealing cylinder (705) slides through the bottom of the tube body (701). The sealing cylinder (705) is a hollow tubular structure with an open bottom. Multiple water inlet holes (708) are provided on the side wall of the sealing cylinder (705). The sealing ring (704) is fixedly connected to the inner wall of the tube body (701). The sealing ring (704) is sleeved on the outside of the sealing cylinder (705). The filter screen (706) The filter screen (706) is located above the sealing cylinder (705) and is fixedly connected to the sealing cylinder (705). The limiting ring (707) is slidably connected to the inner wall of the pipe body (701) to limit the displacement distance of the sealing cylinder (705). The upper side of the sealing ring (704) is provided with multiple locking rods (709), which are located on the water inlet (708). The liquid collection device (800) is located below the processing pipe (700) to receive the filtrate discharged from the processing pipe (700). The injection device (200) includes a lifting mechanism (201), a four-way valve (202), and an injection needle (203). The lifting mechanism (201) is located inside the nucleic acid extractor body (100). The lifting mechanism (201) is equipped with a four-way valve (202). The lifting mechanism (201) is used to control the four-way valve (202) to move up and down. An injection needle (203) is provided at the lower end interface of the four-way valve (202). The lifting mechanism (201) controls the four-way valve (202) to move down, so that the injection needle (203) is inserted into the sealing plug (703). The three upper interfaces of the four-way valve (202) are connected to a pipetting device (300), a positive pressure device (400), and a negative pressure device (500) respectively through pipelines.

2. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, A mounting block (710) is fixedly connected to the bottom of the sealing plug (703) by multiple connecting rods. A spring plate (711) is fixedly connected to the lower side of the mounting block (710). The spring plate (711) is used to lift the sample. An electromagnet (601) is provided on the sample tube rack (600). The electromagnet (601) is used to attract the spring plate (711). A protrusion is provided on the outer wall of the tube cap (702). A positioning groove corresponding to the protrusion is provided on the sample tube rack (600).

3. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The liquid collection device (800) includes a translation mechanism (801), a heating mechanism (802), and a lysis tube (803). The translation mechanism (801) is installed inside the nucleic acid extractor body (100). The heating mechanism (802) is provided on the translation mechanism (801). A blind hole for placing the lysis tube (803) is opened above the heating mechanism (802). The lysis tube (803) is placed on the heating mechanism (802).

4. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 2, characterized in that, The mounting block (710) has a conical structure.

5. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The sample tube rack (600) is installed inside the nucleic acid extractor body (100) via a ball bearing slide rail (602). A fixing mechanism is provided between the sample tube rack (600) and the nucleic acid extractor body (100) to fix the position of the sample tube rack (600).

6. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The bottom of the nucleic acid extractor body (100) is provided with a waste liquid box (804), which is located directly below the processing tube (700). The waste liquid box (804) is used to collect residual liquid dripping from the lower end of the processing tube (700).

7. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The pipetting device (300) employs a peristaltic pump.

8. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The negative pressure device (500) uses a vacuum pump, and the exhaust port of the vacuum pump is equipped with an activated carbon adsorption box, which is used to remove odors.

9. The intelligent extraction system for microbial nucleic acids from fecal and soil samples as described in claim 1, characterized in that, The positive pressure device (400) uses a pressure pump.

Images