Bacterial diagnosis device and bacterial drug resistance detection device

Abstract

The utility model relates to a bacterial diagnosis device and a bacterial drug resistance detection device. The bacterial diagnosis device comprises a sample bin module, a sample culture module, a mechanical cantilever module, a cleaning module, a shaking module, a suction filtration module, an incubation module and a detection module, the sample bin module is used for storing samples; the sample culture module is used for culturing samples; the mechanical cantilever module is used for transferring samples and carrying the sample culture module; the cleaning module is used for cleaning a device for transferring a sample; the shake-up module is used for shaking up the sample; the suction filtration module is used for generating pressure difference in the sample culture module so as to realize substance separation of the sample; the incubation module is used for incubating a sample; the detection module is used for detecting the fluorescence luminescence value of the sample. In the embodiment of the invention, modules for controlling the functions of sample adding, shaking up, suction filtration, incubation, detection and the like are designed, so that full-function automatic integrated bacterial diagnosis and bacterial drug resistance detection are realized. The whole device is high in control precision, low in pollution rate, high in detection speed and high in automation degree.

Description

Technical Field

[0001] The utility model belongs to the technical field of luminescent microorganism detection, in particular to a bacteria diagnosis device and a bacteria drug resistance detection device. Background Art

[0002] Pathogenic bacteria are widely present in our daily lives, and their mutations leading to drug resistance have become a global problem. To make matters worse, since the 1990s, the speed of research and development and marketing of new antibacterial drugs has significantly declined, resulting in a situation where no drugs are available clinically. If left unchecked, the global GDP will decrease by 2-3.5% by 2050, and the losses will reach 100 trillion US dollars. Take tuberculosis (TB) as an example. In recent years, it has become the number one infectious disease, with about 10 million cases each year and about 1 million patients with drug-resistant tuberculosis (TB). However, only three new anti-TB drugs have been approved, and their use is restricted due to their high toxicity. The overall drug resistance rate of tuberculosis (TB) in my country is 36.8%.

[0003] Bacteriophages have the ability to kill drug-resistant bacteria, have strong bactericidal specificity, and can replicate in bacteria, making them unique and the most promising bactericides. Bacteriophages are viruses that invade bacteria and must parasitize in living bacteria. They have strict host specificity. As early as 1925, people proposed the idea of ​​using bacteriophages to identify bacteria. Receptor recombinant phages are a very important diagnostic phage. The bacterial fluorescein gene is embedded in the phage gene, and the modified phage is applied to the bacteria. The infected bacteria can emit fluorescence and then be detected. At present, the existing technology discloses an automated detection platform based on phage technology, which can reduce the manual intervention time to half an hour and improve the efficiency of detection, but the structure is complex and cannot be fully automated.

[0004] Most of the traditional self-luminous bacteria detection equipment based on the autonomous luminescent element bacteriophage (ARP) technology has a single function and can only realize one or several functions such as sample addition, shaking, filtration, incubation, and detection, and cannot realize full-function automatic integrated detection. Utility Model Content

[0005] In order to solve the above technical problems, the present application provides a bacterial diagnosis device and a bacterial resistance detection device.

[0006] In a first aspect, the present application discloses a bacterial diagnostic device, including a sample chamber module, a sample culture module, a mechanical cantilever module, a cleaning module, a shaking module, a filtration module, an incubation module and a detection module;

[0007] The sample chamber module, the cleaning module, the shaking module, the filtration module, the incubation module and the detection module are located at the same level and are all located below the mechanical cantilever module; the sample culture module is movably arranged in the bacterial diagnosis device;

[0008] The sample bin module is used to provide the environment required for storing samples; the sample culture module is used to provide the environment required for culturing samples; the mechanical cantilever module is used to transfer samples in the sample bin module to the sample culture module, and to carry the sample culture module; the cleaning module is used to clean the device used to transfer samples in the mechanical cantilever module; the shaking module is used to shake the samples in the sample culture module; the filtration module is used to generate a pressure difference inside the sample culture module, thereby realizing the material separation of the samples in the sample culture module; the incubation module is used to provide the constant temperature environment required for incubating the samples in the sample culture module; the detection module is used to detect the fluorescence luminescence value of the samples in the sample culture module.

[0009] In some possible embodiments,

[0010] The sample culture module includes a well plate;

[0011] The well plate comprises a well plate body and a plurality of sample loading wells;

[0012] The sample loading well includes a microporous filter membrane at the bottom; the microporous filter membrane is used for material separation of the sample.

[0013] In some possible embodiments,

[0014] The bacterial diagnostic device also includes a hydraulic system, a cleaning liquid storage bin and a waste liquid storage bin;

[0015] The hydraulic system, the cleaning liquid storage bin and the waste liquid storage bin are located at the same level, and are all located below the sample bin module, the cleaning module, the shaking module, the filtration module, the incubation module and the detection module.

[0016] In some possible embodiments,

[0017] The bacteria diagnosis device also includes a rack; the rack includes an upper rack layer, a middle rack layer and a lower rack layer arranged from top to bottom;

[0018] The mechanical cantilever module is located at the upper layer of the rack; the sample chamber module, cleaning module, shaking module, filtration module, incubation module, and detection module are located at the middle layer of the rack; the hydraulic system, cleaning liquid storage bin, and waste liquid storage bin are located at the lower layer of the rack.

[0019] In some possible embodiments,

[0020] The mechanical cantilever module includes a mechanical cantilever fixing frame, a mechanical gripper left and right driving mechanism, a mechanical gripper mechanism, a connecting needle left and right driving mechanism and a connecting needle sample loading mechanism;

[0021] The mechanical cantilever fixing frame is fixed to the middle layer of the frame; the left and right driving mechanisms of the mechanical gripper and the left and right driving mechanisms of the connecting needles are fixed to the mechanical cantilever fixing frame; the mechanical gripper mechanism is connected to the left and right driving mechanisms of the mechanical gripper; the connecting needle loading mechanism is connected to the left and right driving mechanisms of the connecting needles;

[0022] The left and right driving mechanism of the manipulator gripper is used to drive the manipulator gripper mechanism, thereby realizing the transportation of the well plate in the left and right directions; the left and right driving mechanism of the row of needles is used to drive the row of needles loading mechanism, thereby realizing the transfer of samples in the left and right directions.

[0023] In some possible embodiments,

[0024] The manipulator gripper mechanism comprises a manipulator gripper fixing frame, a manipulator gripper front and rear driving mechanism, a manipulator gripper up and down driving mechanism, a manipulator gripper grasping driving mechanism and a manipulator gripper;

[0025] The robot gripper fixing frame is fixed on the left and right driving mechanisms of the robot gripper, and is driven to move by the left and right driving mechanisms of the robot gripper;

[0026] The front and rear driving mechanism of the manipulator gripper is fixed on the moving part of the manipulator gripper fixing frame, and is used to drive the upper and lower driving mechanism of the manipulator gripper to move forward and backward longitudinally;

[0027] The up and down driving mechanism of the manipulator gripper is fixed on the moving parts of the front and rear driving mechanism of the manipulator gripper, and is used to drive the grasping driving mechanism of the manipulator gripper to move up and down;

[0028] The gripping drive mechanism of the manipulator gripper is fixed on the moving parts of the upper and lower drive mechanisms of the manipulator gripper, and is used to drive the movement of the manipulator gripper;

[0029] The manipulator claw is fixed to the end of the manipulator claw grasping drive mechanism.

[0030] In some possible embodiments,

[0031] The coupling needle sample adding mechanism comprises a coupling needle fixing frame, a coupling needle up and down driving mechanism, a coupling needle and a hydraulic pipeline;

[0032] The rear end of the coupling needle fixing frame is fixed to the moving end of the left and right driving mechanism of the coupling needle;

[0033] The coupling needle up and down driving mechanism is fixed at the front end of the coupling needle fixing frame;

[0034] The coupling needle is fixed to the moving end of the coupling needle upper and lower driving mechanism;

[0035] The hydraulic pipeline includes a first end and a second end; the first end of the hydraulic pipeline is connected to the coupling needle, and the second end of the hydraulic pipeline is connected to the cleaning liquid storage bin.

[0036] In some possible embodiments,

[0037] The sample bin module includes a sample bin fixing bracket and a sample bin drawer; the sample bin fixing bracket is a semi-enclosed structure and is fixed to the middle layer of the rack; the sample bin drawer is embedded in the sample bin fixing bracket;

[0038] The sample bin drawer includes at least one drawer compartment;

[0039] The drawer compartment includes an external sealed cover and a cover driving mechanism, as well as an internal temperature control module and at least one material box; the cover driving mechanism is used to close or open the corresponding sealed cover;

[0040] The material box includes a sample adding port and a sampling port at the top, as well as an internal sample storage bin and a small sample storage bin; the sample storage bin is used for storing large-volume samples, and the small sample storage bin is used for storing small-volume samples.

[0041] In some possible embodiments,

[0042] The cleaning module includes a cleaning base, a row of fountain cleaning ports, a waste liquid tank and a waste liquid outlet;

[0043] The cleaning base is fixed on the middle layer of the frame; a trough is arranged on the upper part of the cleaning base;

[0044] The row fountain cleaning port is located in the middle of the tank body and is used to clean the row needles;

[0045] The waste liquid tank is located around the cleaning port of the row fountain, and is used to collect the waste liquid used to clean the row needles, and discharge the waste liquid used to clean the row needles to the waste liquid storage bin through the waste liquid outlet.

[0046] In some possible embodiments,

[0047] The shaking module includes a shaking module base, a shaking station, a cover station, a perforated plate cover, and a shaking drive mechanism;

[0048] The base of the shaking module is fixed to the middle layer of the frame and is used to support the shaking module;

[0049] The shaking station and the cover station are located on the shaking module base and are placed adjacent to each other;

[0050] The well plate cover is placed on the cover station; the well plate cover is used to seal the well plate, thereby preventing the sample in the well plate from spilling out during the shaking process;

[0051] The shaking driving mechanism is located below the shaking station; the shaking driving mechanism is used to drive the shaking station to vibrate on a horizontal plane, thereby shaking the sample in the well plate placed above the shaking station.

[0052] In some possible embodiments,

[0053] The filtration module includes a filtration module fixing frame, a vacuum pump, a pressure regulating plate, a negative pressure chamber and a filtration station;

[0054] The filtration module fixing frame is fixed to the middle layer of the frame and is used to support the filtration module;

[0055] The vacuum pump is used to provide negative pressure power;

[0056] The pressure regulating disk is used to adjust the pressure of the vacuum pump;

[0057] An air suction port is arranged at the lower side of the negative pressure chamber, and the air suction port is connected to the vacuum pump through an air pipe; the interior of the negative pressure chamber includes a filtration waste liquid box, and the bottom surface of the filtration waste liquid box includes a filtration waste liquid hole; the filtration waste liquid box is used to collect the filtration waste liquid, and discharge the filtration waste liquid to the waste liquid storage chamber through the filtration waste liquid hole;

[0058] The suction filtration station is arranged above the negative pressure bin and is connected to the upper side of the negative pressure bin; the suction filtration station comprises an external suction filtration limit block and an internal suction filtration limit groove; the suction filtration limit block is used to place the orifice plate;

[0059] The bottom surface of the orifice plate, the side surface of the filtration limit groove and the negative pressure chamber form a closed space during the filtration process, so as to generate a pressure difference inside the orifice plate with the microporous filter membrane as the boundary, thereby realizing the material separation of the sample.

[0060] In some possible embodiments,

[0061] The incubation module includes an incubation module base, an incubation chamber and a sample transfer robot arm;

[0062] The incubation module base is fixed to the middle layer of the frame, and is used to connect and support the incubation chamber and the sample transfer robot arm located on the incubation module base;

[0063] The incubation chamber includes at least one incubation chamber; the incubation chamber includes a sealed sliding door mechanism at the front and at least one incubation station inside; the incubation station is used to place the well plate and incubate the sample;

[0064] The sample transfer robot arm includes an incubation waiting station and an incubation feeding mechanism, and the incubation waiting station is fixed to the end of the incubation feeding mechanism; the incubation waiting station is used to place the well plate waiting for incubation; the incubation feeding mechanism is used to transfer the well plate on the incubation waiting station to the incubation station.

[0065] In some possible embodiments,

[0066] The detection module includes a detection module fixing frame, a push-pull mechanism and a fluorescence detection device;

[0067] The detection module fixing frame is fixed to the middle layer of the rack, and is used to connect and support the push-pull mechanism and the fluorescence detection device placed adjacently on the detection module fixing frame;

[0068] The push-pull mechanism includes a push-pull mechanism bottom plate, a push-in drive mechanism and a pull-out drive structure; the push-in drive mechanism and the pull-out drive structure are fixed on the push-pull mechanism bottom plate; the push-in drive mechanism is used to push the orifice plate into the fluorescence detection device; the pull-out drive structure is used to pull the orifice plate out of the fluorescence detection device;

[0069] The fluorescence detection device includes a material inlet and outlet device and a photon counting device; the material inlet and outlet device is used for transferring the position of the well plate between the fluorescence detection device and the push-pull mechanism; the photon counting device is used for determining the fluorescence luminescence value of the sample in the sample loading hole of the well plate.

[0070] In a second aspect, an embodiment of the present application discloses a bacterial resistance detection device, including any one of the above-mentioned bacterial diagnostic devices.

[0071] The technical solution provided by the embodiment of the present application has the following technical effects:

[0072] The bacterial diagnosis device of the embodiment of the present application includes a sample chamber module, a sample culture module, a mechanical cantilever module, a cleaning module, a shaking module, a filtration module, an incubation module and a detection module; the sample chamber module is used to store samples; the sample culture module is used to culture samples; the mechanical cantilever module is used to transfer samples and carry the sample culture module; the cleaning module is used to clean the device for transferring samples; the shaking module is used to shake the samples; the filtration module is used to generate a pressure difference inside the sample culture module, thereby achieving material separation of the sample; the incubation module is used to incubate the sample; and the detection module is used to detect the fluorescence luminescence value of the sample. In the embodiment of the present application, by designing modules that control functions such as sample addition, shaking, filtration, incubation, and detection, full-function automatic integrated bacterial diagnosis and bacterial resistance detection are achieved. The entire device has high control accuracy, low contamination rate, fast detection speed and high degree of automation. BRIEF DESCRIPTION OF THE DRAWINGS

[0073] In order to more clearly illustrate the technical solutions and advantages in the embodiments of the present application or the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0074] Figure 1 This is a schematic diagram of a bacterial diagnostic device provided in an embodiment of the present application. Figure 1 ;

[0075] Figure 2 This is a schematic diagram of a bacterial diagnostic device provided in an embodiment of the present application. Figure 2 ;

[0076] Figure 3It is a front schematic diagram of a hole plate provided in an embodiment of the present application;

[0077] Figure 4 is a cross-sectional schematic diagram of an orifice plate provided in an embodiment of the present application;

[0078] Figure 5 is a schematic diagram of a sample chamber module provided in an embodiment of the present application;

[0079] Figure 6 is a schematic diagram of a sample bin drawer provided in an embodiment of the present application;

[0080] Figure 7 is a schematic diagram of a material box provided in an embodiment of the present application;

[0081] Figure 8 is a schematic diagram of a mechanical cantilever module provided in an embodiment of the present application;

[0082] Fig. 9 is a schematic diagram of a mechanical gripper mechanism provided in an embodiment of the present application;

[0083] Fig.10 It is a schematic diagram of a coupled needle sample loading mechanism provided in an embodiment of the present application;

[0084] Fig.11 is a schematic diagram of a cleaning module provided in an embodiment of the present application;

[0085] Fig.12 is a schematic diagram of a shaking module provided in an embodiment of the present application;

[0086] Fig.13 is a schematic diagram of a filtration module provided in an embodiment of the present application;

[0087] Fig.14 It is a schematic diagram of a filtration station provided in an embodiment of the present application;

[0088] Fig.15 is a schematic diagram of an incubation module provided in an embodiment of the present application;

[0089] Fig.16 is a schematic diagram of a detection module provided in an embodiment of the present application;

[0090] Fig.17 is a schematic diagram of a push-pull mechanism provided in an embodiment of the present application;

[0091] Fig.18 is a schematic diagram of a pull-out hopper mechanism provided in an embodiment of the present application;

[0092] Fig.19 It is a schematic diagram of a fluorescence detection device provided in an embodiment of the present application.

[0093] Description of reference numerals:

[0094] Sample bin module 1, sample bin fixing bracket 11, sample bin drawer 12, drawer chamber 121, sealed cover 1211, cover drive mechanism 1212, temperature control module 1213, material box 1214, sample addition port 12141, sampling port 12142, sample storage bin 12143, small sample storage bin 12144; sample culture module 2, orifice plate 21, orifice plate body 211, sample loading hole 212, microporous filter membrane 2121; mechanical cantilever module 3, mechanical cantilever fixing bracket 31, mechanical gripper left and right drive mechanism 32, mechanical gripper mechanism 3 3, mechanical gripper fixing frame 331, mechanical gripper front and rear driving mechanism 332, mechanical gripper up and down driving mechanism 333, mechanical gripper grasping driving mechanism 334, mechanical gripper 335, connecting needle left and right driving mechanism 34, connecting needle sample adding mechanism 35, connecting needle fixing frame 351, connecting needle up and down driving mechanism 352, connecting needle 353, hydraulic pipeline 354; cleaning module 4, cleaning base 41, connecting fountain cleaning port 42, waste liquid tank 43, waste liquid outlet 44; shaking module 5, shaking module base 51, shaking station 52, cover station 53 , orifice plate cover 54, shaking drive mechanism 55; filtration module 6, filtration module fixing frame 61, vacuum pump 62, pressure regulating plate 63, negative pressure bin 64, air suction port 641, filtration waste liquid box 642, filtration waste liquid hole 6421, filtration station 65, filtration limit block 651, filtration limit groove 652, air gap 66, negative pressure bin 64; incubation module 7, incubation module base 71, incubation bin 72, incubation chamber 721, sealed sliding door mechanism 7211, incubation station 7212, sample transfer robot 73, incubation waiting station 731, incubation feeding Mechanism 732; detection module 8, detection module fixing frame 81, push-pull mechanism 82, push-pull mechanism bottom plate 821, push-in drive mechanism 822, pull-out drive structure 823, test station 824, push sheet 825, pull plate 826, pull-out hopper mechanism 827, hopper base 8271, hopper shaft 8272, eccentric pull block 8273, limit protrusion 8274, fluorescence detection device 83, fluorescence detection device 83, feeding and discharging device 831, photon counting device 832; frame 9, upper frame layer 91, middle frame layer 92, lower frame layer 93. DETAILED DESCRIPTION

[0095] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0096] It should be noted that the "one embodiment" or "embodiment" referred to in the specification of the embodiment of the present application refers to a specific feature, structure or characteristic that can be included in at least one implementation of the present application. It should be understood that in the specification and claims of the embodiment of the present application and the above-mentioned drawings, the orientation or position relationship indicated by the terms "upper", "lower", "top", "bottom", etc. is based on the orientation or position relationship shown in the drawings, which is only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. The terms "first" and "second" are only used for descriptive purposes, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, the features defined as "first" and "second" can explicitly or implicitly include one or more of the features. Moreover, the terms "first", "second", etc. are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. It should be understood that the data used in this way can be interchangeable where appropriate, so that the embodiments of the present application described here can be implemented in an order other than those illustrated or described here. In addition, in the description of the present embodiment, unless otherwise specified, "plurality" means two or more. In addition, the terms "including" and "having" and any variations thereof are intended to cover non-exclusive inclusions, for example, a process, method, system or product including a series of steps or units is not necessarily limited to those steps or units clearly listed, but may include other steps or units that are not clearly listed or inherent to these processes, methods, products or devices.

[0097] It should be understood that when a device or component is referred to as "on ...", "adjacent to ...", "connected to" other devices or components, it can be directly on, adjacent to, connected to other devices or components, or there can be intervening devices or components. On the contrary, when a device or component is referred to as "directly on ...", "directly adjacent to ...", "directly connected to" other devices or components, there is no intervening device or component. It should be understood that although the terms first, second, third, etc. can be used to describe various components, areas, layers and / or parts, these components, areas, layers and / or parts should not be limited by these terms. These terms are only used to distinguish one component, area, layer or part from another component, area, layer or part. Therefore, without departing from the teachings of the present application, the first component, area, layer or part discussed below can be expressed as the second component, area, layer or part. And when the second component, area, layer or part is discussed, it does not mean that the present application necessarily has the first component, area, layer or part.

[0098] In order to make the purpose, technical solution and advantages disclosed in the embodiments of the present application more clearly understood, the embodiments of the present application are further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the embodiments of the present application and are not used to limit the embodiments of the present application.

[0099] An embodiment of the present application provides a bacterial diagnosis device. Figure 1 This is a schematic diagram of a bacterial diagnostic device provided in an embodiment of the present application. Figure 1 ,like Figure 1 As shown, the bacterial diagnosis device includes functional modules such as a sample chamber module 1, a sample culture module 2, a mechanical cantilever module 3, a cleaning module 4, a shaking module 5, a suction filtration module 6, an incubation module 7 and a detection module 8. Among them, the sample chamber module 1, the cleaning module 4, the shaking module 5, the suction filtration module 6, the incubation module 7 and the detection module 8 are located at the same level and are all located below the mechanical cantilever module 3; the sample culture module 2 is movably arranged in the bacterial diagnosis device, and the position of the sample culture module 2 can be adjusted according to needs.

[0100] In the embodiment of the present application, the sample chamber module 1 is used to provide an environment required for storing samples; the sample culture module 2 is used to provide an environment required for culture samples; the mechanical cantilever module 3 is used to transfer the sample in the sample chamber module 1 to the sample culture module 2, and to carry the sample culture module 2; the cleaning module 4 is used to clean the device for transferring samples in the mechanical cantilever module 3; the shaking module 5 is used to shake the sample in the sample culture module 2; the filtration module 6 is used to generate a pressure difference inside the sample culture module 2, so as to achieve the material separation of the sample in the sample culture module 2; the incubation module 7 is used to provide a constant temperature environment required for incubating the sample in the sample culture module 2; the detection module 8 is used to detect the fluorescence luminescence value of the sample in the sample culture module 2. Each functional module cooperates with each other in the whole device to realize the functions of sample addition, shaking, filtration, incubation, detection, etc., and realize full-function automatic integrated sample detection.

[0101] In the embodiment of the present application, the bacterial diagnosis device further includes a hydraulic system, a cleaning liquid storage bin and a waste liquid storage bin. The hydraulic system, the cleaning liquid storage bin and the waste liquid storage bin are located at the same level, and are all located below the functional modules such as the sample bin module 1, the cleaning module 4, the shaking module 5, the filtration module 6, the incubation module 7 and the detection module 8, and cooperate with each functional module to play an auxiliary role.

[0102] Figure 2 This is a schematic diagram of a bacterial diagnostic device provided in an embodiment of the present application. Figure 2 ,like Figure 2As shown, the bacterial diagnostic device also includes a frame 9. The frame 9 includes a frame upper layer 91, a frame middle layer 92 and a frame lower layer 93 arranged from top to bottom. The mechanical cantilever module 3 is located in the frame upper layer 91; the sample chamber module 1, the cleaning module 4, the shaking module 5, the filtration module 6, the incubation module 7, and the detection module 8 are located in the frame middle layer 92; the hydraulic system, the cleaning liquid storage bin and the waste liquid storage bin are located in the frame lower layer 93.

[0103] In the embodiment of the present application, the sample culture module 2 includes a well plate 21 . Figure 3 is a front view of a hole plate 21 provided in an embodiment of the present application, Figure 4 is a cross-sectional schematic diagram of a hole plate 21 provided in an embodiment of the present application, such as Figure 3 and Figure 4 As shown, the orifice plate 21 includes an orifice plate body 211 and a plurality of sample-carrying holes 212. The orifice plate body 211 is mainly used for supporting the orifice plate 21, and actions such as clamping, placing, pushing and pulling are all performed on the orifice plate body 211. The orifice plate 21 in the embodiment of the present application is a 96-well plate 21, and the 96-well plate 21 has a total of 96 sample-carrying holes 212 distributed in an 8×12 array in the middle position of the 96-well plate 21, and the sample-carrying holes 212 are used to place samples. The bottom of each sample-carrying hole 212 is a microporous filter membrane 2121, and the microporous filter membrane 2121 has tiny holes for material separation of the sample. In a static state, a suspension composed of culture fluid, bacteriophage, bacteria, etc. cannot penetrate the membrane body due to liquid tension and molecular membrane adsorption force; when the pressure difference between the two sides of the molecular membrane reaches a certain value, other small molecular substances other than microorganisms of the size of bacteria, such as culture fluid, bacteriophages that have not invaded bacteria, etc., can pass through the molecular membrane under pressure to separate from the bacteria.

[0104] Figure 5 is a schematic diagram of a sample chamber module 1 provided in an embodiment of the present application, such as Figure 5 As shown, the sample chamber module 1 includes a sample chamber fixing bracket 11 and a sample chamber drawer 12. The sample chamber fixing bracket 11 is a semi-enclosed structure, fixed to the middle layer of the rack 92. The sample chamber drawer 12 is embedded in the sample chamber fixing bracket 11 and can be pulled forward and backward.

[0105] Figure 6 is a schematic diagram of a sample compartment drawer 12 provided in an embodiment of the present application, such as Figure 6As shown, the sample bin drawer 12 includes at least one drawer chamber 121. In the embodiment of the present application, the sample bin drawer 12 includes two drawer chambers 121. In actual needs, the sample bin drawer 12 can be divided into more or a single drawer chamber 121. A sealed cover 1211 and a cover driving mechanism 1212 are arranged on the top of each drawer chamber 121. The cover driving mechanism 1212 drives the corresponding sealed cover 1211 to close or open the top of each drawer chamber 121, so as to facilitate the individual addition of samples to the corresponding drawer chamber 121. A temperature control module 1213 is arranged inside each drawer chamber 121, so that the independent drawer chamber 121 can adjust its adapted temperature according to different samples. At the same time, the small space structure is conducive to the stability of temperature control and improves the temperature control sensitivity. One or more material boxes 1214 are arranged inside each drawer chamber 121 for storing samples.

[0106] Figure 7 1 is a schematic diagram of a material box 1214 provided in an embodiment of the present application, such as Figure 7 As shown, the material box 1214 adopts a dual gradient storage chamber design, the upper part of the material box 1214 is provided with a sample addition port 12141 and a sampling port 12142, and the interior of the material box 1214 is provided with an upper sample storage chamber 12143 for storing large-volume samples, and a lower small sample storage chamber 12144 for storing small-volume samples. In actual needs, a multi-stage gradient design can be adopted to divide the sample storage area according to the sample volume.

[0107] Figure 8 is a schematic diagram of a mechanical cantilever module 3 provided in an embodiment of the present application, such as Figure 8 As shown, the mechanical cantilever module 3 includes a mechanical cantilever fixing frame 31, a mechanical gripper left and right driving mechanism 32, a mechanical gripper mechanism 33, a row of needles left and right driving mechanism 34 and a row of needles sample loading mechanism 35. The mechanical cantilever fixing frame 31 is fixed to the middle layer 92 of the frame, and the mechanical gripper left and right driving mechanism 32 and the row of needles left and right driving mechanism 34 are fixed on the mechanical cantilever fixing frame 31. The mechanical gripper mechanism 33 is connected to the mechanical gripper left and right driving mechanism 32, and the mechanical gripper left and right driving mechanism 32 is used to drive the mechanical gripper mechanism 33 to carry the 96-well plate 21 horizontally left and right at all stations. The row of needles sample loading mechanism 35 is connected to the row of needles left and right driving mechanism 34, and the row of needles left and right driving mechanism 34 is used to drive the row of needles sample loading mechanism 35 to switch back and forth in the left and right horizontal directions at the stations between the sample chamber module 1, the cleaning module 4, and the shaking module 5 to realize the cycle of sampling, sample loading, cleaning, etc.

[0108] Fig. 9 is a schematic diagram of a mechanical gripper mechanism 33 provided in an embodiment of the present application, such as Fig. 9As shown, the manipulator gripper mechanism 33 includes a manipulator gripper fixing frame 331, a manipulator gripper front and rear driving mechanism 332, a manipulator gripper up and down driving mechanism 333, a manipulator gripper grabbing driving mechanism 334 and a manipulator gripper 335. The manipulator gripper fixing frame 331 is fixed to the manipulator gripper left and right driving mechanism 32, and is driven to move by the manipulator gripper left and right driving mechanism 32; the manipulator gripper front and rear driving mechanism 332 is fixed to the moving part of the manipulator gripper fixing frame 331, and is used to drive the manipulator gripper up and down driving mechanism 333 to move forward and backward longitudinally; the manipulator gripper up and down driving mechanism 333 is fixed to the moving part of the manipulator gripper front and rear driving mechanism 332, and is used to drive the manipulator gripper grabbing driving mechanism 334 to move up and down; the manipulator gripper grabbing driving mechanism 334 is fixed to the moving part of the manipulator gripper up and down driving mechanism 333, and is used to drive the manipulator gripper 335 fixed to its lower end to open and clamp. The driving mechanisms work together to ensure that the robot gripper 335 can grasp and place the 96-well plate 21 at any station.

[0109] Fig.10 is a schematic diagram of a needle-pipette loading mechanism 35 provided in an embodiment of the present application, such as Fig.10 As shown, the coupling needle loading mechanism 35 includes a coupling needle fixing frame 351, a coupling needle up and down driving mechanism 352, a coupling needle 353 and a hydraulic pipeline 354. The rear end of the coupling needle fixing frame 351 is fixed to the moving end of the coupling needle left and right driving mechanism 34; the coupling needle up and down driving mechanism 352 is fixed to the front end of the coupling needle fixing frame 351; and the coupling needle 353 is fixed to the moving end of the coupling needle up and down driving mechanism 352. The hydraulic pipeline 354 is relatively long, one end of the inner channel is connected to the coupling needle 353, and the other end of the peristaltic pump group passing through the lower layer 93 of the frame is connected to the cleaning liquid storage bin (the cleaning liquid is generally distilled water), and the middle pipeline line is fixedly installed along the coupling needle fixing frame 351 and the frame 9. The hydraulic pipeline 354 is a plastic pipe, which has a certain hardness to ensure its vertical support hardness, and has a certain flexibility so that it can also move in the semicircular bending part, thereby realizing the up and down hydraulic pressure of the coupling needle 353. Under the action of each driving mechanism, the row of needles 353 can extract samples from top and bottom, move horizontally to the initial placement station of the 96-well plate 21 for sample addition, and move to the cleaning module 4 station for cleaning. Under the action of the hydraulic system, it can absorb and spray samples, and clean the inner wall of the pipeline and the outer wall of the needle body.

[0110] Fig.11 is a schematic diagram of a cleaning module 4 provided in an embodiment of the present application, such as Fig.11As shown, the cleaning module 4 includes a cleaning base 41, a row of fountain cleaning ports 42, a waste liquid tank 43 and a waste liquid outlet 44. The cleaning base 41 is fixed to the middle layer 92 of the frame, and is used to support the cleaning module 4. A trough body is fixed on the upper part of the cleaning base 41, and a row of fountain cleaning ports 42 for cleaning the row of needles 353 are arranged in the middle of the trough body. The row of fountain cleaning ports 42 include a plurality of spray holes, and the bottom of each spray hole is solid and has a certain depth. After completing a sampling, the row of needles 353 will contain residual samples inside and on the outer wall, which need to be cleaned before the next sampling. The row of needles 353 containing residual samples are moved to the row of fountain cleaning ports 42 under the drive of the mechanical cantilever module 3, and each needle hole in the row of needles 353 corresponds to a spray hole in the row of fountain cleaning ports 42. The row of needles 353 draws cleaning liquid from the cleaning liquid storage bin through a hydraulic system such as a peristaltic pump to clean its interior and exterior walls, and the discharged water hits the bottom surface of each corresponding spray hole; when the drainage pressure is sufficient, the discharged water hits the bottom surface of the spray hole, causing the water column to surge in the direction of the spray hole to form a fountain of a certain height, thereby cleaning the interior and exterior walls of the row of needles 353. The waste liquid after cleaning the row of needles 353 flows into the waste liquid tank 43 located around the row of fountain cleaning ports 42, and then is discharged into the waste liquid storage bin through the waste liquid outlet 44. The entire cleaning module 4 utilizes the backflow principle and makes full use of the hydraulic pipeline system of the row of needles 353, with a simple and compact structure and low cost.

[0111] Fig.12 is a schematic diagram of a shaking module 5 provided in an embodiment of the present application, such as Fig.12As shown, the shaking module 5 includes a shaking module base 51, a shaking station 52, a cover station 53, a well plate cover 54, and a shaking drive mechanism 55. The shaking module base 51 is fixed to the middle layer 92 of the frame, and is used to support the entire shaking module 5. The shaking station 52 is set on the front side of the shaking module base 51. The shaking station 52 is a placement station for the 96-well plate 21 during the shaking process, and is also the initial placement station for the 96-well plate 21, and is also the sample adding station for the 96-well plate 21. The shaking station 52 has multiple functions such as initial placement, sample adding, and shaking, which integrates multiple needs into the same station to achieve reuse, effectively reducing the complexity of the overall structure, saving equipment space, and reducing the size of the equipment. A shaking drive mechanism 55 is fixed at the bottom of the shaking station 52. The shaking drive mechanism 55 adopts an eccentric wheel mechanism to drive the shaking station 52 to vibrate back and forth on the horizontal plane, thereby driving the 96-well plate 21 on the shaking station 52 to move, so as to achieve the purpose of shaking the sample in the 96-well plate 21. A cover station 53 placed in parallel with the shaking station 52 is arranged on the rear side of the shaking module base 51, and the well plate cover 54 of the 96-well plate 21 is stacked on the cover station 53. A plurality of protrusions corresponding to the plurality of sample loading holes 212 of the 96-well plate 21 are arranged on the well plate cover 54. Before shaking the sample, the well plate cover 54 is buckled on the 96-well plate 21 by the mechanical gripper 335, so that the plurality of protrusions seal the corresponding sample loading holes 212 on the 96-well plate 21 one by one, so as to prevent the sample from vibrating and overflowing during the shaking process, thereby preventing the occurrence of problems such as sample crosstalk or contamination of the table.

[0112] Fig.13 is a schematic diagram of a filtration module 6 provided in an embodiment of the present application, such as Fig.13 As shown, the filtration module 6 includes a filtration module fixing frame 61, a vacuum pump 62, a pressure regulating disk 63, a negative pressure bin 64 and a filtration station 65. The filtration module fixing frame 61 is fixed to the middle layer 92 of the frame, and is used to support the entire filtration module 6. The vacuum pump 62 is used to provide negative pressure power. The pressure regulating disk 63 is used to adjust the pressure of the vacuum pump 62. An air suction port 641 is provided on the lower side of the negative pressure bin 64, and the air suction port 641 is connected to the vacuum pump 62 through an air pipe. The filtration station 65 is arranged on the upper side of the negative pressure bin 64 and is connected to the upper side of the negative pressure bin 64.

[0113] Fig.14 is a schematic diagram of a filtration station 65 provided in an embodiment of the present application, such as Fig.14As shown, the filtration station 65 includes an external filtration limit block 651 and an internal filtration limit groove 652. The filtration limit block 651 is used to place the 96-well plate 21, so as to control the position of the 96-well plate 21. The filtration limit groove 652 has no bottom surface, only the side coated with soft glue. During the filtration process, the 96-well plate 21 is placed on the filtration limit block 651, and the bottom surface of the 96-well plate 21, the side of the filtration limit groove 652 and the negative pressure bin 64 form a confined space, so as to generate a pressure difference within the 96-well plate 21 with the microporous filter membrane 2121 as the boundary, thereby realizing the material separation of the sample. A filtration waste liquid box 642 is arranged inside the negative pressure bin 64 and directly below the filtration station 65. There is a certain space between the filtration waste liquid box 642 and the bottom and side surfaces of the 96-well plate 21 on the filtration station 65, and the space forms an air gap 66 during the filtration process, so that the bottom surface of the 96-well plate 21 and the interior of the negative pressure bin 64 are connected, which facilitates the formation of negative pressure on the bottom surface of the 96-well plate 21, and does not hinder the collection of the waste liquid filtered out. The bottom surface of the filtration waste liquid box 642 is provided with a filtration waste liquid hole 6421, which is connected to the waste liquid storage bin of the lower layer 93 of the rack through a water pipe through a one-way throttle valve, so the filtration waste liquid can be discharged to the waste liquid storage bin through the filtration waste liquid hole 6421.

[0114] Fig.15 is a schematic diagram of an incubation module 7 provided in an embodiment of the present application, such as Fig.15 As shown, the incubation module 7 includes an incubation module base 71, an incubation bin 72 and a sample transfer robot 73. The incubation module base 71 is fixed to the middle layer 92 of the frame, and is used to connect and support the incubation bin 72 and the sample transfer robot 73 located above the incubation module base 71. The incubation bin 72 is decomposed into a plurality of incubation chambers 721. In the embodiment of the present application, the incubation bin 72 is decomposed into two incubation chambers 721. In the actual design, it can be adjusted to a single or multiple incubation chambers 721 according to the temperature requirements. A single or multiple incubation stations 7212 are arranged in each incubation chamber 721, and the incubation stations 7212 are used to place the 96-well plate 21 and incubate the samples in the 96-well plate 21. A sealed sliding door mechanism 7211 is separately arranged at the front of each incubation chamber 721, which is convenient for taking and delivering samples of the same incubation temperature and duration. The sample transfer robot 73 is mainly composed of an incubation waiting station 731 and an incubation feeding mechanism 732. The incubation waiting station 731 is fixed to the moving end of the incubation feeding mechanism 732, and is used to place the 96-well plate 21 waiting for incubation. The incubation feeding mechanism 732 is a three-freedom motion module that can move horizontally, vertically, and up and down, and can push the 96-well plate 21 on the incubation waiting station 731 to any incubation station 7212. The 96-well plate 21 to be incubated is placed on the incubation waiting station 731 by the mechanical gripper 335, and then the 96-well plate 21 on the incubation waiting station 731 is transferred to the corresponding incubation station 7212 by the incubation feeding mechanism 732.

[0115] Fig.16 is a schematic diagram of a detection module 8 provided in an embodiment of the present application, such as Fig.16 As shown, the detection module 8 includes a detection module fixing frame 81, a push-pull mechanism 82 and a fluorescence detection device 83. The detection module fixing frame 81 is fixed to the frame middle layer 92, and is used to connect and support the push-pull mechanism 82 and the fluorescence detection device 83 placed adjacently on the detection module fixing frame 81.

[0116] Fig.17 is a schematic diagram of a push-pull mechanism 82 provided in an embodiment of the present application, such as Fig.17 As shown, the push-pull mechanism 82 includes a push-pull mechanism bottom plate 821, a push-in drive mechanism 822, a pull-out drive structure 823, a station to be tested 824, a push sheet 825, a pull plate 826, and a pull-out hopper mechanism 827. The push-in drive mechanism 822 and the pull-out drive structure 823 are fixed on the push-pull mechanism bottom plate 821, the push sheet 825 is fixed on the moving end of the push-in drive mechanism 822, the pull plate 826 is fixed on the moving end of the pull-out drive mechanism, and the pull-out hopper mechanism 827 is fixed on the pull plate 826. During the warehouse entry detection, the manipulator claw 335 places the 96-well plate 21 on the station to be tested 824, and the push-in drive mechanism 822 pushes the 96-well plate 21 into the entrance of the fluorescence detection device 83 through the push sheet 825; after the detection is completed, the pull-out drive structure 823 pushes the 96-well plate 21 out of the fluorescence detection device 83 through the pull plate 826.

[0117] Fig.18 Schematic diagram of a pull-out hopper mechanism 827 provided in an embodiment of the present application, such as Fig.18 As shown, the pull-out hopper mechanism 827 includes a hopper base 8271, a hopper shaft 8272, an eccentric pull block 8273, and a limiting protrusion 8274. When the 96-well plate 21 is pushed out from the fluorescence detection device 83, the front side arm of the 96-well plate 21 will push through the eccentric pull block 8273. During the pushing process, the eccentric pull block 8273 rotates counterclockwise under the thrust, so that the front side wall of the 96-well plate 21 is smoothly pushed out. When the front side wall of the 96-well plate 21 passes through the eccentric pull block 8273, the front side wall of the 96-well plate 21 will stand up to the initial state position under the action of self-generated gravity, and at this time, the pull-out drive structure 823 will drive the pull-out hopper mechanism 827 on the pull plate 826 to move. The eccentric pull block 8273 of the pull-out hopper mechanism 827 cannot rotate clockwise in the initial position due to the limiting protrusion 8274 at the bottom, so it will pull the front side wall of the 96-well plate 21 to move together, pulling the 96-well plate 21 to the test station 824, waiting for the robot claw 335 to take away the sample.

[0118] Fig.19 is a schematic diagram of a fluorescence detection device 83 provided in an embodiment of the present application, such as Fig.19As shown, the fluorescence detection device 83 includes a feeding and discharging device 831 and a photon counting device 832. The feeding and discharging device 831 is used for the movement connection of the 96-well plate 21 between the fluorescence detection device 83 and the push-pull mechanism 82, so as to realize the position transfer of the well plate 21 between the fluorescence detection device 83 and the push-pull mechanism 82. The photon counting device 832 is used to determine the fluorescence luminescence value of the sample in the sample loading well 212 of the well plate 21, so as to achieve the purpose of bacterial diagnosis.

[0119] In the embodiment of the present application, each functional module cooperates with each other in the entire device to realize functions such as sample addition, shaking, filtration, incubation, and detection, and realize full-function automatic integrated sample detection. The mechanical claw 335 in the mechanical cantilever module 3 grabs and moves the 96-well plate 21 to the corresponding operating station, effectively connecting the various functional modules. The row of needles 353 in the mechanical cantilever module 3 realizes the transportation of liquid samples and realizes fully automatic sample addition. The fountain-type structure in the cleaning module 4 realizes the synchronous cleaning of the inner and outer walls of the row of needles 353. The sample bin module 1 adopts an independent closed constant temperature design (adjustable temperature) to ensure sample cleanliness and the activity of bacteria during the detection process. At the same time, the sample material box 1214 adopts a high and low structure to adapt to the storage of samples of different volumes, so that the depth of samples of different volumes is sufficient for the row of needles 353 to absorb. The shaking module 5 adopts a simple crankshaft mechanism to achieve sample shaking. The filtration module 6 uses a vacuum pump 62 combined with a double-layer vacuum tank structure to generate a pressure difference between the top and bottom of the 96-well plate 21, realize negative pressure filtration, and achieve the purpose of material separation. In the incubation module 7, the separately separated incubation chamber 72 facilitates the precise control of the incubation time of each well plate 21 and the precise control of the temperature; the sample transfer robot 73 places the sample to be tested on the incubation station 7212 into the appropriate incubation chamber 72. In the detection module 8, the push-pull mechanism 82 pushes and pulls the 96-well plate 21 of the culture sample into and out of the fluorescence detection device 83, and the fluorescence detection device 83 detects the fluorescence luminescence value of each well position of the 96-well plate 21 to achieve the purpose of bacterial diagnosis. Among them, the pinholes of the row of needles 353 in the row of needles loading mechanism 35, the loading holes of the sample chamber module 13, the cleaning holes in the cleaning module 4, and the holes of the 96-well plate 21 placed horizontally on the shaking station 52 in the shaking module 5, the holes of these modules are in a straight line in the horizontal direction and correspond to each other, so that the row of needles 353 only needs to move horizontally and up and down in two degrees of freedom to complete the movement of its required function, reducing the movement in the front and back directions, making the equipment more concise and efficient. At the same time, the heights of the stations of these modules are similar, reducing the distance of the mechanical cantilever module 3 moving up and down, and improving work efficiency. Compared with the prior art, the present application has a simple structure, high control accuracy, low contamination rate, fast detection speed, high degree of automation, and realizes full-process unmanned automatic sampling. The operator only needs to load the sample and then wait for the result output.

[0120] The present application embodiment provides a method for detecting clinical tuberculosis mycobacterium using the bacterial diagnostic device, the steps of which are as follows:

[0121] S1. Offline pre-treatment, that is, centrifuging the treated clinical samples, removing the supernatant, adding 7H9C culture medium, and generating the samples to be tested;

[0122] S2. Add the sample to be tested, 7H9C culture medium containing Mycobacterium smegmatis, 7H9C culture medium, bacteriophage, bacteriophage inactivator, etc. into five material boxes 1214 respectively, place them in the corresponding sample bin drawers 12, seal the sample bin drawer 12, and adjust the temperature of the corresponding solvent adaptation.

[0123] S3. Set the detection path and corresponding time in the software (usually the factory default settings), and the corresponding mechanism can perform the corresponding movement automatically.

[0124] S4. Place the 96-well plate 21 at the initial placement station (i.e., the shaking station 52); click Start Detection on the software interface; the automatic detection process is as follows.

[0125] S5. The needle row 353 moves to the sample compartment module 1, extends the needle head downward, and absorbs 200 μl of the sample to be tested; the needle row 353 moves horizontally to the shaking station 52, and transfers 200 μl of the sample to be tested to the sample loading well 212 of the 96-well plate 21; then the needle row 353 moves to the cleaning module 4 to clean the pipeline;

[0126] S6. The needle row 353 moves to the position of the sample chamber module 1, extends the needle head downward, and absorbs 20 μl of bacteriophage; the needle row 353 moves horizontally to the shaking station 52, and transfers 20 μl of bacteriophage to the sample loading well 212 of the 96-well plate 21 with the sample to be tested; then the needle row 353 moves to the cleaning module 4 to clean the pipeline, and then returns to the initial position;

[0127] S7. The robot gripper 335 moves, grabs the 96-well plate 21 on the shaking station 52, transports it to the incubation waiting station 731, opens the sealed sliding door mechanism 7211 of the incubation chamber 721, and after the incubation feeding mechanism 732 feeds the 96-well plate 21 to the incubation station 7212, the incubation feeding mechanism 732 returns to its original position, and the sealed sliding door mechanism 7211 of the incubation chamber 721 is closed. Set the temperature and duration (set in advance in the software), and incubate the 96-well plate 21 at 37 degrees Celsius for 6 hours to facilitate the infection of Mycobacterium tuberculosis by bacteriophages;

[0128] S8. After the incubation time is up, the sealed sliding door mechanism 7211 of the incubation chamber 721 opens, and the incubation feeding mechanism 732 takes the 96-well plate 21 out of the incubation station 7212 to the incubation waiting station 731, and takes the 96-well plate 21 out of the warehouse and returns to the original position. At this time, the sealed sliding door mechanism 7211 of the incubation chamber 721 is synchronously closed to reduce external pollution; the robot gripper 335 moves to the incubation waiting station 731, grabs the incubated 96-well plate 21, and transports it to the filtration station 65. After the filtration station 65 detects the 96-well plate 21, it performs negative pressure filtration to remove the culture medium and most of the bacteriophages;

[0129] S9. After the filtration is completed, the manipulator gripper 335 moves to transport the 96-well plate 21 from the filtration station 65 to the shaking station 52. When the manipulator gripper 335 returns to its original position, the row of needles 353 moves to the position of the sample compartment module 1, extends the needle head downward, and absorbs the bacteriophage inactivator; the row of needles 353 moves horizontally to the shaking station 52, transfers the bacteriophage inactivator to the sample loading hole 212 in the 96-well plate 21 with the sample, and then the row of needles 353 moves to the cleaning module 4 to clean the pipeline ... 5 moves to grab the well plate cover 54, covers the 96-well plate 21 with the well plate cover 54, the robot claw 335 opens and rises and detaches from the well plate cover 54, then the shaking module 5 works to mix the sample, let it stand for 1 minute, then the robot grabs and moves to transport the well plate cover 54 to the cover station 53, and then the 96-well plate 21 is transported to the filtration station 65, the robot claw 335 opens and rises and detaches from the 96-well plate 21, after the sample is detected, the filtration module 6 starts to work, and completely inactivates and separates the bacteriophages that are not infected with Mycobacterium tuberculosis;

[0130] S10. After the filtration is completed, the manipulator gripper 335 transports the 96-well plate 21 to the shaking station 52, and then returns to the original position. While the manipulator gripper 335 returns to its original position, the row of needles 353 moves to the position of the sample compartment module 1, extends the needle head downward, and absorbs the 7H9C culture medium containing Mycobacterium smegmatis; the row of needles 353 moves horizontally to the shaking station 52, transfers the 7H9C culture medium containing Mycobacterium smegmatis to the sample loading hole 212 in the 96-well plate 21 with the sample, and then the row of needles 353 moves to the cleaning module 4 to clean the pipeline. While returning to the initial position, the manipulator gripper 335 moves to the shaking station 5 2. Grab the 96-well plate 21 and transport it to the filtration station 65 for filtration to remove the residual phage inactivator; after the filtration is completed, the manipulator gripper 335 transports the 96-well plate 21 to the shaking station 52, and then the manipulator gripper 335 returns to its original position. While it returns to its original position, the row of needles 353 moves to add 200 microliters of 7H9C culture medium to the corresponding sample loading well 212 in the 96-well plate 21 (the sample addition movement process is the same as described above). When the row of needles 353 returns to its original position, the manipulator gripper 335 moves synchronously to transport the 96-well plate 21 to the incubation module 7 for incubation at 30 degrees Celsius (the incubation movement process is the same as described above);

[0131] S11. After the incubation is completed, the robot gripper 335 transports the 96-well plate 21 from the incubation waiting station 731 to the test station 824 of the detection module 8. After the test station 824 detects the incoming material, the door of the photon counting device 832 opens, the loading and unloading device 831 extends, and then the push-pull mechanism 82 pushes the 96-well plate 21 into the loading and unloading device 831. After the loading and unloading device 831 detects that the 96-well plate 21 is in place, it returns to the photon counting device 832, and the photon counting device 832 closes the door. Thereafter, the photon counting device 832 performs fluorescence detection and counting on the sample wells and the control blank wells, and outputs the fluorescence value report of each well in the software. If the luminescence value of the sample well is greater than that of the blank template, the sample is positive, otherwise it is negative.

[0132] S12. After the detection is completed, the photon counting device 832 opens the hatch, the loading and unloading device 831 sends out the 96-well plate 21, the push-pull mechanism 82 moves the 96-well plate 21 to the test station 824, and then the loading and unloading device 831 returns to the photon counting device 832, and the photon counting device 832 closes the hatch. At the same time, the robot claw 335 moves to move the 96-well plate 21 on the test station 824 to the shaking station 52, and then returns to its original position. The staff will take out the 96-well plate 21 from the equipment for post-processing.

[0133] An embodiment of the present application provides a bacterial resistance detection device, including any one of the above-mentioned bacterial diagnostic devices, for quantitatively detecting bacterial resistance.

[0134] The present application embodiment also provides a method for screening drug-resistant bacteria using the bacterial resistance detection device, and the steps are as follows: (the specific movement processes such as adding samples, incubating, filtering, detecting, and shaking are the same as the above-mentioned method for detecting clinical tuberculosis mycobacterium using a bacterial diagnostic device, the difference being that the reagents added and the order are different)

[0135] S1. Cultivate clinical drug-resistant bacteria, add 7H9C medium, disperse, and add 200 μl of bacterial solution to each loading well 212 in the 96-well plate 21;

[0136] S2. 10 μl of different concentrations and types of anti-tuberculosis drugs were added to each loading well 212, and 20 μl of bacteriophage was added. The loading well 212 without adding antimicrobial drugs was used as a positive reference;

[0137] S3. Incubate the 96-well plate 21 at 37 degrees Celsius for 6 hours to allow the phage to infect Mycobacterium tuberculosis;

[0138] S4. The incubated 96-well plate 21 is subjected to negative pressure filtration to remove the culture medium and most of the phages and drugs;

[0139] S5. Add a bacteriophage inactivator to the 96-well plate 21, mix well, let stand for 1 minute, and filter to completely inactivate the bacteriophage that is not infected with Mycobacterium tuberculosis and wash the antibacterial drug;

[0140] S6. Add 7H9C medium containing Mycobacterium smegmatis to the 96-well plate 21, and filter to remove the phage inactivating agent;

[0141] S7. Add 200 μl of 7H9C medium to the 96-well plate 21 and transfer to the instrument for incubation at 30 degrees Celsius;

[0142] S8. Perform optical detection on the 96-well plate 21, compare the detected luminescence value with the positive reference, and use the calculation formula to obtain the final drug resistance result of Mycobacterium tuberculosis.

[0143] It should be noted that the above-mentioned sequence of the embodiments of the present application is for description only and does not represent the advantages and disadvantages of the embodiments. The above-mentioned specific embodiments of this specification are described. Other embodiments are within the scope of the attached claims. In some cases, the actions or steps recorded in the claims can be performed in an order different from that in the embodiments and still achieve the desired results. In addition, the processes depicted in the drawings do not necessarily require the specific order or continuous order shown to achieve the desired results. In some embodiments, multitasking and parallel processing are also possible or may be advantageous.

[0144] Each embodiment in this specification is described in a progressive manner, and the same or similar parts between the embodiments can be referred to each other, and each embodiment focuses on the differences from other embodiments. In particular, for the device embodiment, since it is basically similar to the method embodiment, the description is relatively simple, and the relevant parts can be referred to the partial description of the method embodiment.

[0145] A person skilled in the art will understand that all or part of the steps to implement the above embodiments may be accomplished by hardware or by instructing related hardware through a program, and the program may be stored in a computer-readable storage medium, and the above-mentioned storage medium may be a read-only memory, a disk or an optical disk, etc.

[0146] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the protection scope of the present application.

Claims

1. A bacterial diagnostic device, characterized in that: It includes a sample chamber module, a sample culture module, a mechanical cantilever module, a cleaning module, a shaking module, a filtration module, an incubation module and a detection module; The sample chamber module, the cleaning module, the shaking module, the filtration module, the incubation module and the detection module are located at the same level and are all located below the mechanical cantilever module; the sample culture module is movably arranged in the bacterial diagnosis device; The sample bin module is used to provide an environment required for storing samples; the sample culture module is used to provide an environment required for culturing the samples; the mechanical cantilever module is used to transfer the samples in the sample bin module to the sample culture module, and to carry the sample culture module; the cleaning module is used to clean the device for transferring samples in the mechanical cantilever module; the shaking module is used to shake the samples in the sample culture module; the filtration module is used to generate a pressure difference inside the sample culture module, thereby realizing the material separation of the samples in the sample culture module; the incubation module is used to provide a constant temperature environment required for incubating the samples in the sample culture module; the detection module is used to detect the fluorescence luminescence value of the samples in the sample culture module.

2. The bacterial diagnostic device according to claim 1, characterized in that: The sample culture module includes a well plate; The orifice plate comprises an orifice plate body and a plurality of sample loading holes; The sample loading hole comprises a microporous filter membrane at the bottom; the microporous filter membrane is used for substance separation of the sample.

3. The bacterial diagnostic device according to claim 2, characterized in that: It also includes a hydraulic system, a cleaning fluid storage bin, and a waste fluid storage bin; The hydraulic system, the cleaning liquid storage bin and the waste liquid storage bin are located at the same level, and are all located below the sample bin module, the cleaning module, the shaking module, the filtration module, the incubation module and the detection module.

4. The bacterial diagnostic device according to claim 3, characterized in that: It also includes a rack; the rack includes an upper rack layer, a middle rack layer and a lower rack layer arranged from top to bottom; The mechanical cantilever module is located at the upper layer of the rack; the sample chamber module, the cleaning module, the shaking module, the filtration module, the incubation module, and the detection module are located at the middle layer of the rack; the hydraulic system, the cleaning liquid storage chamber, and the waste liquid storage chamber are located at the lower layer of the rack.

5. The bacterial diagnostic device according to claim 4, characterized in that: The mechanical cantilever module includes a mechanical cantilever fixing frame, a mechanical gripper left and right driving mechanism, a mechanical gripper mechanism, a connecting needle left and right driving mechanism and a connecting needle sample loading mechanism; The mechanical cantilever fixing frame is fixed to the middle layer of the frame; the mechanical gripper left and right driving mechanism and the row of needles left and right driving mechanism are fixed to the mechanical cantilever fixing frame; the mechanical gripper mechanism is connected to the mechanical gripper left and right driving mechanism; the row of needles sample loading mechanism is connected to the row of needles left and right driving mechanism; The left and right driving mechanism of the manipulator gripper is used to drive the manipulator gripper mechanism, thereby realizing the transportation of the well plate in the left and right directions; the left and right driving mechanism of the connecting needle is used to drive the connecting needle loading mechanism, thereby realizing the transfer of the sample in the left and right directions.

6. The bacterial diagnostic device according to claim 5, characterized in that: The mechanical gripper mechanism comprises a mechanical gripper fixing frame, a mechanical gripper front and rear driving mechanism, a mechanical gripper up and down driving mechanism, a mechanical gripper grasping driving mechanism and a mechanical gripper; The mechanical gripper fixing frame is fixed on the left and right driving mechanism of the mechanical gripper, and is driven to move by the left and right driving mechanism of the mechanical gripper; The front and rear driving mechanism of the manipulator gripper is fixed on the moving part of the manipulator gripper fixing frame, and is used to drive the upper and lower driving mechanism of the manipulator gripper to move longitudinally forward and backward; The mechanical gripper up and down driving mechanism is fixed to the moving part of the mechanical gripper front and back driving mechanism, and is used to drive the mechanical gripper grasping driving mechanism to move up and down; The mechanical gripper grasping driving mechanism is fixed to the moving part of the mechanical gripper up and down driving mechanism, and is used to drive the movement of the mechanical gripper; The mechanical gripper is fixed to the end of the mechanical gripper grabbing driving mechanism.

7. The bacterial diagnostic device according to claim 5, characterized in that: The coupling needle sample loading mechanism comprises a coupling needle fixing frame, a coupling needle up and down driving mechanism, a coupling needle and a hydraulic pipeline; The rear end of the coupling needle fixing frame is fixed to the moving end of the left and right driving mechanism of the coupling needle; The coupling needle up and down driving mechanism is fixed at the front end of the coupling needle fixing frame; The coupling needle is fixed to the moving end of the coupling needle upper and lower driving mechanism; The hydraulic pipeline includes a first end and a second end; the first end of the hydraulic pipeline is connected to the coupling needle, and the second end of the hydraulic pipeline is connected to the cleaning liquid storage bin.

8. The bacterial diagnostic device according to claim 4, characterized in that: The sample bin module comprises a sample bin fixing bracket and a sample bin drawer; the sample bin fixing bracket is a semi-enclosed structure and is fixed to the middle layer of the rack; the sample bin drawer is embedded in the interior of the sample bin fixing bracket; The sample bin drawer comprises at least one drawer compartment; The drawer compartment includes an external sealed cover and a cover driving mechanism, as well as an internal temperature control module and at least one material box; the cover driving mechanism is used to close or open the corresponding sealed cover; The material box includes a sample addition port and a sampling port at the top, and an internal sample storage compartment and a small sample storage compartment; the sample storage compartment is used to store large-volume samples, and the small-sample storage compartment is used to store small-volume samples.

9. The bacterial diagnostic device according to claim 7, characterized in that: The cleaning module includes a cleaning base, a row of fountain cleaning ports, a waste liquid tank and a waste liquid outlet; The cleaning base is fixed to the middle layer of the frame; a trough is arranged on the upper part of the cleaning base; The row fountain cleaning port is located in the middle of the tank body and is used to clean the row needles; The waste liquid tank is located around the cleaning port of the row fountain, and is used to collect the waste liquid used to clean the row needles, and discharge the waste liquid used to clean the row needles to the waste liquid storage bin through the waste liquid outlet.

10. The bacterial diagnostic device according to claim 4, characterized in that: The shaking module includes a shaking module base, a shaking station, a cover station, a perforated plate cover, and a shaking drive mechanism; The shaking module base is fixed to the middle layer of the frame and is used to support the shaking module; The shaking station and the cover station are located on the shaking module base and are placed adjacent to each other; The well plate cover is placed on the cover station; the well plate cover is used to seal the well plate, thereby preventing the sample in the well plate from overflowing during the shaking process; The shaking drive mechanism is located below the shaking station; the shaking drive mechanism is used to drive the shaking station to vibrate on a horizontal plane, thereby shaking the sample in the well plate placed above the shaking station.

11. The bacterial diagnostic device according to claim 4, characterized in that: The filtration module includes a filtration module fixing frame, a vacuum pump, a pressure regulating plate, a negative pressure chamber and a filtration station; The filtration module fixing frame is fixed to the middle layer of the frame and is used to support the filtration module; The vacuum pump is used to provide negative pressure power; The pressure regulating disk is used to adjust the pressure of the vacuum pump; An air suction port is arranged at the lower side of the negative pressure chamber, and the air suction port is connected to the vacuum pump through an air pipe; the interior of the negative pressure chamber includes a filtration waste liquid box, and the bottom surface of the filtration waste liquid box includes a filtration waste liquid hole; the filtration waste liquid box is used to collect filtration waste liquid, and discharge the filtration waste liquid to the waste liquid storage chamber through the filtration waste liquid hole; The suction filtration station is arranged above the negative pressure bin and is in communication with the upper side of the negative pressure bin; the suction filtration station comprises an external suction filtration limit block and an internal suction filtration limit groove; the suction filtration limit block is used to place the orifice plate; The bottom surface of the orifice plate, the side surface of the filtration limiting groove and the negative pressure chamber form a closed space during the filtration process, so as to generate a pressure difference inside the orifice plate with the microporous filter membrane as the boundary, thereby realizing the material separation of the sample.

12. The bacterial diagnostic device according to claim 4, characterized in that: The incubation module comprises an incubation module base, an incubation chamber and a sample transfer robot arm; The incubation module base is fixed to the middle layer of the frame, and is used to connect and support the incubation chamber and the sample transfer robot arm located above the incubation module base; The incubation chamber includes at least one incubation chamber; the incubation chamber includes a sealed sliding door mechanism at the front and at least one incubation station inside; the incubation station is used to place the well plate and incubate the sample; The sample transfer robot arm includes an incubation waiting station and an incubation feeding mechanism, and the incubation waiting station is fixed to the end of the incubation feeding mechanism; the incubation waiting station is used to place the well plate waiting for incubation; the incubation feeding mechanism is used to transfer the well plate on the incubation waiting station to the incubation station.

13. The bacterial diagnostic device according to claim 4, characterized in that: The detection module includes a detection module fixing frame, a push-pull mechanism and a fluorescence detection device; The detection module fixing frame is fixed to the middle layer of the frame, and is used to connect and support the push-pull mechanism and the fluorescence detection device which are placed adjacently on the detection module fixing frame; The push-pull mechanism comprises a push-pull mechanism bottom plate, a push-in drive mechanism and a pull-out drive structure; the push-in drive mechanism and the pull-out drive structure are fixed on the push-pull mechanism bottom plate; the push-in drive mechanism is used to push the orifice plate into the fluorescence detection device; The pull-out driving structure is used to pull the orifice plate out of the fluorescence detection device; The fluorescence detection device includes a material loading and unloading device and a photon counting device; the material loading and unloading device is used to transfer the position of the well plate between the fluorescence detection device and the push-pull mechanism; the photon counting device is used to determine the fluorescence luminescence value of the sample in the sample loading hole of the well plate.

14. A bacterial resistance detection device, characterized in that: A bacterial diagnostic device comprising the device described in any one of claims 1 to 13.

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