Centrifugal machine and application thereof

Abstract

The invention discloses a centrifugal machine and application thereof, the centrifugal machine comprises a main body frame, a partition plate is arranged in the middle of the main body frame, and the centrifugal machine is divided into a centrifugal chamber and an accessory chamber by the partition plate; the rotor assembly is arranged above the partition plate, is positioned in the centrifugal chamber and is used for placing a sample; a refrigeration assembly is arranged on the outer wall of the centrifugal chamber and used for cooling or refrigerating the centrifugal chamber; the positioning assembly is arranged below the partition plate and located in the accessory chamber. When the centrifugal machine is used for carrying out separation and enrichment treatment on infectious samples, most of human cells, proteins, saccharides, salts and other impurities in uncultured or cultured clinical samples such as blood, cerebrospinal fluid, urine, hydrothorax and ascites and the like can be effectively removed, and relatively pure living pathogens are obtained.

Description

Technical Field

[0001] The present invention relates to the technical field of microbial detection, and in particular to a centrifuge and its application, and more particularly to a centrifuge and its application in extracting and enriching trace microorganisms from complex matrices. Background Art

[0002] Infectious diseases are common clinical diseases. Every year, 31.5 million people globally suffer from bloodstream infections, 5 million people suffer from meningitis, and half of women and one-twentieth of men globally suffer from urinary tract infections. In addition, abdominal and thoracic infections are also very common. Pathogen diagnosis is a key link in its diagnosis and treatment, and the conventional pathogen detection method is the microbial culture method. However, this method is not an ideal gold standard because it takes a long time (1 - 5 days), the results are often too late, the positive rate is low (the detection rate is less than 15%), the results are incomplete or not sensitive enough, and it may be misleading and relatively labor-intensive. Therefore, it is difficult to meet the clinical needs.

[0003] For infectious diseases, especially bloodstream infections, since the pathogen content in the sample is relatively low (1 - 10 CFU / mL), it usually takes 1 - 5 days for the diagnostic time from sampling from the patient to providing the infection result of the sample. Therefore, 19.4 million people globally experience severe bloodstream infections every year, and the average mortality rate reaches 16 - 40%. It is estimated that 3 million cases of neonatal sepsis occur globally every year, resulting in 1.2 million child deaths every year, with a mortality rate of 11 - 19%. In addition, more than 75,000 women die from postpartum sepsis every year. In American hospitals, sepsis is not only the most expensive disease to treat but also the main cause of death. There are up to 3.1 million cases of sepsis in the United States every year, with a cost of $24 billion, and the mortality rate is between 20% and 50%. In Europe, the probability of death due to sepsis is even higher, about 10 - 15% higher than that in the United States. Recent studies have shown that for patients with severe bloodstream infections or septic shock, without antibiotic treatment, the likelihood of death increases by 7.6% per hour. If antibiotics are not used within the first 6 hours of recognizing hypotension, the survival rate will also be greatly reduced. Therefore, shortening the diagnostic time can greatly improve the survival rate of patients. Therefore, a rapid early diagnosis method is the most reliable hope for improving the survival rate of patients with infectious diseases and limiting the disability of survivors, and a high-sensitivity, small-sample-demand product that can provide diagnostic-related information within a few hours is an urgent need in clinical practice.

[0004] Drug susceptibility testing is a method used to detect the drug resistance and sensitivity of bacteria or other microorganisms to antibiotics or other drugs. It is an important means to guide rational clinical drug use, reduce drug abuse, and improve treatment effects. Traditional drug susceptibility testing mainly includes the disk diffusion method, dilution method, and automated drug susceptibility testing systems, etc. To meet the needs of rapid clinical diagnosis, drug susceptibility testing needs to develop in a faster and more convenient direction. By adopting new technologies and methods, the time cycle of drug susceptibility testing can be shortened, providing more timely drug use guidance for clinicians. However, the results of drug susceptibility testing are often affected by various factors such as culture conditions and microbial species. Moreover, infection samples often need to be cultured before drug susceptibility testing can be carried out. The time for sample enrichment, isolation, and culture is relatively long, and it is impossible to quickly select the appropriate antibacterial drugs and reasonable antibacterial drug doses.

[0005] MALDI-TOF MS is a new type of soft ionization bio-mass spectrometry developed in recent years. Clinical microbiology laboratories have begun to possess this equipment and widely use it to identify a large number of bacteria and fungi after cultivation. The identification results are automatically interpreted, analyzed, reported, and the specimens are automatically unloaded throughout the process. Moreover, the detection cost is low. The consumables for the instrument only require sample plates and special mass spectrometry matrices, without any other additional reagents, and the technical requirements for staff are not high. Due to its high efficiency and accuracy in identification, and the low cost of each experiment, it has become a popular method for microbial identification.

[0006] Pathogen culture is still the gold standard for detecting infection samples. However, pathogen culture has many disadvantages. First, the diagnostic time is relatively long. Usually, it takes 1 to 5 days from sampling from the patient to obtaining the infection results of the sample. Then, performing drug susceptibility testing on the purified pathogen takes even longer. During this period, the common treatment method is to use broad-spectrum non-targeted antibiotics to treat the patient. This may achieve effects in treating the disease, but it will cause many microorganisms to develop multiple drug resistances. Second, the differences in pathogens in the sample lead to very different requirements for culture conditions. The dominant bacteria after cultivation are very likely not in the original state in the sample. Third, if the obtained infectious samples are used for drug susceptibility testing or MALDI-TOF MS detection, the sample pretreatment process is too complex. Pathogen culture requires a large amount of manpower, material resources, time, and money, with a relatively high cost, and also has relatively high requirements for operators, making it not easy to promote. Finally, the bacteria cultured after a long time are very likely to be contaminated bacteria, resulting in the inability to provide correct reference results for clinicians.

[0007] The MALDI-TOF MS mass spectrometer cannot currently be used for the direct detection of clinical specimens and mixed cultures. The analysis object must be a single, pure microorganism. Its highly accurate and reproducible identification of microorganisms is based on the detection of microbial ribosomal proteins (relative molecular mass 2000 - 20000), which are highly abundant proteins that are constitutively expressed intracellularly in microorganisms. Therefore, theoretically, different culture conditions will not cause significant differences in the identification results of MALDI-TOF MS. However, to achieve more ideal identification results, the laboratory currently needs to culture twice and then pick monoclonal colonies for MALDI-TOF MS identification, which is time-consuming and laborious. In addition, MALDI-TOF MS identification of bacteria usually requires a bacterial amount of 105 - 107 colony-forming units (CFU). Therefore, if direct detection is performed, samples in the initial stage of bloodstream infection or meningitis infection are likely to not enrich enough bacteria for detection or require too many samples. Moreover, if there is a mixed infection of more than two types in the sample, the detection results usually fail or are incorrect. Summary of the Invention

[0008] To address the above technical problems, the present invention provides a centrifuge, which includes a main body frame 6-1. A partition plate is provided in the middle of the main body frame, and the partition plate divides the centrifuge into a centrifugation chamber and an accessory chamber;

[0009] A rotor assembly 6-3 is provided above the partition plate, and the rotor assembly is located in the centrifugation chamber and is used for placing samples;

[0010] The outer wall of the centrifugation chamber is provided with a refrigeration assembly 6-4 for cooling or refrigerating the centrifugation chamber;

[0011] A positioning assembly 6-6 is provided below the partition plate, and the positioning assembly 6-6 is located in the accessory chamber.

[0012] According to an embodiment of the present invention, the centrifuge further includes a top cover assembly 6-2, and the top cover assembly is used for sealing or covering the centrifuge.

[0013] Preferably, one end of the top cover assembly is fixed to the main body frame 6-1.

[0014] According to an embodiment of the present invention, the centrifuge further includes a main shaft motor assembly 6-5, and the main shaft motor assembly is rotatably connected to the partition plate.

[0015] According to an embodiment of the present invention, the rotor assembly 6-3 includes a centrifuge head 6-8 and a cup hanging assembly 6-7. The centrifuge head is fixedly connected to the upper output shaft of the main shaft motor assembly 6-5, and the cup hanging assembly 6-7 is arranged at equal angles away from the center of the centrifuge head. The cup hanging assembly 6-7 is used for placing centrifuge tubes or sample tubes.

[0016] According to an embodiment of the present invention, the connection between the positioning component 6-6 and the main body frame 6-1 belongs to a known solution in the art.

[0017] The working principle of the centrifuge of the present invention is as follows:

[0018] First, the automatic door of the top cover component 6-2 is opened, then the positioning component 6-6 positions the empty hanging cup component on the rotor component 6-3 at the automatic door according to requirements, and then the consumables to be centrifuged (or centrifuge tubes) are placed into the designated hanging cup component 6-7. Then, the top cover component 6-2 operates to close the automatic door. Then, the main shaft motor component 6-5 starts to rotate and operate according to the setting. During this process, due to the friction between the rotor component 6-3 and the air in the cavity, the temperature will rise rapidly. At this time, the refrigeration component 6-4 starts to operate to reduce the temperature of the cavity until the set temperature is reached. When the centrifugation program is completed, the positioning component 6-6 will rotate the consumables to be taken out to the automatic door opening position according to the setting (wherein, the consumables to be taken out are controlled by the upper computer software), and then the automatic door on the top cover component 6-2 is opened to take out the centrifuged consumables.

[0019] The present invention also provides a workstation for extracting and enriching trace microorganisms from complex matrices. The workstation includes a main frame 1, a robotic arm, a mixing component, the above-mentioned centrifuge 6, a sample position component 17, a workbench component 15, and a reagent position component 14;

[0020] The robotic arm is located at the upper part of the main frame, and the robotic arm is controlled by a control system;

[0021] The mixing component, the sample position component 17, the workbench component 15, and the reagent position component 14 are arranged side by side in sequence in the middle of the main frame and are located below the robotic arm;

[0022] The centrifuge is located at the lower part of the main frame.

[0023] According to an embodiment of the present invention, the robotic arm includes a pipetting robotic arm 4 and a tube transferring and lid opening / closing robotic arm 5, and the pipetting robotic arm and the tube transferring and lid opening / closing robotic arm are arranged side by side. The pipetting robotic arm is used to add or remove liquid into or from a reagent tube. For example, the reagent tube is a sample tube or a centrifuge tube. The tube transferring and lid opening / closing robotic arm is used to move the reagent tube or to open or close the lid of the reagent tube.

[0024] According to an embodiment of the present invention, the mixing component includes an inversion mixing component 8 and a heating component 9.

[0025] Preferably, the mixing component further includes a shaking component 10.

[0026] Preferably, the inversion mixing assembly 8, the heating assembly 9, and the oscillation assembly 10 are placed side by side in sequence.

[0027] According to an embodiment of the present invention, a weighing platform assembly 11 is further provided in the middle of the main frame, and the weighing platform assembly 11 is placed side by side with the reagent position assembly 14. The weighing platform assembly 11 is used for balancing when centrifuge tubes enter the centrifuge.

[0028] The reagent position assembly 14 is used for placing additives. For example, the additives include at least one of lysis solution, microbial co-precipitant, washing solution, etc.

[0029] According to an embodiment of the present invention, at least one set of pipette tip position assemblies 12 is further provided in the middle of the main frame, and the pipette tip position assemblies 12 are placed side by side with the weighing platform assembly 11; the pipette tip position assemblies are used for placing consumable pipette tips. Preferably, there are at least two sets of pipette tip position assemblies in the middle of the main frame, including a 200 μL pipette tip position assembly and a 5 mL pipette tip position assembly 13. Preferably, the at least two sets of pipette tip position assemblies are respectively used for placing consumable 200 μL pipette tips and consumable 5 mL pipette tips.

[0030] According to an embodiment of the present invention, the workstation further includes a waste liquid and waste disposal position assembly 16, and the waste liquid and waste disposal position assembly 16 is located in the middle of the main frame 1. Preferably, the waste liquid and waste disposal position assembly 16 is placed side by side with the workbench assembly 15.

[0031] According to an embodiment of the present invention, the workstation further includes a waste liquid and waste collection bucket 7, which is located at the lower part of the main frame, and the waste liquid and waste collection bucket 7 is placed side by side with the centrifuge assembly 6 for collecting waste liquid or waste.

[0032] According to an embodiment of the present invention, an air filtration assembly 2 is further provided on the top of the main frame, and the air filtration assembly is used for filtering air.

[0033] According to an embodiment of the present invention, an operation screen 3 is further provided outside the main frame, and the operation screen 3 is used for controlling the robotic arm for subsequent experimental operations. Preferably, the operation screen 3 is arranged on the side outside the main frame 1, such as on the right side or the left side.

[0034] According to an embodiment of the present invention, a barcode scanner assembly 18 is further provided in the middle of the main frame, and the barcode scanner assembly is placed side by side with the mixing assembly. Preferably, the barcode scanner assembly is placed side by side with the inversion mixing assembly 8.

[0035] The present invention also provides a method for extracting and enriching trace microorganisms, and the method is implemented by using the above workstation. The method includes:

[0036] (1) Use a robotic arm to move the sample tube containing the blood sample to the mixing assembly to mix the sample. After mixing, use the robotic arm to move the mixed sample tube and the centrifuge tube to the workbench assembly 5 respectively;

[0037] (2) Use a robotic arm to transfer the sample in the sample tube to the centrifuge tube to obtain a centrifuge tube containing the blood sample;

[0038] (3) Use a robotic arm to pick up the auxiliary agent from the reagent position assembly and add it to the centrifuge tube containing the blood sample, and continue to use the robotic arm to move the centrifuge tube to the mixing assembly to mix the sample evenly;

[0039] (4) Use a robotic arm to move the centrifuge tube to the centrifuge assembly for centrifugation;

[0040] (5) After centrifugation, use a robotic arm to move the centrifuge tube to the workbench assembly, remove the supernatant, and complete the extraction and enrichment of trace microorganisms.

[0041] According to an embodiment of the present invention, in step (1), the robotic arm includes a pipetting robotic arm and a tube moving and lid opening / closing robotic arm; the pipetting robotic arm is used to add or remove liquid into or from the centrifuge tube. The tube moving and lid opening / closing robotic arm is used to move the centrifuge tube or to open or close the lid of the centrifuge tube.

[0042] According to an embodiment of the present invention, in step (1), the mixing assembly includes an inversion mixing assembly 8 and a heating assembly 9.

[0043] Preferably, the mixing assembly further includes a shaking assembly 10.

[0044] According to an embodiment of the present invention, step (1) further includes the process of collecting waste pipette tips; after the robotic arm picks up the blood sample from the reagent position assembly through the pipette tip and adds it to the centrifuge tube, the waste pipette tip is placed into the waste liquid and waste material collection bucket; the waste liquid and waste material collection bucket is arranged in the waste liquid and waste material throwing position assembly.

[0045] According to an embodiment of the present invention, in step (3), the robotic arm picks up the auxiliary agent from the reagent position assembly through a consumable pipette tip. Preferably, the pipette tip is placed in the pipette tip position assembly 12.

[0046] According to an embodiment of the present invention, step (3) further includes the process of collecting waste pipette tips; after the robotic arm picks up the auxiliary agent from the reagent position assembly through the pipette tip and adds it to the centrifuge tube containing the blood sample, the waste pipette tip is placed into the waste liquid and waste material collection bucket; the waste liquid and waste material collection bucket is arranged in the waste liquid and waste material throwing position assembly.

[0047] According to an embodiment of the present invention, steps (1) and (2) are specifically:

[0048] Use a robotic arm to move the sample tube containing the blood sample into the mixing component to mix the sample. After mixing, use the robotic arm to move the centrifuge tube and the sample tube containing the blood sample into the workbench component. Use the robotic arm to open the lids of the centrifuge tube and the sample tube, and then use the robotic arm to pick up a consumable pipette tip from the pipette tip position component 12. Transfer the blood sample from the sample tube to the centrifuge tube through the pipette tip to obtain a centrifuge tube containing the blood sample.

[0049] According to an embodiment of the present invention, in step (1), a scanning operation can also be performed on the centrifuge tube and the sample tube first; the scanning operation is realized by the barcode scanner component 18.

[0050] According to an embodiment of the present invention, in step (1), before performing the operation, a pumping and air exchange operation can also be performed on the workstation first to achieve aseptic operation; the pumping and air exchange operation is realized by the air filtration component 2.

[0051] According to an embodiment of the present invention, in step (4), weight balancing can also be performed on the centrifuge tube containing the sample, and the balancing operation is realized by the balancing platform component 11.

[0052] According to an embodiment of the present invention, in step (5), the robotic arm removes the supernatant from the centrifuge tube through the consumable pipette tip.

[0053] According to an embodiment of the present invention, the robotic arm is controlled by an operation screen, and the operation screen is externally connected to a control system.

[0054] Advantages of the present invention:

[0055] 1. The centrifuge of the present invention can achieve high-speed centrifugation and quickly reduce the temperature raised during operation through its refrigeration function. Compared with similar products, it has a higher rotation speed, more accurate positioning, and more precise temperature control. The overall structure is compact, with low noise and reliable operation; and it does not require manual operation, complex personnel training, and is time-saving, labor-saving, and labor-saving.

[0056] 2. Using the centrifuge of the present invention to separate and enrich infectious samples can effectively remove most impurities such as human cells, proteins, sugars, salts, etc. in clinical samples such as blood, cerebrospinal fluid, urine, pleural effusion, and ascites, whether uncultured or cultured, to obtain relatively pure live pathogens.

[0057] 3. The pathogen products isolated from uncultured samples can be subjected to susceptibility testing or MALDI-TOF MS detection after simple culture (only one-step liquid culture) and bacterial collection, while the products of cultured samples such as blood, cerebrospinal fluid, pleural effusion, and ascites can be directly used for susceptibility testing or MALDI-TOF MS detection after simple concentration. This protocol shortens the culture process for the diagnosis and identification of pathogens in clinical infection samples, enabling faster acquisition of susceptibility test results and mass spectrometry microbial identification results, and providing more timely diagnostic and treatment references for clinicians.

[0058] 4. The workstation of the present invention is used to extract and enrich trace microorganisms from complex matrices, and its workflow includes sample identification, lid opening and closing, consumable handling, sample addition, mixing (including oscillation, heating, etc.), centrifugation, liquid aspiration and discard, etc., realizing a fully automatic separation and processing process. Among them, the lid opening and closing mechanism is more compact in structure, has a lower failure rate, and operates more stably compared to other similar products; the pipetting robotic arm adopts a piston rod type liquid aspiration / drainage mode with stronger stability, higher precision, and stronger stability; the centrifuge component combines high speed, positioning, and refrigeration. While achieving high-speed centrifugation, it quickly reduces the temperature raised during operation through its refrigeration function, with a higher rotation speed, more accurate positioning, and more precise temperature control compared to similar products. The workstation of this application is also equipped with an air filtration system, which can keep the test environment sterile throughout the operation of the instrument, making the test more accurate and effectively avoiding contamination.

[0059] 5. The workstation of the present invention can enrich microorganisms with extremely low abundance, significantly improving the accuracy and reliability of detection. Combining artificial intelligence technology, it realizes high-throughput sample processing and data analysis, is applicable to the detection of trace microorganisms in multiple fields, and has broad application prospects.

[0060] 6. The method for extracting and enriching trace microorganisms of the present invention can effectively remove most impurities such as human cells, proteins, sugars, salts, etc. in uncultured or cultured clinical samples such as blood, cerebrospinal fluid, urine, pleural effusion, and ascites, obtaining relatively pure live pathogens.

[0061] 7. In the method of the present invention, the pathogen products isolated from uncultured samples can be subjected to susceptibility testing or MALDI-TOF MS detection after simple culture (only one-step liquid culture) and bacterial collection, while the products of cultured samples such as blood, cerebrospinal fluid, pleural effusion, and ascites can be directly used for susceptibility testing or MALDI-TOF MS detection after simple concentration. The method of the present invention shortens the culture time for the diagnosis and identification of pathogens in clinical infection samples, enabling faster acquisition of susceptibility test results and mass spectrometry microbial identification results, and providing more timely diagnostic and treatment references for clinicians.

[0062] 8. The workstation of the present invention uses conventional consumables and pre-packaged reagents to separate microorganisms in blood samples. It requires no manual operation, no complex personnel training and exercise, saving time, effort and labor. Description of the Drawings

[0063] Figure 1 is a schematic structural view of the centrifuge of the present invention;

[0064] Figure 2 is a top view of the rotor assembly of the present invention;

[0065] In the figure, 6-1, main body frame; 6-2, top cover assembly; 6-3, rotor assembly; 6-4, refrigeration assembly; 6-5, main shaft motor assembly; 6-6, positioning assembly; 6-7, cup hanging assembly; 6-8, centrifuge head.

[0066] Figure 3 is the front view of the workstation of the present invention.

[0067] Figure 4 is a top view of cross-section A of the workstation of the present invention.

[0068] 1, main frame; 2, air filtration assembly; 3, operation screen; 4, pipetting robotic arm; 5, pipette and lid opening / closing robotic arm; 6, centrifuge assembly; 7, waste liquid and waste collection bucket; 8, inversion mixing assembly; 9, heating assembly; 10, oscillation assembly; 11, matching platform assembly; 12, 200 μL pipette tip position assembly; 13, 5 mL pipette tip position assembly; 14, reagent position assembly; 15, workbench assembly; 16, waste liquid and waste throwing position assembly; 17, sample position assembly; 18, barcode scanner assembly.

[0069] Figure 5 is the process of separating and enriching microorganisms by the workstation in Example 2. Detailed Embodiments

[0070] The technical solutions of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are only for illustrative and explanatory purposes of the present invention, and should not be construed as limiting the protection scope of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection of the present invention.

[0071] Unless otherwise specified, the raw materials and reagents used in the following embodiments are all commercially available products, or can be prepared by known methods.

[0072] Example 1

[0073] A centrifuge, which includes: a main body frame 6-1, a partition plate is provided in the middle of the main body frame, and the partition plate divides the centrifuge into a centrifugation chamber and a fitting chamber;

[0074] A rotor assembly 6-3 disposed above the partition plate, and the rotor assembly is located in the centrifugation chamber for placing samples;

[0075] The outer wall of the centrifugation chamber is provided with a refrigeration assembly 6-4 for cooling or refrigerating the centrifugation chamber;

[0076] A positioning assembly 6-6 disposed below the partition plate, and the positioning assembly 6-6 is located in the accessory chamber.

[0077] The centrifuge further includes a top cover assembly 6-2 for sealing or covering the centrifuge, and one end of the top cover assembly is fixed to the main body frame 6-1.

[0078] The centrifuge further includes a main shaft motor assembly 6-5, and the main shaft motor assembly is rotatably connected to the partition plate.

[0079] The rotor assembly 6-3 includes a centrifuge head 6-8 and a cup hanging assembly 6-7. The centrifuge head is fixedly connected to the upper output shaft of the main shaft motor assembly 6-5, and the cup hanging assembly 6-7 is arranged at equal angles away from the center of the centrifuge head. The cup hanging assembly 6-7 is used for placing centrifuge tubes or sample tubes.

[0080] The connection between the positioning assembly 6-6 and the main body frame 6-1 is a known solution in the art.

[0081] The working principle of the centrifuge is as follows:

[0082] First, the automatic door of the top cover assembly 6-2 is opened, then the positioning assembly 6-6 positions the empty cup hanging assembly on the rotor assembly 6-3 to the automatic door according to requirements, and then the consumables to be centrifuged (or centrifuge tubes) are placed into the specified cup hanging assembly 6-7. Then the top cover assembly 6-2 runs to close the automatic door; then the main shaft motor assembly 6-5 starts to rotate and run according to the setting. During this process, due to the friction between the rotor assembly 6-3 and the air in the cavity, the temperature will rise sharply. At this time, the refrigeration assembly 6-4 starts to run to lower the cavity temperature until the set temperature is reached; when the centrifugation program is completed, the positioning assembly 6-6 will rotate the consumables to be taken out to the automatic door opening position according to the setting (wherein, the consumables to be taken out are controlled by the upper computer software), and then the automatic door on the top cover assembly 6-2 is opened to take out the centrifuged consumables.

[0083] A workstation for extracting and enriching trace microorganisms from complex matrices, which includes a main frame 1, a pipetting robotic arm 4, a pipette and lid opening / closing robotic arm 5, an inversion mixing assembly 8, a heating assembly 9, a shaking assembly 10, the above-mentioned centrifuge 6, a sample position assembly 17, a workbench assembly 15, and a reagent position assembly 14;

[0084] The robotic arm (including the pipetting robotic arm 4, the tube transferring and lid opening / closing robotic arm 5) is located at the upper part of the main frame, and the robotic arm is controlled by a control system;

[0085] In the middle of the main frame, there are also a dispensing platform assembly 11 and two groups of tip positions assemblies 12.

[0086] The inversion mixing assembly 8, the heating assembly 9, the shaking assembly 10, the dispensing platform assembly 11, and the two groups of tip positions assemblies 12 are arranged side by side in sequence in the middle of the main frame and are located below the robotic arm;

[0087] The sample position assembly 17 is arranged side by side with the heating assembly 9;

[0088] The workstation further includes a waste liquid and waste disposal position assembly 16, which is located in the middle of the main frame 1; the shaking assembly 10, the workbench assembly 15, and the waste liquid and waste disposal position assembly 16 are arranged side by side in sequence;

[0089] The dispensing platform assembly 11 and the reagent position assembly 14 are arranged side by side; the dispensing platform assembly 11 is used for balancing when the centrifuge tube enters the centrifuge. The reagent position assembly 14 is used for placing reagents or additives, for example, including lysis solution, microbial settling agent, washing solution, etc.

[0090] The centrifuge assembly is located at the lower part of the main frame.

[0091] The pipetting robotic arm is used to add or remove liquid from a reagent tube (sample tube or centrifuge tube), and the tube transferring and lid opening / closing robotic arm is used to move the reagent tube or to open or close the lid of the reagent tube.

[0092] Preferably, there are two groups of tip positions assemblies 12 in the middle of the main frame, including a 200 μL tip position assembly and a 5 mL tip position assembly 13. Preferably, the two groups of tip positions assemblies are respectively used for placing consumable 200 μL pipette tips and consumable 5 mL pipette tips.

[0093] The workstation further includes a waste liquid and waste collection bucket 7, which is located at the lower part of the main frame. The waste liquid and waste collection bucket 7 is arranged side by side with the centrifuge 6 and is used for collecting waste liquid or waste.

[0094] At the top of the main frame, there is also an air filtration assembly 2, and the air filtration assembly is used for filtering air.

[0095] Outside the main frame, there is also an operation screen 3, and the operation screen 3 is used to control the robotic arm for subsequent experimental operations. Preferably, the operation screen 3 is set on the side outside the main frame 1, for example, on the right side or the left side.

[0096] In the middle of the main frame, a barcode scanner assembly 18 is also provided, and the barcode scanner assembly is placed side by side with the mixing assembly. Preferably, the barcode scanner assembly is placed side by side with the inversion mixing assembly 8.

[0097] The working principle of the workstation of the present invention is as follows:

[0098] (1) Sample tubes and centrifuge tube consumables are placed at the corresponding positions of the sample position assembly 17, the reagent kit is placed at the corresponding position of the reagent position assembly 14, 200 μL pipette tip consumables are placed at the corresponding positions of the 200 μL pipette tip position assembly 12, and 5 mL pipette tip consumables are placed at the corresponding positions of the 5 mL pipette tip position assembly 13. Then, the instrument is operated through the operation screen 3 to start the test process.

[0099] (2) First, the pipetting and lid opening / closing robotic arm 5 transfers the sample tubes and centrifuge tube consumables in the sample position assembly 17 to the barcode scanner assembly 18 for barcode scanning. Then, the pipetting and lid opening / closing robotic arm 5 transfers the sample tubes to the inversion mixing assembly 8 for mixing the sample blood. After mixing, the pipetting and lid opening / closing robotic arm 5 transfers the sample tubes and centrifuge tubes to the corresponding positions of the workbench assembly 15, and opens the lids of the sample tubes and centrifuge tubes. At the same time, the pipetting robotic arm 4 moves above the 5 mL pipette tip position assembly 13 to pick up a 5 mL pipette tip, and then moves to the workbench assembly 15 to dispense the sample in the sample tube into the centrifuge tube. Finally, the pipetting robotic arm 4 runs above the waste liquid and waste disposal position assembly 16 to discard the used 5 mL pipette tip;

[0100] (3) The pipetting and lid opening / closing robotic arm 5 closes the lids of the sample tubes on the workbench assembly 15, and clamps and returns them to the corresponding positions of the sample position assembly 17. Then, the pipetting robotic arm 4 runs above the 200 μL pipette tip position assembly 12 to pick up a 200 μL pipette tip, then runs above the reagent position assembly 14 to aspirate the corresponding reagent, then runs above the workbench assembly 15 to add the corresponding auxiliary agent into the centrifuge tube. Finally, the pipetting robotic arm 4 runs above the waste liquid and waste disposal position assembly 16 to discard the used 200 μL pipette tip.

[0101] (4) Then, the pipetting robotic arm 4 moves above the 5 mL pipette tip position assembly 13 to pick up a 5 mL pipette tip, then runs above the reagent position assembly 14 to aspirate the corresponding auxiliary agent, then runs above the workbench assembly 15 to add the corresponding auxiliary agent into the corresponding centrifuge tube. Finally, the pipetting robotic arm 4 runs above the waste liquid and waste disposal position assembly 16 to discard the used 5 mL pipette tip;

[0102] (5) Then, the pipetting and lid-opening / closing robotic arm 5 moves above the workbench assembly 15 to perform the lid-closing operation on the centrifuge tube, clamps and transfers it to the corresponding position of the shaking assembly 10 for shaking operation; after the shaking is completed, the pipetting and lid-opening / closing robotic arm 5 sequentially places the centrifuge tubes into the centrifuge assembly 6, and automatically calculates whether balancing is required according to the number of centrifuge tubes placed.

[0103] (6) If balancing is required, the pipetting and lid-opening / closing robotic arm 5 moves to the balancing platform assembly 11 for balancing. After balancing, the centrifuge tubes are placed at the corresponding positions of the centrifuge assembly 6, and then the centrifuge assembly 6 starts to run to perform the centrifugation operation.

[0104] (7) After centrifugation, the pipetting and lid-opening / closing robotic arm 5 picks up the centrifuge tubes and places them on the workbench assembly 15 for lid-opening operation. Then, the pipetting robotic arm 4 inserts a 5 ml pipette tip from the 5 ml pipette tip assembly and runs to the centrifuge tube with the lid opened to perform the supernatant extraction operation. After completion, it runs to the waste liquid / waste material disposal assembly 16 to discard the waste liquid / discard the pipette tip; finally, the pipetting and lid-opening / closing robotic arm 5 performs the lid-closing operation on the centrifuge tubes on the workbench assembly 15.

[0105] (8) Repeat steps (5)-(7) six times. After the shaking is completed, the pipetting and lid-opening / closing robotic arm 5 transfers the centrifuge tubes to the heating assembly 9 for heating. After heating is completed, the centrifuge tubes are placed back at the corresponding positions of the sample position assembly 17.

[0106] Example 2

[0107] S1. Scanning and identification: Under the action of the pipetting and lid-opening / closing robotic arm 5, the sample tube and the centrifuge tube are transferred to the barcode scanner assembly 18 for barcode scanning and identification / recording;

[0108] S2. Sample mixing: The pipetting and lid-opening / closing robotic arm 5 transfers the sample tube to the mixing assembly for inversion mixing. After completion, it is transferred to the workbench assembly 15 together with the centrifuge tube;

[0109] S3. First step of blood sample aliquoting and reagent addition - lysis: The pipetting and lid-opening / closing robotic arm 5 opens the lids of the sample tube and the centrifuge tube. The pipetting robotic arm 4 inserts a 5 ml pipette tip from the 5 mL pipette tip assembly 13 to quantitatively aliquot the blood sample into the corresponding centrifuge tubes, and sends the discarded pipette tip to the waste liquid and waste material disposal assembly 16 to complete the pipette tip discarding operation; then the pipetting robotic arm 4 inserts the corresponding consumable pipette tip to quantitatively add lysis solution A and microbial co-precipitant to the centrifuge tubes that have just been aliquoted with blood sample, and then sends the discarded pipette tip to the waste liquid and waste material disposal assembly 16; and the pipetting and lid-opening / closing robotic arm 5 closes the lids of the sample tube and the centrifuge tube, and the sample tube is placed back in place;

[0110] S4. Transfer the centrifuge tube to the oscillation module 10 by the pipetting and lid-opening / closing robot arm 5 for 1 minute of oscillation, and then transfer it to the centrifuge module 6 for 2 - 3 minutes of centrifugation (automatic balancing during this process);

[0111] S5. Discard the supernatant: After centrifugation, the centrifuge tube is transferred from the centrifuge 6 to the workbench 15 by the pipetting and lid-opening / closing robot arm 5, and the lid is opened. Then, a 5 ml pipette tip is inserted by the pipetting robot arm to quantitatively remove the supernatant from the centrifuge tube. Subsequently, the supernatant is discarded at the waste liquid outlet of the waste liquid and waste material disposal module, and the used pipette tip is discarded at the waste material outlet of the waste liquid and waste material disposal module;

[0112] S6. Second to fourth lysis of the blood sample: A 5 ml pipette tip is inserted by the pipetting robot arm to aspirate a quantitative amount of lysis buffer B, C, or D and add it to the centrifuge tube from which the supernatant has just been removed. Then, the centrifuge tube is capped by the pipetting and lid-opening / closing robot arm and transferred to the oscillation module for 1 minute of oscillation, and then transferred to the centrifuge for 2 - 3 minutes of centrifugation (automatic balancing during this process); The above operations are repeated 3 times (the lysis buffers added are B, C, and D in sequence);

[0113] S7. Washing: The centrifuge tube after the fourth lysis is taken out of the centrifuge by the pipetting and lid-opening / closing robot arm and transferred to the workbench module 15, and the lid is opened; Then, a 5 ml pipette tip is inserted by the pipetting robot arm to aspirate washing buffer A, B, or C and add it to the centrifuge tube. The pipetting and lid-opening / closing robot arm 5 caps the centrifuge tube and transfers it to the oscillation module for 1 minute of oscillation, and then transfers it to the centrifuge for 2 minutes of centrifugation (automatic balancing during this process). After centrifugation, perform the supernatant discarding operation of S5; The above operations are repeated 3 times (the washing buffers added are A, B, and C in sequence)

[0114] S8. Collection: Transfer the product after the last S7 washing to the designated position at the sample position by the pipetting and lid-opening / closing robot arm.

[0115] In summary, the above is the full-process operation flow of the workstation for separating and enriching microorganisms, which saves about 25% of the time compared to the manual full-process operation; Depending on the sample volume, the bacteria can be enriched to 5 - 10 times the original concentration.

[0116] The above has given an exemplary description of the embodiments of the present invention. However, the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent replacements, improvements, etc. made by those skilled in the art within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A centrifuge, characterized in that, The centrifuge includes a main body frame 1, and a partition plate is provided in the middle of the main body frame. The partition plate divides the centrifuge into a centrifugation chamber and an accessory chamber; A rotor assembly disposed above the partition plate, and the rotor assembly is located in the centrifugation chamber for placing samples; A refrigeration assembly is provided on the outer wall of the centrifugation chamber for cooling or refrigerating the centrifugation chamber; A positioning assembly disposed below the partition plate, and the positioning assembly is located in the accessory chamber.

2. The centrifuge according to claim 1, wherein The centrifuge further includes a top cover assembly for sealing or covering the centrifuge.

3. The centrifuge according to claim 2, characterized in that, One end of the top cover assembly is fixed to the main body frame 1.

4. The centrifuge according to claim 1, characterized in that, The rotor assembly includes a centrifuge head and a cup hanging assembly. The centrifuge head is fixedly connected to the upper output shaft of the main shaft motor assembly. The cup hanging assemblies are arranged at equal angles away from the center of the centrifuge head; the cup hanging assemblies are used for placing centrifuge tubes or sample tubes. Preferably, the connection between the positioning assembly and the main body frame 1 is a known solution in the art.

5. A workstation for extracting and enriching trace microorganisms from a complex matrix, characterized in that, The workstation includes a main frame (1), a robotic arm, a mixing assembly, the centrifuge (6) according to any one of claims 1-4, a sample position assembly (17), a workbench assembly (15), and a reagent position assembly (14); The robotic arm is located at the upper part of the main frame, and the robotic arm is controlled by a control system; The mixing assembly, the sample position assembly (17), the workbench assembly (15), and the reagent position assembly (14) are arranged side by side in sequence in the middle of the main frame and are located below the robotic arm; The centrifuge is located at the lower part of the main frame.

6. The workstation according to claim 5, characterized in that The robotic arm includes a pipetting robotic arm (4) and a tube transferring and lid opening / closing robotic arm (5), and the pipetting robotic arm and the tube transferring and lid opening / closing robotic arm are arranged side by side.

7. The workstation according to claim 5, wherein, The mixing assembly includes an inversion mixing assembly (8) and a heating assembly (9). Preferably, the mixing assembly further includes a shaking assembly (10). Preferably, the inversion mixing assembly, the heating assembly, and the shaking assembly are arranged side by side in sequence.

8. The workstation according to claim 5, wherein A dispensing platform assembly (11) is further provided in the middle of the main frame, and the dispensing platform assembly is arranged side by side with the reagent position assembly (14). Preferably, at least one set of pipette tip position assemblies (12) is further provided in the middle of the main frame, and the pipette tip position assemblies are arranged side by side with the dispensing platform assembly (11); the pipette tip position assemblies are used for placing consumable pipette tips.

9. The workstation according to claim 5, wherein The workstation further includes a waste liquid and waste disposal position assembly (16), and the waste liquid and waste disposal position assembly is located in the middle of the main frame. Preferably, the waste liquid and waste disposal position assembly is arranged side by side with the workbench assembly. Preferably, the workstation further includes a waste liquid and waste collection bucket (7) located at the lower part of the main frame. The waste liquid and waste collection bucket is arranged side by side with the centrifuge assembly for collecting waste liquid or waste.

10. A method for extracting and enriching trace microorganisms, characterized in that, The method is implemented by using the workstation according to any one of claims 5-9, and the method includes: (1) Using the robotic arm to move the sample tube containing the blood sample to the mixing assembly to mix the sample. After mixing, use the robotic arm to move the mixed sample tube and the centrifuge tube to the workbench assembly respectively; (2) Using the robotic arm to transfer the sample in the sample tube to the centrifuge tube to obtain a centrifuge tube containing the blood sample; (3) Use a robotic arm to transfer the auxiliary agent from the reagent position component to the centrifuge tube containing the blood sample, and then continue to use the robotic arm to move the centrifuge tube to the mixing component to evenly mix the sample; (4) Use a robotic arm to move the centrifuge tube to the centrifuge component for centrifugation; (5) After centrifugation, use a robotic arm to move the centrifuge tube to the workbench component to remove the supernatant, completing the extraction and enrichment of trace microorganisms.

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