Automatic liquid sample microorganism rapid enrichment and concentration device

Abstract

The utility model belongs to the field of biological detection, and relates to a biological enrichment and detection technology, in particular to a device for enriching and concentrating microorganisms in a liquid sample. The automatic microorganism enrichment and concentration device mainly comprises a power supply module, a suction filtration module, a flushing module, a gas-liquid control module and an input / output display module.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological detection, and particularly relates to a device for enriching and concentrating microorganisms in a liquid sample. BACKGROUND

[0002] When detecting biological particles such as bacteria and viruses in a liquid sample, whether the target can be detected depends on its concentration. When the concentration of the target substance in the liquid sample is too low, the target substance cannot be detected.

[0003] The biological enrichment technology is to use different methods to enrich the target substance, so that the concentration of the target substance is increased to be detectable. Common enrichment methods include magnetic bead method, chromatography method, filtration method, etc. The filtration method uses a filter membrane with micro-nano size to intercept the target substance, and under the action of external force such as gravity, air pressure, mechanical force, etc., the liquid sample passes through the filter membrane, and the target substance is intercepted on one side of the filter membrane. Finally, a small amount of eluent is used to elute the target substance, so as to achieve the purpose of concentration and enrichment.

[0004] In the prior art, the enrichment device compresses the eluent and carbon dioxide in one gas cylinder during the elution process. The eluent is squeezed out in the form of bubbles in a short opening process of carbon dioxide. Although bubbles can be generated, the amount of eluent and the flushing strength (air pressure) cannot be accurately controlled. At the same time, the use of high-pressure carbon dioxide and eluent for co-storage has safety hazards in high-temperature, low-temperature environments and under impact.

[0005] In addition, in the prior art, the flushing liquid / high-pressure carbon dioxide is preloaded with a foam flushing liquid. This method cannot accurately control the amount of flushing liquid and the flushing strength, and the target recovery may not be complete or the activity may be reduced. The flushing strength will decrease as the gas pressure in the bottle decreases. The flushing module in the device of the present application includes an eluent pump, an eluent in-place sensor, an air pump, a gas storage cylinder, a gas pressure sensor (integrated on the control panel), and a bubbler. The volume of the flushing liquid can be set and automatically controlled. After the flushing liquid flows into the flushing pipeline, the high-pressure gas pushes the flushing liquid along the pipeline to flush and recover the target substance. The volume and pressure of the flushing liquid can be accurately controlled to ensure the enrichment effect.

[0006] In the prior art, the sample is manually / mechanically flushed repeatedly during the recovery process, and only a filter membrane is used as the interception method. This method may cause problems such as incomplete interception, embedding in filter holes, and rupture of the target recovery shape, resulting in low recovery rate of the target recovery. In order to solve these problems, self-developed magnetic beads or enrichment magnetic particles are added to the sample, so that the target recovery is adsorbed on the magnetic beads and wrapped by the magnetic beads, the volume is increased to prevent interception failure, and embedding in filter holes and rupture of the target recovery shape are avoided. The recovery rate can be improved and the activity can be maintained. SUMMARY

[0007] The present application aims to provide a liquid automatic liquid sample microbial rapid enrichment and concentration device.

[0008] The microbial automatic enrichment and concentration device provided by the present application mainly comprises a power supply module, a filtration module, a flushing module, a gas-liquid control module and an input-output display module.

[0009] The power supply module comprises an AC power supply plug or a rechargeable battery.

[0010] The power supply module is connected with the gas-liquid control module through a circuit, and the gas-liquid control module is connected with the filtration module, the flushing module and the input-output module through a circuit.

[0011] The filtration module comprises two versions.

[0012] (1) The negative pressure bottle version filtration module comprises a filtration pump (2), a negative pressure bottle (1), a first electromagnetic valve (11) and a second electromagnetic valve (12).

[0013] (2) The direct filtration version filtration module comprises a filtration pump (2).

[0014] The connection modes of the components are as follows:

[0015] The negative pressure bottle version filtration module: the air inlet of the filtration pump (2) is connected with the air outlet of the negative pressure bottle (1) through an air pipe, the water inlet of the negative pressure bottle (1) is connected with the water outlet of the second electromagnetic valve (12), and the air outlet of the first electromagnetic valve (11), the water inlet of the second electromagnetic valve (12) and the filtration port of the ultrafiltration tube (3) are connected through a three-way joint.

[0016] The direct filtration version filtration module: the water inlet of the filtration pump (2) is connected with the filtration port of the ultrafiltration tube (3) through a water pipe; the water outlet of the filtration pump (2) is connected with a water pipe, and the water pipe is placed in the waste liquid bottle (1).

[0017] The flushing module comprises three parts: a liquid feeding part, a high-pressure gas part and a mixing part.

[0018] The flushing module of the negative pressure bottle version and the direct filtration version is the same, and comprises three parts: a liquid feeding part, a high-pressure gas part and a mixing part.

[0019] The liquid feeding part comprises a flushing liquid bottle (4) and a liquid feeding pump (5).

[0020] The high-pressure gas part comprises a gas charging pump (8), a second sensor (9), a gas storage bottle (10) and a third electromagnetic valve (13).

[0021] The mixing part comprises a bubbler (6).

[0022] The connection modes of the components are as follows:

[0023] The liquid inlet part: the liquid inlet pump (5) is connected with the liquid inlet of the liquid bottle (4) through a hose.

[0024] The high-pressure gas part: the air charging pump (8), the second sensor (9) and the gas inlet of the gas cylinder (10) are connected with the three-way joint through a pipeline.

[0025] The liquid outlet of the liquid inlet pump (5), the gas outlet of the third electromagnetic valve (13) and the inlet end of the bubbler (6) are connected with the three-way joint, so that the liquid inlet part, the high-pressure gas part and the mixing part of the flushing module are connected.

[0026] The outlet end of the bubbler (6) is connected with the flushing port of the ultrafiltration tube (3) through a pipeline.

[0027] The gas-liquid control module: a control circuit board.

[0028] The connection modes of the components are as follows:

[0029] The gas-liquid control module is connected with the suction filter pump (2), the first electromagnetic valve (11), the second electromagnetic valve (12), the liquid inlet pump (5), the air charging pump (8), the third electromagnetic valve (13) and the first sensor (7) through a circuit.

[0030] The input-output display module: a touch display screen (17).

[0031] The touch display screen (17) is connected with the gas-liquid control module through a circuit.

[0032] The automatic microbial rapid enrichment and concentration device comprises a negative pressure bottle version automatic liquid sample microbial enrichment and concentration device and a direct suction filter version automatic liquid sample microbial enrichment and concentration device.

[0033] The negative pressure bottle version automatic liquid sample microbial enrichment and concentration device is connected by the following components:

[0034] The air inlet of the suction filter pump (2) is connected with the air outlet of the negative pressure bottle (1) through an air pipe, the water inlet of the negative pressure bottle (1) is connected with the water outlet of the second electromagnetic valve (12), the air outlet of the first electromagnetic valve (11), the water inlet of the second electromagnetic valve (12) and the suction filter port of the ultrafiltration tube (3) are connected with the three-way joint,

[0035] The liquid inlet part: the liquid inlet pump (5) is connected with the liquid inlet of the liquid bottle (4) through a hose.

[0036] The high-pressure gas part: the air charging pump (8), the second sensor (9) and the gas inlet of the gas cylinder (10) are connected with the three-way joint through pipelines, the gas outlet of the gas cylinder (10) is connected with the gas inlet of the third electromagnetic valve (13) through a pipeline,

[0037] The liquid outlet of the liquid feeding pump (5), the gas outlet of the third electromagnetic valve (13) and the entering end of the bubbler (6) are connected with the three-way joint, so that the liquid feeding part, the high-pressure gas part and the mixing part of the flushing module are connected, and the first sensor (7) is located between the three-way joint and the bubbler (6),

[0038] The discharging end of the bubbler (6) is connected with the flushing port of the ultrafiltration tube (3) through a pipeline.

[0039] The enrichment method of the negative pressure bottle version automatic enrichment and concentration device is as follows:

[0040] The power is turned on to start the instrument, the filtration pump (2) is started, and the negative pressure bottle (1) is in a negative pressure state; under the action of the negative pressure, the liquid sample is introduced into the negative pressure bottle (1) from the liquid outlet of the ultrafiltration tube (3) through a pipeline; after the filtration is completed, the filtration pump (2) is stopped, the first electromagnetic valve (11) is opened to remove the negative pressure of the negative pressure bottle (1), and then the first electromagnetic valve (11) is closed; the liquid feeding pump (5) is started, the flushing liquid is pumped into the pipeline from the flushing liquid bottle (4) through the liquid feeding pump (5); the flushing liquid reaches the first sensor (7), the liquid feeding pump (5) is stopped, the air charging pump (8) is started to charge air, the pressure of the gas cylinder (10) is increased, and the second sensor (9) senses that the pressure in the gas cylinder (10) reaches a set value; after that, the air charging pump (8) is controlled to stop by a control program, the second electromagnetic valve (12) is closed and the third electromagnetic valve (13) is opened, the high-pressure gas pushes the flushing liquid in the pipeline to pass through the bubbler (6), the foam-shaped flushing liquid is formed in the bubbler (6), enters the inside of the ultrafiltration tube through the flushing port of the ultrafiltration tube, and flushes the target substances retained by the ultrafiltration membrane, the flushing liquid is discharged from the sample inlet of the ultrafiltration tube and is recovered; after the gas pressure of the gas cylinder (10) returns to normal pressure, the third electromagnetic valve (13) is closed.

[0041] The direct filtration version automatic liquid sample microbial enrichment and concentration device comprises the following components which are connected:

[0042] The water inlet of the filtration pump (2) is connected with the filtration port of the ultrafiltration tube (3) through a water pipe, the water outlet of the filtration pump (2) is connected with the water pipe, and the water pipe is placed in the waste liquid bottle (1),

[0043] The liquid feeding part: the flushing liquid bottle (4) is connected with the liquid inlet of the liquid feeding pump (5) through a hose, the liquid outlet of the liquid feeding pump (5) is connected with the three-way joint through a hose,

[0044] The high-pressure gas part: the air charging pump (8), the second sensor (9) and the gas inlet of the gas cylinder (10) are connected with the three-way joint through pipelines, the gas outlet of the gas cylinder (10) is connected with the gas inlet of the third electromagnetic valve (13) through a pipeline,

[0045] The liquid outlet of the liquid pumping pump (5), the gas outlet of the third electromagnetic valve (13) and the entering end of the bubbler (6) are connected with the three-way joint, so that the liquid pumping part, the high-pressure gas part and the mixing part of the flushing module are connected, and the first sensor (7) is located between the three-way joint and the bubbler (6),

[0046] The discharging end of the bubbler (6) is connected with the flushing port of the ultrafiltration tube (3) through a pipeline.

[0047] The enrichment method of the automatic enrichment and concentration device in the direct suction filtration version is as follows:

[0048] The power is turned on to start the instrument, the suction filtration pump (2) is started, the liquid sample is discharged from the liquid outlet of the ultrafiltration tube (3) and enters the suction filtration pump (2), and then is discharged into the waste liquid bottle (1) through the liquid outlet of the suction filtration pump (2); after the filtration is completed, the suction filtration pump (2) is stopped; the liquid pumping pump (5) is started, the flushing liquid is pumped into the pipeline from the flushing liquid bottle (4) through the liquid pumping pump (5); the flushing liquid reaches the first sensor (7), the liquid pumping pump (5) is stopped, the air charging pump (8) is started to charge air, the pressure of the gas cylinder (10) is increased, the second sensor (9) senses that the pressure in the gas cylinder (10) reaches the set value, and then the air charging pump (8) is stopped under the control of the control program; the third electromagnetic valve (13) is opened, the high-pressure gas pushes the flushing liquid in the pipeline to pass through the bubbler (6), and the foam-shaped flushing liquid formed in the bubbler (6) enters the inside of the ultrafiltration tube through the flushing port of the ultrafiltration tube (3) to flush the target substances intercepted by the ultrafiltration membrane, and the flushing liquid is discharged from the sample inlet of the ultrafiltration tube and is recovered; after the gas pressure of the gas cylinder (10) returns to normal pressure, the third electromagnetic valve (13) is closed.

[0049] Compared with the prior art, the present application has the following beneficial effects:

[0050] 1) The pressure of the high-pressure gas for pushing the flushing can be adjusted, so that the damage to the structure of pathogenic microorganisms caused by a large impact is avoided, the activity of the recovered target substances is improved, and the phenomenon that the gas pressure decreases with the flushing process is avoided when the high-pressure carbon dioxide preloaded flushing liquid is used, so that the consistency of the flushing intensity each time is ensured.

[0051] 2) The flushing liquid contains a surfactant and a defoaming agent, under the pushing of the high-pressure gas, the flushing liquid is mixed in the bubbler to generate foam, and the flushing liquid flushes the inside of the filter membrane in the form of foam, so that the contact area and contact time of the flushing liquid and the filter membrane are increased.

[0052] 3) High-pressure gas is not stored with the flushing liquid, which is safer; and the flushing liquid dosage and flushing pressure can be set according to the use requirements, suitable for different types of samples, which can improve the enrichment recovery rate and the activity of the sample after enrichment.

[0053] 4) Independent bubble generator, close air pressure inside and outside the foam, soft when breaking, reduce energy release when breaking, reduce microbial activity, and reduce aerosol generation.

[0054] 5) The filter has multiple different specifications, the filter membrane pore size, filter membrane filter diameter, filter membrane surface treatment form, and filter membrane interception area are different, the filter membrane form can be hollow fiber membrane and flat membrane, which can be selected according to different sample types and sample amounts, etc. Requirements to ensure enrichment efficiency.

[0055] 6) Use a negative pressure bottle to hold the sample filter waste liquid, the float rises with the liquid level, and when it is too high, the liquid will not enter the negative pressure bottle, preventing the negative pressure bottle from being too full and spilling onto the test table.

[0056] 7) The device has self-developed control software and hardware, simple operation, stable operation, high safety, and supports function expansion. BRIEF DESCRIPTION OF DRAWINGS

[0057] Figure 1 , negative pressure bottle version of automatic liquid sample microbial rapid enrichment and concentration device gas-liquid pipeline connection diagram

[0058] Figure 2 , direct filtration version of automatic liquid sample microbial rapid enrichment and concentration device gas-liquid pipeline connection diagram

[0059] Figure 3 , automatic liquid sample microbial rapid enrichment and concentration device schematic diagram

[0060] Figure 4 , automatic liquid sample microbial rapid enrichment and concentration device schematic diagram

[0061] Figure 5 , automatic liquid sample microbial rapid enrichment and concentration device schematic diagram

[0062] Figure 6 , negative pressure bottle version of automatic liquid sample microbial rapid enrichment and concentration device module relationship diagram

[0063] Figure 7 , direct filtration version of automatic liquid sample microbial rapid enrichment and concentration device module relationship diagram

[0064] In the figure:

[0065] 1. Negative pressure bottle, 2. Vacuum pump, 3. Ultrafiltration tube, 4. Rinse solution bottle, 5. Liquid filling pump, 6. Aerator, 7. First sensor, 8. Air pump, 9. Second sensor, 10. Gas storage bottle, 11. First solenoid valve, 12. Second solenoid valve, 13. Third solenoid valve, 14. Control circuit board, 15. Power supply module, 16. Switch, 17. Touch screen. Detailed Implementation

[0066] The present invention will be further illustrated by the following examples.

[0067] The product and its preparation method according to the present invention will be further described below with reference to specific embodiments, but the present invention is not limited to the following embodiments. Unless otherwise specified, the methods described are conventional methods. Unless otherwise specified, the raw materials can be obtained from publicly available commercial sources.

[0068] Example 1: Automated Liquid Sample Microbial Rapid Enrichment and Concentration Device in Negative Pressure Bottles

[0069] like Figure 1 As shown, the automated liquid sample microbial rapid enrichment and concentration device with negative pressure bottle is composed of the following connected components:

[0070] The air inlet of the filtration pump (2) is connected to the exhaust port of the negative pressure bottle (1) through an air pipe. The water inlet of the negative pressure bottle (1) is connected to the water outlet of the second solenoid valve (12). The air outlet of the first solenoid valve (11), the water inlet of the second solenoid valve (12), and the filtration port of the ultrafiltration tube (3) are connected through a three-way connector.

[0071] Liquid supply section: The flushing solution bottle (4) is connected to the inlet of the liquid supply pump (5) via a hose, and the outlet of the liquid supply pump (5) is connected to a three-way connector via a hose.

[0072] High-pressure gas section: The air pump (8), the second sensor (9), and the air inlet of the gas cylinder (10) are connected to a three-way connector via pipelines. The air outlet of the gas cylinder (10) is connected to the air inlet of the third solenoid valve (13) via pipelines.

[0073] The liquid outlet of the liquid pump (5), the air outlet of the third solenoid valve (13), and the inlet of the aerator (6) are connected by a three-way connector, thereby connecting the liquid supply part, the high-pressure gas part, and the mixing part of the flushing module. The first sensor (7) is located between the three-way connector and the aerator (6).

[0074] The outlet of the aerator (6) is connected to the flushing port of the ultrafiltration tube (3) via a pipeline.

[0075] Example 2: Automated Microbial Rapid Enrichment and Concentration Device for Direct Filtration Samples

[0076] like Figure 2As shown, the automated liquid sample microbial rapid enrichment and concentration device with direct filtration is composed of the following connected components:

[0077] The inlet of the filtration pump (2) is connected to the filtration port of the ultrafiltration tube (3) via a water pipe, and the outlet of the filtration pump (2) is connected to a water pipe, which is then placed into the waste liquid bottle (1).

[0078] Liquid supply section: The flushing solution bottle (4) is connected to the inlet of the liquid supply pump (5) via a hose, and the outlet of the liquid supply pump (5) is connected to a three-way connector via a hose.

[0079] High-pressure gas section: The air pump (8), the second sensor (9), and the air inlet of the gas cylinder (10) are connected to a three-way connector via pipelines. The air outlet of the gas cylinder (10) is connected to the air inlet of the third solenoid valve (13) via pipelines.

[0080] The liquid outlet of the liquid pump (5), the air outlet of the third solenoid valve (13), and the inlet of the aerator (6) are connected by a three-way connector, thereby connecting the liquid supply part, the high-pressure gas part, and the mixing part of the flushing module. The first sensor (7) is located between the three-way connector and the aerator (6).

[0081] The outlet of the aerator (6) is connected to the flushing port of the ultrafiltration tube (3) via a pipeline.

[0082] Example 3: Circuit connection of the automated liquid sample microbial rapid enrichment and concentration device using a negative pressure bottle.

[0083] like Figure 6 As shown, the power supply module is connected to the switch and the gas-liquid control module through the circuit. The gas-liquid control module is connected to the first sensor 7, the second sensor 9, the filtration pump 2, the air pump 8, the touch screen 17, the liquid pump 5, the first solenoid valve 11, the second solenoid valve 12, and the third solenoid valve 13 through the circuit.

[0084] Example 4: Circuit connection of the direct filtration automated body sample enrichment and concentration device

[0085] like Figure 7 As shown, the power supply module is connected to the switch and the gas-liquid control module through the circuit. The gas-liquid control module is connected to the first sensor 7, the second sensor 9, the liquid pump 5, the filtration pump 2, the air pump 8, the touch screen 17, and the third solenoid valve 13 through the circuit.

Claims

1. An automated liquid sample microbial rapid enrichment and concentration device, mainly composed of a power supply module, a filtration module, a rinsing module, a gas-liquid control module, and an input / output display module. in, Power supply module, including AC power plug or rechargeable battery; The power supply module is connected to the gas-liquid control module via wires, and the gas-liquid control module is connected to the filtration module, the rinsing module, and the input / output module via wires. The filtration module includes two versions: The negative pressure bottle version of the vacuum filtration module includes: a vacuum filtration pump (2), a negative pressure bottle (1), a first solenoid valve (11), and a second solenoid valve (12). The direct filtration module includes: a filtration pump (2); The connection methods for each component are as follows: Negative pressure bottle version of the suction filtration module: the air inlet of the suction pump (2) is connected to the exhaust port of the negative pressure bottle (1) through the air pipe, the water inlet of the negative pressure bottle (1) is connected to the water outlet of the second solenoid valve (12), and the air outlet of the first solenoid valve (11), the water inlet of the second solenoid valve (12) and the suction port of the ultrafiltration tube (3) are connected through a three-way connector. Direct filtration module: The inlet of the filtration pump (2) is connected to the filtration port of the ultrafiltration tube (3) through a water pipe; the outlet of the filtration pump (2) is connected to a water pipe, and the water pipe is placed into the waste liquid bottle (1). Among them, the flushing module: The rinsing module of the negative pressure bottle version is the same as that of the direct suction filtration version, including three parts: liquid feeding section, high-pressure gas section, and mixing section; The liquid filling section includes: a flushing solution bottle (4) and a liquid filling pump (5); High-pressure gas section: gas pump (8), second sensor (9), gas storage cylinder (10), third solenoid valve (13); Mixing section: foamer (6); The connection methods for each component are as follows: Liquid filling section: The flushing liquid bottle (4) is connected to the inlet of the liquid filling pump (5) through a hose, and the outlet of the liquid filling pump (5) is connected to the three-way connector through a hose; High-pressure gas section: The air pump (8), the second sensor (9), and the air inlet of the gas cylinder (10) are connected to the three-way connector through the pipeline. The air outlet of the gas cylinder (10) is connected to the air inlet of the third solenoid valve (13) through the pipeline. The liquid pump (5) outlet, the third solenoid valve (13) air outlet, and the aerator (6) inlet are connected by a three-way connector, thereby connecting the liquid supply part, high-pressure gas part, and mixing part of the flushing module; the sensor 1 is located between the three-way connector and the aerator.

2. The apparatus according to claim 1, characterized in that, The automated liquid sample enrichment and concentration device with negative pressure bottle is composed of the following connected components: The air inlet of the filtration pump (2) is connected to the exhaust port of the negative pressure bottle (1) through an air pipe. The water inlet of the negative pressure bottle (1) is connected to the water outlet of the second solenoid valve (12). The air outlet of the first solenoid valve (11), the water inlet of the second solenoid valve (12), and the filtration port of the ultrafiltration tube (3) are connected through a three-way connector. Liquid supply section: The flushing solution bottle (4) is connected to the inlet of the liquid supply pump (5) via a hose, and the outlet of the liquid supply pump (5) is connected to a three-way connector via a hose. High-pressure gas section: The air pump (8), the second sensor (9), and the air inlet of the gas cylinder (10) are connected to a three-way connector via pipelines. The air outlet of the gas cylinder (10) is connected to the air inlet of the third solenoid valve (13) via pipelines. The liquid pump (5) outlet, the third solenoid valve (13) air outlet, and the aerator (6) inlet are connected by a three-way connector, thereby connecting the liquid supply part, high-pressure gas part, and mixing part of the flushing module. The first sensor (7) is located between the three-way connector and the aerator (6). The outlet of the aerator (6) is connected to the flushing port of the ultrafiltration tube (3) through a pipeline.

3. The apparatus according to claim 1, characterized in that, The direct filtration automated liquid sample enrichment and concentration device consists of the following connected components: The inlet of the filtration pump (2) is connected to the filtration port of the ultrafiltration tube (3) through a water pipe, and the outlet of the filtration pump (2) is connected to a water pipe, which is then placed into the waste liquid bottle (1). Liquid supply section: The flushing solution bottle (4) is connected to the inlet of the liquid supply pump (5) via a hose, and the outlet of the liquid supply pump (5) is connected to a three-way connector via a hose. High-pressure gas section: The air pump (8), the second sensor (9), and the air inlet of the gas cylinder (10) are connected to a three-way connector via pipelines. The air outlet of the gas cylinder (10) is connected to the air inlet of the third solenoid valve (13) via pipelines. The liquid pump (5) outlet, the third solenoid valve (13) air outlet, and the aerator (6) inlet are connected by a three-way connector, thereby connecting the liquid supply part, high-pressure gas part, and mixing part of the flushing module. The first sensor (7) is located between the three-way connector and the aerator (6). The outlet of the aerator (6) is connected to the flushing port of the ultrafiltration tube (3) through a pipeline.

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