Sterility checking device and detection method thereof

Abstract

The application discloses a sterile examination device, which comprises a double-layer tank body, a microporous filter film, a sterile absorption pad and an exhaust filtering assembly, wherein the double-layer tank body comprises an inner cavity and an outer cavity outside the inner cavity, and a lower chamber of the inner cavity is communicated with the outer cavity through a small hole; the microporous filter film is fixed in the middle part of the inner cavity and divides the inner cavity into an upper chamber and the lower chamber; the sterile absorption pad is located below the microporous filter film, and a neutralizing agent is prearranged in the sterile absorption pad; the exhaust filtering assembly is communicated with the upper chamber of the inner cavity, and a hydrophobic membrane is arranged in the exhaust filtering assembly. The sterile examination device and the detection method thereof can first realize bidirectional flushing and neutralization of the absorption pad, can efficiently remove bacteriostatic substances on both sides of the filter film, and can avoid false negative; secondly, the device can realize online integrity test, can prevent false positive caused by leakage of the device, and can realize full-closed operation, so that the risk of external source pollution is reduced; and meanwhile, the device is suitable for sterile examination of difficult-to-filter samples containing preservatives, antibiotics and the like.

Description

Technical Field

[0001] This invention relates to the field of microbial detection technology, specifically to a sterility testing device and its detection method. Background Technology

[0002] Membrane filtration is one of the legally mandated methods for sterility testing. According to the pharmacopoeias of various countries, membrane filtration should be the first choice as long as the properties of the test sample allow it. The core principle of this method is to filter the test sample solution through a microporous membrane with a pore size of no more than 0.45 μm, so that microorganisms are trapped on the membrane. Then, the antimicrobial components are removed by rinsing with sterile rinsing solution. Finally, the membrane is transferred to a culture medium for incubation to determine whether the test sample is sterile. However, the following problems exist in actual operation. First, there is the risk of external contamination. Puncture of the injection needle, exposure of the vent hole, and poor sealing of the pipeline connection may all lead to the entry of exogenous microorganisms, resulting in false positives. Second, the rinsing is insufficient. Samples containing antibacterial components, such as antibiotics and preservatives, require a large amount of rinsing solution to remove the antibacterial substances, but one-way rinsing cannot effectively remove the antibacterial agents remaining on the lower surface of the filter membrane. Furthermore, the integrity of the device cannot be checked online. Before incubation, it is impossible to confirm whether the filter membrane is damaged or whether the device is leaking. If a positive result is found after several days of incubation, it is impossible to distinguish whether the contamination is due to sample contamination or device contamination, indicating room for improvement. Summary of the Invention

[0003] (a) Technical problems to be solved To address the shortcomings of existing technologies, this invention provides a sterile inspection device and its detection method to solve the aforementioned problems.

[0004] (II) Technical Solution To achieve the above objectives, at least one embodiment of this disclosure provides a sterility testing device, comprising: A double-walled tank, comprising an inner cavity and an outer cavity located outside the inner cavity, wherein the lower chamber of the inner cavity is connected to the outer cavity through a small hole; A microporous filter membrane is fixed in the middle of the inner cavity, dividing the inner cavity into an upper chamber and a lower chamber; A sterile absorbent pad is located below the microporous filter membrane, and a neutralizing agent is pre-placed inside the sterile absorbent pad; Both the sample injection valve and the culture medium injection valve are connected to the upper chamber of the inner cavity; An exhaust filter assembly is connected to the upper chamber of the inner cavity, and a hydrophobic membrane is provided inside the exhaust filter assembly; An exhaust on / off valve is disposed between the exhaust filter assembly and the upper chamber of the inner cavity; Positive and negative pressure gas inlets are connected to the upper chamber of the inner cavity and are used to alternately apply positive and negative pressure to allow liquid to pass through the microporous filter membrane in both directions; A waste liquid discharge valve is located in the lower chamber of the inner cavity; A pressure sensor is used to monitor the pressure in the cavity; and An on / off valve is installed between the positive and negative pressure air inlet and the upper chamber of the inner cavity to seal the inner cavity.

[0005] For example, in a sterile inspection device provided in at least one embodiment of this disclosure, the sterile absorbent pad is disposed close to the underside of the microporous filter membrane, and the neutralizing agent is in a lyophilized state, and the neutralizing agent is selected from one or more of lecithin, Tween 80, histidine, and sodium thiosulfate.

[0006] For example, in a sterile testing device provided in at least one embodiment of this disclosure, both the sample injection valve and the culture medium injection valve are needle-free puncture valves and are equipped with self-sealing silicone pads.

[0007] For example, in a sterile inspection device provided in at least one embodiment of this disclosure, the hydrophobic membrane pore size in the exhaust filtration assembly is 0.22 μm; and the pore size of the microporous filter membrane is 0.45 μm.

[0008] For example, in a sterile inspection device provided in at least one embodiment of this disclosure, the inner cavity is a culture chamber and the outer cavity is a negative pressure buffer chamber.

[0009] According to another aspect of the present invention, a method for sterility testing is also provided, comprising the following steps: S1 Integrity Test: Open the on / off valve, close the exhaust on / off valve, and fill the inner cavity with sterile air to a predetermined pressure through the positive and negative pressure air circuit interface. Then close the on / off valve and use the pressure sensor to monitor the pressure change in the inner cavity. If the pressure drop does not exceed the set threshold during the pressure holding period, the device is deemed to be in good condition. S2 Sample Filtration: Open the on / off valve and the exhaust on / off valve, inject the test sample into the upper chamber of the inner cavity through the sample injection valve to exhaust the gas in the upper chamber, then close the exhaust on / off valve, apply negative pressure through the positive and negative pressure gas path interface, so that the test sample passes through the filter membrane and enters the lower chamber, the filtrate is absorbed by the sterile absorption pad and the antibacterial substance is neutralized by the neutralizing agent therein; S3 bidirectional flushing: Open the exhaust on / off valve, inject flushing fluid into the upper chamber of the inner cavity through the culture medium injection valve to discharge the gas in the upper chamber, then close the exhaust on / off valve, keep the on / off valve open, and alternately apply positive and negative pressure through the positive and negative pressure gas circuit interface to make the flushing fluid flow back and forth on the microporous filter membrane at least 3 times; S4 Waste liquid discharge: Keep the exhaust valve closed, apply positive pressure to force the flushed liquid into the lower chamber of the inner cavity, and open the waste liquid discharge valve to discharge the waste liquid; S5 Culture Medium Injection and Cultivation: Close the waste liquid discharge valve and the on / off valve, open the exhaust on / off valve, inject the culture medium through the culture medium injection valve, then close the exhaust on / off valve, seal the culture, and observe the results.

[0010] For example, in the sterility testing method provided in at least one embodiment of this disclosure, the positive pressure gauge pressure applied alternately in step S3 is 0.02~0.10 MPa, the negative pressure gauge pressure is -0.02~-0.10 MPa, and the pressure holding time is 2~10 seconds each time; the predetermined pressure in step S1 is 5~10 kPa, and the set threshold is a pressure drop ≤0.5 kPa within 5 minutes.

[0011] For example, in the sterility testing method provided in at least one embodiment of this disclosure, the rinsing solution in step S3 is an aqueous solution containing 0.1% peptone and 0.1% Tween 80, or a 0.9% sterile sodium chloride solution.

[0012] For example, in the sterility testing method provided in at least one embodiment of this disclosure, the culture medium in step S5 is thioglycolate fluid culture medium, and the culture conditions are 35°C for 14 days.

[0013] For example, in at least one embodiment of the sterility testing method provided in this disclosure, the method further includes: Positive control: Using the same batch of the sterile testing device, add known positive microorganisms and operate according to steps S1 to S5 as a positive control; Negative control: Use sterile water instead of the test sample, and follow steps S1 to S5 as a negative control; Result determination: If the sample device becomes turbid after incubation, and the positive control is turbid while the negative control is clear, it is judged as positive; if the sample device is clear while the positive control is turbid, it is judged as negative; if the negative control is turbid, the test is invalid.

[0014] (III) Beneficial Effects Compared with the prior art, the present invention provides a sterility inspection device and its detection method, which has the following beneficial effects: Firstly, bidirectional rinsing combined with absorption pad neutralization can efficiently remove antibacterial substances from both sides of the filter membrane, avoiding false negatives. Secondly, online integrity testing can be performed to prevent false positives caused by device leakage, and the fully enclosed operation reduces the risk of external contamination. At the same time, it is suitable for sterility testing of samples that are difficult to filter, such as those containing preservatives and antibiotics. Attached Figure Description

[0015] To more clearly illustrate the technical solutions of the embodiments of this disclosure, the accompanying drawings of the embodiments will be briefly described below. Obviously, the drawings described below only relate to some embodiments of this disclosure and are not intended to limit this disclosure.

[0016] Figure 1 This is a schematic diagram of the structure of a sterile inspection device according to the present invention.

[0017] In the diagram: 1. Double-walled tank; 2. Microporous filter membrane; 3. Sterile absorption pad; 4. Sample injection valve; 5. Culture medium injection valve; 6. Exhaust filter assembly; 7. Positive and negative pressure gas path interface; 8. Waste liquid discharge valve; 9. Pressure sensor. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] It should be understood that in the various embodiments of the present invention, the sequence number of each process does not imply the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present invention.

[0020] It should be understood that in this invention, "comprising" and "having," and any variations thereof, are intended to cover non-exclusive inclusion, for example, a process, method, system, product, or device that includes a series of steps or units is not necessarily limited to those steps or units that are explicitly listed, but may include other steps or units that are not explicitly listed or that are inherent to such process, method, product, or device.

[0021] The applicant's research revealed the following problems in actual operation: First, there is the risk of external contamination. Puncture of the injection needle, exposure of the vent hole, and poor sealing of pipeline connections can all lead to the entry of exogenous microorganisms, resulting in false positives. Second, there is insufficient rinsing. Samples containing antibacterial components, such as antibiotics and preservatives, require a large amount of rinsing solution to remove the antibacterial substances, but one-way rinsing cannot effectively remove the antibacterial agents remaining on the lower surface of the filter membrane. Furthermore, the integrity of the device cannot be checked online. Before incubation, it is impossible to confirm whether the filter membrane is damaged or whether the device is leaking. If a positive result is found after several days of incubation, it is impossible to distinguish whether the contamination is due to sample contamination or device contamination, indicating room for improvement.

[0022] Based on the above ideas, at least one embodiment of this disclosure provides a sterility inspection device and its detection method. The following describes a sterility inspection device and its detection method provided by this disclosure through several specific embodiments.

[0023] Example 1: Figure 1An embodiment of the aseptic testing device of the present invention is shown. The device includes a double-walled container 1, which has an inner cavity and an outer cavity located outside the inner cavity. The inner cavity is used as a culture chamber, and the outer cavity is used as a negative pressure buffer chamber. The lower chamber of the inner cavity is connected to the outer cavity through a small hole. This structure can establish a negative pressure in the outer cavity to balance the pressure on both sides of the filter membrane. When an operational error causes the pressure on one side of the filter membrane to be too high, the outer cavity can play a buffering role to prevent the microporous filter membrane from rupturing due to excessive pressure difference.

[0024] The microporous filter membrane 2 is fixedly installed in the middle of the inner cavity, thereby dividing the inner cavity into an upper chamber and a lower chamber. The pore size of the microporous filter membrane 2 is preferably 0.45 μm to trap microorganisms and allow liquid to pass through. A sterile absorbent pad 3 is provided close to the bottom of the microporous filter membrane 2. The sterile absorbent pad 3 is pre-filled with a neutralizing agent. The neutralizing agent exists in the absorbent pad in a lyophilized state. The neutralizing agent is selected from one or more of lecithin, Tween 80, histidine, and sodium thiosulfate. For example, a combination of lecithin and Tween 80 can be used. During the filtration process, the liquid in the sample passes through the filter membrane into the lower chamber and is absorbed by the sterile absorbent pad. The residual antibacterial substances can be effectively neutralized by the neutralizing agent in the sterile absorbent pad, thereby eliminating the antibacterial properties.

[0025] The components connected to the upper chamber of the inner cavity include: sample injection valve 4, culture medium injection valve 5, exhaust filter assembly 6, and positive and negative pressure gas path interface 7. The sample injection valve 4 and culture medium injection valve 5 are preferably needleless puncture valves with self-sealing silicone pads, which can be used to inject liquid by puncture with a sterile syringe without introducing external microorganisms. The exhaust filter assembly 6 is connected to the upper chamber of the inner cavity and has a hydrophobic membrane inside. The pore size of the hydrophobic membrane is preferably 0.22 μm, which allows gas to pass through but prevents microorganisms from entering, thereby maintaining a sterile environment while exhausting gas. An exhaust on / off valve is also provided between the exhaust filter assembly 6 and the upper chamber of the inner cavity to control the opening and closing of this passage.

[0026] The positive and negative pressure gas interface 7 is used to connect an external positive or negative pressure source to alternately apply positive and negative pressure to the upper chamber of the inner cavity, thereby enabling the liquid to pass through the microporous filter membrane in both directions, realizing the bidirectional flushing function unique to this invention. An on / off valve is provided between the positive and negative pressure gas interface 7 and the upper chamber of the inner cavity. This on / off valve is used to control the opening and closing of the gas path between the gas interface and the upper chamber of the inner cavity, and to seal the gas path when closed. Specifically, during the device integrity test, gas needs to be filled through the gas interface and then the on / off valve is closed to form a sealed space in the inner cavity. The pressure drop is monitored by a pressure sensor to accurately determine whether there is a leak in the device. When performing bidirectional flushing or sample filtration, the on / off valve needs to be opened to allow the external positive or negative pressure source to communicate with the inner cavity, so as to drive the fluid to pass through the filter membrane in both directions. The on / off valve can be in the form of a pinch valve, plug valve, etc.

[0027] At the bottom of the device, in the lower chamber of the inner cavity, there is a waste liquid discharge valve 8, which is used to discharge waste liquid after rinsing. A pressure sensor 9 is connected to the inner cavity to monitor the pressure changes in the inner cavity in real time, especially to provide pressure data during integrity testing and rinsing operations.

[0028] Example 2: Method for performing sterility testing using the above-described device. This example uses eye drops containing preservatives as an example to illustrate the complete process of performing sterility testing using the device.

[0029] It should be noted that this method is also applicable to other liquid medicines.

[0030] Step S1: Integrity test. First, remove the device from the aseptic packaging, connect the positive and negative pressure air circuit interface 7 to an external pressure source such as a peristaltic pump or syringe, open the on / off valve, close the exhaust on / off valve, and fill the inner cavity with sterile air through the positive and negative pressure air circuit interface 7 until the display value of the pressure sensor 9 reaches the predetermined pressure, for example, 5 kPa to 10 kPa, preferably 5 kPa. Then close the on / off valve and use the pressure sensor 9 to monitor the pressure change in the inner cavity. If the pressure drop does not exceed 0.5 kPa within 5 minutes, the device is deemed to be in good condition and can continue to be used. If the pressure drop exceeds this threshold, a new device needs to be replaced and the test repeated.

[0031] Step S2: Sample filtration. After the device is properly sealed, open the on / off valve and the exhaust on / off valve. Use a sterile syringe to draw the test sample and inject it into the upper chamber of the inner cavity through the sample injection valve 4. During the injection, the air in the upper chamber is discharged through the exhaust filter assembly 6. After the injection is completed, close the exhaust on / off valve and apply negative pressure through the positive and negative pressure air circuit interface 7. The test sample is driven by the negative pressure to pass through the filter membrane 2 and enter the lower chamber. The filtration flow rate is controlled at about 10 mL / min. After the filtrate enters the lower chamber, it is absorbed by the sterile absorption pad 3. The residual antibacterial substances such as benzalkonium chloride are neutralized by the neutralizing agent in the absorption pad, thereby avoiding the inhibition of subsequent microbial growth by the antibacterial substances.

[0032] Step S3: Bidirectional rinsing. After filtration, open the exhaust valve and inject rinsing fluid into the upper chamber of the inner cavity through the culture medium injection valve 5 to expel the gas in the upper chamber. Then close the exhaust valve and keep it open. Apply positive and negative pressure alternately through the positive and negative pressure gas interface 7. The absolute value of each pressure can be 0.02~0.10 MPa, for example, positive pressure 0.05 MPa and negative pressure -0.05 MPa. Hold the pressure for 2~10 seconds each time, for example, 3 seconds. Repeat this cycle at least 3 times, preferably 5 times. Through this alternating pressure application, the rinsing fluid is repeatedly drawn and passed through the microporous filter membrane 2, thoroughly flushing away the antibacterial substances remaining on the upper and lower surfaces of the filter membrane. Furthermore, some of the antibacterial substances that have passed through the membrane pores are further inactivated by the neutralizing agent in the sterile absorbent pad 3. This bidirectional rinsing step can significantly improve the removal efficiency of antibacterial substances and reduce the risk of false negatives.

[0033] Step S4: Waste liquid discharge. After rinsing, keep the exhaust valve closed and the on / off valve open. Apply positive pressure to force all the flushed liquid into the lower chamber of the inner cavity. Then open the waste liquid discharge valve 8 to completely discharge the waste liquid.

[0034] Step S5: Culture medium injection and incubation. Close the waste liquid discharge valve 8 and the on / off valve. Open the exhaust on / off valve and inject 100 mL of thioglycolate fluid culture medium through the culture medium injection valve 5 to submerge the filter membrane. During the injection process, air is discharged from the exhaust filter assembly 6. After the injection is completed, close the exhaust on / off valve. At this time, the inner cavity is in a fully closed state. Place the device in an incubator and incubate at 35°C for 14 days. Observe the turbidity of the culture medium in the device daily.

[0035] Positive and negative controls are required to verify the validity of the test results. Positive control: Use the same batch of sterile testing equipment. In step S2, add 10 CFU of Staphylococcus aureus or other specified positive bacteria to the test sample. The remaining steps are exactly the same as S1 to S5 above. Negative control: Use sterile water instead of the test sample and operate according to steps S1 to S5.

[0036] Result interpretation: If the test sample becomes turbid after incubation, and the positive control is turbid while the negative control is clear, the sterility test result of the test sample is considered positive, i.e. unqualified. If the test sample is clear while the positive control is turbid, it is considered negative, i.e. qualified. If the negative control is turbid, it indicates that there is exogenous contamination in the entire operation process, and this test is invalid and needs to be repeated.

[0037] The method described in this embodiment was used to test eye drops containing benzalkonium chloride. The results were as follows: the test sample was clear, the positive control became turbid within 48 hours, the negative control was clear, and the sterility test was qualified. In the comparative test, when the same batch of samples was treated with a traditional one-way flushing device without an absorbent pad, no Staphylococcus aureus was observed to grow in the positive control within 72 hours, showing a false negative. This proves that the two-way flushing and absorbent pad neutralization design of the present invention can efficiently remove antibacterial substances and ensure the accuracy of the test.

[0038] Example 3: Sterility test of large-volume injection solution. This example is basically the same as Example 2, except that the test sample is 500 mL glucose injection solution without antibacterial agent. In order to optimize efficiency, the rinsing solution can be replaced with 0.9% sterile sodium chloride solution. The number of alternating positive and negative pressure cycles in step S3 can be reduced to 3 times. Other operations and culture conditions remain unchanged. After 14 days, the test sample device is clear, and the positive control is turbid. The result is qualified.

[0039] Example 4: Application of online integrity testing. This example emphasizes the important role of integrity. In the test, after inflating the device to 5 kPa according to step S1 and closing the on / off valve, it was observed that the pressure dropped by 1.2 kPa within 5 minutes, exceeding the threshold of 0.5 kPa. The device was immediately abandoned, replaced with a new device, and the test was repeated. The sealing was qualified. Subsequent incubation results showed that the test sample was clear, the positive control was turbid, and the negative control was clear, indicating that the test was effective. This example avoids false positive results that may be caused by contaminants introduced due to leakage of the device itself, demonstrating the practical value of online integrity testing.

[0040] In summary, the aseptic testing device and method proposed in this invention, through a fully enclosed aseptic interface, a pressure-buffered double-layered tank, a sterile absorbent pad with a neutralizing agent, at least one on / off valve for sealing the inner cavity, and a specific bidirectional flushing procedure, achieves the following beneficial effects: First, the online integrity testing function can confirm the device's sealing performance before incubation, preventing false positives caused by device leakage; Second, bidirectional flushing, combined with the sterile absorbent pad under the filter membrane, can deeply remove residual antibacterial substances on both sides and inside the filter membrane, thereby effectively avoiding false negatives; Third, the fully enclosed operation throughout the process greatly reduces the risk of external contamination and improves the accuracy and reliability of aseptic test results. This device and method are particularly suitable for the aseptic testing of test samples containing preservatives, antibiotics, and other antibacterial substances.

[0041] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A sterile inspection device, characterized in that, include: A double-walled tank, comprising an inner cavity and an outer cavity located outside the inner cavity, wherein the lower chamber of the inner cavity is connected to the outer cavity through a small hole; A microporous filter membrane is fixed in the middle of the inner cavity, dividing the inner cavity into an upper chamber and a lower chamber; A sterile absorbent pad is located below the microporous filter membrane, and a neutralizing agent is pre-placed inside the sterile absorbent pad; Both the sample injection valve and the culture medium injection valve are connected to the upper chamber of the inner cavity; An exhaust filter assembly is connected to the upper chamber of the inner cavity, and a hydrophobic membrane is provided inside the exhaust filter assembly; An exhaust on / off valve is disposed between the exhaust filter assembly and the upper chamber of the inner cavity; Positive and negative pressure gas inlets are connected to the upper chamber of the inner cavity and are used to alternately apply positive and negative pressure to allow liquid to pass through the microporous filter membrane in both directions; A waste liquid discharge valve is located in the lower chamber of the inner cavity; A pressure sensor is used to monitor the pressure in the cavity; and An on / off valve is installed between the positive and negative pressure air inlet and the upper chamber of the inner cavity to seal the inner cavity.

2. The aseptic testing device according to claim 1, characterized in that, The sterile absorbent pad is placed in close contact with the underside of the microporous filter membrane, and the neutralizing agent is in a lyophilized state and is selected from one or more of lecithin, Tween 80, histidine, and sodium thiosulfate.

3. The aseptic testing device according to claim 1, characterized in that, Both the sample injection valve and the culture medium injection valve are needle-free puncture valves and are equipped with self-sealing silicone gaskets.

4. The aseptic testing device according to claim 1, characterized in that, The hydrophobic membrane in the exhaust filtration assembly has a pore size of 0.22 μm; the microporous filter membrane has a pore size of 0.45 μm.

5. The aseptic testing device according to claim 1, characterized in that, The inner cavity is a culture chamber, and the outer cavity is a negative pressure buffer chamber.

6. A method for performing aseptic testing using the aseptic testing apparatus according to any one of claims 1-5, characterized in that, Includes the following steps: S1 Integrity Test: Open the on / off valve, close the exhaust on / off valve, and fill the inner cavity with sterile air to a predetermined pressure through the positive and negative pressure air circuit interface. Then close the on / off valve and use the pressure sensor to monitor the pressure change in the inner cavity. If the pressure drop does not exceed the set threshold during the pressure holding period, the device is deemed to be in good condition. S2 Sample Filtration: Open the on / off valve and the exhaust on / off valve, inject the test sample into the upper chamber of the inner cavity through the sample injection valve to exhaust the gas in the upper chamber, then close the exhaust on / off valve, apply negative pressure through the positive and negative pressure gas path interface, so that the test sample passes through the filter membrane and enters the lower chamber, the filtrate is absorbed by the sterile absorption pad and the antibacterial substance is neutralized by the neutralizing agent therein; S3 bidirectional flushing: Open the exhaust on / off valve, inject flushing fluid into the upper chamber of the inner cavity through the culture medium injection valve to discharge the gas in the upper chamber, then close the exhaust on / off valve, keep the on / off valve open, and alternately apply positive and negative pressure through the positive and negative pressure gas circuit interface to make the flushing fluid flow back and forth on the microporous filter membrane at least 3 times; S4 Waste liquid discharge: Keep the exhaust valve closed, apply positive pressure to force the flushed liquid into the lower chamber of the inner cavity, and open the waste liquid discharge valve to discharge the waste liquid; S5 Culture Medium Injection and Cultivation: Close the waste liquid discharge valve and the on / off valve, open the exhaust on / off valve, inject the culture medium through the culture medium injection valve, then close the exhaust on / off valve, seal the culture, and observe the results.

7. The method for aseptic testing according to claim 6, characterized in that, In step S3, the positive pressure gauge pressure applied alternately is 0.02~0.10 MPa, and the negative pressure gauge pressure is -0.02~-0.10 MPa, with each pressure holding time being 2~10 seconds; the predetermined pressure mentioned in step S1 is 5~10 kPa, and the set threshold is a pressure drop ≤0.5 kPa within 5 minutes.

8. The method for aseptic testing according to claim 6, characterized in that, The rinsing solution in step S3 is an aqueous solution containing 0.1% peptone and 0.1% Tween 80, or a 0.9% sterile sodium chloride solution.

9. The method for aseptic testing according to claim 6, characterized in that, The culture medium mentioned in step S5 is thioglycolate fluid medium, and the culture conditions are 35°C for 14 days.

10. The method for aseptic testing according to claim 6, characterized in that, The method further includes: Positive control: Using the same batch of the sterile testing device, add known positive microorganisms and operate according to steps S1 to S5 as a positive control; Negative control: Use sterile water instead of the test sample, and follow steps S1 to S5 as a negative control; Result determination: If the sample device becomes turbid after incubation, and the positive control is turbid while the negative control is clear, it is judged as positive; if the sample device is clear while the positive control is turbid, it is judged as negative; if the negative control is turbid, the test is invalid.

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