A method and device for detecting bacteria in water

Abstract

The present invention provides a method and device for detecting bacteria in water. By setting the first through-hole membrane and the second through-hole membrane in the enrichment module, rapid enrichment of Escherichia coli in water can be achieved, and the enrichment process is stable and not prone to clogging. Then, a reaction reagent reacts specifically with Escherichia coli to obtain an enrichment solution containing Escherichia coli. Subsequently, the enrichment solution is mixed with an oil solution to generate micro-droplets and amplified. The reaction reagent is converted into a fluorescent substance under the enzymatic action of Escherichia coli. After statistical analysis, the content of Escherichia coli in water can be determined. Its detection time is short, and the detection result can be obtained in only one hour. The detection sensitivity is high, the detection specificity is good, and on-site detection can be carried out anytime and anywhere. In addition, by replacing different reaction reagents, different types of bacteria can be detected, thus greatly expanding the range of detected bacteria types. Moreover, the device for detecting bacteria in water can be reused repeatedly for a long time, thereby reducing the cost.

Description

Technical Field

[0001] The present invention relates to the technical field of environmental monitoring, and particularly to a method and device for detecting bacteria in water. Background Art

[0002] The life and production of the people are inseparable from the water environment, and water is the basis for the survival of all organisms in the world. With the increasingly serious problem of water pollution, the water quality safety of drinking water has attracted more and more attention from people. Humans often come into contact with bacteria in water through drinking, touching, etc., especially Escherichia coli in domestic drinking water, which causes serious harm to human health. In order to ensure the hygienic quality of domestic drinking water, it is necessary to conduct water quality detection to obtain the Escherichia coli pollution index, so as to judge whether the domestic drinking water source is contaminated by Escherichia coli.

[0003] The main detection methods for Escherichia coli are the enzyme substrate method and the ATP fluorescence detection method. For example, the Colilert-18 detection instrument developed by IDEXX Company in the United States only needs to put the sample to be detected into a multi-well cavity test bag and incubate it for 18 to 24 hours to achieve the detection of Escherichia coli. Although the WTF series products launched by Jingxiang Company have a high degree of automation, they still rely on bacterial culture, resulting in a relatively long detection time and a large instrument volume. The detection instrument developed by Hengmei Company based on the ATP fluorescence method is simple to operate and portable, with a fast detection speed, and only takes 15 seconds to complete the detection, but it has low sensitivity and poor detection specificity. It can be seen that the above detection methods need to culture Escherichia coli for a long time to achieve highly sensitive detection, so it will lead to problems such as too long detection time and inability to sample on-site.

[0004] At present, with the continuous occurrence of domestic drinking water source pollution incidents in various places, new requirements have been put forward for the detection of bacteria in water, that is, on the premise of ensuring the reliability and accuracy of the test results, a large number of tests need to be completed more quickly and efficiently. At the same time, due to the high requirements for the storage conditions of water samples in the detection of water body bacteria, on-site detection is often required at the water sample collection point in the wild. Therefore, the conventional detection methods are difficult to meet the above detection requirements. Summary of the Invention

[0005] In view of the above-mentioned disadvantages of the prior art, the purpose of the present invention is to provide a method and device for detecting bacteria in water, which are used to solve the problems of long detection time, low detection sensitivity, poor detection specificity, single type of detected bacteria and inability to perform on-site detection in the prior art.

[0006] To achieve the above purpose and other related purposes, the present invention provides a device for detecting bacteria in water, and the detection device includes:

[0007] A micro-droplet module for generating micro-droplets, the micro-droplet module comprising an enrichment liquid inlet, an oil liquid inlet and a micro-droplet outlet, and the enrichment liquid inlet being sequentially connected to a first control valve and a first pump body, and the oil liquid inlet being sequentially connected to a second control valve and a second pump body;

[0008] An enrichment module, the enrichment module comprising a water source inlet, a waste liquid outlet, a reaction reagent inlet, an enrichment liquid outlet, and a first through-hole membrane and a second through-hole membrane that are connected to each port and allow liquid to pass through in sequence, and the waste liquid outlet being sequentially connected to a third control valve and a third pump body, the reaction reagent inlet being sequentially connected to a fourth control valve and a fourth pump body, the water source inlet being connected to a fifth control valve, and the enrichment liquid outlet being connected to a sixth control valve;

[0009] A water source storage unit connected to the fifth control valve;

[0010] A waste liquid collection unit connected to the third pump body and the micro-droplet outlet respectively;

[0011] An enrichment liquid collection unit connected to the sixth control valve and the enrichment liquid outlet respectively;

[0012] A reaction reagent supply unit connected to the fourth pump body;

[0013] An oil liquid supply unit connected to the second pump body.

[0014] Optionally, the micro-droplet module includes a micro-droplet generation part and a micro-droplet storage part connected to the micro-droplet generation part, wherein the enrichment liquid inlet and the oil liquid inlet are arranged on the micro-droplet generation part.

[0015] Optionally, the first pump body, the second pump body and the fourth pump body are positive pressure pumps, and the third pump body is a negative pressure pump.

[0016] Optionally, the first through-hole membrane is arranged above the second through-hole membrane.

[0017] Optionally, the pore diameter of the first through-hole membrane is larger than that of the second through-hole membrane.

[0018] Optionally, the pore diameter of the first through-hole membrane is 2 μm to 20 μm, and the pore diameter of the second through-hole membrane is 0.1 μm to 0.45 μm.

[0019] The present invention provides a method for detecting bacteria in water, the detection method comprising the following steps:

[0020] Providing the detection device according to any one of claims 1 to 6;

[0021] Close the fourth control valve and the sixth control valve, open the third control valve and the fifth control valve, start the third pump body so that the water source to be detected enters the enrichment module, and sequentially pass through the first through-hole membrane and the second through-hole membrane, and the enriched bacteria are obtained in the second through-hole membrane;

[0022] Close the third control valve and the fifth control valve, open the fourth control valve and the sixth control valve, start the fourth pump body so that the reaction reagent enters the second through-hole membrane, specifically react with the enriched bacteria to obtain an enrichment solution, and store the enrichment solution in the enrichment solution collection unit;

[0023] Open the second control valve and the first control valve, start the second pump body and the first pump body, and pump the enrichment solution and the oil liquid into the micro-droplet module to obtain water-in-oil micro-droplets;

[0024] Set a specific temperature to amplify the micro-droplets in the micro-droplet module and perform an enzymatic reaction for a certain period of time, thereby generating a fluorescent substance, and statistically calculate the fluorescent substance to obtain the concentration of bacteria in the water source to be detected.

[0025] Optionally, the diameter of the micro-droplets is 10 μm to 200 μm.

[0026] Optionally, use a CCD camera to take pictures to identify whether the micro-droplets have fluorescence characteristics to determine whether the micro-droplets contain bacteria.

[0027] Optionally, the temperature of the enzymatic reaction is 44 degrees, and the time of the enzymatic reaction is 40 min.

[0028] Optionally, the type of the bacteria is Escherichia coli.

[0029] As described above, a method and device for detecting bacteria in water according to the present invention have the following beneficial effects: Through the setting of the double through-hole membrane in the enrichment module, rapid enrichment of bacteria in the water source to be detected can be achieved, and the bacteria enrichment process is stable and not easily blocked. The reaction reagent reacts specifically with the bacteria to generate an enrichment solution with bacteria. Subsequently, the enrichment solution and the oil liquid are mixed to generate micro-droplets and amplified and subjected to an enzymatic reaction for 40 minutes, thereby converting the reaction reagent into a fluorescent substance. By taking pictures, analyzing and statistically calculating, the content of bacteria in the water source to be detected is finally determined. The detection time is short, the detection sensitivity is high, the detection specificity is good, and on-site detection can be carried out anytime and anywhere. In addition, by replacing different reaction reagents, different types of bacteria can be detected, thereby greatly expanding the scope of bacteria detection. In addition, the detection device can be reused for a long time, thereby reducing the maintenance cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Shown is a schematic flow diagram of a method for detecting bacteria in water according to the present invention.

[0031] Figure 2 Shown is a schematic structural diagram of a device for detecting bacteria in water according to the present invention.

[0032] Figure 3 Shown is a schematic structural diagram of an enrichment module in a device for detecting bacteria in water according to the present invention.

[0033] Figure 4 Shown is a schematic structural diagram of a microdroplet module in a device for detecting bacteria in water according to the present invention.

[0034] Figure 5 Shown is a schematic working diagram of the detection device when enriching bacteria in a method for detecting bacteria in water according to the present invention.

[0035] Figure 6 Shown is a schematic working diagram of the detection device when obtaining an enrichment solution in a method for detecting bacteria in water according to the present invention.

[0036] Figure 7 Shown is a schematic working diagram of the detection device when generating microdroplets in a method for detecting bacteria in water according to the present invention.

[0037] Description of component labels

[0038] 101 Enrichment module

[0039] 1011 Water source inlet

[0040] 1012 Waste liquid outlet

[0041] 1013 Reagent inlet

[0042] 1014 Enrichment solution outlet

[0043] 1015 First through-hole membrane

[0044] 1016 Second through-hole membrane

[0045] 102 Microdroplet module

[0046] 1021 Microdroplet generation part

[0047] 1022 Microdroplet storage part

[0048] 1023 Enrichment solution inlet

[0049] 1024 Oil liquid inlet

[0050] 1025 Microdroplet outlet

[0051] 103 Water source storage unit

[0052] 1031 Water source to be detected

[0053] 104 Fifth control valve

[0054] 105 Waste liquid collection unit

[0055] 1051 Waste liquid

[0056] 106 Third pump body

[0057] 107 Third control valve

[0058] 108 Enriched liquid collection unit

[0059] 1081 Enriched liquid

[0060] 109 Sixth control valve

[0061] 110 Reaction reagent supply unit

[0062] 1101 Reaction reagent

[0063] 111 Fourth pump body

[0064] 112 Fourth control valve

[0065] 113 Oil supply unit

[0066] 1131 Oil

[0067] 114 Second pump body

[0068] 115 Second control valve

[0069] 116 First pump body

[0070] 117 First control valve

[0071] Steps S1 to S5 Specific implementation manners

[0072] The following uses specific specific examples to illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners, and various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0073] For ease of description, spatial relationship terms such as "below", "beneath", "lower", "under", "above", "on" etc. may be used herein to describe the relationship of one element or feature shown in the drawings to other elements or features. It will be understood that these spatial relationship terms are intended to encompass other directions of the device in use or operation, in addition to the directions depicted in the drawings. Further, when a layer is referred to as being "between" two layers, it can be the only layer between the two layers, or there can be one or more intervening layers.

[0074] It should be understood that the use of terms such as "first", "second" etc. to define components is only for the convenience of differentiating the above-mentioned components. Without additional statements, these terms have no special meaning, and thus should not be construed as limiting the scope of protection of the present invention.

[0075] In this application, unless otherwise clearly specified and defined, terms such as "mounted", "connected", "coupled", "fixed" etc. should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or integrated; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the internal communication of two components or the interaction relationship between two components, unless otherwise clearly defined. For those of ordinary skill in the art, the specific meanings of the above terms in this application can be understood according to specific circumstances.

[0076] Please refer to Figures 1 to 7 . It should be noted that the illustrations provided in this embodiment only schematically illustrate the basic concept of the present invention. Therefore, only the components related to the present invention are shown in the illustrations, rather than being drawn according to the number, shape and size of the components in actual implementation. The type, quantity and proportion of each component in actual implementation can be arbitrarily changed, and the component layout type may also be more complex.

[0077] In one embodiment of the present invention, a detection device for bacteria in water is provided. The detection device includes: an enrichment module 101, a microdroplet module 102, a water source storage unit 103, a waste liquid collection unit 105, an enrichment liquid collection unit 108, a reaction reagent supply unit 110, and an oil liquid supply unit 113.

[0078] As Figure 3 shown, the enrichment module 101 includes a water source inlet 1011, a waste liquid outlet 1012, a reaction reagent inlet 1013, an enrichment liquid outlet 1014, and a first through-hole membrane 1015 and a second through-hole membrane 1016 that are in communication with each port so that liquid can pass through in sequence. As Figure 2As shown, the water source inlet 1011 is connected to an external water source storage unit 103, and a fifth control valve 104 is connected at the water source inlet 1011. The water source storage unit 103 includes a water source storage container and a water source inlet pipeline. Among them, the volume of the water source storage container is generally 100 ml, so as to quantitatively sample the water source 1031 to be detected. The water source inlet pipeline is communicated with the fifth control valve 104 to control the on-off state of the water source inlet pipeline through the fifth control valve 104.

[0079] As Figure 2 and Figure 3 shown, the waste liquid outlet 1012 is sequentially connected to a third control valve 107 and a third pump body 106. A waste liquid collection unit 105 is also connected to the third pump body 106. The waste liquid collection unit 105 includes a first waste liquid pipeline and a waste liquid storage container. The waste liquid generated during the bacteria enrichment process flows through the first waste liquid pipeline to the waste liquid storage container. Since the water source storage container and the water source inlet 1011 of the enrichment module 101 are communicated through the water source inlet pipeline and the water source inlet 1011 is provided with a fifth control valve 104, when performing bacteria enrichment, the reaction reagent inlet 1013 and the enrichment liquid outlet 1014 are kept in a closed state. At this time, the third control valve 107 and the fifth control valve 104 are kept in an open state, and the third pump body 106 is started. Since the third pump body 106 is a negative pressure pump, negative pressure can be generated so that the water source 1031 to be detected can enter the enrichment module 101 through the water source inlet 1011. Since the water source inlet 1011, the first through-hole membrane 1015, the second through-hole membrane 1016 and the waste liquid outlet 1012 are communicated with each other, after the water source 1031 to be detected enters the enrichment module 101, it first passes through the first through-hole membrane 1015. The pore diameter of the first through-hole membrane 1015 is 2 μm to 20 μm. For example, it can be 2 μm, 10 μm or 20 μm. There is no excessive limitation here. A large amount of impurities in the water source 1031 to be detected will be filtered out in the first through-hole membrane 1015. Then the water source 1031 to be detected passes through the second through-hole membrane 1016. The pore diameter of the second through-hole membrane 1016 is 0.1 μm to 0.45 μm. For example, it can be 0.1 μm, 0.25 μm or 0.45 μm. There is no excessive limitation here. The bacteria in the water source 1031 to be detected will be retained in the second through-hole membrane 1016 and enriched in the second through-hole membrane 1016. In this embodiment, the type of the bacteria is Escherichia coli. After the water source 1031 to be detected passes through the second through-hole membrane 1016, the excess waste liquid will pass through the waste liquid outlet 1012 and flow through the first waste liquid pipeline to the waste liquid storage container of the waste liquid collection unit 108.

[0080] As Figure 2 and Figure 3As shown, a fourth control valve 112 and a fourth pump body 111 are connected to the reaction reagent inlet 1013. Also connected to the fourth pump body 111 is a reaction reagent supply unit 110, which includes a reaction reagent storage and a reaction reagent supply pipeline. A certain amount of reaction reagent 1101 is stored in the reaction reagent storage. The type of the reaction reagent 1101 is 4-methylumbelliferyl-β-D-glucuronide (4-MUG). The reaction reagent 1101 can be hydrolyzed by β-glucuronidase (GUS) in Escherichia coli into 4-methylumbelliferone (4-MU). A sixth control valve 109 is connected to the enrichment liquid outlet 1014. Also connected to the sixth control valve 109 is an enrichment liquid collection unit 108, which includes an enrichment liquid storage and an enrichment liquid collection pipeline. The enrichment liquid storage is used to collect the enrichment liquid 1081. In this embodiment, when enriching bacteria for a specific reaction to generate the enrichment liquid, the water source inlet 1011 and the waste liquid outlet 1012 are in a closed state, that is, the third control valve 107 and the fifth control valve 104 remain closed to prevent the reaction reagent 1101 from flowing back to the waste liquid collection unit 105 and the water source storage unit 103. Then, the fourth control valve 112 and the sixth control valve 109 are kept open, and the fourth pump body 111 is started. Since the fourth pump body 111 is a positive pressure pump, it can generate power to enable the reaction reagent 1101 in the reaction reagent storage to enter the enrichment module 101 and react specifically with the bacteria enriched in the second through-hole membrane 1016 to generate the enrichment liquid 1081. The generated enrichment liquid 1081 will then pass through the enrichment liquid outlet 1014, flow through the enrichment liquid collection pipeline to the enrichment liquid collection unit 108, and be stored in the enrichment liquid storage.

[0081] As Figure 4As shown in the figure, the micro-droplet module 102 includes a micro-droplet generating unit 1021. An enrichment liquid inlet 1023 and an oil liquid inlet 1024 are provided on the micro-droplet generating unit 1021. Among them, an enrichment liquid supply pipeline is provided between the enrichment liquid inlet 1023 and the enrichment liquid collection unit 108. A first control valve 117 and a first pump body 116 are respectively connected to the enrichment liquid inlet 1023, and the first pump body 116 is a positive pressure pump. Thus, the enrichment liquid 1081 is pumped into the micro-droplet generating unit 1021 by the power provided by the first pump body 116. The oil liquid supply unit 113 contains an oil liquid with different characteristics from the enrichment liquid 1081. An oil liquid supply pipeline is provided between the oil liquid inlet 1024 and the oily solution supply unit 113. A second control valve 115 and a second pump body 114 are successively connected to the oil liquid inlet 1024, and the second pump body 114 is a positive pressure pump. Thus, the oil liquid 1131 is pumped into the micro-droplet generating unit 1021 by the power provided by the second pump body 114. The enrichment liquid 1081 and the oil liquid 1131 can generate water-in-oil micro-droplets in the micro-droplet generating unit 1021. The micro-droplet module 102 further includes a micro-droplet storage unit 1022. Among them, the micro-droplet storage unit 1022 is interconnected with the micro-droplet generating unit 1021. Thus, the micro-droplets can enter the micro-droplet storage unit 1022 and undergo amplification and enzymatic reactions in the micro-droplet storage unit 1022.

[0082] In another embodiment of the present invention, a method for detecting bacteria in water is further provided. The detection method includes the following steps:

[0083] S1: Provide the detection device according to any one of claims 1 to 6;

[0084] S2: Close the fourth control valve and the sixth control valve, open the third control valve and the fifth control valve, start the third pump body to make the water source to be detected enter the enrichment module, and successively pass through the first through-hole membrane and the second through-hole membrane, and the enriched bacteria are obtained in the second through-hole membrane;

[0085] S3: Close the third control valve and the fifth control valve, open the fourth control valve and the sixth control valve, start the fourth pump body to make the reaction reagent enter the second through-hole membrane, specifically react with the enriched bacteria to obtain an enrichment liquid, and store the enrichment liquid in the enrichment liquid collection unit;

[0086] S4: Open the second control valve and the first control valve, start the second pump body and the first pump body, and pump the enrichment liquid and the oil liquid into the micro-droplet module to obtain water-in-oil micro-droplets;

[0087] S5: Set a specific temperature to amplify the microdroplets in the microdroplet module and perform an enzymatic reaction for a certain period of time, thereby generating a fluorescent substance, and statistically calculate the fluorescent substance to obtain the concentration of bacteria in the water source to be detected.

[0088] Specifically, provide the detection device according to any one of claims 1 to 6. As Figure 5 shown, in step S2, place the wastewater containing bacteria in the water source storage unit 103 as the water source 1031 to be detected. The type of the bacteria is Escherichia coli. Therefore, when enriching Escherichia coli, keep the reaction reagent inlet 1013 and the enrichment liquid outlet 1014 closed, that is, close the fourth control valve 112 and the sixth control valve 109, keep the third control valve 107 and the fifth control valve 104 open, and start the third pump body 106. Since the third pump body 106 is a negative pressure pump, negative pressure can be generated so that the water source 1031 to be detected can enter the enrichment module 101 through the water source inlet 1011. Since the water source inlet 1011, the first through-hole membrane 1015, the second through-hole membrane 1016 and the waste liquid outlet 1012 are interconnected, after the water source 1031 to be detected enters the enrichment module 101, it first passes through the first through-hole membrane 1015. Since the aperture of the first through-hole membrane 1015 is 2 μm to 20 μm and the aperture of the second through-hole membrane 1016 is 0.1 μm to 0.45 μm, a large amount of impurities in the water source 1031 to be detected are filtered out in the first through-hole membrane 1015, while Escherichia coli will be retained in the second through-hole membrane 1016 and enriched in the second through-hole membrane 1016.

[0089] Specifically, as Figure 6 shown, in step S3, when generating the enrichment liquid, keep the water source inlet 1011 and the waste liquid outlet 1012 closed, that is, close the third control valve 107 and the fifth control valve 104 to prevent the reaction reagent 1101 from flowing back to the waste liquid collection unit 105 and the water source storage unit 103. Then keep the fourth control valve 112 and the sixth control valve 109 open, and start the fourth pump body 111. Since the reaction reagent inlet 1013, the enrichment liquid outlet 1014 and the first through-hole membrane 1015 are interconnected and the fourth pump body 111 is a positive pressure pump, the power generated by the fourth pump body 111 can be used to enable the reaction reagent 1101 in the reaction reagent supply unit 110 to enter the enrichment module 101 and react specifically with the bacteria enriched in the second through-hole membrane 1016 to generate the enrichment liquid 1081. The enrichment liquid 1081 will flow through the enrichment liquid outlet 1014 to the enrichment liquid collection unit 108 and be stored in the enrichment liquid storage.

[0090] Specifically, as Figure 7As shown, in step S4, the third control valve 107, the fourth control valve 112, the fifth control valve 104, and the sixth control valve 109 are closed, the second control valve 115 and the first control valve 117 are opened, the second pump body 114 is started to pump the enriched liquid 1081 in the enriched liquid collection unit 108 into the micro-droplet module 102, and the first pump body 116 is started to pump the oil liquid 1131 in the oil liquid supply unit 113 into the micro-droplet module 102 as well. The enriched liquid 1081 and the oil liquid 1131 are dispersed with each other in the micro-droplet generation part 1021 to generate water-in-oil micro-droplets. The diameter of the generated micro-droplets is 10 μm to 200 μm. For example, it can be 10 μm, 50 μm, 100 μm, 150 μm, or 200 μm. The diameter size of the micro-droplets can be selected according to actual needs and is not limited here.

[0091] In step S5, the micro-droplets generated in the micro-droplet generation part 1021 will then enter the micro-droplet storage part 1022 and spread out in the micro-droplet storage part 1022. A specific temperature is set to enable the micro-droplets to amplify and undergo an enzymatic reaction for a certain period of time. Specifically, the temperature of the enzymatic reaction is set to 44 degrees, so that the β-glucuronidase (GUS) in Escherichia coli has the highest activity. Since the reaction reagent used in step S2 is 4-methylumbelliferyl-β-D-glucuronide (4-MUG), and the enriched bacterial type is Escherichia coli, and 4-methylumbelliferyl-β-D-glucuronide can be hydrolyzed by the β-glucuronidase (GUS) in Escherichia coli into 4-methylumbelliferone (4-MU). Conducting an enzymatic reaction for 40 minutes can make the production amount of 4-methylumbelliferone reach the maximum. Since 4-methylumbelliferone is a fluorescent substance, the CCD is used to take pictures to identify whether the micro-droplets have fluorescence characteristics to determine whether the micro-droplets contain bacteria, and then by counting and calculating the number of fluorescences, the content of Escherichia coli in the water source to be detected can be determined.

[0092] In summary, the present invention provides a method and device for detecting bacteria in water. Through the arrangement of the first through-hole membrane and the second through-hole membrane in the enrichment module, rapid enrichment of Escherichia coli in water can be achieved, and the enrichment process is stable and not easily blocked. Then, a specific reaction occurs between the reaction reagent and Escherichia coli to obtain an enrichment solution containing Escherichia coli. Subsequently, the enrichment solution is mixed with an oil solution to generate microdroplets, which are amplified and subjected to an enzymatic reaction for about one hour, enabling the reaction reagent to be converted into a fluorescent substance under the action of Escherichia coli. After statistical analysis, the content of Escherichia coli in water can be determined. Compared with the prior art, the detection time is short, and the detection result can be obtained in only one hour. The detection sensitivity is high, the detection specificity is good, and on-site detection can be carried out anytime and anywhere. In addition, by replacing different reaction reagents, different types of bacteria can be detected, thus greatly expanding the range of bacteria detection types. Moreover, the device for detecting bacteria in water can be reused repeatedly for a long time, thereby reducing the cost. Therefore, the present invention effectively overcomes various shortcomings in the prior art and has high industrial utilization value.

[0093] The above embodiments are only illustrative of the principles and effects of the present invention and are not intended to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes made by those with ordinary knowledge in the technical field without departing from the spirit and technical idea disclosed by the present invention should still be covered by the claims of the present invention.

Claims

1. A method for detecting bacteria in water, characterized in that, The detection method includes the following steps: Provide a detection device, which at least includes: A micro-droplet module for generating micro-droplets. The micro-droplet module includes an enrichment liquid inlet, an oil liquid inlet, and a micro-droplet outlet. The enrichment liquid inlet is sequentially connected with a first control valve and a first pump body, and the oil liquid inlet is sequentially connected with a second control valve and a second pump body; An enrichment module including a water source inlet, a waste liquid outlet, a reaction reagent inlet, an enrichment liquid outlet, and a first through-hole membrane and a second through-hole membrane connected to each port to allow the liquid to pass through in sequence. The waste liquid outlet is sequentially connected with a third control valve and a third pump body, the reaction reagent inlet is sequentially connected with a fourth control valve and a fourth pump body, the water source inlet is connected with a fifth control valve, and the enrichment liquid outlet is connected with a sixth control valve; A water source storage unit connected to the fifth control valve; A waste liquid collection unit connected to the third pump body and the micro-droplet outlet respectively; An enrichment liquid collection unit connected to the sixth control valve and the enrichment liquid outlet respectively; A reaction reagent supply unit connected to the fourth pump body; An oil liquid supply unit connected to the second pump body; Close the fourth control valve and the sixth control valve, open the third control valve and the fifth control valve, and start the third pump body to make the water source to be detected enter the enrichment module and pass through the first through-hole membrane and the second through-hole membrane in sequence, and the enriched bacteria are obtained in the second through-hole membrane; Close the third control valve and the fifth control valve, open the fourth control valve and the sixth control valve, and start the fourth pump body to make the reaction reagent enter the second through-hole membrane. The reaction reagent reacts specifically with the enriched bacteria to generate an enrichment liquid, and store the enrichment liquid in the enrichment liquid collection unit; Open the second control valve and the first control valve, start the second pump body and the first pump body, and pump the enrichment liquid and the oil liquid into the micro-droplet module to obtain water-in-oil micro-droplets; Set a specific temperature to perform amplification and an enzymatic reaction for a certain period of time on the micro-droplets in the micro-droplet module, thereby generating a fluorescent substance, and perform statistical calculation on the fluorescent substance to obtain the concentration of bacteria in the water source to be detected.

2. The detection method according to claim 1, characterized in that: The diameter of the micro-droplets is 10μm - 200μm.

3. The detection method according to claim 1, characterized in that: Use a CCD camera to take pictures to identify whether the micro-droplets have fluorescence characteristics to determine whether the micro-droplets contain bacteria.

4. The detection method according to claim 1, wherein: The temperature of the enzymatic reaction is 44 degrees, and the time of the enzymatic reaction is 40 min.

5. The detection method according to claim 1, characterized in that: The type of the bacteria is Escherichia coli.

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