Bacterial identification microfluidic chip and use method
By designing a microfluidic chip for bacterial identification, centrifugal force is used to distribute bacterial solution, and a bent microchannel is combined to achieve sealed sample addition and color development. This solves the problems of inconvenience and cross-contamination in the existing microplate method, improves identification efficiency and accuracy, and is suitable for culture in various environments.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- JIANG SU LING XIN SHENG WU KE JI YOU XIAN GONG SI
- Filing Date
- 2023-05-25
- Publication Date
- 2026-07-21
AI Technical Summary
Existing microplate methods for microbial identification are inconvenient to operate, involve complicated sample loading processes, are prone to cross-contamination, have inaccurate identification efficiency, cannot meet the requirements for detecting multiple biochemical characteristics, and cannot observe gas changes under sealed conditions, thus affecting the accuracy of the results.
A microfluidic chip for bacterial identification was designed, which uses a reagent tray in the shape of a CD disc. It uses centrifugal force to distribute bacterial samples into multiple reaction chambers. Combined with a bent microchannel and an annular guide channel, it can achieve sealed sample addition and automatic addition of colorimetric reagents. It is suitable for culturing in both caustic and anaerobic environments.
It simplifies the equipment structure, reduces costs, improves identification efficiency and accuracy, avoids aerosol cross-contamination, meets the needs of various biochemical trait detection, and is suitable for cultivation in caustic and anaerobic environments.
Smart Images
Figure CN116689051B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of bacterial identification technology, specifically to a microfluidic chip for bacterial identification and its usage method. Background Technology
[0002] Bacterial identification is a component of taxonomy. Clinical bacterial identification can classify bacteria down to the genus and species level, and it is increasingly widely used in rapid bacterial detection, bacterial resistance testing, and epidemiological surveys of bacterial infections. Bacterial identification methods include biochemical identification, nucleic acid detection, serological identification, automated instrument identification, and mass spectrometry. Among these, biochemical identification is the most classic and widely used method in clinical bacterial identification. Biochemical identification primarily utilizes the differences in bacteria's ability to break down nutrients and the differences in their metabolic products to identify bacteria. This includes tests for protein breakdown products, catalases, sugar breakdown products, oxidases, and coagulating enzymes. Currently, micro-biochemical identification tubes and bacterial identification plates are commonly used in clinical practice. These require manual sample addition. For example, Chinese patent CN110270391A discloses a micro-biochemical identification tube rack, including a support frame. The support frame includes a base plate, a first upright plate fixedly installed at the left end of the base plate, and a second upright plate fixedly installed at the end of the base plate away from the first upright plate. Side plates are fixedly installed on opposite sides of the first and second upright plates. A first test tube placement layer is engaged with the opposite side of the two side plates. A second test tube placement layer is fixedly installed on the top of the side plates, located on the opposite side of the first and second upright plates. Both the first and second test tube placement layers have through holes and round holes at their tops. A fixing block is fixedly installed on the top of the base plate, with a limiting groove on its top. A first locking groove is formed on the front surface of the side plates. The test tubes are engaged within the limiting groove, improving stability and preventing spillage of the solution in the test tubes. Furthermore, one test tube rack can simultaneously complete forty biochemical identification items, resulting in high identification efficiency and ease of use. However, since several or dozens of biochemical identification tubes are required for identification, the sample addition process is time-consuming and labor-intensive, and the complicated sample addition process increases the chance of aerosols and cross-contamination.
[0003] For another example, Chinese Patent CN101413876A discloses a microplate bioassay method for rapidly detecting the antibacterial activity of antibacterial substances. This method utilizes the high antibacterial properties of a test sample against sensitive indicator bacteria, and uses a microplate with good standardization and parallelization characteristics as a high-throughput detection platform. After the sensitive bacteria indicator solution and the test sample are fully mixed in the microplate wells and cultured for an appropriate time, the antibacterial activity of the sample is rapidly detected by measuring the change in the turbidity of the culture solution in each well of the microplate using an enzyme-labeled instrument. This transformation overcomes the drawbacks of existing bioassay methods, such as cumbersome steps and significant interference from human factors. However, for the fully automated microbial identification supporting the current microbial identification microplate method, due to the complex sample addition process, the instrument's mechanical structure correspondingly requires an XYZ robotic arm and a microplate cover grabbing device, resulting in a relatively complex structure and a large volume of the device, thus leading to a high manufacturing cost.
[0004] In addition, the current microbial identification microplate method uses an unsealed cover. When certain microorganisms, such as Enterobacteriaceae bacteria, ferment carbon source substrates such as glucose and lactose to produce acid and gas, the generated gas will escape into the air and cannot be observed, which will lead to a relatively large amount of liquid evaporation during the cultivation process, thus affecting the accuracy of result identification. Moreover, when colorimetric reagents need to be added to judge the growth state of bacteria in some bacterial identification biochemical reactions, the current microbial identification microplate method is also inconvenient to operate. Summary of the Invention
[0005] Aiming at the problems of the current microbial identification microplate method, such as inconvenient operation, cross-contamination during sample addition or cultivation, inaccurate identification efficiency, and inability to meet the requirements of detecting multiple biochemical traits of bacteria, this application provides a bacterial identification microfluidic chip and a usage method to achieve improving the efficiency of bacterial identification while ensuring no contamination and accurate identification during the bacterial identification process, and meeting the needs of changes in bacterial biochemical traits during the bacterial identification process.
[0006] The specific technical solutions include:
[0007] A bacterial identification microfluidic chip includes a bacterial identification reagent disk in the shape of a CD disc.
[0008] Centered on the center of the bacterial identification reagent disk, a shaft hole, a bacterial liquid sample addition groove, and an annular diversion flow channel are sequentially arranged from the center to the outside of the bacterial identification reagent disk.
[0009] The bacterial liquid sample addition groove is arranged outside the shaft hole.
[0010] The annular diversion flow channel is arranged outside the bacterial liquid sample addition groove, and the bacterial liquid sample addition groove is connected to the annular diversion flow channel through a bent microchannel.
[0011] A bacterial identification reaction chamber is provided on the outside of the annular guide channel, and the bacterial identification reaction chamber is connected to the annular guide channel through the bent microchannel.
[0012] Preferably, a colorimetric reagent addition chamber is provided on the outer side of the shaft hole, and the colorimetric reagent addition chamber is connected to the annular guide channel through a flow channel;
[0013] A colorimetric reaction chamber is provided on the outside of the annular guide channel. The colorimetric reaction chamber is matched with the colorimetric reagent addition chamber and is connected to the colorimetric reagent addition chamber through a wedge-shaped microchannel.
[0014] Preferably, the bacterial identification reaction chambers are evenly distributed on the outside of the annular guide channel and are connected to the annular guide channel through the bent microchannel.
[0015] Preferably, the bacterial identification reaction chamber is larger than the colorimetric reagent reaction chamber.
[0016] Preferably, the bacterial identification reaction chamber is pre-filled with a bacterial biochemical identification drying agent.
[0017] Preferably, the bacterial solution loading tank has an arc-shaped structure, with one end closed and the other end connected to the annular guide channel through the bent microfluidic channel;
[0018] After adding chemical reagents into the bacterial solution sample loading tank, the sample loading hole of the bacterial solution sample loading tank is sealed with a film.
[0019] Preferably, a dynamic balancing groove is also provided on the outer side of the shaft hole.
[0020] A method for using a microfluidic chip for bacterial identification includes the following steps:
[0021] Step 1: Add the bacterial solution sample to be tested into the bacterial solution sample tank;
[0022] Step 2: Under the action of centrifugal force, the bacterial liquid sample to be tested enters the annular guide channel through the bent microchannel, and then flows into the bacterial identification reaction chamber through the bent microchannel.
[0023] Step 3: After the bacterial identification reaction chamber interacts with the bacterial biochemical identification drying agent and is cultured for a certain period of time, the bacterial growth and metabolic state is determined by using microscopic photography combined with artificial intelligence analysis, and the bacterial identification result is obtained by combining the microbial biochemical identification table or database.
[0024] Preferably, in step 3, after determining the growth and metabolic state of bacteria, if some bacterial identification reaction chambers need to add chromogenic reagents after culture, chromogenic reagents can be added to the chromogenic reagent addition chamber. With the action of centrifugal force, the chromogenic reagents flow from the chromogenic reagent addition chamber into the chromogenic reagent reaction chamber, and the results are determined after the chromogenic reaction.
[0025] A method for using a microfluidic chip for bacterial identification, used for the identification of certain microorganisms cultured in caustic environments, includes the following steps:
[0026] Step 1: Add the bacterial solution sample to be tested into the bacterial solution sample tank;
[0027] Step 2: Under the action of centrifugal force, the bacterial liquid sample to be tested enters the annular guide channel through the bent microchannel, and then flows into the bacterial identification reaction chamber through the bent microchannel.
[0028] Step 3: After the bacterial liquid sample to be tested enters the bacterial identification reaction chamber, chemical reagents are added to the bacterial sample loading tank to create an aerobic microenvironment, and the sample loading hole on the bacterial sample loading tank is sealed with a film.
[0029] Step 4: Observe the growth and metabolic state of bacteria in the bacterial identification reaction chamber under caustic microenvironment culture, and obtain the bacterial identification results by combining the microbial biochemical identification table or database.
[0030] This invention has at least the following advantages:
[0031] 1) The bacterial identification microfluidic chip designed in this invention has a simple structure. It only requires a centrifugation module to replace the complex XYZ robotic arm and microplate cover gripping device, which greatly reduces the equipment cost. It uses centrifugal force to fill the bacterial solution of the bacterial sample to be tested into multiple bacterial identification reaction chambers at one time for biochemical reaction, which can detect multiple biochemical characteristics of bacteria. The operation is simple. The sample addition method is convenient. The sample addition process is carried out inside the microfluidic chip. It is sealed with a membrane before the reaction. The inside of the chip is sealed. Therefore, no aerosols or cross-contamination will be generated after sample addition or during the culture process, which greatly improves the identification efficiency and accuracy.
[0032] 2) This invention sets up a chromogenic reagent addition chamber and a chromogenic reaction chamber on the microfluidic chip for bacterial identification. When it is necessary to add a chromogenic reagent to determine the bacterial reaction result, the chromogenic reagent is directly centrifuged from the chromogenic reagent addition chamber into the chromogenic reaction chamber by centrifugation. The result is determined after the chromogenic reaction. The operation is simple.
[0033] 3) The microfluidic chip for bacterial identification designed in this invention can also be applied to certain microorganisms that require a caustic environment for culture. After the bacterial culture is centrifuged and enters the bacterial identification reaction chamber, chemical reagents are added to the bacterial culture sample tank to create a caustic microenvironment. The sample loading hole on the bacterial culture sample tank is then sealed with a membrane. This can meet the needs of some microorganism identification experiments that require a caustic environment for culture, greatly simplifying the culture environment for caustic bacteria and ordinary microorganisms and saving resources.
[0034] The above description is only an overview of the technical solution of this application. In order to better understand the technical means of this application and implement it in accordance with the contents of the specification, and to make the above and other objects, features and advantages of this application more obvious and understandable, the preferred embodiments of this application are described in detail below with reference to the accompanying drawings.
[0035] The above and other objects, advantages and features of this application will become more apparent to those skilled in the art from the following detailed description of specific embodiments in conjunction with the accompanying drawings. Attached Figure Description
[0036] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort. In all drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the elements or parts are not necessarily drawn to scale.
[0037] Figure 1 This is a schematic diagram of the structure of a microfluidic chip for bacterial identification provided by the present invention;
[0038] Figure 2 A plan view of a microfluidic chip for bacterial identification provided by the present invention;
[0039] The attached diagrams are labeled as follows: 1. Bacterial identification reagent tray; 2. Shaft hole; 3. Bacterial solution sample loading groove; 4. Chromogenic reagent sample loading chamber; 5. Annular guide channel; 6. Bent microchannel; 7. Bacterial identification reaction chamber; 8. Chromogenic reaction chamber; 9. Wedge-shaped microchannel; 10. Dynamic balance concave. Detailed Implementation
[0040] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. In the following description, specific details such as specific configurations and components are provided merely to help fully understand the embodiments of this application. Therefore, those skilled in the art should understand that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of this application. In addition, for clarity and brevity, descriptions of known functions and structures are omitted in the embodiments.
[0041] In this article, the term "and / or" is merely a description of the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can mean: A exists alone, B exists alone, and A and B exist simultaneously. The term " / and" in this article describes another type of relationship between related objects, indicating that two relationships can exist. For example, A / and B can mean: A exists alone, and A and B exist alone. In addition, the character " / " in this article generally indicates that the related objects before and after it are in an "or" relationship.
[0042] It should also be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion.
[0043] Example 1
[0044] This embodiment describes the structure of a microfluidic chip for bacterial identification.
[0045] As attached Figure 1 , 2 As shown, Figure 1 This is a schematic diagram of the structure of a microfluidic chip for bacterial identification provided by the present invention. Figure 2 A plan view of a microfluidic chip for bacterial identification provided by the present invention.
[0046] The microfluidic chip for bacterial identification includes a CD-shaped bacterial identification reagent tray 1. Centered on the center of the bacterial identification reagent tray 1, an axial hole 2, a bacterial solution loading groove 3, and an annular flow channel 5 are arranged sequentially from the center outwards from the outer side of the bacterial identification reagent tray 1.
[0047] The bacterial solution addition groove 3 is located on the outside of the shaft hole 2;
[0048] The annular guide channel 5 is disposed outside the bacterial solution sample loading tank 3, and the bacterial solution sample loading tank 3 is connected to the annular guide channel 5 through a bent microchannel 6.
[0049] A bacterial identification reaction chamber 7 is provided on the outside of the annular guide channel 5, and the bacterial identification reaction chamber 7 is connected to the annular guide channel 5 through the bent microchannel 6.
[0050] Furthermore, a colorimetric reagent addition chamber 4 is provided on the outer side of the shaft hole 2, and the colorimetric reagent addition chamber 4 is connected to the annular guide channel 5 through a flow channel;
[0051] A colorimetric reaction chamber 8 is provided on the outside of the annular guide channel 5. The colorimetric reaction chamber 8 is matched with the colorimetric reagent addition chamber 4 and is connected to the colorimetric reagent addition chamber 4 through a wedge-shaped microchannel 9.
[0052] Furthermore, the bacterial identification reaction chamber 7 is evenly distributed on the outside of the annular guide channel 5 and is connected to the annular guide channel 5 through the bent microchannel 6.
[0053] In this embodiment, 37 bacterial identification reaction chambers and 3 colorimetric reaction chambers are provided, with corresponding bent microfluidic channels 6 and wedge-shaped microfluidic channels 9.
[0054] Furthermore, the size of the bacterial identification reaction chamber 7 is larger than that of the colorimetric reagent reaction chamber 8.
[0055] Furthermore, the bacterial identification reaction chamber 7 is pre-filled with a bacterial biochemical identification drying agent.
[0056] Furthermore, a dynamic balancing groove 10 is provided on the outer side of the shaft hole 2.
[0057] Furthermore, the bacterial solution loading tank 3 has an arc-shaped structure, with one end closed and the other end connected to the annular guide channel via a bent microfluidic channel 6. After chemical reagents are added to the bacterial solution loading tank 3, the loading port is sealed with a membrane. Because microbial identification takes a long time, generally 4 to 24 hours, preventing the evaporation of the culture medium during this extended incubation period is a crucial issue. Current microplate methods for microbial identification use unsealed covers, resulting in significant liquid evaporation during incubation, which affects the accuracy of the results. This application employs a membrane seal, greatly reducing the evaporation of the culture medium.
[0058] Furthermore, the bent microfluidic channel 6 is an S-shaped microfluidic channel. Compared to microfluidic straight channels and microplates, the tortuous S-shaped microfluidic channel designed in this application helps to delay the evaporation of certain culture media. The following table shows the test experiments conducted on the S-shaped microfluidic channel, microfluidic straight channel, and microplate, respectively. The microorganisms to be identified were placed at 37°C for 4, 8, 16, and 24 hours, and the evaporation rate of the microorganisms under each channel structure was observed.
[0059] Table 1. Comparison of evaporation rates of microbial identification solutions under different structures.
[0060]
[0061] Experimental data comparison clearly shows that: with a direct-flow microfluidic channel, the liquid evaporation rate is approximately 5% after 16 hours of incubation at 37°C; the evaporation rate using the conventional microplate method with a cap exceeds 30%; while using a tortuous S-shaped microfluidic channel, the liquid evaporation rate is less than 1% after 16 hours of incubation at 37°C. Therefore, by setting the microfluidic channel to an S-shaped structure in this application, it will greatly help to delay the evaporation of certain microbial culture media during the identification process, thus affecting the identification effect and accuracy.
[0062] This bacterial identification structure eliminates the need for amplification and culturing, shortening the identification time. In this embodiment, taking 20 bacterial identification reaction chambers as an example, with each chamber having a volume of 25 μL, only about 1 mL of bacterial solution sample is needed per chamber. The reduced bacterial solution means a smaller number of bacteria are required to prepare a bacterial solution of a certain concentration.
[0063] Current microplate methods for microbial identification require 100-200 μL of bacterial suspension per well. If 20 wells are used, at least 2-4 mL of bacterial suspension is needed in each well. Moreover, it is usually necessary to pick colonies from solid plate culture and perform amplification culture again to obtain a sufficient number of bacteria, making the identification process cumbersome.
[0064] In this embodiment, only 2 to 5 colonies need to be picked directly to meet the identification needs of 20 bacterial identification reaction chambers, reducing the amplification culture process and time, and improving identification efficiency.
[0065] The bacterial identification microfluidic chip described in this embodiment has a simple structure. It only requires a centrifugation module to replace the complex XYZ robotic arm and microplate cover gripping device, simplifying the design and greatly reducing equipment costs. Utilizing the principle of centrifugal force, the bacterial sample to be tested is distributed into dozens of bacterial identification reaction chambers at once for biochemical reaction. It can detect multiple biochemical characteristics of bacteria. The operation is simple and the sample addition process is fast. The sample addition process is carried out inside the microfluidic chip, which is sealed with a membrane before the reaction. The inside of the chip is sealed, so no aerosols or cross-contamination will be generated after sample addition or during the culture process, which greatly improves the identification efficiency and accuracy.
[0066] In addition, in this embodiment, the microfluidic chip for bacterial identification is equipped with a chromogenic reagent addition chamber and a chromogenic reaction chamber. When it is necessary to add a chromogenic reagent to determine the bacterial reaction result, the chromogenic reagent is directly centrifuged from the chromogenic reagent addition chamber into the chromogenic reaction chamber using centrifugal force. The result is then determined after the chromogenic reaction, making the operation simple.
[0067] Example 2
[0068] Based on Example 1, this example introduces a method for using a microfluidic chip for bacterial identification.
[0069] A method for using a microfluidic chip for bacterial identification includes the following steps:
[0070] Step 1: Add the bacterial solution sample to be tested into the bacterial solution sample tank 3;
[0071] Step 2: Under the action of centrifugal force, the bacterial liquid sample to be tested enters the annular guide channel 5 through the bent microchannel 6, and then flows into each bacterial identification reaction chamber 7 through the bent microchannel 5 respectively.
[0072] Step 3: After the bacterial identification reaction chamber 7 is incubated with the bacterial biochemical identification drying agent for 4-18 hours, the bacterial growth and metabolic status is determined by using photomicrography and artificial intelligence analysis, and the bacterial identification results are obtained by combining the microbial biochemical identification table or database.
[0073] In step 3, after determining the growth and metabolic state of bacteria, if some bacterial identification reaction chambers 7 need to be filled with chromogenic reagent after culture, the chromogenic reagent can be added to the chromogenic reagent addition chamber 4. With the action of centrifugal force, the chromogenic reagent flows from the chromogenic reagent addition chamber 4 into the chromogenic reagent reaction chamber 8. After the chromogenic reaction, the result is judged. The operation is simple.
[0074] The bacterial species to be identified are categorized into Staphylococcus identification, Streptococcus identification, Enterobacter identification, non-fermenting bacteria identification, and yeast identification cards. The bacterial biochemical identification drying reagents pre-placed in the bacterial identification reaction chamber 7 may include different combinations of drying reagents such as lysine decarboxylase test, ornithine decarboxylase test, arginine dihydrolase test, urease test, malonate utilization test, indole production test, phenylalanine deaminase test, β-galactosidase test, aesculin hydrolysis test, glucose fermentation test, euonymus alcohol fermentation test, lactose fermentation test, calendula alcohol fermentation test, maltose fermentation test, D-mannitol fermentation test, sucrose fermentation test, cellobiose fermentation test, disaccharide fermentation test, L-rhamnose fermentation test, inositol fermentation test, α-methyl-D-glucose fermentation test, D-sorbitol fermentation test, raffinose fermentation test, trehalose fermentation test, 3-hydroxy-2-butanone production test, potassium cyanide growth test, hydrogen sulfide production test, and gelatin liquefaction test.
[0075] The usage method described in this embodiment is convenient to operate and takes into account the judgment of microbial reaction results that require the addition of colorimetric reagents. It is also convenient to operate and reflects the high applicability of the microfluidic chip for bacterial identification, which can meet various needs of microbial identification.
[0076] Example 3
[0077] Based on the above embodiments 1-2, this embodiment takes the CO2 culture environment and the anaerobic culture environment as examples of culturing microorganisms in a caustic environment to introduce a method for bacterial identification using the microfluidic chip designed in this application:
[0078] (1) Taking Neisseria meningitidis cultured in a CO2 environment as an example, a CO2 caustic microenvironment was established:
[0079] Step 1: Add meningococcal bacteria to bacterial solution sample tank 3;
[0080] Step 2: Under the action of centrifugal force, meningococci enter the annular guide channel 5 through the bent microchannel 6, and then flow into each bacterial identification reaction chamber 7 through the bent microchannel 5 respectively.
[0081] Step 3: After the meningococcus enters the bacterial identification reaction chamber 7, sodium bicarbonate and hydrochloric acid are added to the bacterial sample loading tank 3 in proportion. The sample loading hole on the bacterial sample loading tank 3 is sealed with a film. The sodium bicarbonate and hydrochloric acid come into contact to generate carbon dioxide and establish a CO2 culture environment.
[0082] Step 4: Observe the growth and metabolic state of Neisseria meningitidis in the bacterial identification reaction chamber 7 under CO2 caustic microenvironment culture using photomicrography and artificial intelligence analysis, and obtain the bacterial identification results by combining the microbial biochemical identification table or database.
[0083] (2) Taking anaerobic bacteria that can only grow in an anaerobic culture environment as an example, an anaerobic-caustic microenvironment was established:
[0084] Step 1: Add the bacterial solution to be tested into the bacterial solution sample tank 3;
[0085] Step 2: Under the action of centrifugal force, the bacterial solution to be tested enters the annular guide channel 5 through the bent microchannel 6, and then flows into each bacterial identification reaction chamber 7 through the bent microchannel 5 respectively.
[0086] Step 3: After the bacterial solution enters the bacterial identification reaction chamber 7, add an appropriate amount of mercaptoethanol or pyrogallol and sodium hydroxide solution to the bacterial solution sample loading tank 3, and seal the sample loading hole on the bacterial sample loading tank 3 with a film to absorb oxygen and create an anaerobic culture environment inside the chip.
[0087] Step 4: Observe the growth and metabolic state of the bacterial culture in the bacterial identification reaction chamber 7 under anaerobic and caustic microenvironment culture using photomicrography and artificial intelligence analysis, and obtain the bacterial identification results by combining the microbial biochemical identification table or database.
[0088] Whether establishing a CO2 caustic microenvironment or an anaerobic caustic microenvironment, this bacterial identification microfluidic chip can maintain a caustic microenvironment throughout the entire culture and identification reaction cycle. The biochemical reaction results of bacteria under this caustic microenvironment can be observed at any time, and it can be cultured in the same incubator as conventional culture chips. Current microplate methods, however, require dedicated caustic environment incubators (such as CO2 incubators or anaerobic incubators). In such environments, if an identification plate needs to be removed for observation, the entire caustic environment in the incubator must be re-established. This results in high resource consumption and cumbersome operation, significantly limiting the clinical application of caustic bacteria identification.
[0089] The establishment of the caustic environment in this embodiment demonstrates that using the microfluidic chip for bacterial identification in this application greatly simplifies the culture environment for caustic bacteria and common microorganisms, saves resources, is easier to operate, and provides excellent identification efficiency and results.
[0090] This application provides a microfluidic chip for bacterial identification and a method for using it, which improves the efficiency of bacterial identification while ensuring no contamination and high accuracy in the bacterial identification process. It meets the needs of bacterial biochemical changes during bacterial identification, has high applicability, is easy to operate, and has low manufacturing cost.
[0091] The above description is merely a preferred embodiment of this application and does not limit the scope of protection of this application. Various modifications and variations can be made to this application by those skilled in the art. Any changes, modifications, substitutions, integrations, and parameter alterations to these embodiments within the spirit and principles of this application, achieved through conventional substitutions or by realizing the same function without departing from the principles and spirit of this application, fall within the scope of protection of this application.
Claims
1. A method for using a microfluidic chip for bacterial identification, characterized in that, The identification of microorganisms cultured in certain caustic environments includes the following steps: Step 1: Add the bacterial solution sample to be tested into the bacterial solution sample tank (3); Step 2: Under the action of centrifugal force, the bacterial liquid sample to be tested enters the annular guide channel (5) through the bent microchannel (6), and then flows into the bacterial identification reaction chamber (7) through the bent microchannel (6); Step 3: After the bacterial liquid sample to be tested enters the bacterial identification reaction chamber (7), chemical reagents are added to the bacterial sample loading tank (3) to create an aerobic microenvironment, and the sample loading hole on the bacterial sample loading tank (3) is sealed with a film. Step 4: Observe the growth and metabolic state of bacteria in the bacterial identification reaction chamber (7) under caustic microenvironment culture, and obtain the bacterial identification results by combining the microbial biochemical identification table or database. A microfluidic chip for bacterial identification includes a CD-shaped bacterial identification reagent tray (1). Centered on the center of the bacterial identification reagent tray (1), a shaft hole (2), a bacterial solution addition groove (3), and an annular guide channel (5) are arranged sequentially from the center outwards from the outside of the bacterial identification reagent tray (1). The bacterial solution sample loading tank (3) is located outside the shaft hole (2); The annular guide channel (5) is located outside the bacterial solution sample loading tank (3), and the bacterial solution sample loading tank (3) and the annular guide channel (5) are connected by a bent microchannel (6); A bacterial identification reaction chamber (7) is provided on the outside of the annular guide channel (5). The bacterial identification reaction chamber (7) is connected to the annular guide channel (5) through a bent microchannel (6). A bacterial biochemical identification drying agent is pre-placed in the bacterial identification reaction chamber (7). A colorimetric reagent addition chamber (4) is provided on the outside of the shaft hole (2), and the colorimetric reagent addition chamber (4) is connected to the annular guide channel (5) through the flow channel; A colorimetric reaction chamber (8) is provided on the outside of the annular guide channel (5). The colorimetric reaction chamber (8) is matched with the colorimetric reagent addition chamber (4) and is connected to the colorimetric reagent addition chamber (4) through a wedge-shaped microchannel (9). The bent microchannel (6) is an S-shaped microfluidic channel; The bacterial solution loading tank (3) has an arc-shaped structure. One end of the bacterial solution loading tank (3) is closed, and the other end is connected to the annular guide channel (5) through the bent microchannel (6). After adding chemical reagents into the bacterial solution loading tank (3), the loading hole of the bacterial solution loading tank (3) is sealed with a film.
2. The method of using a microfluidic chip for bacterial identification according to claim 1, characterized in that, The bacterial identification reaction chamber (7) is evenly distributed outside the annular guide channel (5) and is connected to the annular guide channel (5) through the bent microchannel (6).
3. The method of using a microfluidic chip for bacterial identification according to claim 2, characterized in that, The bacterial identification reaction chamber (7) is larger than the colorimetric reaction chamber (8).
4. The method of using a microfluidic chip for bacterial identification according to claim 1, characterized in that, A dynamic balancing groove (10) is also provided on the outer side of the shaft hole (2).