Integrated micro-fluidic chip device and system for microorganism enrichment and detection

Abstract

The invention relates to an integrated micro-fluidic chip device and system for microorganism enrichment and detection, the integrated micro-fluidic chip device comprises a micro-fluidic chip, the micro-fluidic chip comprises a micro-channel, the micro-channel comprises a liquid inlet and a liquid outlet, and the micro-channel allows to-be-detected liquid to flow; the micro-channel comprises a detection section; the filter membrane layer covers the inner wall of the detection section, and the filter membrane layer is used for capturing and screening microorganisms with specific sizes; the laser irradiates exciting light to the detection section, so that the specific microorganisms in the detection section are subjected to fluorescence reaction; the receiving assembly and the laser are arranged on the upper side and the lower side of the micro-fluidic chip respectively, and the receiving assembly is used for receiving fluorescence generated by the fluorescence reaction in the detection section so as to evaluate the content of microorganisms in the to-be-detected liquid. The filter membrane layer is arranged in the detection section to capture the microorganisms, so that the content of the microorganisms in the detection section is increased, the detection efficiency is improved, and the sensitivity and accuracy during low-concentration detection are improved.

Description

Technical Field

[0001] The present application relates to the technical field of microbial detection, and in particular to an integrated microfluidic chip device and system for microbial enrichment and detection. Background Art

[0002] In the fields of environmental monitoring, food safety, medical diagnosis and biosafety, it is crucial to detect the presence and concentration of microorganisms quickly and accurately. Traditional microbial detection methods include culture, immunoassay and molecular biology methods. Although these methods are widely used, they are usually complicated to operate, time-consuming, and lack sensitivity and specificity. In recent years, microfluidic technology has developed rapidly in the field of microbial detection due to its high efficiency, low cost and high-throughput analysis.

[0003] Among related technologies, microfluidic systems can simulate laboratory operations on tiny chips, making sample processing and analysis automated and integrated, significantly improving operational efficiency and detection sensitivity. In particular, combining microfluidic technology with fluorescence spectroscopy, electrochemical detection and other methods provides a new solution for detecting microorganisms.

[0004] However, traditional methods such as culture methods are usually time-consuming and require several days or even longer to obtain results; although immunological methods and molecular biology techniques have increased the speed of detection, they often require complex sample pretreatment and professional operations, which are not efficient enough in rapid response and on-site detection scenarios. In addition, many existing microfluidic systems, although partially automated, are still limited in capture efficiency and detection sensitivity for low-concentration targets. These systems often fail to fully utilize the potential of microfluidic technology, especially in continuous monitoring and real-time data analysis.

[0005] Therefore, the existing microfluidic systems have low capture efficiency in microbial detection and insufficient detection sensitivity for low-concentration targets. Summary of the invention

[0006] The embodiments of the present application provide an integrated microfluidic chip device and system for microbial enrichment and detection to solve the technical problems in the related art that the microfluidic system has low efficiency in microbial detection and capture and insufficient detection sensitivity for low-concentration targets.

[0007] In a first aspect, an integrated microfluidic chip device for microbial enrichment and detection is provided, comprising:

[0008] A microfluidic chip, wherein the microfluidic chip comprises a microchannel, wherein the microchannel comprises a liquid inlet and a liquid outlet, and wherein the microchannel is used for the flow of a liquid to be detected; and the microchannel comprises a detection section;

[0009] A filter membrane layer, the filter membrane layer covers the inner wall of the detection section, and the filter membrane layer is used to capture and screen microorganisms of a specific size;

[0010] a laser, wherein the laser irradiates an excitation light toward the detection section so as to cause a fluorescent reaction of specific microorganisms in the detection section;

[0011] A receiving component, wherein the receiving component and the laser are respectively arranged on the upper and lower sides of the microfluidic chip, and the receiving component is used to receive the fluorescence generated by the fluorescence reaction in the detection section to evaluate the content of microorganisms in the liquid to be detected.

[0012] In some embodiments, the surface of the filter membrane layer is modified with biorecognition molecules.

[0013] 3. The integrated microfluidic chip device for microorganism enrichment and detection according to claim 2, characterized in that the biorecognition molecules include antibodies or oligonucleotide probes.

[0014] In some embodiments, the pore size of the filter membrane layer surface is 1-15 microns.

[0015] In some embodiments, the filter membrane layer is made of polytetrafluoroethylene, polycarbonate or polymethyl methacrylate.

[0016] In some embodiments, the detection section is located within the irradiation area of ​​the laser, and the detection section is located within the receiving area of ​​the receiving component;

[0017] The detection section has multiple bends or curves to increase the length of the detection section.

[0018] In some embodiments, the detection segment includes a plurality of connected unit segments, and the unit segments include straight segments or curved segments.

[0019] In some embodiments, the detection section is arranged in the shape of an annular channel, and the fluid to be detected forms a reflux in the detection section.

[0020] In some embodiments, the receiving component includes a receiving objective lens and a receiving detector, wherein the receiving objective lens is arranged between the receiving detector and the microfluidic chip, and the receiving objective lens converges the fluorescence generated by the fluorescence reaction in the detection segment onto the photosensitive surface of the receiving detector.

[0021] The beneficial effects of the technical solution provided by this application include:

[0022] The embodiment of the present application provides an integrated microfluidic chip device for microorganism enrichment and detection. During detection, the liquid to be detected enters the microchannel from the liquid inlet and leaves the microchannel from the liquid outlet, so that the liquid to be detected flows in the microchannel. When the liquid to be detected flows to the detection section, the excitation light emitted by the laser causes the specific microorganisms in the detection section to produce a fluorescent reaction, and the fluorescence generated by the fluorescent reaction in the detection section is received by the receiving component to evaluate the content of the specific microorganisms in the liquid to be detected.

[0023] When the liquid to be tested flows to the detection section, the filter layer captures specific microorganisms, and as the liquid to be tested flows, the microorganisms are gathered at the filter layer in the detection section, which improves the sensitivity of the detection. Even if the concentration of microorganisms in the liquid to be tested is low, low-concentration microorganisms can still be accurately detected. In addition, since the microorganisms are detected after being gathered, the detection efficiency is improved.

[0024] In a second aspect, an integrated microfluidic chip system for microbial enrichment and detection is provided, comprising the integrated microfluidic chip device for microbial enrichment and detection as described above.

[0025] Another embodiment of the present application provides an integrated microfluidic chip system for microbial enrichment and detection. Since the integrated microfluidic chip system for microbial enrichment and detection includes the above-mentioned integrated microfluidic chip device for microbial enrichment and detection, the beneficial effects of the integrated microfluidic chip system for microbial enrichment and detection are consistent with the beneficial effects of the above-mentioned integrated microfluidic chip device for microbial enrichment and detection, which will not be repeated here. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0027] Figure 1 A schematic diagram of a microfluidic chip provided in an embodiment of the present application;

[0028] Figure 2 Schematic diagram of an integrated microfluidic chip device for microbial enrichment and detection provided in an embodiment of the present application.

[0029] In the figure: 1, microfluidic chip; 1a, microchannel; 1a1, detection section; 1b, liquid inlet; 1c, liquid outlet; 2, laser; 3, receiving objective lens; 4, receiving detector. DETAILED DESCRIPTION

[0030] In order to make the purpose, technical solution and advantages of the embodiments of the present application clearer, the technical solution in the embodiments of the present application will be clearly and completely described below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0031] The embodiment of the present application provides an integrated microfluidic chip device and system for microorganism enrichment and detection, which increases the content of microorganisms in the detection section by arranging a filter membrane layer in the detection section to capture microorganisms, improves the detection efficiency, and improves the sensitivity and accuracy of low-concentration detection. The present application solves the technical problem that the microfluidic system in the related art has low efficiency in microorganism detection and capture and insufficient detection sensitivity for low-concentration targets.

[0032] Reference Figure 1 and Figure 2 , an integrated microfluidic chip device for microorganism enrichment and detection, comprising a microfluidic chip 1, a laser 2 and a receiving component. The microfluidic chip 1 comprises a microchannel 1a, the microchannel 1a comprises a liquid inlet 1b and a liquid outlet 1c, and the microchannel 1a is for the flow of the liquid to be detected. The microchannel 1a comprises a detection section 1a1, and the microorganisms in the liquid to be detected enter the detection section 1a1 and are detected. In this embodiment, the material of the microfluidic chip 1 comprises plastic polydimethylsiloxane (PDMS) or polymethyl methacrylate (PMMA).

[0033] Reference Figure 1 and Figure 2 , the laser 2 irradiates the excitation light toward the detection section 1a1 to make the specific microorganisms in the detection section 1a1 produce a fluorescent reaction. The receiving component and the laser 2 are respectively arranged on the upper and lower sides of the microfluidic chip 1. The receiving component is used to receive the fluorescence generated by the fluorescent reaction in the detection section 1a1 to evaluate the content of the microorganisms in the liquid to be detected.

[0034] It should be noted that when detecting different microorganisms, the wavelength of the excitation light generated by the laser 2 is different, so as to achieve the detection of multiple microorganisms, as well as the detection of only specific microorganisms.

[0035] Furthermore, the integrated microfluidic chip device for microorganism enrichment and detection also includes a filter membrane layer, which covers the inner wall of the detection section 1a1. The filter membrane layer is used to capture and screen microorganisms of a specific size.

[0036] Specifically, the filter membrane layer is a porous filter membrane, which captures microorganisms of different sizes by arranging the sizes of its pore sizes.

[0037] Reference Figure 1 and Figure 2 When the liquid to be detected flows to the detection section 1a1, the filter membrane layer captures specific microorganisms, and as the liquid to be detected flows, the microorganisms are gathered at the filter membrane layer in the detection section 1a1, which improves the sensitivity of the detection. Even if the concentration of microorganisms in the liquid to be detected is low, low-concentration microorganisms can still be accurately detected. In addition, since the microorganisms are detected after being gathered, the detection efficiency is improved.

[0038] Furthermore, the surface of the filter membrane layer is modified with biorecognition molecules.

[0039] This arrangement enables the filter layer to specifically adsorb target microorganisms, thereby realizing the detection of specific microorganisms and improving the detection accuracy. In addition, the filter layer ensures that the microorganisms can be effectively captured when the water sample flowing through the detection section 1a1 passes through the filter layer, so that the microorganisms are effectively enriched, the concentration of the microorganisms is increased, and the sensitivity of the detection is improved.

[0040] Among them, the biorecognition molecules include antibodies or oligonucleotide probes.

[0041] The filter layer can not only capture microorganisms through physical screening, but also enhance the capture effect through the specific binding of biorecognition molecules to target microorganisms. Biorecognition molecules (such as antibodies, oligonucleotide probes or other affinity molecules) are modified to the surface of the filter layer. These biorecognition molecules can specifically recognize and bind to target microbial surface molecules (such as antigens or other markers). In this way, the filter layer can capture specific microorganisms, such as Escherichia coli or Salmonella.

[0042] Furthermore, the pore size of the filter membrane layer surface is 1-15 microns.

[0043] In this way, a filter membrane layer with a suitable pore size is arranged according to the size of the microorganism to facilitate physical screening of specific microorganisms. Among them, the size of general microorganisms is between 1 and 10 microns, and the pore size of the filter membrane layer surface is arranged according to the size of the different microorganisms to achieve specific capture.

[0044] In this embodiment, the material of the filter membrane layer includes polytetrafluoroethylene, polycarbonate or polymethyl methacrylate. Since the filter membrane layer needs to run for a long time in the experiment, the selection of the material needs to ensure that it will not degrade or chemically react under contact with the biological sample or water sample, thereby affecting the experimental results. Polymer materials such as polytetrafluoroethylene, polycarbonate or polymethyl methacrylate have excellent chemical stability and mechanical strength, ensuring that the filter membrane layer can be in a stable state for a long time without causing adverse effects on the detection.

[0045] In addition, the capture rate of microorganisms in the detection section 1a1 can be increased by optimizing the flow rate of the fluid to be detected. For example, the flow rate of the liquid to be detected can be controlled within 50 μL / min.

[0046] The detection section 1a1 is located in the irradiation area of ​​the laser 2, and the detection section 1a1 is located in the receiving area of ​​the receiving component, that is, the detection section 1a1 is arranged in the area where the laser 2 and the receiving component act.

[0047] Reference Figure 1 and Figure 2 , wherein the detection section 1a1 is bent or curved in multiple sections to increase the length of the detection section 1a1. By increasing the length of the detection section 1a1 in a limited area, the distance that the liquid to be detected flows in the detection section 1a1 is increased to increase the capture rate of microorganisms and effectively enrich microorganisms. It is suitable for high-sensitivity, high-precision and high-efficiency detection of low-concentration microorganisms.

[0048] Specifically, the detection segment 1a1 includes a plurality of connected unit segments, and the unit segments include straight segments or curved segments. That is, the specific formation of the detection segment 1a1 is not limited, and the detection segment 1a1 can be a plurality of straight segments, a plurality of curved segments, or a combination of at least one straight segment and at least one curved segment.

[0049] Reference Figure 1 and Figure 2 In this embodiment, the detection section 1a1 is arranged in the shape of an annular channel, and the fluid to be detected forms a reflux in the detection section 1a1.

[0050] In this way, the reflux design of the detection section 1a1 ensures that the uncaptured microorganisms can flow back through the filter membrane layer with a high capture probability. This design can significantly increase the capture rate of low-concentration microorganisms, especially when the concentration of microorganisms is low, the reflux design of the detection section 1a1 provides additional opportunities for effective enrichment.

[0051] The receiving component includes a receiving lens 3 and a receiving detector 4. The receiving lens 3 is arranged between the receiving detector 4 and the microfluidic chip 1. The receiving lens 3 gathers the fluorescence generated by the fluorescence reaction in the detection section 1a1 onto the photosensitive surface of the receiving detector 4. In this way, after the microorganisms are captured and fluorescently labeled by fluorescence spectroscopy, the content of microorganisms in the liquid to be detected can be quickly and accurately evaluated.

[0052] The embodiment of the present application provides an integrated microfluidic chip device for microorganism enrichment and detection. During detection, the liquid to be detected enters the microchannel 1a from the liquid inlet 1b and leaves the microchannel 1a from the liquid outlet 1c, so that the liquid to be detected flows in the microchannel 1a. When the liquid to be detected flows to the detection section 1a1, the excitation light emitted by the laser 2 causes the specific microorganisms in the detection section 1a1 to produce a fluorescent reaction, and the fluorescence generated by the fluorescent reaction in the detection section 1a1 is received by the receiving component to evaluate the content of the specific microorganisms in the liquid to be detected.

[0053] When the liquid to be detected flows to the detection section 1a1, the filter membrane layer captures specific microorganisms, and as the liquid to be detected flows, the microorganisms are gathered at the filter membrane layer in the detection section 1a1, which improves the sensitivity of the detection. Even if the concentration of microorganisms in the liquid to be detected is low, low-concentration microorganisms can still be accurately detected. In addition, since the microorganisms are detected after being gathered, the detection efficiency is improved.

[0054] Another embodiment of the present application provides an integrated microfluidic chip system for microorganism enrichment and detection, including the integrated microfluidic chip device for microorganism enrichment and detection as described above.

[0055] Another embodiment of the present application provides an integrated microfluidic chip system for microbial enrichment and detection. Since the integrated microfluidic chip system for microbial enrichment and detection includes the above-mentioned integrated microfluidic chip device for microbial enrichment and detection, the beneficial effects of the integrated microfluidic chip system for microbial enrichment and detection are consistent with the beneficial effects of the above-mentioned integrated microfluidic chip device for microbial enrichment and detection, which will not be repeated here.

[0056] In the description of the present application, it should be noted that the terms "upper", "lower", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. Unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be an indirect connection through an intermediate medium, or it can be a connection between the two elements. For those of ordinary skill in the art, the specific meanings of the above terms in the present application can be understood according to the specific circumstances.

[0057] It should be noted that, in this application, relational terms such as "first" and "second" are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the presence of other identical elements in the process, method, article or device including the elements.

[0058] The above description is only a specific implementation of the present application, so that those skilled in the art can understand or implement the present application. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present application. Therefore, the present application will not be limited to the embodiments shown herein, but will conform to the widest range consistent with the principles and novel features applied for herein.

Claims

1. An integrated microfluidic chip device for microbial enrichment and detection, characterized in that: It includes: A microfluidic chip, wherein the microfluidic chip comprises a microchannel, wherein the microchannel comprises a liquid inlet and a liquid outlet, and wherein the microchannel is used for the flow of a liquid to be detected; and the microchannel comprises a detection section; A filter membrane layer, the filter membrane layer covers the inner wall of the detection section, and the filter membrane layer is used to capture and screen microorganisms of a specific size; a laser, wherein the laser irradiates an excitation light toward the detection section so as to cause a fluorescent reaction of specific microorganisms in the detection section; A receiving component, wherein the receiving component and the laser are respectively arranged on the upper and lower sides of the microfluidic chip, and the receiving component is used to receive the fluorescence generated by the fluorescence reaction in the detection section to evaluate the content of microorganisms in the liquid to be detected.

2. The integrated microfluidic chip device for microbial enrichment and detection according to claim 1, characterized in that: The surface of the filter membrane layer is modified with biological recognition molecules.

3. The integrated microfluidic chip device for microbial enrichment and detection according to claim 2, characterized in that: The biorecognition molecules include antibodies or oligonucleotide probes.

4. The integrated microfluidic chip device for microbial enrichment and detection according to any one of claims 1 to 3, characterized in that: The pore size of the filter membrane layer surface is 1-15 microns.

5. The integrated microfluidic chip device for microbial enrichment and detection according to claim 1, characterized in that: The material of the filter membrane layer includes polytetrafluoroethylene, polycarbonate or polymethyl methacrylate.

6. The integrated microfluidic chip device for microbial enrichment and detection according to claim 1, characterized in that: The detection section is located in the irradiation area of ​​the laser, and the detection section is located in the receiving area of ​​the receiving component; The detection section has multiple bends or curves to increase the length of the detection section.

7. The integrated microfluidic chip device for microbial enrichment and detection according to claim 6, characterized in that: The detection section includes a plurality of connected unit sections, and the unit sections include straight line sections or curved sections.

8. The integrated microfluidic chip device for microbial enrichment and detection according to claim 6, characterized in that: The detection section is arranged in the shape of an annular channel, and the fluid to be detected forms a reflux in the detection section.

9. The integrated microfluidic chip device for microbial enrichment and detection according to claim 1, characterized in that: The receiving component includes a receiving objective lens and a receiving detector. The receiving objective lens is arranged between the receiving detector and the microfluidic chip. The receiving objective lens converges the fluorescence generated by the fluorescence reaction in the detection section onto the photosensitive surface of the receiving detector.

10. An integrated microfluidic chip system for microorganism enrichment and detection, comprising the integrated microfluidic chip device for microorganism enrichment and detection as claimed in any one of claims 1 to 9.

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