Apparatus and method for classifying and measuring airborne microorganisms

Abstract

An embodiment of the present invention provides a technology relating to an apparatus capable of accurately performing real-time analysis of types of microorganisms in bioaerosols in the air, and the like. The apparatus for classifying and measuring airborne microorganisms, according to an embodiment of the present invention, comprises: a measurement unit including a microfluidic chip having microchannels formed therein and an optical sensor configured to measure light generated when microorganisms passing through the microchannels react with a luminescent agent; a supply operation unit configured to supply, to the microfluidic chip, a feed solution containing microorganisms, a lysis buffer which lyses microbial cells to extract adenosine triphosphate (ATP), and a luminescent agent; and an analysis control unit configured to receive signals from the optical sensor of the measurement unit and classify the types of microorganisms.

Description

Apparatus and method for measuring airborne microorganism classification

[0001] The present invention relates to a device and method for measuring airborne microorganism classification, and more particularly, to a technology related to a device capable of accurately performing real-time analysis of the types of microorganisms in bioaerosols in the air.

[0002] The COVID-19 pandemic has fueled public interest in bioaerosols. In particular, in Korea, the shutdown of industrial operations during the pandemic has led to increased opportunities for clear skies, leading to increased interest in air quality.

[0003] In response to the growing interest, research and development on bioaerosol measurement technology is increasing. However, in response to the increased interest in air quality, rapid bioaerosol measurement technology that can directly affect humans still needs to be improved from a user-friendly perspective.

[0004] Methods for measuring airborne microorganisms include a culture method in which biological particles floating in a sample gas are captured on a solid or liquid surface suitable for growth, cultured under an appropriate temperature and humidity environment for a certain period of time, and then the number of captured microorganisms is calculated from the number of colonies that appear on the surface, and a staining method using a fluorescence microscope after staining.

[0005] Recently, the ATP bioluminescence method, which utilizes the principle of ATP (adenosine triphosphate) and luciferin / luciferase reacting to emit light, has enabled rapid work by reducing the series of processes required for ATP scavenging, ATP extraction, and luminescence measurement to about 30 minutes.

[0006] However, with the above methods, not only is it impossible to measure floating microorganisms existing in the air in real time, but it is also difficult to quickly determine the type of microorganism, i.e., whether the microorganism is a bacterium or a fungus.

[0007] In Korean Patent No. 10-1936156 (Title of the Invention: Sampling and Detection Device and Manufacturing Method Capable of Preventing Luminescence and Fluorescence Noise Signals), a method is disclosed in which, when detecting microorganisms (bacteria) using a high-density aggregated luminescent substance-fixed substrate and a nuclease, only those that react with the nuclease are separated from the luminescent substance-fixed substrate and detected, thereby enabling quantitative analysis of nuclease without noise signals and easily analyzing enzyme activity according to the type of nuclease.

[0008] The purpose of the present invention to solve the above problems is to provide a technology related to a device that can accurately perform real-time analysis of the types of microorganisms in bioaerosols in the air.

[0009] The technical problems to be solved by the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0010] The present invention for achieving the above purpose comprises a measurement unit having a microfluidic chip having a microfluidic channel formed therein and a light sensor for measuring light generated when microorganisms passing through the microfluidic channel react with a luminescent agent; a supply operation unit for supplying a supply solution containing microorganisms, a lysis buffer for lysing microbial cells to extract adenosine triphosphate (ATP), and the luminescent agent to the microfluidic chip; and an analysis control unit for receiving a signal of the light sensor from the measurement unit and selecting the type of microorganism.

[0011] In an embodiment of the present invention, the supply operation unit may be equipped with a buffer injector that injects the lysis buffer into the microfluidic chip.

[0012] In an embodiment of the present invention, the buffer injector can inject at least one lysis buffer among a plurality of lysis buffers into the microfluidic chip.

[0013] In an embodiment of the present invention, the analysis control unit may store analysis data, which is data on the type of microorganism matching the RLU and the type of the lyse buffer by the signal of the optical sensor.

[0014] In an embodiment of the present invention, the supply operation unit may be equipped with a supply liquid injector that injects the supply liquid into the microfluidic chip.

[0015] In an embodiment of the present invention, the supply operation unit may be equipped with a luminescent agent injector that injects the luminescent agent into the microfluidic chip.

[0016] In an embodiment of the present invention, the light sensor may be a PMT (photomultiplier tube) sensor.

[0017] In an embodiment of the present invention, the signal of the optical sensor can be transmitted to the analysis control unit via an external network.

[0018] In an embodiment of the present invention, the analysis control unit may be installed in an electronic device used by a user.

[0019] The present invention for achieving the above object comprises a supply liquid injection step in which the supply liquid is generated in the supply operation unit and the supply liquid is injected from the supply operation unit into the first microchannel of the microchannel chip; a buffer injection step in which the lysis buffer is injected from the supply operation unit into the second microchannel of the microchannel chip; a luminescent agent injection step in which the luminescent agent is injected from the supply operation unit into the third microchannel of the microchannel chip; a measuring step in which the measuring unit measures a fluid passing through the measuring channel of the microchannel chip and transmits a measuring signal to the analysis control unit; and an analysis step in which the analysis control unit selects and stores the type of microorganism.

[0020] The effect of the present invention according to the above configuration is that information on the type and concentration of microorganisms can be quantified and organized, and information on such microorganisms can be obtained in real time.

[0021] In addition, the effect of the present invention is that information on the above-mentioned microorganisms can be confirmed in real time using electronic devices, etc., thereby increasing the efficiency of classification and measurement of microorganisms.

[0022] The effects of the present invention are not limited to the effects described above, and should be understood to include all effects that can be inferred from the detailed description of the present invention or the composition of the invention described in the claims.

[0023] FIG. 1 is a block diagram of a configuration of a classification measuring device according to an embodiment of the present invention.

[0024] Figure 2 is a schematic diagram of the configuration of a supply operation unit and a measurement unit according to one embodiment of the present invention.

[0025] Hereinafter, the present invention will be described with reference to the attached drawings. However, the present invention can be implemented in various different forms and is therefore not limited to the embodiments described herein. In the drawings, irrelevant parts have been omitted for clarity of description, and similar parts have been designated with similar reference numerals throughout the specification.

[0026] Throughout the specification, when a part is said to be "connected (connected, contacted, or coupled)" to another part, this includes not only cases where it is "directly connected," but also cases where it is "indirectly connected" with another part in between. Furthermore, when a part is said to "include" a component, this does not exclude other components, but rather implies that it may include other components, unless otherwise specifically stated.

[0027] The terminology used herein is merely used to describe specific embodiments and is not intended to limit the present invention. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, it should be understood that the terms "comprises" or "has" indicate the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but do not exclude in advance the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0028] Hereinafter, the present invention will be described in detail with reference to the attached drawings.

[0029]

[0030] Fig. 1 is a block diagram of a configuration of a classification measuring device according to an embodiment of the present invention, and Fig. 2 is a schematic diagram of a configuration of a supply operation unit (100) and a measuring unit (200) according to an embodiment of the present invention. In Fig. 2, a is an enlarged view showing light measurement by a light sensor (220).

[0031] As shown in FIGS. 1 and 2, the classification measurement device of the present invention includes a measuring unit (200) having a microfluidic chip (210) in which microfluidic channels are formed and a light sensor (220) that measures light generated when microorganisms passing through the microfluidic channel react with a luminescent agent; a supply operation unit (100) that supplies a supply solution containing microorganisms, a solution that dissolves microbial cells to extract adenosine triphosphate (ATP), and a luminescent agent to the microfluidic chip (210); and an analysis control unit (300) that receives a signal from the light sensor (220) from the measuring unit (200) and selects the type of microorganism.

[0032] Microorganisms may be viruses, bacteria, fungi, etc., and when multiple microorganisms are floating in the air, aerosols and bioaerosols containing microorganisms move together in the air. The classification and measurement device of the present invention can be used to select the type of microorganism.

[0033] To this end, the classification measuring device of the present invention may include a configuration as described above, and a first microchannel, a second microchannel, a third microchannel, and a measuring channel (211) may be formed as microchannels in the microchannel chip (210), and each of the first microchannel, the second microchannel, the third microchannel, and the measuring channel (211) may be sequentially connected.

[0034]

[0035] The supply operation unit (100) may be equipped with a supply solution injector (120) that injects the supply solution into the microfluidic chip (210). At this time, the supply solution may be injected from the supply solution injector (120) into the first microfluidic channel of the microfluidic chip (210).

[0036] The supply liquid injector (120) may be provided with a supply liquid generation tank (121) having an internal space and generating a supply liquid; and a supply pump (122), which is a pump that provides pressure to the supply liquid discharged from the supply liquid generation tank (121) to cause the supply liquid to flow to the microfluidic chip (210).

[0037] In order to inject a supply solution into the microfluidic chip (210), external air and water can be supplied to the supply solution generation tank (121), and external air can be delivered to the supply solution generation tank (121) using an air pump, and water can be supplied with pressure by a water pump (101) and delivered to the supply solution generation tank (121).

[0038] In the internal space of the supply liquid generation tank (121), water and air can come into contact, and a cyclonic flow of water and air can be performed in the internal space of the supply liquid generation tank by the structure of the supply liquid generation tank (121) or an internal vortex generating device.

[0039] At this time, gases such as oxygen and nitrogen that are not mixed with water are discharged to the outside, and microorganisms existing in the air can mix with water to form a supply liquid.

[0040] A supply pump (122) is formed between the microfluidic chip (210) and the supply liquid generation tank (121), and the supply liquid formed as described above is subjected to pressure while passing through the supply pump (122), and the supply liquid can be injected into the microfluidic chip (210).

[0041] At this time, a control signal is transmitted from the analysis control unit (300) to the supply pump (122), and thereby the pressure of the supply pump (122) is controlled during the injection process of the supply liquid, thereby controlling the hydraulic pressure and flow rate of the liquid passing through the microchannel of the microchannel chip (210).

[0042]

[0043] The supply operation unit (100) may be equipped with a buffer injector (110) that injects a lysis buffer into the microfluidic chip (210). At this time, the buffer injector (110) may inject at least one lysis buffer among a plurality of lysis buffers into the microfluidic chip (210).

[0044] To this end, the buffer injector (110) may be provided with a plurality of buffer tanks (111) for storing lysis buffer; a buffer pump (112), which is a pump connected to the plurality of buffer tanks (111) and provides pressure to the lysis buffer discharged from at least one of the plurality of buffer tanks (111) to cause the lysis buffer to flow to the microfluidic chip (210); and a buffer valve, which is an electronic valve formed in a path between the plurality of buffer tanks (111) and the buffer pump (112) to control the flow of the lysis buffer.

[0045] When the supply liquid is injected into the first microchannel from the supply liquid injector (120) as described above, the analysis control unit (300) can transmit a control signal to one or more of the plurality of buffer valves and the buffer pump (112).

[0046] Then, the buffer valve, which has been commanded to open by the control signal, opens the flow path within the buffer valve, and the buffer pump (112), which has been commanded to operate by the control signal, starts operating, so that the lyse buffer in the buffer tank (111) connected to the opened buffer valve can pass through the buffer pump (112) and be injected into the second micro-flow path.

[0047] At this time, the supply liquid passing through the second microchannel and the lysis buffer are mixed, and through this mixing, a solution that dissolves microorganisms is created, and this solution can flow into the third microchannel.

[0048] Here, various types of lysis buffers can be used, and specifically, one or more substances selected from among various substances such as alcohol, Tris, EDTA, SDS, CTAB, Triton X100, MgCl2, KCl, NaCl, and other detergents can be used as the lysis buffer, and each buffer tank (111) can store a different lysis buffer.

[0049]

[0050] The supply operation unit (100) may be equipped with a luminescent agent injector (130) that injects luminescent agent into the microchannel chip (210). At this time, the luminescent agent may be injected from the luminescent agent injector (130) into the third microchannel of the microchannel chip (210).

[0051] To this end, the luminescent injector (130) may be equipped with a luminescent tank (131) that stores luminescent agent; and a luminescent pump (132), which is connected to the luminescent tank (131) and provides pressure to the luminescent agent discharged from the luminescent tank (131) to cause the luminescent agent to flow into the microfluidic chip (210).

[0052] Here, the ATP reaction luminescent agent may include luciferin and luciferase. At this time, the luminescent agent is magnesium ion (Mg 2+ ) may be included.

[0053] While the above-described solution passes through the second microchannel and the third microchannel, dissolution of microorganisms occurs within the solution, and accordingly, ATP can be extracted from the microorganisms.

[0054] The ATP extracted in this manner then reacts with a luminescent agent mixed with the solution, causing the microorganism to glow and produce light. The process of ATP reacting with a luminescent agent to produce light is already a well-known technology, so a detailed description will be omitted.

[0055] As described above, the solution in which light is generated by the reaction between the luminescent agent and ATP flows into the measuring channel (211) connected to the third microchannel, and a light sensor (220) can be installed on the measuring channel (211).

[0056] Here, the light sensor (220) may be a PMT (photomultiplier tube) sensor. However, the type of light sensor (220) is not limited to this, and other sensors capable of precise light measurement may be used.

[0057] The light sensor (220) senses light in the solution passing through the measuring path (211), and accordingly, a signal is generated from the light sensor (220), and the signal from the light sensor (220) can be transmitted to the analysis control unit (300).

[0058]

[0059] The analysis control unit (300) can store analysis data, which is data on the type of microorganisms that match the type of RLU and lysis buffer based on the signal of the light sensor (220). Here, information on the intensity of light can be generated based on the signal of the light sensor (220).

[0060] Among microorganisms contained in a solution, bacteria, germs, and fungi may exhibit varying degrees of lysis depending on the lysis buffer due to structural differences in their cells. Furthermore, the same source (microorganism) may exhibit different signal values, i.e., light intensity values, during luminescence measurements, depending on the type of lysis buffer.

[0061] Using this principle, the analysis control unit (300) can analyze the signal of the light sensor (220) to derive the RLU (Relative Light Unit) value measured using various lysis buffer solutions using bacteria / fungi.

[0062] In addition, the analysis control unit (300) can determine the type of lysine buffer injected into the microfluidic chip (210) by detecting an open buffer valve, and can determine the intensity of light and the RLU value derived using the same by the signal of the light sensor (220).

[0063] In addition, the analysis control unit (300) can search for the type of microorganism matching the analysis data using the RLU value and the type of lysis buffer, and accordingly, the analysis control unit (300) can analyze whether the microorganism is a bacterium, germ, or fungus and derive a result value for the type of microorganism.

[0064] In addition, the analysis data matches not only the types of microorganisms as described above, but also the concentration of microorganisms according to the RLU value and the type of lysis buffer, so that the concentration of microorganisms can be identified according to the RLU value and the type of lysis buffer.

[0065] In relation to analysis data, analysis data can be generated by providing microorganisms whose types are known in advance to a supply injector (120), then injecting a selected lysis buffer into a microfluidic chip (210), and organizing the RLU value obtained by the signal of the optical sensor (220).

[0066] The above air and water can be supplied multiple times at predetermined time intervals to the supply liquid generation tank (121), and the classification measurement device of the present invention can perform multiple classification measurements while changing the type of lyse buffer during each supply among the multiple supplies.

[0067] In addition, the types of microorganisms existing in the air can be derived by performing multiple classification measurements as described above, and in particular, the accuracy of the microbial classification measurement can be improved by determining the types of microorganisms derived in large numbers during multiple classification measurements as the types of microorganisms existing in the air.

[0068] That is, the phenomenon of deviation occurring by dissolving some fungi while dissolving bacteria by the lysis buffer can be minimized.

[0069]

[0070] The analysis control unit (300) can be installed in an electronic device (400) used by a user. In addition, the signal of the optical sensor (220) can be transmitted to the analysis control unit (300) via an external network.

[0071] Here, various devices such as smartphones, tablet PCs, and computers can be used as electronic devices (400), and various methods such as the Internet, Wi-Fi, and Bluetooth can be used as external networks.

[0072] As described above, using the analysis data, the analysis control unit (300) can quantify and organize information on the type and concentration of microorganisms, and the resulting information generated thereby can be immediately checked by the user in real time according to the Internet of Things (IoT) configuration described above.

[0073]

[0074] Hereinafter, the classification measurement method of the present invention using the classification measurement device of the present invention as described above will be described. The classification measurement method of the present invention includes a supply solution injection step; a buffer injection step; a luminescent agent injection step; a measurement step; and an analysis step; and each step can be performed sequentially.

[0075] In the supply liquid injection step, the supply liquid is generated in the supply operating unit (100), and the supply liquid can be injected from the supply operating unit (100) into the first microchannel of the microchannel chip (210).

[0076] In the buffer injection step, the lysis buffer can be injected from the supply operation unit (100) into the second microchannel of the microchannel chip (210).

[0077] In the luminescent agent injection step, the luminescent agent can be injected from the supply operation unit (100) into the third microchannel of the microchannel chip (210).

[0078] In the measurement step, the measuring unit (200) can measure the fluid passing through the measuring channel (211) of the microfluidic chip (210) and transmit the measurement signal to the analysis control unit (300).

[0079] In the analysis stage, the analysis control unit (300) can select and store the types of microorganisms.

[0080] The remaining details of the classification measuring device of the present invention are the same as those disclosed in the description of the classification measuring device of the present invention described above.

[0081]

[0082] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive. For example, each component described as a single entity may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined manner.

[0083] The scope of the present invention is indicated by the claims described below, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims

1. A measuring unit comprising a microchannel chip having a microchannel formed therein and a light sensor that measures light generated when microorganisms passing through the microchannel react with a luminescent agent; A supply solution containing microorganisms, a lysis buffer that lyses the cells of the microorganisms to extract adenosine triphosphate (ATP), and a supply operation unit that supplies the luminescent agent to the microfluidic chip; and An airborne microorganism classification and measurement device characterized by including an analysis control unit that receives a signal from the optical sensor from the measuring unit and selects the type of microorganism.

2. In claim 1, An airborne microorganism classification and measurement device characterized in that the supply operation unit comprises a buffer injector that injects the lysis buffer into the microfluidic chip.

3. In claim 2, An airborne microorganism classification and measurement device characterized in that the buffer injector injects at least one lysis buffer among a plurality of lysis buffers into the microfluidic chip.

4. In claim 1, An airborne microorganism classification and measurement device characterized in that the analysis control unit stores analysis data, which is data on the type of microorganism matching the RLU by the signal of the optical sensor and the type of the lyse buffer.

5. In claim 1, An airborne microorganism classification and measurement device characterized in that the above supply operation unit comprises a supply liquid injector that injects the supply liquid into the microfluidic chip.

6. In claim 1, An airborne microorganism classification and measurement device characterized in that the supply operation unit comprises a luminescent agent injector that injects the luminescent agent into the microfluidic chip.

7. In claim 1, An airborne microorganism classification measuring device characterized in that the above optical sensor is a PMT (photomultiplier tube) sensor.

8. In claim 1, An airborne microorganism classification measuring device, characterized in that the signal of the above optical sensor is transmitted to the analysis control unit via an external network.

9. In claim 1, An airborne microorganism classification and measurement device characterized in that the above analysis control unit is installed in an electronic device used by a user.

10. In a method for measuring airborne microorganism classification using the airborne microorganism classification measuring device of claim 1, A supply liquid injection step in which the supply liquid is generated in the supply operating unit and the supply liquid is injected from the supply operating unit into the first microchannel of the microchannel chip; A buffer injection step of injecting the lyse buffer from the supply operation unit into the second microchannel of the microchannel chip; A luminescent agent injection step for injecting the luminescent agent into the third microchannel of the microchannel chip from the above supply operation unit; A measuring step in which the measuring unit measures the fluid passing through the measuring channel of the microfluidic chip and transmits a measurement signal to the analysis control unit; and A method for measuring airborne microorganism classification, characterized in that it includes an analysis step of selecting and storing the type of microorganism in the above analysis control unit.

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