KR20110128600A - Airborne microbial measurement apparatus and measurement method using the microorganism dissolution system - Google Patents

Abstract

PURPOSE: An apparatus for measuring airborne microbes using a microorganism dissolution system is provided to quickly measure in real time with safety. CONSTITUTION: An apparatus for measuring airborne microbes using a microorganism dissolution system comprises: a filter(10) for collecting airborne microbes and absorbing ATP reaction luminescent agent; a microorganism dissolution system(20) for dissolving microbes by electromagnetic force and extracting ATP(adenosine triphosphate); a photodetector(30) for detecting luminous intensity of light generated by reaction ATP and ATP reaction luminescent agent and measuring the concentration of the microbes. The ATP reaction luminescent agent is luciferin. The microorganism dissolution system is an ion generator for extracting APT.

Description

Airborne microbial measurement apparatus and measurement method using the microorganism dissolution system}

The present invention relates to an apparatus and method for measuring suspended microorganisms present in the gas phase, and real-time measuring apparatus and measurement method of suspended microorganisms in the gas phase to quickly measure the suspended microorganisms in the gas phase using the ATP bioluminescence measuring method. It is about.

Recently, avian influenza, swine flu, etc. have been raised, the problem of air infection has emerged, and thus airborne microbial measurement is more important, and the biosensor market is growing rapidly accordingly.

In the conventional method of measuring airborne microorganisms, the biological particles suspended in the sample gas are collected on a solid or liquid surface suitable for propagation, incubated in a suitable temperature and humidity environment for a certain period of time, and then collected in the colony water on the surface. And culture methods for obtaining and staining using a fluorescence microscope after staining.

Recently, ATP bioluminescence method uses the principle that ATP (adenosine triphosphate) and luciferin / luciferase shine to produce light. The process has been reduced to about 30 minutes, allowing for quick work.

However, according to the above methods, it is not possible to measure the floating microorganisms present in the weather in real time, and a series of manual operations including separate sampling and pretreatment are required. There was a limit not to develop.

In fact, conventional biosensors require a minimum sampling time of 20 minutes to a maximum of 2 hours because they need to be applied after a separate sampling process when measuring airborne microorganisms. UV-APS of US TSI However, as it is more than 200 million won in Hanwha, only a part of professional research institutes can be used, but it is impossible to disseminate it in reality.

In addition, in order to apply the ATP bioluminescence method, ATP extractant is basically required, but when it is used in the meteorological floating microbial measurement system, it may adversely affect the human body, and the ATP extractant may be applied to the automatic system. It must be supplied continuously, but there is a problem in that it is expensive to continuously supply ATP extractant which is currently commercially available.

The present invention devised to solve the problems described above, by using the ATP bioluminescence measuring method to quickly measure the airborne microorganisms present in the gas phase does not need to go through a series of manual operations, as well as real-time automatic measurement, The present invention relates to a real-time measurement device and a measurement method of suspended microorganisms in the weather, which can realize safety and low cost.

The present invention for achieving the object as described above, the filter 10 is trapped in the airborne microorganisms, the ATP reaction light emitting agent is absorbed; A microbial lysis system 20 for extracting ATP (adenosine triphosphate) by dissolving microorganisms using electromagnetic force; And a light receiving device which detects luminous intensity of light generated when the ATP extracted by the microbial lysis system 20 reacts with the ATP reactive light emitting agent of the filter 10 to extract the concentration of the microorganism. (30); the real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system configured to include the technical gist.

Here, the filter 10 is installed in a state in which the ATP reactive light emitting agent is absorbed, or the ATP reactive light emitting agent supply device 11 for supplying the ATP reactive light emitting agent to the filter 10; The ATP reactive light emitter is preferably luciferin.

In addition, the filter 10 preferably has a collection efficiency of 90% or more with respect to 1 µm particles.

In addition, the microbial dissolution system 20, the ion generator for damaging the cell wall of the microorganism by the repulsion between the charged ions attached to the microorganism; preferably, the ion generator, the ion generator, the diameter of the discharge tip 10 It is preferable that it is an ozone-free ion generator which uses the carbon brush of micrometer or less.

The microbial dissolution system 20 may be a plasma discharger that extracts ATP while damaging the cell wall of the microorganism by collision of high concentration ions and electrons generated by high voltage discharge.

In addition, the light receiving element 30 preferably has a sensitivity capable of receiving a wavelength band of 400 nm or more and 700 nm or less.

And, the display device 40 for displaying in real time the concentration of the microorganisms extracted by the light intensity detected by the light receiving element 30; preferably further comprises a.

In addition, it is preferable that the flow generating means 50 is installed to generate a pneumatic pressure difference forcing the flow of the atmosphere toward the filter 10 side.

In addition, the present invention, while sampling the suspended microorganisms in the gas phase in the filter 10 absorbed by the ATP reaction light emitting agent, while dissolving the microorganisms by the continuous microbial dissolution system 20 is caught on the filter 10 By extracting the ATP (adenosine triphosphate) of the microorganism, induces the luminous reaction between the ATP reaction light emitting agent and the ATP of the filter 10 in real time, using a microbial dissolution system for measuring the concentration of microorganisms using the light receiving element 30 Real-time measurement of suspended microorganisms in the weather is another technical subject.

In addition, the present invention, the microorganism collecting step of collecting the suspended microorganisms in the filter 10; ATP extraction step of operating the microbial dissolution system 20 to dissolve the microorganisms collected in the filter 10 to extract ATP (adenosine triphosphate); And real-time detection in which the ATP is extracted in the ATP extraction step and the light intensity of the light generated by reacting with the ATP reactive light emitter absorbed in the filter 10 is measured in real time with the light receiving element 30. It is another technical gist of the method for real-time measurement of suspended microorganisms in the gas phase using a microbial dissolution system configured to include.

Here, the real-time display step of converting the data detected by the light receiving element 30 in the real-time detection step in real time to display in real time; preferably further comprises a.

According to the present invention having the above-described configuration, while sampling the suspended microorganisms in the gaseous phase to the filter absorbed by the ATP reaction light emitting agent, while dissolving the microorganisms by the continuous microbial dissolution system and extracting the ATP of the microorganisms caught in the filter As a result, there is an effect of inducing a light-emitting reaction between the ATP-reactive light emitting agent of the filter and ATP in real time.

The microbial lysis system enables rapid measurement of suspended microorganisms in the gas phase within 5 minutes by ATP bioluminescence measurement, while the process from sampling to ATP extraction and bioluminescence is performed automatically without a series of manual tasks. Another effect is the real-time automatic measurement of heavy suspended microorganisms.

In addition, by applying semi-permanently available devices such as ion generators and plasma dischargers to the microbial dissolution system, the high cost, management, and cost of the continuous supply and control of reagents such as lysis-buffer to dissolve microorganisms It can be safely used at low cost without the difficulty of maintenance and toxicity to human body, and can be controlled easily by electric method.

Existing biosensors not only are expensive but also require a lot of manpower and cost due to a series of manual operations, but according to the present invention, real-time measurement of floating microorganisms is realized by real-time automatic measurement, low cost, and stabilization of floating microorganisms in the weather. There is another effect of making the device universal and pervasive.

As a result, it is possible to easily measure harmful microbial propagation of mad cow disease, swine cholera, bird flu, or food in barns and food factories, effectively preventing social and economic losses caused by air infection, and rapidly growing biosensors. Another effect is that it can contribute to the improvement of human welfare by spreading biosensors by satisfying market demand.

1-conceptual diagram showing a first embodiment of a real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system according to the present invention
2 is a conceptual diagram showing a second embodiment of a real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system according to the present invention
3-A graph showing the results of meteorological suspended bacteria measurement according to operating time
4-flowchart of a method for real-time measurement of suspended microorganisms in the gas phase using a microbial lysis system according to the present invention

With reference to the drawings will be described in more detail with respect to the real-time measuring apparatus and measurement method of suspended microorganisms in the gas phase using a microbial dissolution system according to the present invention.

Floating microorganism real-time measurement device in the gas phase using a microbial dissolution system according to the present invention, as shown in Figure 1, 2, consisting of a filter 10, microbial dissolution system 20, a light receiving element 30, Simultaneous sampling of the suspended microorganisms in the filter 10 and the microbial dissolution system 20 (described below) continuously dissolves the microorganisms and extracts ATP (adenosine triphosphate) to an automatic measurement system for bioluminescence.

The filter 10, the microorganisms suspended in the atmosphere passing through the filter 10 is filtered by the filter 10 while passing through the filter 10 is collected, the filter 10 that the microorganisms are collected ) Absorbs the ATP reactive light emitting agent required for ATP bioluminescence.

In the state in which the ATP-reactive light emitting agent is absorbed in the filter 10, as shown in FIG. 1, the filter 10 in which the ATP-reactive light-emitting agent is already applied or absorbed is provided, or in FIG. 2. As shown, the ATP reaction light emitting agent supply device 11 for spraying or supplying the ATP reactive light emitting agent as often as necessary to the filter 10 may be configured separately from the mesh member constituting the filter 10. .

In general, visible pollen, mold, microorganisms, fiber dust, etc. have a particle size of 100 μm or more, and in the case of bacteria, the filter 10 has a size of 0.1 μm or more and 100 μm. In view of the appropriateness of the collection efficiency, such as the initial investment cost, maintenance costs, etc., it is preferable to select one having a collection efficiency of 90% or more for 1 µm particles.

The ATP reaction light emitting agent supply device 11 is not limited to a specific structure and form as long as it can supply and absorb the liquid ATP reactive light emitting agent to the filter 10 side, the use environment of the configuration of a known liquid supply device, Since it is preferable to apply more suitable things in consideration of conditions, such as an apparatus specification, comprehensively, the detailed description is abbreviate | omitted.

The microbial lysis system 20 dissolves the microorganisms collected in the filter 10 or flows toward the filter 10 by using an electromagnetic force of ions or electrons, and thus ATP (adenosine triphosphate), DNA, and RNA in the microorganism. In general, a device component for extracting and the like, wherein dissolving the microorganisms does not dissolve the microorganisms into liquid state, but decomposes one microorganism into a plurality of elements.

When the microbial dissolution system 20 is configured as an ion generator, the larger the diameter of the discharge tip provided in the ion generator, the greater the power consumption. When the power consumption is high, not only ions but also ozone harmful to the human body may occur. Therefore, it is preferable to apply an ozone-free ion generator using a carbon brush having a diameter of the discharge tip of 10 µm or less.

According to the ozone-free ion generator using a carbon brush having a discharge tip diameter of 10 μm or less, it has a low power consumption of 4 W or less, and ozone is generated at less than 0.01 ppm. Ozone management standards of 0.06 ppm or less under Article 27 (1) of the Industrial Safety and Health Act can be satisfactorily satisfied.

When the microbial dissolution system 20 is configured as an ion generator, the microbial cell wall of the microorganism is damaged by repulsion between charged ions attached to the microorganisms, and ATP is extracted. When the microbial dissolution system is configured as a plasma discharger, a high concentration of ions generated by high voltage discharge is generated. As a result of the collision of electrons, ATP is extracted while damaging the cell wall of the microorganism.

The ATP extracted by the microbial lysis system 20 is exposed to the outside of the cell of the microorganism and at the same time reacts with the ATP reaction light emitting adsorbed on the filter 10 to generate light, and converts the light into electricity. A light receiving element 30, such as a photodiode (PD) or an avalanche photodiode (APD), detects luminous intensity of light generated by ATP bioluminescence to detect microorganisms. Extract the concentration.

All organisms store the energy from the oxidation of organic matter in a compound called ATP, and then hydrolyze it as needed to exercise and maintain body temperature using the energy released at that time. It also causes bioluminescence.

The light receiving element measures the photon flux or the optical power by converting the energy of photons absorbed by the element into a measurable form. It is widely used as a device for detecting optical signals in optical fiber communication systems operating in the near infrared region (0.8 ~ 1.6 μm).

In particular, photoelectric detectors of light-receiving elements generate carriers such as electrons and holes in a material constituting the element by photons absorbed by the element, and by the flow of the carriers A device in which a measurable current is generated, ie based on photoeffects, is suitable for application in the present invention.

The wavelength of the electromagnetic wave, which is felt brightly by the human eye, ranges from about 380 nm to 780 nm. For monochromatic light, the wavelength is short to celadon 400 to 500 nm, blue 450 to 500 nm, green 500 to 570 nm, sulfur 570 to 590 nm, and orange 590 to At 610 nm and red 610 to 700 nm, the light receiving element 30 has a sensitivity capable of receiving a wavelength band of 400 nm or more and 700 nm or less.

In collecting the airborne microorganisms in the filter 10, a pneumatic difference is generated by forcibly flowing the atmosphere of one side to the other side based on the filter 10 using a flow generating means 50 such as a blower or a pump. The microbial dissolution system 20 and the light receiving element 30 are installed on a path through which air flows to the filter 10, that is, on one side of the filter 10. 50 is installed on the other side of the filter 10.

The higher the concentration of microorganisms, the greater the amount of ATP extracted, and the greater the degree of luminous intensity. The luminous intensity detected by the light-receiving element 30 is subjected to a signal processing process that finally converts the concentration of the microorganism into a concentration of the display device. 40) displays the concentration of microorganisms in real time.

Bioluminescence is a kind of photochemical reaction in which an organic compound is oxidized by the action of an enzyme, and the energy released by the body is in the form of light energy. Luciferin, a luminescent substance, combines with ATP to form a complex of luciferin-ATP. Produce two molecules. Luciferin is here reduced and is represented as LH2 (LH2 + ATP → LH2-AMP + 2H3PO4).

The LH2-AMP generated in the reaction is in an unstable energy state as it reacts with oxygen and oxidizes, and thus, the unstable oxidized product decomposes to generate light (hv) while generating oxidized luciferin and AMP. Where L is the oxidized luciferin and L-AMP * indicates the luciferin-AMP complex in an unstable energy state. (LH2-AMP + 1/2 O2 → L-AMP * + H2O) (L-AMP * → L + AMP + hv (light energy))

 Since LH2-AMP is reacted with oxygen (1/2 O2) and oxidized by the catalysis of an enzyme called luciferase, bioluminescence occurs in the presence of luciferin, ATP, luciferase and oxygen. It is calculated that 1 photon is emitted from oxidation of.

When the ATP reaction light-emitting agent is composed of luciferin, suspended microorganisms can be rapidly measured within 5 minutes by the above-described process, and the graph shown in FIG. 3 is the agent of the present invention shown in FIG. After configuring the first embodiment, the change in the measured value of the gaseous floating bacteria according to the system operation time is displayed, and the maximum brightness is measured within 3 minutes (180sec), and it is confirmed that the measurement time is taken within 3 minutes from this. Can be.

In the experiment of the graph shown in Figure 3, the ozone free ion generator was used as the microbial dissolution system 20, air flow rate: 3 l / min, temperature 23 ℃,-ion 9 × 10 ^ 6 number / cm 3, bio aerosol The experiment was conducted under the condition of 93000 CFU / ㎥, and the intensity unit is RLU (relative luminescent unit).

The real-time measurement method of suspended microorganisms in the gas phase using the microbial lysis system according to the present invention relates to a method for automatically measuring the concentration of microorganisms in real time using the real-time measuring apparatus of suspended microorganisms in the gas phase according to the present invention having the configuration as described above. .

Sampling the microorganisms in the gas phase to the filter 10 absorbed by luciferin, while dissolving the microorganisms by the continuously operating microbial dissolution system 20 and ATP ( Adenosine triphosphate) is extracted to induce a luminescence reaction between luciferin and ATP of the filter 10 in real time, and the concentration of microorganisms is measured using the light receiving element 30.

As shown in FIG. 4, the microbial collection step, the ATP extraction step, the real time detection step, and the real time display step are sequentially performed, but the whole process is performed in a short time within 5 minutes, and each step is performed. The continuous operation on each component has the same effect as the whole process at the same time.

In the microbial collection step, suspended microorganisms are collected in the filter 10, and in the ATP extraction step, the microbial dissolution system 20 is operated to dissolve the microorganisms collected in the filter 10 to form ATP (adenosine triphosphate). Extract.

In the real-time detection step, the ATP is extracted in the ATP extraction step and the intensity of light generated by reacting with luciferin absorbed by the filter 10 is measured in real time by the light receiving element 30. In the real-time detection step, the data detected by the light receiving element 30 is converted into a concentration of microorganisms and displayed in real time.

According to the real-time measuring device and measurement method of suspended microorganisms in the gas phase using the microbial dissolution system according to the present invention having the configuration as described above, and while sampling the suspended microorganisms in the gas phase on the filter 10 absorbed luciferin, continuously By dissolving the microorganisms by the microbial lysis system 20 in operation and extracting the ATP of the microorganisms caught in the filter 10, the luminescent reaction between luciferin and the ATP of the filter 10 is induced in real time.

In the past, ion generators, plasma dischargers, and related technologies were used only to remove harmful substances such as bio aerosols, particles, and gases, and were limited to methods using reagents such as lysis-buffers to dissolve microorganisms. In the present invention, a device that can be used semi-permanently such as an ion generator and a plasma discharger is removed from the present invention and applied to the microbial dissolution system.

As a result, airborne microorganisms present in the gas phase can be quickly measured within 5 minutes by ATP bioluminescence measurement, and the process from sampling to ATP extraction and bioluminescence is automatically performed without a series of manual operations. Real-time automatic measurement

By applying semi-permanent devices such as ion generators and plasma dischargers to the microbial dissolution system, the high cost, management, maintenance and maintenance of the conventional reagents such as lysis-buffer It can be safely used at low cost without concern for toxicity, and can be controlled easily by electric method.

Existing biosensors not only are expensive, but also have a large burden on human resources and costs due to a series of manual operations, but according to the present invention, real-time measurement of floating microorganisms by real-time automatic measurement, low cost, and stabilization of suspended microorganisms in the weather. Make the device universal and popularized.

As a result, it is possible to easily measure harmful microbial propagation of mad cow disease, swine cholera, bird flu, or food in barns and food factories, effectively preventing social and economic losses caused by air infection, and rapidly growing biosensors. It can meet the demand of the market and contribute to the improvement of human welfare due to the diffusion of biosensors.

The present invention has been described above with reference to preferred embodiments of the present invention, but the present invention is not limited to these embodiments, and the claims and detailed description of the present invention together with the embodiments in which the above embodiments are simply combined with existing known technologies. In the present invention, it can be seen that the technology that can be modified and used by those skilled in the art are naturally included in the technical scope of the present invention.

10 filter 11: ATP reaction light emitting device supply device
20: microbial dissolution system 30: light receiving element
40: display device 50: flow generating means

Claims (14)

A filter 10 in which suspended microorganisms are collected in the gas phase and the ATP reaction light emitting agent is absorbed;
A microbial lysis system 20 for extracting ATP (adenosine triphosphate) by dissolving microorganisms using electromagnetic force; And
A light receiving device for extracting the concentration of microorganisms by detecting the luminous intensity of light generated by the ATP extracted by the microbial lysis system 20 reacts with the ATP reactive light emitting agent of the filter 10 ( 30);
Real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that comprising a.
The method of claim 1, wherein the filter 10,
Apparatus for real-time measurement of suspended microorganisms in the gas phase using a microbial lysis system, characterized in that the ATP reaction light emitting agent is installed in a state of being absorbed.
The method of claim 1,
An ATP reactive light emitting agent supply device (11) for supplying the ATP reactive light emitting agent to the filter (10);
Real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that the configuration further comprises.
The method of claim 1, wherein the ATP reactive light emitting agent,
Real-time measurement of suspended microorganisms in the gas phase using a microbial lysis system, characterized in that luciferin (luciferin).
The method of claim 1, wherein the filter 10,
Real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that the collection efficiency of more than 90% for 1㎛ particles.
The method of claim 1, wherein the microbial lysis system 20,
An ion generator extracting ATP while damaging the cell wall of the microorganism by repulsion between charged ions attached to the microorganism;
Real-time measuring device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that.
The method of claim 6, wherein the ion generator,
An apparatus for real-time measurement of suspended microorganisms in a gas phase using a microbial dissolution system, characterized in that it is an ozone-free ion generator using a carbon brush having a diameter of a discharge tip of 10 μm or less.
The method of claim 1, wherein the microbial lysis system 20,
A plasma discharger for extracting ATP while damaging the cell wall of the microorganism by collision of high concentration of ions and electrons generated by high voltage discharge;
Real-time measuring device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that.
The method of claim 1, wherein the light receiving element 30,
A real-time measurement device for suspended microorganisms in the gas phase using a microbial lysis system, characterized in that it has a sensitivity capable of receiving a wavelength band of 400nm to 700nm or less.
The method of claim 1,
A display device 40 which displays the concentration of the microorganism extracted by the light intensity detected by the light receiving element 30 in real time;
Real-time measurement device of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that the configuration further comprises.
The method of claim 1,
Flow generating means (50) installed to generate a pneumatic differential forcing the atmosphere to flow toward the filter (10);
Real-time measurement device for suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that the configuration further comprises.
Sampling the suspended microorganisms in the gaseous phase to the filter 10 absorbed by the ATP reaction light emitting agent, while dissolving the microorganisms by the continuously operating microbial dissolution system 20 and the ATP (adenosine triphosphate) of the microorganisms caught in the filter 10 ) To induce the luminescence reaction between the ATP reaction light emitting agent and ATP in real time of the filter 10, and to measure the concentration of microorganisms using the light receiving element 30 in the gas phase using a microbial dissolution system. Real time microbial measurement method.
A microbial collecting step of collecting the suspended microorganisms in the filter 10;
ATP extraction step of operating the microbial dissolution system 20 to dissolve the microorganisms to extract ATP (adenosine triphosphate); And
Real-time detection step of measuring the luminous intensity of the light generated by the light receiving element 30 in real time at the same time the ATP is extracted in the ATP extraction step and reacted with the ATP reaction light-emitting agent absorbed in the filter 10 ;
Real-time measurement method of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that comprises a.
The method of claim 13,
A real-time display step of converting the data detected by the light receiving element 30 in the real-time detection step into a microorganism concentration and displaying in real time;
Real-time measurement method of suspended microorganisms in the gas phase using a microbial dissolution system, characterized in that further comprises.

Images