CN115015203A - Exhaled aerosol microorganism rapid detection device - Google Patents

Exhaled aerosol microorganism rapid detection device Download PDF

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Publication number
CN115015203A
CN115015203A CN202210699079.6A CN202210699079A CN115015203A CN 115015203 A CN115015203 A CN 115015203A CN 202210699079 A CN202210699079 A CN 202210699079A CN 115015203 A CN115015203 A CN 115015203A
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China
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spectrum
aerosol particles
aerosol
fluorescence
exhaled
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Inventor
许锋
王镝
潘宇祥
曹庆朋
钱利滨
郑旭彬
程晨
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Zhejiang Lab
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Zhejiang Lab
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6402—Atomic fluorescence; Laser induced fluorescence
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/22—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material
    • G01N23/223—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by measuring secondary emission from the material by irradiating the sample with X-rays or gamma-rays and by measuring X-ray fluorescence

Abstract

The invention discloses an exhaled aerosol microorganism rapid detection device. The device comprises a spectrum collection cavity, a collection carrier, an optical detection assembly and a radiation source, wherein the collection carrier is arranged in the spectrum collection cavity and used for bearing aerosol particles, the optical detection assembly generates light beams to irradiate the aerosol particles to excite the generated fluorescence for detection to obtain a two-dimensional fluorescence spectrum, and the radiation source is arranged in the spectrum collection cavity and irradiates towards the aerosol particles, so that the detected two-dimensional fluorescence spectrum is changed into a three-dimensional fluorescence spectrum. The invention adds a radiation source for irradiation on a detection structure for detecting the two-dimensional fluorescence spectrum, is used for scanning the conformational change of the protein on the surface of the pathogen to obtain the three-dimensional fluorescence spectrum reflecting the microbial information, and further can realize the rapid extraction and detection on site.

Description

Exhaled aerosol microorganism rapid detection device
Technical Field
The invention relates to a component detection device, belonging to the technical field of exhaled gas detection, in particular to an exhaled aerosol microorganism rapid detection device based on continuous irradiation three-dimensional fluorescence.
Background
Respiratory infections are transmitted by Exhaled Breath Aerosols (EBAs) and are the focus of global attention. Current research indicates that aerosols are the predominant form of transmission of respiratory infectious pathogens. Patients breathe, speak, cough, and sneeze, which produce aerosols that can be exposed to human infection. The on-site detection of pathogens in the exhaled breath of the human body is the preferred solution to eliminate the current monitoring hysteresis.
Currently, most exhaled aerosol microorganism identification methods are offline detection methods, such as: microbial culture method, molecular microbiology method, and immunoassay method. However, the traditional laboratory offline detection device has the defects of long detection period, incapability of being applied on site, complex nucleic acid extraction, dependence on desktop equipment, easiness in failure of sensors and the like. Therefore, offline detection is difficult to identify the sudden pathogenic aerosol exposure event in situ, accurately and quickly. For the on-line monitoring method, laser-induced fluorescence was used as early as 1994 for identifying microbial warfare agents by the army of the united states: the built fluorescence laser radar system can detect microbial aerosol on 0.6-3.0 km high altitude.
Currently, fluorescent microbial aerosol sensors (WIBS) are used in the study of the distribution characteristics of microbial aerosols. The research shows that the aerosol in the city has microbial fluorescent particles and the concentration of the fluorescent particles is positively correlated with the concentration of the carbon particles. Still other researchers have probed bacillus anthracis microbial agents with ultraviolet laser induced radar detection systems.
In conclusion, the fluorescence spectrum technology has the advantages of high detection sensitivity, strong specificity and simplicity and convenience in operation, but the existing microbial aerosol fluorescence identification only obtains intensity information, and cannot realize accurate identification of microbial components and concentrations in exhaled aerosols on site. The components and the concentration of microorganisms in the exhaled aerosol are difficult to detect in strong interference only by the conventional fluorescence technology, and the species and the concentration of the microorganisms in the exhaled aerosol cannot be accurately identified on site.
Disclosure of Invention
The invention aims to provide a device for rapidly detecting microorganisms in exhaled aerosol based on continuous irradiation three-dimensional fluorescence, which aims to overcome the technical problems that the conventional fluorescence technology is difficult to detect the components and the concentration of the microorganisms in exhaled aerosol under the conditions of strong interference, low concentration and the like, and the technical problem that the types and the concentrations of the microorganisms in exhaled aerosol cannot be identified accurately in real time on site.
The technical scheme of the invention is as follows:
comprises a spectrum acquisition cavity;
comprises a collecting carrier, a spectrum collecting cavity and a spectrum collecting cavity, wherein the collecting carrier is arranged in the spectrum collecting cavity and carries aerosol particles obtained by human body exhalation;
the device comprises a light detection assembly, a light source and a control assembly, wherein the light detection assembly generates light beams to irradiate aerosol particles, and detects fluorescence generated by excitation of microorganisms on the aerosol particles to obtain a two-dimensional fluorescence spectrum;
the device comprises a radiation source, wherein the radiation source is arranged in a spectrum collection cavity and faces to aerosol particles on a collection carrier, the radiation source irradiates on the aerosol particles to influence fluorescence generated by excitation of microorganisms on the aerosol particles, and a detected two-dimensional fluorescence spectrum is changed into a three-dimensional fluorescence spectrum.
The aerosol particles are obtained by sampling from human breath through a sampling assembly, and the sampling assembly comprises an expired gas sampler and a breath sampling nozzle;
the breath sampling nozzle is arranged on the human mouth to receive the breath of the human body and discharge the breath to the expired gas sampler;
the expired gas sampler receives the human breath from the breath sampling nozzle, forms aerosol particles through electrodeposition treatment and is arranged on the collecting carrier.
The optical detection component comprises a laser, a convex lens, a fluorescent filter, a spectrum shooting instrument and an optical trap; the laser emits laser, the laser irradiates aerosol particles on the collecting carrier to excite fluorescence, light beams transmitted through the aerosol particles are absorbed by the light trap, and the fluorescence excited on the aerosol particles is detected and received by the spectral shooting instrument after passing through the convex lens and the fluorescence filter.
The convex lens and the fluorescent filter are arranged in the spectrum acquisition cavity, and the laser, the reflector, the spectrum shooting instrument and the light trap are arranged outside the spectrum acquisition cavity.
In the optical detection assembly, light beams irradiated onto aerosol particles are kept continuous, and detection and reception of fluorescence generated by excitation of microorganisms on the aerosol particles are discontinuous; meanwhile, the irradiation of the aerosol particles by the radiation source is discontinuous.
The detection receiving of the optical detection component for generating fluorescence by exciting microorganisms on the aerosol particles and the irradiation of the aerosol particles by the radiation source are asynchronous in time sequence.
The collecting carrier specifically comprises a collecting plate which is used as a carrier for bearing the exhaled aerosol particles.
The collecting carrier is also provided with a temperature control device and an ultrasonic device;
the temperature control device is arranged on the bottom surface of the collecting carrier and is used for controlling the temperature of the collecting carrier and the aerosol particles on the collecting carrier;
and the ultrasonic device is arranged on the bottom surface of the collecting carrier and used for generating ultrasonic vibration to remove aerosol particles on the collecting carrier after identification and detection.
The irradiation type emitted by the radiation source comprises one or more of microwave, ultrasound and (x) ray.
The irradiation parameters of the radiation source comprise resolution, maximum treatment intensity and integration time.
The invention utilizes the device to carry out the following detection processes:
s1, depositing human exhaled breath sol particles on the irradiated collecting carrier;
s2, setting the type and irradiation parameters of irradiation treatment;
s4, continuously irradiating the aerosol particles on the collecting carrier through a radiation source, and scanning a fluorescence spectrum for detecting the whole process that the pathogen surface protein conformation of the microorganisms on the aerosol particles is irradiated to obtain a three-dimensional fluorescence spectrum;
in the three-dimensional fluorescence spectrum, the fluorescence intensity is related to the wavelength and the irradiation intensity.
S5, analyzing the three-dimensional fluorescence spectrum to extract contour characteristic spectrum as fluorescence fingerprint, and performing characteristic extraction and modeling;
and S6, processing according to the established model and the extracted characteristics to realize the detection of the microorganisms in the aerosol exhaled by the human body.
The collecting carrier is also provided with a temperature control device, and the following steps S3 are carried out after S2 and before S4: the temperature control device is turned on to keep the temperature on the collection carrier constant. The temperature is controlled within the range of 0-36 ℃.
And S6, specifically, processing the extracted features according to the established model to obtain the species and the content of the microorganisms in the exhaled breath of the human body.
The three-dimensional fluorescence spectrum is obtained by continuous discontinuous detection and uptake in the discontinuous radiation treatment process.
The obtained three-dimensional fluorescence spectrum contains variables generated in the process that microorganism particles in the aerosol are radiated, and the species and concentration information of the microorganisms in the aerosol of the human respiratory gas are analyzed by utilizing the specificity of the microorganisms expressed in the radiation treatment through spectral treatment and modeling analysis.
The microorganism to which the present invention is directed is a microorganism that fluoresces when excited by laser irradiation, and is generally a microorganism such as a respiratory infectious agent including a protein structure, for example, severe acute respiratory syndrome coronavirus, influenza virus type ii and iii, measles, and the like.
The invention utilizes the fluorescent on-line detection mode in combination with the optimization of irradiation, and can effectively, quickly and accurately analyze and detect to obtain the information of the types and the concentrations of microorganisms in the aerosol.
Generally, aerosol microorganism particles can also generate fluorescence by laser irradiation, but due to the low concentration of microorganisms in exhaled aerosol and the complex components of exhaled aerosol, the problems of weak fluorescence signal and easy interference of the signal are caused, and effective identification and detection of aerosol microorganisms cannot be realized.
The invention adds continuous irradiation, changes the protein conformation of the surface of the microbial pathogen, obtains the fluorescence spectrum of the whole irradiation process in the changing process, forms a three-dimensional fluorescence spectrum from the two-dimensional fluorescence spectrum, extracts a contour line characteristic spectrum from the three-dimensional fluorescence, analyzes the spectral information of the three-dimensional fluorescence, such as the peak position, the height, the change rate, the trough length, the peak ridge length, the width and the like, and further judges and obtains the identification and detection results of the microorganism.
Compared with the prior art, the invention has the following beneficial effects:
the device can be used for processing aerosol particles through continuous irradiation to obtain fluorescence fingerprints, a structure for irradiation (such as microwave, ultraviolet ray and X-ray irradiation) is added on a light detection assembly of a two-dimensional fluorescence spectrum, and a three-dimensional fluorescence spectrum containing microorganism information is obtained through rapidly scanning the fluorescence spectrum of the conformational change process of the pathogen surface protein of a microorganism under irradiation, so that the device can be used for obtaining the type and concentration information of the microorganism through inversion by utilizing the specificity of the microorganism expressed in the irradiation processing, and the breakthrough of the field detection of the exhaled aerosol microorganisms and the pathogen is realized.
By introducing the radiation, the invention not only can realize the screening of the microorganism information, but also can weaken the requirement of the detection system on the light source intensity, improve the stability of the detection system, realize the real-time and rapid extraction of the fluorescence spectrum of the microorganism in the aerosol on site, and is beneficial to improving the efficiency and the accuracy of the field detection of the exhaled aerosol microorganism.
Drawings
FIG. 1 is a flow chart of one embodiment of the present invention;
FIG. 2 is a schematic view of an apparatus embodying the present invention;
FIG. 3 is a schematic diagram of an apparatus according to an embodiment of the present invention;
FIG. 4 is a cross-sectional view A-A of FIG. 3;
FIG. 5 is a graph showing the results of continuous irradiation of exhaled breath sol microorganisms with three-dimensional fluorescence.
FIG. 6 is a result graph of contour line characteristic spectrum of exhaled breath sol microorganism continuously irradiated with three-dimensional fluorescence.
In the figure: the device comprises a spectrum shooting instrument 1, a spectrum collecting cavity 2, an expired gas sampler 3, a laser 4, a convex lens 5, a fluorescent filter 6, a radiation source 7, a light trap 8, a temperature control device 9, an ultrasonic device 10, a collecting plate 11, a breathing sampling nozzle 12 and a reflector 13; a sampling and filtering link 14, a composite sampling link 15, a spectrum acquisition cavity link 16, a beam-collecting air outlet link 17, a spectrum incident optical fiber 18, a spectrum emergent optical fiber 19, a radiation source 20 and an aerosol deposition component 21; a collision cutter segment 22, a corona discharge segment 23, a quartz collecting plate 24, a temperature controller 25 and an ultrasonic device 26.
Detailed Description
In order to make the objects, features and advantages of the present invention more apparent and understandable, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making creative efforts based on the embodiments of the present invention, belong to the protection scope of the present invention.
The invention is explained in more detail below with reference to the figures and examples.
As shown in FIG. 2, the device mainly comprises a spectrum collection cavity 2, a collection carrier, an optical detection assembly and a radiation source 7.
The device comprises a spectrum acquisition cavity 2, a spectrum detection cavity and a spectrum detection cavity, wherein the spectrum acquisition cavity is used for accommodating aerosol particles and related components to form a detection environment;
comprises a collecting carrier, which is arranged in a spectrum collecting cavity 2 and carries aerosol particles obtained by the exhalation of a human body;
the device comprises an optical detection assembly for detecting a two-dimensional fluorescence spectrum, wherein the optical detection assembly generates a light beam to irradiate the aerosol particles, and detects fluorescence generated by the excitation of microorganisms on the aerosol particles to obtain the two-dimensional fluorescence spectrum;
the device comprises a radiation source 7 which is arranged in a spectrum collection cavity 2 and faces to aerosol particles on a collection carrier, wherein the radiation source 7 irradiates the aerosol particles to influence fluorescence generated by excitation of microorganisms on the aerosol particles, so that the protein conformation of the aerosol microorganisms is changed, a detected two-dimensional fluorescence spectrum for recording the whole change process is formed, a three-dimensional fluorescence spectrum is formed, and the microorganisms are rapidly detected by using the three-dimensional fluorescence spectrum.
In specific implementation, aerosol particles are obtained by sampling from human respiratory gas through a sampling assembly, wherein the sampling assembly comprises an expired gas sampler 3 and a respiratory sampling nozzle 12;
the breath sampling nozzle 12 is arranged on the mouth of the human body to receive the breath of the human body and discharge the breath to the expired gas sampler 3;
the expired gas sampler 3 receives the human breath from the breath sampling nozzle 12, collects aerosol particles through electrodeposition and places the aerosol particles on a collection carrier. Expired gas sample thief 3 can effectively gather the aerosol particulate matter of expiration, and the front end has set up a breathing sampling mouth 12, still contains basic functions such as drying and filtration.
The optical detection component comprises a laser 4, a convex lens 5, a fluorescent filter 6, a spectrum shooting instrument 1 and an optical trap 8; the laser 4 emits laser light, which is ultraviolet light. The laser irradiates on the aerosol particles on the collecting carrier to excite fluorescence, the light beam transmitted through the aerosol particles is absorbed by the light trap 8, and the fluorescence excited on the aerosol particles is detected and received by the spectral shooting instrument 1 after passing through the convex lens 5 and the fluorescence filter 6.
The spectrum shooting instrument 1 has the function of shooting the fluorescence spectrum in the collection cavity, has certain requirements on sensitivity, signal-to-noise ratio and response time, and can finish spectrum collection within the continuous irradiation treatment discontinuous time. The laser 4 acts as a fluorescence excitation light source and the light beam is directed through a mirror 13 onto the exhaled aerosol particles in the collector plate 11.
In specific implementation, the optical detection assembly may further include a reflector 13, and laser light emitted by the laser 4 is reflected by the reflector 13 and then irradiates the aerosol particles on the collecting carrier.
In the specific implementation, two convex lenses 5 are arranged between the aerosol particles and the spectrum uptake instrument 1, and a fluorescence filter 6 is arranged between the two convex lenses 5.
In specific implementation, the convex lens 5 and the fluorescence filter 6 are both arranged in the spectrum acquisition cavity 2, and the laser 4, the reflector 13, the spectrum shooting instrument 1 and the optical trap 8 are all arranged outside the spectrum acquisition cavity 2.
In the optical detection assembly, light beams irradiated onto aerosol particles are kept continuous, and detection and reception of fluorescence generated by excitation of microorganisms on the aerosol particles are discontinuous; while the irradiation of the aerosol particles by the radiation source 7 is discontinued.
The detection receiving of fluorescence generated by the excitation of microorganisms on the aerosol particles in the optical detection assembly and the irradiation of the aerosol particles by the radiation source 7 are asynchronous in time sequence, and the time action is staggered, so that the optical detection assembly does not detect and receive the fluorescence when the radiation source 7 irradiates the aerosol particles, and the radiation source 7 does not irradiate the aerosol particles when the optical detection assembly detects and receives the fluorescence.
The collection carrier comprises in particular a collection plate 11 as a carrier for the exhaled aerosol particles.
The collecting carrier is also provided with a temperature control device 9 and an ultrasonic device 10;
the temperature control device 9 is arranged on the bottom surface of the collecting plate 11 for collecting the carriers and is used for controlling the temperature of the collecting plate 11 for collecting the carriers and the aerosol particles on the collecting plate to ensure that the temperature of the particles is constant and the temperature quenching of fluorescence is prevented;
the temperature control device 9 is capable of heating and cooling, keeping the temperature on the collecting plate 11 of the collecting carrier constant.
The ultrasonic device 10 is arranged on the bottom surface of the collecting plate 11 for collecting the carrier, and is used for generating ultrasonic vibration to remove aerosol particles on the collecting plate 11 for collecting the carrier after identification and detection, so that the function of removing the aerosol particles is achieved.
The spectrum collection cavity 2 comprises main functional components of a convex lens 5, a fluorescent filter 6, a radiation source 7, an optical trap 8, a temperature control device 9, an ultrasonic device 10 and a collection plate 11, wherein the convex lens 5, the fluorescent filter 6 and the optical trap 8 are optical path components, the radiation source 7 has a function of emitting specific radiation to exhaled aerosol particles, the temperature control device 9 is used for maintaining the temperature of the collection plate 11 to be constant, and the ultrasonic device 10 is used for transmitting ultrasonic waves to the collection plate 11 and the exhaled aerosol and has the functions of auxiliary treatment and cleaning.
The type of irradiation emitted by the radiation source 7 comprises one or more of microwaves, ultrasound, x-rays. The radiation source is capable of continuous irradiation in one or more irradiation modes.
The irradiation parameters of the radiation source 7 include resolution, maximum treatment intensity, and integration time; irradiation can be controlled by setting irradiation parameters.
The settings of the irradiation parameters are stored and implemented by the controller or switch of the radiation source 7.
Laser emitted by the laser 4 is reflected by aerosol particles to form intermediate reflected light, the intermediate reflected light and excited fluorescence with different wavelengths transmit through the convex lens 5, but the laser emitted by the laser 4 is filtered through the fluorescence filter 6, and only the excited fluorescence with different wavelengths is remained to be incident into the spectrum shooting instrument 1.
The exhaled gas sampler 3 is provided with two electrodes with positive and negative polarities respectively, the electrode with one polarity is connected with external voltage, and the electrode with the other polarity is positioned on the bottom surface of the collecting carrier.
In particular below the quartz collection plate 24.
The processing procedure of the embodiment of the invention is as follows:
step S1: human body exhaled breath sol is deposited on a collecting plate 11 with irradiation and temperature control functions, the collecting plate 11 is a carrier for bearing exhaled breath sol particles, and a temperature control device 9 is arranged on the collecting plate to eliminate temperature quenching caused by continuous irradiation; the particles on the aerosol will deposit on the collecting plate 11 to form aerosol particles, which will adhere with microorganisms.
Step S2: the kind of the irradiation treatment, the kind of the embodiment was determined as the microwave,
the irradiation parameters were set as: the maximum treatment intensity is 100mW/m 2 The resolution was 1/s and the total integration time was 10 s.
Step S3: opening the temperature control device 9, setting a target temperature T to be 4 ℃, keeping the temperature on the collecting plate 11 constant, adjusting and controlling the temperature of the collecting plate 11 to be constant at T through the temperature control device, and enabling the error to be as small as possible and not more than 0.1 ℃;
step S4: according to the arrangement, the aerosol particles on the collecting plate 11 are continuously irradiated by the radiation source 7, short intermittent irradiation is set according to the resolution, the maximum processing intensity and the integration time, the laser 4 in the optical detection assembly emits ultraviolet light to irradiate the aerosol particles, the fluorescence spectrum of the whole process of pathogen surface protein conformation irradiation of the microorganisms is rapidly scanned and detected by the spectrum ingestion instrument 1, and the three-dimensional fluorescence spectrum is obtained, wherein the fluorescence intensity is a function of the wavelength and the continuous irradiation intensity.
The irradiation time and the scanning detection time of the spectrum taking device 1 are staggered and do not overlap in time. Usually intermittent irradiation, and scanning detection by the optical spectrum taking device 1 is carried out in the period between two adjacent irradiations.
Step S5: drawing a three-dimensional fluorescence surface according to the obtained three-dimensional fluorescence spectrum, analyzing the three-dimensional fluorescence surface, extracting a contour line characteristic spectrum as a fluorescence fingerprint, and performing characteristic extraction and modeling;
the embodied models are support vector classification and support vector regression models, which are used to identify the microbial species and microbial content, respectively.
Step S6: and carrying out inversion processing on the extracted features according to the established model to obtain the types and the contents of the microorganisms in the aerosol exhaled by the human body.
And for the types of the microorganisms, extracting peak positions, quantity, gradient and volume characteristics from the contour line characteristic spectrum, and further processing by a support vector classification model to obtain the types of the microorganisms.
And for the content of the microorganisms, the content of the microorganisms is obtained by processing through a support vector regression model through peak height, height difference, gradient and volume characteristics extracted from the contour line characteristic spectrum.
Fig. 3 and 4 are schematic structural views of an embodiment of the device according to the present invention. As shown in fig. 3 and 4, the pipeline assembly mainly includes a sampling filtering link 14, a composite sampling link 15, a spectrum collecting cavity link 16, and a bundling air outlet link 17, which are connected in sequence, and importantly, the spectrum collecting cavity link 16 includes a spectrum incident optical fiber 18, a spectrum emergent optical fiber 19, a radiation source 20, and an aerosol deposition assembly 21 including ultrasound and temperature control, the aerosol deposition assembly 21 includes a quartz collecting plate 24, a temperature controller 25, and an ultrasound device 26, and the composite sampling link 15 includes a collision cutter link 22 and a corona discharge link 23.
The combined sampling link 15 comprises a collision cutter link 22 and a corona discharge link 23, the aerosol enters the combined sampling link 15 from the sampling and filtering link 14, particles with specific diameters in the aerosol are intercepted through the collision cutter link 22, the corona discharge link 23 is provided with a negative plate connected with external voltage, and the negative plate of the corona discharge link 23 is processed and then charged with negative electricity, so that the aerosol flows to the spectrum acquisition cavity link 16.
A quartz collecting plate 24 is arranged in a pipeline of the spectrum collecting cavity link 16, aerosol particles are deposited on the quartz collecting plate 24, a negative plate is arranged beside the quartz collecting plate 24, and the negatively charged aerosol is electrodeposited and falls onto the quartz collecting plate 24 when being close to a positive plate of the quartz collecting plate 24.
The radiation source 20 is inserted on the pipeline of the spectrum acquisition cavity link 16 and extends into the inner cavity of the pipeline, the spectrum incident optical fiber 18 of the laser 4 and the spectrum emergent optical fiber 19 of the receiving end of the spectrum ingesting instrument 1 are also inserted on the pipeline of the spectrum acquisition cavity link 16 and extend into the inner cavity of the pipeline, the spectrum incident optical fiber 18 and the spectrum emergent optical fiber 19 are arranged vertically and non-coaxially, the spectrum incident optical fiber 18 is used for emitting laser to irradiate aerosol particles, and the spectrum emergent optical fiber 19 is used for detecting and receiving fluorescence which penetrates through the aerosol particles.
A temperature controller 25 and an ultrasonic device 26 are arranged below the bottom surface of the quartz collecting plate 24, and finally three-dimensional fluorescence of aerosol particles under continuous irradiation is detected and received through the spectrum outgoing optical fiber 19 under the combined action of the radiation source 20, the temperature controller 25 and other components. After the detection is finished, the ultrasonic device 26 is turned on to drive the quartz collecting plate 24 to vibrate to remove aerosol particles, and the next detection is carried out.
FIGS. 5 and 6 are results of one example of continuous irradiation of three-dimensional fluorescence by exhaled breath sol microorganisms.
Contour line characteristic spectrograms of the exhaled aerosol microorganism continuously irradiated three-dimensional fluorescence and the exhaled aerosol microorganism continuously irradiated three-dimensional fluorescence are respectively shown in fig. 5 and fig. 6.
As can be seen in FIG. 5, the continuous irradiation three-dimensional fluorescence adds a new dimension of continuous microwave irradiation treatment on the basis of the two-dimensional fluorescence spectrum, and the initial fluorescence spectrum has an obvious peak near 360nm, which gradually decreases with the microwave treatment and does not change at 4 treatment intensity units. After 6 units of treatment intensity, a peak around 790nm appears, which is a signal for non-microbial impurities in the exhaled breath sol.
As can be seen in FIG. 6, the continuous irradiation of three-dimensional fluorescence can be used to obtain clear fingerprint by extracting contour characteristic spectrum. The peak position, the number, the gradient and the volume as well as the elevation and the height difference of the peak are easy to extract. The peaks corresponding to fig. 5 have distinct peaks around 360nm and 790nm, respectively, and clearly record the change process of the peak and the valley.
In the above example, the types and contents of the microorganisms are obtained by extracting the peak positions, the number, the gradient, the volume and the elevations and the height differences of the peaks on the continuously irradiated three-dimensional fluorescence surface and processing the three-dimensional fluorescence surface through the support vector classification and the support vector regression model, and the microorganisms are identified from the exhaled aerosol as severe acute respiratory syndrome coronavirus with the type of the microorganisms being 10 5 virus/m 3 。
The results show that the peak position, the number, the gradient and the volume on the continuous irradiation three-dimensional fluorescence surface, the elevation and the height difference of the peaks and the like contain a great amount of information of microorganisms in the exhaled aerosol, and the method becomes a novel method for identifying the microorganisms in the exhaled aerosol.
In the foregoing embodiments, the descriptions of the embodiments have respective emphasis, and for parts that are not described in detail in a certain embodiment, reference may be made to the related descriptions of other embodiments.
While the invention has been described with reference to an apparatus for detecting exhaled breath sol microorganisms by continuous irradiation of three-dimensional fluorescence, it will be understood by those skilled in the art that various changes in the embodiments and applications of the apparatus can be made without departing from the spirit and scope of the invention.

Claims (10)

1. The utility model provides a quick detection device of exhalation aerosol microorganism which characterized in that:
comprises a spectrum collection cavity (2);
comprises a collecting carrier, which is arranged in a spectrum collecting cavity (2) and carries aerosol particles obtained by the exhalation of a human body;
the device comprises a light detection assembly, a light source and a control assembly, wherein the light detection assembly generates light beams to irradiate aerosol particles, and detects fluorescence generated by excitation of microorganisms on the aerosol particles to obtain a two-dimensional fluorescence spectrum;
the device comprises a radiation source (7) which is arranged in a spectrum collection cavity (2) and faces to aerosol particles on a collection carrier, wherein the radiation source (7) irradiates the aerosol particles to influence fluorescence generated by excitation of microorganisms on the aerosol particles, so that a detected two-dimensional fluorescence spectrum is changed into a three-dimensional fluorescence spectrum.
2. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the aerosol particles are obtained by sampling from human breath through a sampling assembly, and the sampling assembly comprises an expired gas sampler (3) and a breath sampling nozzle (12);
the breath sampling nozzle (12) is arranged on the human mouth to receive the breath of the human body and discharge the breath to the expired gas sampler (3);
the expired gas sampler (3) receives the human breath from the breath sampling nozzle (12), forms aerosol particles through electrodeposition treatment and is arranged on the collecting carrier.
3. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the optical detection component comprises a laser (4), a convex lens (5), a fluorescence filter (6), a spectrum shooting instrument (1) and an optical trap (8); the laser (4) emits laser, the laser irradiates aerosol particles on the collecting carrier to excite fluorescence, light beams transmitted through the aerosol particles are absorbed by the light trap (8), and the fluorescence excited on the aerosol particles is detected and received by the spectrum shooting instrument (1) after passing through the convex lens (5) and the fluorescence filter (6).
4. The exhaled aerosol microorganism rapid detection apparatus according to claim 3, characterized in that: the convex lens (5) and the fluorescence filter (6) are arranged in the spectrum acquisition cavity (2), and the laser (4), the reflector (13), the spectrum shooting instrument (1) and the light trap (8) are arranged outside the spectrum acquisition cavity (2).
5. The device for rapidly detecting microorganisms in exhaled aerosols according to claim 1 or 3, wherein: in the optical detection assembly, light beams irradiated onto aerosol particles are kept continuous, and detection and reception of fluorescence generated by excitation of microorganisms on the aerosol particles are discontinuous; meanwhile, the radiation source (7) does not continuously irradiate the aerosol particles.
6. The exhaled breath sol microorganism rapid detection apparatus according to claim 5, wherein: the detection receiving of the optical detection component for exciting the microorganisms on the aerosol particles to generate fluorescence and the irradiation of the radiation source (7) on the aerosol particles are asynchronous in time sequence.
7. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the collecting carrier specifically comprises a collecting plate (11) which is used as a carrier for bearing exhaled aerosol particles.
8. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the collecting carrier is also provided with a temperature control device (9) and an ultrasonic device (10);
the temperature control device (9) is arranged on the bottom surface of the collecting carrier and is used for controlling the temperature of the collecting carrier and aerosol particles on the collecting carrier;
and the ultrasonic device (10) is arranged on the bottom surface of the collecting carrier and is used for generating ultrasonic vibration to remove aerosol particles on the collecting carrier after identification and detection.
9. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the irradiation type emitted by the radiation source (7) comprises one or more of microwave, ultrasound and (x) ray.
10. The exhaled aerosol microorganism rapid detection apparatus according to claim 1, characterized in that: the irradiation parameters of the radiation source (7) comprise resolution, maximum treatment intensity and integration time.
CN202210699079.6A 2022-06-20 2022-06-20 Exhaled aerosol microorganism rapid detection device Pending CN115015203A (en)

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