CN112945934A - Rapid screening system, device and method for field epidemic prevention detection - Google Patents
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Abstract
The invention relates to a rapid screening system, a rapid screening device and a rapid screening method for field epidemic prevention detection. The invention directly detects viruses or other pathogens related to a respiratory system by using a nano material or a substrate material with a specific configuration and a surface enhanced Raman scattering technology, and can achieve higher accuracy and sensitivity by combining with a corresponding classification recognition algorithm, distinguish different pathogens and have ultrahigh detection sensitivity. The method can feed back the screening result in real time, and effectively promotes the sharing of epidemic prevention information and the control of suspected cases. The device provided by the invention is simple and convenient to operate for the detection personnel and use for the detected personnel; the invention can transform the screened target virus through the establishment of the database. The invention is suitable for rapidly screening novel coronavirus carriers on site, and can also play a role in other epidemic prevention works appearing in the future.
Description
Technical Field
The invention relates to the technical field of Raman spectrum detection and pathogen screening, in particular to a rapid screening method based on a surface enhanced Raman scattering principle and applicable to field epidemic prevention detection.
Background
The novel coronaviruses are very infectious and spread mainly by droplets and contact. Pneumonia often occurs in infected persons, and acute respiratory distress syndrome with severe respiratory complications, even leading to death, occurs in a small number. Early screening diagnosis and timely quarantine control are still the key to prevent further spread of epidemic. One problem faced in epidemic prevention work is that a part of infected persons are asymptomatic infected persons, and the part of people have no obvious clinical manifestations such as fever, weakness and the like, but are positive when virus nucleic acid detection is carried out, so that the virus is proved to be infected with the novel coronavirus, and has strong infectivity, which adds a great burden to the control of epidemic situation. Therefore, how to provide a rapid field detection and epidemic prevention method with low cost and high accuracy, which is suitable for different virus samples, is one of the problems to be solved urgently under the current global severe epidemic situation.
Through a specially prepared metal micro-nano structure, when the metal micro-nano structure is excited by proper light, an electromagnetic field on the surface of the metal micro-nano structure is enhanced, so that a Raman signal generated by molecules adsorbed nearby the metal micro-nano structure is greatly enhanced, namely Surface Enhanced Raman Scattering (SERS). SERS overcomes the defect of low sensitivity of Raman spectrum, and can obtain structural information which is not easy to obtain by conventional Raman spectrum. Although the direct detection of viruses related to respiratory systems by introducing noble metal nano materials or substrate materials with specific configurations and utilizing the SERS technology has been reported, the detection still depends on expensive Raman equipment and complex laboratory steps, and is not suitable for epidemic prevention work needing rapid field detection.
Disclosure of Invention
The invention aims to provide a rapid detection system and a screening method based on a surface enhanced Raman scattering principle and capable of being used for field epidemic prevention detection.
The screening method provided by the invention can realize in-situ detection on the nano chip, the Raman signal after sample excitation is rapidly collected, and the artificial intelligence algorithm model is compared with the existing database to perform accurate and rapid judgment, so that the method is suitable for on-site rapid screening of respiratory tract related pathogens.
In a first aspect, the present invention provides a rapid screening system, comprising:
the nano chip is used for enriching a sample to be detected and enhancing an optical signal of the sample to be detected;
the optical detector is used for detecting and outputting the optical signal of the sample to be detected;
the optical signal intelligent algorithm processing module is used for identifying the types and the quantity of pathogens contained in the sample to be detected based on the optical signal output by the optical detector;
and the cloud management platform is used for receiving, storing and sharing the real-time data acquired by rapidly screening the site, and assisting epidemic prevention management work.
In the rapid screening system, the optical signal intelligent algorithm processing module comprises an optical signal correction module, a to-be-tested sample Raman signal library and a cloud uploading module.
In the rapid screening system of the present invention, the optical detector uses an ultraviolet absorption spectroscopy technology, a visible spectroscopy technology, an infrared absorption spectroscopy technology, a surface plasmon resonance technology, or a raman spectroscopy technology.
Specifically, in the rapid screening system of the present invention, the optical detector may be a portable optical instrument with rapid response, the obtained optical signal data has certain fingerprint information, and can distinguish biochemical molecules, germs and other microorganisms without labels, the optical detection technology mainly includes a raman spectroscopy technology, and other technologies such as an ultraviolet, visible and infrared absorption spectroscopy technology, surface or local surface plasmon resonance and the like can be used instead of or in addition to the raman spectroscopy technology.
In the rapid screening system of the present invention, the nano chip is made of a nano material having a certain raman signal enhancement effect. The material is generally prepared from noble metal nano particles, rods and wires with a local surface plasma resonance phenomenon through self-assembly, micro-nano processing or in-situ chemical synthesis.
Preferably, the nano chip is formed by vertically arranging gold nanowires. The nano chip formed by the vertically arranged gold nano wires with the Raman enhancement effect can effectively improve the Raman signal intensity, and meanwhile, nano-level pores are formed among the nano wires to adsorb and enrich the detection object.
Specifically, in the rapid screening system of the present invention, taking a nano chip formed by vertically arranged gold nanowires as an example, the preparation method includes:
(1) preparing a gold seed particle dispersion by using a tetrachloroauric acid aqueous solution and a citrate aqueous solution as raw materials;
(2) preparing piranha solution by using sulfuric acid and hydrogen peroxide in a ratio of 3:1, heating by using a magnetic stirrer, soaking a silicon wafer, ultrasonically cleaning the silicon wafer by using deionized water, and drying by using nitrogen;
(3) adding 3-aminopropyl-3-methoxysilane into an absolute ethanol solution, adding a silicon wafer to perform amination, soaking, and drying with nitrogen;
(4) immersing the aminated silicon wafer into a gold seed solution, soaking, and drying with nitrogen to obtain a silicon wafer adsorbed by gold particles;
(5) sequentially adding absolute ethyl alcohol, deionized water and HAuCl4The (tetrachloroauric acid), 4-mercaptobenzoic acid ethanol solution and ascorbic acid water solution are fully and evenly blown by a liquid transfer gun, then silicon wafers are added, the silicon wafers are soaked to grow the nanowire array, finally the nanowire array is taken out by tweezers, and nitrogen is addedAnd (5) air-drying to obtain the gold nano chip.
In the rapid screening system of the present invention, the rapid screening system further comprises a carrier for carrying the nano chip.
In epidemic prevention detection, the carrier can be co-located with detection personnel for a long time, such as a disposable mask which is worn on the face of the detection personnel for a long time; the test carrier may also be in temporary contact with the subject person, such as a gas blower.
In the invention, the carrier is made of a macroporous material with good gas permeability and is used for filtering exhaled gas and simultaneously carrying the nano chip. Can be an existing epidemic prevention article such as a mask, and can also be formed by modifying an air blowing cylinder. The carrier can ensure that the nano chip directly faces the mouth and the nose of a person to be detected, so that the nano chip can adsorb potential germs carried in the exhaled air of the person to be detected as much as possible.
As a specific embodiment of the invention, a mask is used as a carrier in field epidemic prevention screening. When the mask is worn, the melt-blown fabric filter screen in the mask can intercept and capture possible germs to play an enrichment role. And the cost of the disposable mask is very low, and the disposable mask can be frequently replaced and is convenient to wear. Therefore, the mask is very suitable for being used as a detection carrier of a nano chip to carry out rapid detection on the novel coronavirus.
Specifically, the rapid screening system comprises a nano chip for enriching a detection object and enhancing a detection signal; a carrier for carrying the nano-chip and filtering exhaled air; raman detection technology for rapid detection on a nano-chip; and the optical signal intelligent algorithm processing module is used for judging the exhaled gas information of the detected person and the cloud management platform.
In a second aspect, the present invention provides an apparatus comprising the above rapid screening system.
In a third aspect, the present invention provides a method for on-site rapid screening by using the rapid screening system or the apparatus, including:
detecting a Raman signal of the nano chip for enriching the expired gas of the examined person by using a Raman detector;
and normalizing, checking and denoising the Raman signal based on an optical signal intelligent algorithm processing module, comparing the Raman signal with the pathogen optical signal recorded in a database, outputting a pathogen identification result and uploading the pathogen identification result to a cloud for sharing. The rapid screening method for the on-site epidemic prevention detection provided by the invention can distinguish suspected pathogen carriers from normal persons, assist the epidemic prevention work and have no diagnosis confirming function.
The pathogen of the invention is respiratory tract related virus; the novel coronavirus and influenza virus are preferred.
Specifically, in field epidemic prevention, the flow when the rapid screening system or the device or the method is used for rapid screening is as follows: the exhaled air possibly carrying the germs to be detected is filtered, adhered and enriched on the nanometer chip by the examined person through normal breathing, coughing, sneezing and the like for a period of time.
The beneficial effects of the invention at least comprise:
(1) the rapid screening system provided by the invention can achieve higher accuracy and sensitivity, and can distinguish different pathogen types;
(2) the rapid screening system provided by the invention uses the advanced SERS nanometer chip material and an intelligent analysis algorithm, can feed back results in real time, and is suitable for on-site rapid detection and screening in various scenes;
(3) in the rapid screening system provided by the invention, detection data is acquired by the portable Raman detector in real time, whether pathogens exist or not and classification is instantly judged by intelligently comparing the existing database, and a result is fed back to field detection personnel. Meanwhile, data results are uploaded to a cloud epidemic prevention management system so as to be convenient for overall management and early warning;
(4) the rapid screening system provided by the invention integrates the collection of expired gas of a person to be inspected, the real-time acquisition of detection signals, the output of virus identification results and the real-time uploading of epidemic prevention information to a cloud platform based on the requirements of the prior art and on-site epidemic prevention, and can effectively promote the sharing of epidemic prevention information and the control of suspected cases.
Drawings
FIG. 1 is a structural diagram of the resulting nano-chip prepared in example 1.
FIG. 2 is a flowchart showing the use of a mask as a detection carrier in example 2.
Fig. 3 is a flowchart of the intelligent algorithm processing module in the field test in embodiment 2.
FIG. 4 is a schematic view showing the structure of the detecting carrier in example 3 using a gas blower.
FIG. 5 is a Raman spectrum of the novel coronavirus from different droplet samples of example 4.
FIG. 6 is a comparison of Raman spectra of the novel coronavirus and saliva detected in the sample of example 5.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
EXAMPLE 1 preparation of the Nanocells
The embodiment provides a preparation method of an SERS (gold nano chip), which comprises the following steps:
1) preparing a 16nm gold seed particle dispersion by using a tetrachloroauric acid aqueous solution (25mM) and a citrate aqueous solution as raw materials;
2) preparing piranha solution according to the ratio of 3:1 of sulfuric acid to hydrogen peroxide, heating the solution to 65 ℃ by using a magnetic stirrer, soaking a silicon wafer for 30min, then ultrasonically cleaning the silicon wafer for 15min by using deionized water, and drying the silicon wafer by using nitrogen;
3) a liquid transfer gun sucks 18.5 mu L of 3-aminopropyl-3-methoxysilane and adds the 3-aminopropyl-3-methoxysilane into 20mL of absolute ethyl alcohol solution, then silicon chips are added to carry out amination, and the silicon chips are soaked for 1h and dried by nitrogen;
4) immersing the aminated silicon wafer into 10mL of gold seed solution for 2h, and drying by nitrogen to obtain a silicon wafer adsorbed by gold particles;
5) preparing 4-mercaptobenzoic acid ethanol solution and ascorbic acid aqueous solution, weighing 0.0026g of 4-mercaptobenzoic acid powder, and adding into 1mL of absolute ethanol to prepare 4-mercaptobenzoic acid ethanol solution; weighing 0.0219g of ascorbic acid powder and adding the ascorbic acid powder into 2mL of deionized water;
6) 21.5mL of absolute ethanol were added to a 10cm petri dish in sequence3.45mL deionized water, 2.03mL HAuCl4And fully and uniformly blowing 1mL of prepared 4-mercaptobenzoic acid ethanol solution and 2mL of prepared ascorbic acid aqueous solution by using a liquid transfer gun, then adding a silicon wafer, soaking for 30min to grow a nanowire array, taking out the nanowire array by using a pair of tweezers, and drying by using nitrogen.
The nano chip prepared by the embodiment can be placed in a mask, as shown in fig. 1; the structure of the prepared nano chip in this example is shown in FIG. 1.
Example 2 use of masks for on-site epidemic prevention detection and screening
This example presents a method for using a mask for on-site epidemic prevention detection and screening.
Firstly, a specially prepared SERS nano chip is added to a mask filter membrane and distributed to a tested person for normal breathing for use for a plurality of hours, and virus particles or related substances possibly contained in exhaled air are adsorbed and enriched. Then, the mask is recovered at an epidemic situation monitoring point, the probe of the portable Raman device is used for carrying out on-site collection of Raman spectrum on the nano chip on the mask, analysis is carried out through an intelligent algorithm model, the mask is disinfected and sterilized after the analysis is finished, and the replaced mask is safely treated. The field test flow chart is shown in figure 2.
And feeding back results in time according to the output information of the intelligent algorithm processing module, and performing control on suspected cases in time and uploading the results to the cloud. The data uploading and the field epidemic prevention taking measures flow chart is shown in figure 3.
Example 3 insufflation cartridge for in situ epidemic detection and screening
This example utilizes a gas blower similar to an alcohol detector for on-site epidemic prevention testing and screening.
The head of the air blowing cylinder is required to be additionally provided with a filter element filled with porous fiber structure materials, the filter element is used as a carrier of the SERS nanometer chip and is used for attaching and enriching virus particles and the like possibly contained in exhaled air, and the structural schematic diagram of the air blowing cylinder is shown in figure 4. The other end of the air blowing cylinder is connected with a probe of a Raman testing device, so that Raman spectrum collection and result feedback can be carried out immediately after the exhaled air of the testee is collected. Meanwhile, the possibility of probe pollution is also avoided through the spatial isolation of the blowing cylinder. In actual detection, the optical signal intelligent algorithm module can further optimize according to specific conditions and collected test data.
Example 4 detection of novel coronavirus in different droplet samples
In this example, the nano-chip and the optical detector prepared in example 1 were used to obtain raman spectrograms of 118 novel coronavirus samples, which are shown in fig. 5, and fig. 5 shows the consistency of raman spectrums of the novel coronavirus samples in different droplet samples.
Different chemical molecules and biological macromolecules including viruses and the like have fingerprint Raman spectrograms, and different molecules can be qualitatively analyzed and distinguished; the nanometer chip has ultrahigh detection sensitivity, and the enhancement factor of the surface enhanced Raman scattering is 10 at most14-1015The detection of the virus can be realized; the novel coronavirus optical signals obtained by the virus screening system are stable and consistent, and the novel coronavirus Raman signals can be accurately obtained by the virus screening system.
EXAMPLE 5 saliva and novel coronavirus in samples
This example provides a comparison of raman spectra of saliva with a novel coronavirus.
In this example, the nano-chip prepared in example 1 was used to enrich a sample mixed with human saliva and a novel coronavirus; the enriched nano-chip is optically detected, and the obtained Raman spectrum is compared and shown in figure 6. Fig. 6 shows that the nano-chip provided by the present invention can accurately and respectively produce saliva and virus samples, and further confirms that the rapid screening system provided by the present invention can accurately determine whether the person to be examined is a virus carrier.
Example 6 on-site real-time screening of examinees
The present embodiments provide for the use of the rapid screening system for in-situ testing.
In this embodiment, after the examinee wears the mask containing the nano-chip for 5 to 240 minutes, the raman spectrum information of the particles enriched in the nano-chip is detected by using the portable raman spectrometer, and compared with fig. 6, it is determined whether the examinee is a carrier of the novel coronavirus.
After the Raman spectrum is collected, automatic comparison and analysis are carried out on analysis software carried by the instrument, the analysis software is fed back to detection personnel in real time for field prevention and control, and meanwhile, the detection result is uploaded to a cloud platform, so that uniform epidemic prevention management is facilitated.
EXAMPLE 7 device for on-site epidemic prevention screening
The present embodiment provides a device comprising the rapid screening system of the present invention. The device that this embodiment provided collects the expired gas of the person of examining in the short time, the droplet that coughs sneeze and go out etc. carries out raman spectrum's collection and comparison, accomplishes the high in the clouds and uploads and information sharing.
The device provided by the embodiment can collect one or more times of expired gas of a person to be inspected during field epidemic prevention, and enrich possible virus particles in the expired gas in the nano chip; the Raman detector detects the nano chip, the Raman signal of the expired gas of the detected person is processed in the intelligent algorithm module, whether the detected person is a carrier of certain viruses or not is displayed in real time, and the number of the viruses carried by the detected person is also displayed; and meanwhile, the detection result is reported to the cloud end for sharing through the intelligent algorithm module.
In specific implementation, in order to ensure the safety of the detection environment, the expired air is discharged after being filtered and sterilized by using a filter membrane after exhaled particles in the expired air of a detected person are collected.
The device provided by the invention can efficiently and accurately acquire the information whether the inspected person carries the virus or not in a short time, timely give the decision of passing or isolation, and can realize centralized management of epidemic prevention and information disclosure through the intelligent algorithm module.
Example 8 Rapid screening System for Multi-pathogen screening
The present embodiment provides an application of the rapid screening system to multi-pathogen screening.
The rapid screening system has flexibility, and can rapidly complete conversion only by sampling, establishing a database and optimizing an intelligent recognition algorithm at the initial stage of epidemic situation outbreak, thereby achieving the purpose of rapidly screening new epidemic situations.
When different pathogens are confronted, the rapid screening system provided by the invention does not need secondary investment, all hardware and software in the system do not need to be adjusted, and only the intelligent identification algorithm is updated. The rapid screening system can integrate intelligent algorithms for identifying different germs, thereby realizing the identification capability of multiple pathogens.
EXAMPLE 9 Rapid screening System for patient Condition monitoring
The rapid screening system provided by the invention can also be used for disease condition management of sick personnel, and the rapid screening system provided by the invention contains an intelligent recognition algorithm and a cloud management platform, can be applied to continuous reading dynamic monitoring of a single sample, can assist research and medical personnel in monitoring the dynamic disease condition of a patient, and can better know the characteristics and disease law of pathogens causing epidemic situations.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A rapid screening system, the system comprising:
the nano chip is used for enriching a sample to be detected and enhancing an optical signal of the sample to be detected;
the optical detector is used for detecting and outputting the optical signal of the sample to be detected;
the optical signal intelligent algorithm processing module is used for identifying the pathogen type and the number of the samples to be detected based on the optical signals output by the optical detector;
and the cloud management platform is used for receiving, storing and sharing the real-time data acquired by rapidly screening the site, and assisting epidemic prevention management work.
2. The rapid screening system according to claim 1, wherein the optical signal intelligent algorithm processing module comprises an optical signal correction module, an optical signal library of the sample to be tested, and a cloud uploading module.
3. The rapid screening system of claim 1, wherein the optical detector uses ultraviolet absorption spectroscopy, visible spectroscopy, infrared absorption spectroscopy, or surface plasmon resonance, raman spectroscopy;
preferably the optical detector uses raman spectroscopy.
4. The rapid screening system of claim 1, wherein the nanochip is comprised of a noble metal having a localized surface plasmon resonance phenomenon;
preferably, the nano chip is formed by vertically arranging gold nanowires.
5. The rapid screening system according to claim 4, wherein the preparation method of the nano chip with the gold nano wire vertically arranged structure comprises:
(1) preparing a gold seed particle dispersion by using a tetrachloroauric acid aqueous solution and a citrate aqueous solution as raw materials;
(2) preparing piranha solution by using sulfuric acid and hydrogen peroxide in a ratio of 3:1, heating by using a magnetic stirrer, soaking a silicon wafer, ultrasonically cleaning the silicon wafer by using deionized water, and drying by using nitrogen;
(3) adding 3-aminopropyl-3-methoxysilane into an absolute ethanol solution, adding a silicon wafer to perform amination, soaking, and drying with nitrogen;
(4) immersing the aminated silicon wafer into a gold seed solution, soaking, and drying with nitrogen to obtain a silicon wafer adsorbed by gold particles;
(5) and sequentially adding absolute ethyl alcohol, deionized water, tetrachloroauric acid, a 4-mercaptobenzoic acid ethanol solution and an ascorbic acid aqueous solution, fully and uniformly blowing by using a liquid transfer gun, adding a silicon wafer, soaking to grow a nanowire array, and drying by using nitrogen to obtain the nano chip.
6. The rapid screening system according to any one of claims 1 to 5, further comprising a carrier for carrying the nanochip.
7. The rapid screening system of claim 6, wherein the carrier is comprised of a gas permeable macroporous material;
preferably, the carrier is a mask or a gas blower.
8. The rapid screening system of claim 7, wherein the object being screened by the rapid screening system is a pathogen transmitted through the respiratory tract.
9. A device comprising the rapid screening system of any one of claims 1-8.
10. A method for real-time screening of pathogens of interest using the rapid screening system of any one of claims 1-8 or the device of claim 9, comprising:
detecting the nano chip enriched with samples such as expired gas and droplets of the detected person by using a Raman detector, and outputting a Raman signal;
and the Raman signal is normalized, verified and denoised based on an optical signal intelligent algorithm processing module, is compared with the pathogen Raman signal recorded in the database, and is output and uploaded with a cloud terminal for sharing.
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