Monkey pox virus instant detection method based on combination of resonance SERS probe and test strip

Abstract

The invention belongs to the technical field of analysis and testing, and particularly discloses a monkey pox virus instant detection method based on combination of a resonance SERS probe and a test strip, and the method comprises the following steps: preparation of an Au NPs (at) IR808 (at) Ab1 resonance SERS probe, preparation of a biotin antibody Ab2 (at) Bio, preparation of an SA test strip, SERS signal sensing of monkey pox virus protein, and SERS qualitative and quantitative analysis. The sensitivity is improved by solving the root problem of probe sensitivity, the reaction of a double-antibody sandwich method in a test strip system is easily interfered by a matrix and the relationship between the binding efficiency and the sensitivity is researched, and by means of the external force of a biotin-streptavidin system which is higher in reaction rate and binding efficiency, the sensitivity of the biotin-streptavidin system can be detected, so that the sensitivity of the biotin-streptavidin system can be detected, and the sensitivity of the biotin-streptavidin system is improved. The efficiency of capturing the antibody probe by the paper strip is enhanced, so that the sensitivity is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of analysis and testing, and specifically relates to a method for real-time detection of monkeypox virus based on a resonance SERS probe combined with a test strip. Background Art

[0002] On August 14, 2024, the World Health Organization announced that the monkeypox epidemic once again constituted a "Public Health Emergency of International Concern" (PHEIC), the highest level of alert issued by the organization for global public health emergencies. The severity of this epidemic lies in the rapid increase in monkeypox cases and the high risk of global spread. According to the Africa Centers for Disease Control and Prevention, since 2024, at least 13 countries in Africa have reported 17,541 cases of monkeypox and 517 deaths. Because the monkeypox virus (MPXV) is a zoonotic viral disease, this variant can be transmitted through a variety of routes, including sexual contact, such as contact with contaminated objects, and through mucous membranes and broken skin, droplets, etc. Even pregnant women may transmit it to the fetus through the placenta, birth canal or breastfeeding. In addition, monkeypox is a self-limiting disease, and severe cases may have complications such as secondary bacterial infection in the skin lesions, bronchopneumonia, encephalitis, corneal infection, sepsis, etc.

[0003] At present, there is no specific anti-monkeypox virus drug in China, and the main treatment is symptomatic support and treatment of complications. The recommended specimen type for hospital diagnosis of monkeypox is skin lesion material, including lesion exudate swabs, multiple lesion surfaces or lesion scabs. Medical confirmation of specimens of suspected cases basically adopts nucleic acid amplification test (NAAT), such as real-time or conventional polymerase chain reaction (PCR). These commonly used biological diagnostic methods rely on expensive large instruments, are prone to contamination, and are cumbersome and time-consuming. Therefore, the development of new biological diagnostic technologies with high sensitivity, low cost, portability and rapidity for on-site POCT rapid detection of monkeypox virus is very critical for the early screening of the virus and effective control of its spread.

[0004] As the most successfully developed POCT detection technology, test strips have been widely used in the early screening and diagnosis of various pandemic disease viruses based on their technical advantages in rapid and convenient on-site diagnosis. For example, in recent years, corresponding commercial colloidal gold test strips have been developed for POCT detection of pandemic diseases such as the new coronavirus, influenza A and B. However, this colorimetric test strip based on visual identification has low sensitivity and accuracy, making it difficult to achieve early diagnosis of viral infection and cannot be used as a "gold standard" method. Therefore, solving the problem of low sensitivity of test strips can, to a certain extent, promote the application and development of test strips, a minimalist POCT diagnostic technology, from an auxiliary technology to a gold standard technology.

[0005] As a highly sensitive analytical detection method, SERS technology has a lot of work combining SERS technology with test strips to reduce the identification sensitivity of test strips due to its special advantages such as fingerprint spectrum, high accuracy, high sensitivity, multi-target, resistance to photobleaching and photodegradation. For example, Ren et al. reported a highly sensitive and accurate test strip based on Au-NPs antibody synthesis for the detection of cardiac troponin I, a major biomarker for acute myocardial infarction. The detection limit is as low as 10pg / mL, which is almost equivalent to the detection sensitivity of PCR. Choi et al. achieved quantitative detection of thyroid stimulating hormone (TSH) by using a highly sensitive SERS signal probe. Both hyperthyroidism and hypothyroidism can be diagnosed. Therefore, the technology of SERS combined with test strips combines the advantages of simple, fast, and low-cost POCT of test strips with the ultra-sensitive advantages of SERS probes, making it expected to develop into a new type of accurate, fast, and reliable POCT diagnostic technology. The key to this technology is to give full play to the high sensitivity of SERS biological probes and to ensure the high affinity of antigen antibodies in the test strip system.

[0006] The SERS bioprobe used in test strips is generally an antibody gold assembled with Raman signal molecules, and its SERS performance is the key to improving sensitivity. In addition to the enhanced performance of gold nanoparticles, the main reason affecting the SERS performance of the probe is the particularity of the Raman signal molecule. For example, whether the scattering interface of the Raman signal molecule is large enough to ensure a superior SERS signal and whether it can match the resonance with the excitation light wavelength to further amplify its SERS signal. A large number of studies are dedicated to overcoming the defects of existing SERS biosensors by solving the root problem of probe sensitivity. In addition, since the technology of SERS combined with test strips for POCT diagnosis uses the "double antibody sandwich method" to react in the test strip system, the sensitivity is easily affected by factors such as binding efficiency, reaction system and matrix interference, especially in complex systems such as throat swabs. Summary of the invention

[0007] The purpose of the present invention is to overcome the defects of the prior art and provide a method for instant detection of monkeypox virus based on a resonant SERS probe combined with a test strip, which improves the sensitivity by solving the root problem of probe sensitivity, and through studying the susceptibility of the "double antibody sandwich method" to matrix interference in the test strip system, and the relationship between binding efficiency and sensitivity, by using the external force of the biotin-streptavidin system with faster reaction rate and stronger binding efficiency, the efficiency of the paper strip in capturing the antibody probe is enhanced, thereby improving the sensitivity.

[0008] In order to achieve the above objectives, the technical solution of the present invention is: a method for instant detection of monkeypox virus based on a resonant SERS probe combined with a test strip, specifically comprising the following steps:

[0009] (1) Preparation of Au NPs@IR808@Ab1 resonant SERS probe: The Raman signal molecule IR808, which has both resonance and antibody binding functions, is modified on the surface of gold nanoparticles to form AuNPs@IR808. The carboxyl group of the IR808 molecule is activated to link with the antibody Ab1 to form the resonant SERS biosensor probe AuNPs@IR808@Ab1.

[0010] (2) Preparation of biotin antibody Ab2@Bio: Assemble biotin on antibody Ab2 and prepare biotin antibody Ab2@Bio by dialysis in PBS buffer;

[0011] (3) Preparation of streptavidin (SA) test strips: Glue a nitrocellulose membrane (NC membrane) onto a substrate, and then modify streptavidin (SA) onto the NC membrane using a gold spraying film streaking instrument to form a detection line T line. Then, assemble a sample pad, a conjugate pad, and absorbent paper at the other end of the NC membrane in sequence, and then cut the strips to prepare SA test strips with SA as the T line.

[0012] (4) SERS signal sensing of monkeypox virus protein: The Au NPs@IR808@Ab1 resonant SERS probe prepared in step (1), the biotin antibody Ab2@Bio prepared in step (2) and the monkeypox virus protein (MXPV-protein) sample were mixed in proportion and then dropped onto the sample pad of the SA-test strip prepared in step (3). After 10-15 minutes, the SERS signal of the T line of the test strip was collected using a portable Raman spectrometer with a 785-strip closed spot;

[0013] (5) SERS qualitative and quantitative analysis: Fit the curve of the characteristic peak area in the SERS spectrum as it changes with the monkeypox virus protein concentration, and identify the positive and negative nature of the sample to be tested and quantify the virus concentration of the positive sample based on the characteristic peak area.

[0014] In a preferred embodiment of the present invention, the gold nanoparticles in step (1) are prepared by sodium citrate reduction method, and the particle size of the gold nanoparticles is 30-60 nm.

[0015] In a preferred embodiment of the present invention, the Raman signal molecule in step (1) is an IR808 molecule which carries a carboxyl group and can bind to an antibody and resonate with a 785 laser.

[0016] In a preferred embodiment of the present invention, the concentration of the Raman signal molecule IR808 in step (1) is 0.3-0.7 mM, and the added amount is 4-8 uL / mL.

[0017] In a preferred embodiment of the present invention, the antibody in step (1) and the antibody in step (2) are two antibodies that recognize different antigen surface determinants, and the two antibodies can be mixed in any proportion.

[0018] In a preferred embodiment of the present invention, the biotin antibody Ab2@Bio in step (2) is prepared by mixing the antibody Ab2 and biotin Bio, placing the mixture in a dialysis bag and dialyzing it in a 1.5-2.5 mM PBS buffer for 16-24 hours.

[0019] In a preferred embodiment of the present invention, the SA test strip in step (3) is designed to be 7-8 cm long, 3.5-4.5 mm wide, with a SA concentration of 1-2 mg / mL and a T line width of 1.5-2.5 mm.

[0020] In a preferred embodiment of the present invention, in step (3), the sample pad is pre-modified with a hydrophobic surfactant, and the composition of the surfactant is 0.5 wt % S9 or 0.01-0.05 wt % TW-20.

[0021] In a preferred embodiment of the present invention, in step (4), the volume ratio of Au NPs@IR808@Ab1 probe, Ab2@Bio and monkeypox virus protein (MXPV-protein) sample is (3-5):1:(70-80), the mixing time is 10-60s, and the test is performed after the test strip flows for 12-16min.

[0022] In a preferred embodiment of the present invention, when the portable Raman spectrometer with 785 strip-shaped spots collects the SERS signal of the T line of the test paper in step (4), the width of the laser spot is adjusted to match the width of the T line.

[0023] In a preferred embodiment of the present invention, when the SERS signal of the T line of the test paper strip is collected by a portable Raman spectrometer with 785 strip-shaped light spots in step (4), the collection time is 100 ms and the power is 100 mw.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. The present invention The detection system for monkeypox virus protein of the present invention is composed of a resonant SERS probe modified with a bifunctional signal molecule IR808 and a large number of antibodies, a biotin antibody, and a SA test strip for capturing biotin. The sensitivity and stability of the probe ensure the reliability of the detection.

[0026] 2. The present invention uses a portable Raman instrument with a strip-shaped light spot for detection. The width of the strip-shaped light spot just matches the width of the T line of the test paper, and the detection improves the stability of the detection result.

[0027] 3. The SA test strip of the present invention, after the sample pad is treated with a hydrophobic surfactant, effectively suppresses the false positive problem caused by particle retention in the SERS test strip system, thereby greatly improving the detection sensitivity in the throat swab system.

[0028] 4. The SA test strip of the present invention is detected by capturing the SERS probe through the Bio-capturing antibody. Based on the principle that one SA can bind to four Bios, the Bio-SA system is used to greatly improve the reaction efficiency of the antigen-antibody "double antibody sandwich method", allowing the test strip to quickly and sensitively capture the probe in the presence of viral antigen protein.

[0029] 5. The detection method of the present invention has been verified by a large number of throat swab system samples from different individuals. The detection time using a portable Raman instrument usually does not exceed 15 minutes, which can meet the needs of high-sensitivity real-time detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 It is a schematic diagram of the preparation and detection of the SA test strip of the present invention;

[0031] Figure 2 It is a schematic diagram of the mixture of the MXPV sample and Au NPs@IR808@Ab1 and Ab2@Bio in Example 1, which is captured to the T line by the SA test strip and then detected by a portable Raman spectrometer matching the T line;

[0032] Figure 3 is the UV absorption spectra of Au NPs, AuNPs@IR808 and Au NPs@IR808@Ab1 in Example 1;

[0033] Figure 4 is the SERS spectra corresponding to the MXPV proteins at different concentration gradients in the standard solution in Example 1;

[0034] Figure 5 is the curve of the change of SERS characteristic peak area in the throat swab system with the concentration of MXPV protein in Example 2;

[0035] Figure 6 These are the results of MXPV viral protein detection in throat swabs from 30 different people in Example 3. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is described in more detail below with reference to the accompanying drawings and specific embodiments, but the protection scope of the present invention is not limited to these embodiments.

[0037] The synthesis method of AuNPs gold nanoparticles in the following examples is as follows: Prepared by sodium citrate reduction method, first measure 200mL ultrapure water and place it in a 250mL single-mouth original bottom flask. Place the flask on a magnetic stirring heater, inject 2.4mL 0.825wt% HAuCl4 solution, heat the solution to boiling (about 20 minutes), then inject 1.5mL 1wt% trisodium citrate (CA) solution, and continue boiling for 30 minutes. After stopping the reaction, naturally cool to room temperature.

[0038] Example 1

[0039] This example uses a standard buffer solution that does not contain MXPV, and adds different concentrations of MPXV viral protein to make its final concentration 1pg / mL-100pg / mL. Mix AuNPs@IR808@Ab1, Ab2@Bio and virus solutions of different concentrations in a volume ratio of 4:1:75. Place the test strip horizontally in the card slot, take 75μL of the mixed liquid and drop it on the sample pad of the test strip, and after 13 minutes, use the designed 785 strip portable Raman spectrometer for testing. During the detection, the strip light spot is aligned with the T line, and the test is performed under the conditions of 100ms and 100mW. The specific detection steps are as follows:

[0040] (1) Preparation of Au NPs@IR808@Ab1 resonant SERS probe: 8 mL of gold colloid solution with a particle size of 55 nm was placed in a glass sample tube and stirred on a magnetic stirrer. When the colloidal particles rotated evenly at high speed and formed a vortex, 32 uL of 0.5 mM Raman signal molecule IR808 solution was quickly added. After stirring for 5 minutes, the molecule modification was completed to form an Au@IR808 system. Then, the Au-IR808 added to the PBS (pH 7.4) buffer system was activated with activators EDC and NHS at concentrations of 5 mM for 30 minutes. Finally, the antibody Ab1 was added to the activated Au NPs@IR8088 system at a concentration of 20 μg / mL. After Au NPs@IR808 modified antibody Ab1, it was incubated overnight in a refrigerator at 4°C with low speed rotation. After centrifugation, the excess Ab1 antibody was washed away with PBS buffer to obtain Au NPs@IR808@Ab1. Au NPs@IR808@Ab1 was blocked with PBS blocking solution containing 0.05wt% BSA for 1 hour. After blocking, the blocking solution was removed by centrifugation, and finally Au NPs@IR808@Ab1 was stored for a long time with the blocking solution. Figure 3 UV-visible spectra of AuNPs, Au NPs@IR808, and Au NPs@IR808@Ab1.

[0041] (2) Preparation of biotin antibody Ab2@Bio: Add 0.1μM biotin to 500μL of antibody Ab2 at a concentration of 1mg / mL and incubate at low speed for 2h. Then transfer the liquid to a dialysis membrane and dialyze in 2mM PBS. The dialyzed PBS solution needs to be changed once at 16h and 18h, and then the liquid is transferred to a centrifuge tube to prepare Ab2@Bio. Finally, add an equal volume of glycerol to Ab2@Bio and store it in a refrigerator at -20℃.

[0042] (3) Preparation of streptavidin (SA) test strips: After pasting a 2 cm long CN140 nitrocellulose membrane on the purchased PVC board base, a 0.5 mg / mL streptavidin SA was sprayed on the CN140 nitrocellulose membrane using a test strip gold spraying film meter to form a detection line T line, and then dried in a constant temperature and humidity oven at a temperature of 45°C and a humidity of 20% for 2 h. Then, a 1.5 cm long SB09 sample pad (previously modified with a hydrophobic surfactant of 0.5 wt% S9), a 1 cm long binding pad, and a 3 cm long absorbent pad were pasted in sequence on the other end of the CN140 nitrocellulose membrane according to the standard test strip process. Finally, a strip cutter was used to cut the SA test strips of streptavidin (SA) with a length of 7 cm, a width of 3.9 mm, and a T line width of 2 mm. The preparation and detection process of the SA test strips are as follows: Figure 1 shown.

[0043] (4) SERS detection of monkeypox virus protein: In a standard buffer solution without MXPV, add MPXV virus protein of different concentrations to make the final concentration of 1pg / mL-100pg / mL. Mix Au NPs@IR808@Ab1, Ab2@Bio and monkeypox virus protein samples of different concentrations in a volume ratio of 4:1:75 for 10s to obtain a mixed liquid; at this time, based on the antigen-antibody binding reaction, an Au NPs@IR808@Ab1-MXPV-Ab2@Bio structure will be formed in the above mixed liquid; place the test strip horizontally in the card slot, take 75μL of the above mixed liquid and drop it on the sample pad of the test strip. When the mixed liquid flows through the T line of the test strip, the streptavidin (SA) on the T line will bind Au to the sample pad through the efficient binding advantage of SA and Bio. NPs@IR808@Ab1-MXPV-Ab2@Bio was firmly fixed on the T line of the test strip to form a double antibody sandwich structure based on the SA-Bio system. After the liquid flowed in the test strip for 13 minutes, it was tested with the designed 785 strip portable Raman spectrometer. During the detection, the strip spot was aligned with the T line, the acquisition time was 100ms, the laser power was 100mW, and the average value was recorded for three parallel tests. The sampling and detection process is as follows: Figure 2 As shown, the SERS spectra of MPXV protein at different concentrations on the T line are as follows Figure 4 shown.

[0044] (5) Qualitative and quantitative analysis of SERS: The characteristic peak intensities of the SERS spectra obtained by the T-line of the test strips with different concentrations of MPXV protein are different. By fitting a curve showing the change of the characteristic peak area with the MPXV protein concentration, the semi-quantitative detection of MPXV protein can be achieved through this curve, and negative and positive samples can be accurately distinguished within the detection limit.

[0045] Example 2

[0046] This example adopts the method of adding different concentrations of MPXV virus protein to the throat swab buffer solution without MXPV so that the final concentration is 1pg / mL-100pg / mL. The specific detection method is the same as that in Example 1.

[0047] Figure 5 This is the curve showing the change of characteristic peak intensity of the SERS spectrum corresponding to different concentrations of MPXV protein in the throat swab system on the T line as a function of concentration.

[0048] Example 3

[0049] This example uses 30 throat swab sample solutions that do not contain MXPV, adding MPXV viral proteins at concentrations of 1 pg / mL and 0 pg / mL, and distinguishing the spectra of negative and positive samples to verify the sensitivity and reliability of the scheme in different throat swab systems. The specific detection method is the same as in Example 1.

[0050] Figure 6 The figure is a comparison of the discrimination between positive and negative samples when 1pg / mL and 0pg / mL concentrations of MPXV protein were added to throat swabs from 30 different individuals.

[0051] The present invention develops a highly sensitive surface enhanced resonance Raman spectroscopy (SERRS) biosensor based on antigen-antibody recognition, uses special probe molecules that can match the wavelength of excitation light to produce a resonance effect, prepares highly sensitive SERRS nanoprobes, and uses SERS combined with test strip technology to detect monkeypox virus protein. By improving the test strip preparation process and designing a portable Raman spectrometer that matches the test strip, the stability of the acquired spectrum is ensured; based on the preparation method of regulating and optimizing the SERRS sensor probe, the system reaction mode is explored to improve the sensitivity of viral protein detection. Further, in the throat swab system, the matrix interference is reduced by functionalizing the paper strip with a surfactant, and the "sandwich method" reaction mode is accelerated with the help of the biotin-streptavidin system (Biotin-SA). Finally, through the throat swab detection of a large number of monkeypox virus proteins, the detection sensitivity and accuracy of the method are evaluated and the feasibility of the developed system in responding to the monkeypox pandemic infectious disease is proved. The resonance advantage of the bifunctional signal molecule and the combined advantage of the antibody avoid the defects of weak enhancement performance and insufficient protein capture ability of the traditional probe. In addition, the antigen-antibody binding efficiency was greatly enhanced by the Biotin-SA system, and 0.5% S9 was used to effectively suppress the false positives of the throat swab system, solving the problem that some SERS test strip systems are susceptible to false positive interference. This method is easy to operate, low cost, fast, and easy to carry using a portable Raman spectrometer. It has the advantages of high sensitivity of SERS and instant detection of test strips, and is a SERS method that is very suitable for rapid on-site detection.

[0052] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some or all of the technical features thereof may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip, characterized in that: The following steps are involved: (1) Preparation of Au NPs@IR808@Ab1 resonant SERS probe: The Raman signal molecule IR808, which has both resonance and antibody binding functions, is modified onto the surface of gold nanoparticles to form Au NPs@IR808. The carboxyl group of the IR808 molecule is activated to link to the antibody Ab1, forming the resonant SERS biosensor probe AuNPs@IR808@Ab1. (2) Preparation of biotin antibody Ab2@Bio: Assemble biotin on antibody Ab2 and prepare biotin antibody Ab2@Bio by dialysis in PBS buffer; (3) Preparation of SA test strips: NC membrane is pasted on the bottom plate, and SA is modified on the NC membrane by a gold spraying film scribing instrument to form a detection line T line. Then, a sample pad, a binding pad and a water-absorbing paper are assembled on the other end of the NC membrane in sequence, and then the strips are cut to prepare SA test strips with SA as the T line. (4) SERS signal sensing of monkeypox virus protein: The Au NPs@IR808@Ab1 resonance SERS probe prepared in step (1), the biotin antibody Ab2@Bio prepared in step (2) and the monkeypox virus protein sample were mixed in proportion and then dropped onto the sample pad of the SA-test strip prepared in step (3). After 10-15 minutes, the SERS signal of the T line of the test strip was collected using a portable Raman spectrometer; (5) SERS qualitative and quantitative analysis: Fit the curve of the characteristic peak area in the SERS spectrum as it changes with the monkeypox virus protein concentration, and identify the positive and negative nature of the sample to be tested and quantify the virus concentration of the positive sample based on the characteristic peak area.

2. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (1), the gold nanoparticles are prepared by sodium citrate reduction method, and the particle size of the gold nanoparticles is 30-60nm.

3. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: The concentration of the Raman signal molecule IR808 in step (1) is 0.3-0.7 mM, and the added amount is 4-8 uL / mL.

4. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: The antibody in step (1) and the antibody in step (2) are two antibodies that recognize different antigen surface determinants, and the two antibodies can be mixed in any proportion.

5. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (2), the concentration of PBS buffer is 1.5-2.5 m, and the dialysis time is 16-24 h.

6. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (3), the SA test strip is designed to be 7-8 cm long, 3.5-4.5 mm wide, with a SA concentration of 1-2 mg / mL and a T line width of 1.5-2.5 mm.

7. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (3), the sample pad is pre-modified with a hydrophobic surfactant, wherein the composition of the surfactant is 0.5 wt % S9 or 0.01-0.05 wt % TW-20.

8. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (4), the volume ratio of Au NPs@IR808@Ab1 probe, Ab2@Bio and monkeypox virus protein sample is (3-5):1:(70-80), the mixing time is 10-60s, and the test is performed after the test strip flows for 12-16min.

9. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip as claimed in claim 1, characterized in that: In the step (4), when the portable Raman spectrometer with 785 strip-shaped light spots collects the SERS signal of the T line of the test paper strip, the width of the laser spot is adjusted to match the width of the T line.

10. The method for instant detection of monkeypox virus based on a resonance SERS probe combined with a test strip according to claim 1, characterized in that: In the step (4), when the SERS signal of the T line of the test paper strip is collected using a portable Raman spectrometer with 785 strip-shaped light spots, the collection time is 100 ms and the power is 100 mw.

Images