CN111812321A - Novel quantitative detection method for coronavirus particles based on nano-plasmon resonance - Google Patents
Novel quantitative detection method for coronavirus particles based on nano-plasmon resonance Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
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- 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
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/55—Specular reflectivity
- G01N21/552—Attenuated total reflection
- G01N21/553—Attenuated total reflection and using surface plasmons
- G01N21/554—Attenuated total reflection and using surface plasmons detecting the surface plasmon resonance of nanostructured metals, e.g. localised surface plasmon resonance
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/08—RNA viruses
- G01N2333/165—Coronaviridae, e.g. avian infectious bronchitis virus
Abstract
A novel quantitative detection method of coronavirus particles based on nano-plasmon resonance comprises the following steps: preparing plasma photon resonance coupling effect equipment and gold nanoparticles for amplifying detection signals; modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance sensing chip, and marking the nano gold particles with SARS-COV-2 labeled antibody; adding SARS-CoV-2 virus particle samples with different titers into the marked nano gold particles; adding the mixed solution into a pore plate to be detected to complete the capture of the SARS-CoV-2 virus particle and marked nano gold particle compound; the quantitative detection of SARS-CoV-2 virus particle is realized by detecting the variation curve of the absorbance difference value under specific wavelength before and after sample loading along with time.
Description
Technical Field
The invention relates to the technical field of detection, in particular to a novel quantitative detection method for coronavirus particles based on nano-plasmon resonance.
Background
Measurement of molecular interaction kinetics is becoming increasingly important in drug discovery, genetic screening and clinical diagnostics, as dynamic binding information can improve understanding of disease and can provide new ideas for treatment. Surface Plasmon Resonance (SPR) sensors, such as the commercial Biacore SPR biosensor system, can monitor kinetic biomolecular interactions in real time, without labeling, and are not affected by a large background. Since the commercial Biacore SPR biosensor system belongs to the conventional SPR apparatus, it requires a specially-made apparatus, a specially-assigned person for its operation, so that a large amount of money is required to purchase or use the biosensor system.
The novel coronavirus is the seventh coronavirus that has been discovered so far to infect humans. Is a single-stranded positive-strand RNA virus, about 30Kb in length, and belongs to one of the largest genomic RNA viruses. The spurt glycoprotein S-protein of the novel coronavirus can be combined with angiotensin converting enzyme 2(ACE2) of human respiratory epithelial cells and lung tissue cells so as to enter the cells, and a virus RNA polymerase is expressed by taking a self RNA chain as a translation template. Then a series of transcription synthesis, synthesis and replication of various structural protein mRNA are carried out, and the new coronavirus assembled and secreted to the outside of the cell is assembled. Currently, there is no specific antiviral drug to treat new coronaviruses, requiring patients to stimulate autoimmunity to combat the virus. The existing developed SARS-COV-2 virus nucleic acid detection kit has the limitations of long period, high requirements for detection personnel and environment, low detection accuracy, and the like, and the protein detection kit aiming at SARS-COV-2 virus also has the problem of low detection sensitivity.
Disclosure of Invention
The invention aims to solve the technical problem of the prior art, and provides a quantitative detection method of novel coronavirus particles (SARS-CoV-2) based on a nano plasma resonance sensing chip to solve the problem of higher detection cost of a commercial Biacore SPR biosensor system and improve a virus particle detection mode.
According to the invention, the quantitative detection method of the novel coronavirus particles based on the nano-plasmon resonance sensing chip comprises the following steps:
the first step is as follows: preparing a pore plate and a nano plasma resonance sensing chip, and arranging the nano plasma resonance sensing chip in a pore of the pore plate to prepare a plasma photon resonance coupling effect device;
the second step is as follows: preparing nano gold particles for amplifying detection signals;
the third step: modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance sensing chip for specifically detecting the titer of SARS-CoV-2 virus particles;
the fourth step: marking the nano gold particles by using a SARS-COV-2 marked antibody;
the fifth step: adding SARS-CoV-2 virus particle samples with different titers into the marked nano gold particles to prepare a mixed solution;
a sixth step: adding the mixed solution into a pore plate to be detected to complete the capture of the SARS-CoV-2 virus particle and marked nano gold particle compound;
a seventh step of: the quantitative detection of SARS-CoV-2 virus particle is realized by detecting the variation curve of the absorbance difference value under specific wavelength before and after sample loading along with time.
Preferably, the specific wavelength is one or more of a wavelength of 580nm, a wavelength of 605nm, a wavelength of 620nm, and a wavelength of 640 nm.
Preferably, the nano gold particles are 15-80 nm, and preferably 20-70 nm.
Preferably, the method of preparing the gold nanoparticles for amplifying a detection signal includes: adding a chloroauric acid solution into deionized water, heating to boil, then adding a trisodium citrate solution, continuing to heat, stirring by using a magnetic stirrer, changing the color of the solution from black to orange red, heating for a preset time, stopping heating, then standing the solution, and cooling to room temperature to obtain an Au nanoparticle dispersed solution as a gold nanoparticle.
Preferably, the method for modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance chip comprises the following steps: placing the nano plasma resonance chip in 11-mercaptoundecanoic acid solution at room temperature, cleaning with ethanol, and drying with nitrogen; adding 400 × 10-3M 1-ethyl-3-3-dimethylaminopropyl carbodiimide and 100 × 10-3M N-hydroxysuccinimide into a 1:1 mixed solution for incubation; after incubation, putting monoclonal antibody SARS-COV-2 capture antibody into the cell, incubating at room temperature, cleaning, and drying; adding bovine serum albumin blocking solution for incubation at room temperature, removing the bovine serum albumin blocking solution, adding ethanolamine solution for incubation at room temperature to cover unreacted NHS ester; then the glass is cleaned by deionized water, dried by nitrogen and placed in a dry environment.
Preferably, the method for labeling the gold nanoparticles with the SARS-COV-2 labeled antibody comprises: regulating the pH of the gold nanoparticle solution to the isoelectric point of SARS-COV-2 labeled antibody or ACE2 protein by using K2CO3 solution, adding the SARS-COV-2 labeled antibody or ACE2 protein antibody, mixing uniformly, incubating at room temperature, adding bovine serum albumin blocking solution with final concentration of 0.5-1.5%, and incubating at room temperature; setting the rotation speed of the centrifuge at 5000-.
Preferably, the capturing of the SARS-CoV-2 viral particle and the labeled gold nanoparticle complex comprises: adding SARS-COV-2 virus particles into SARS-COV-2 virus S-protein antibody or ACE2 protein labeled nano gold particle complex solution, mixing, adding the mixture into a chip orifice plate covered with SARS-COV-2 capture antibody or protein, and placing the orifice plate into an enzyme labeling instrument for oscillation reaction.
Preferably, the curve obtained by measuring the change of the difference in absorbance at a specific wavelength before and after loading the sample with time comprises: before sample adding, taking out the chip pore plate covered with the SARS-COV-2 capture antibody or protein, adding 100 mul deionized water into the hole to be tested, and detecting the first absorbance value of the hole to be tested under the specific wavelength by using an enzyme-labeling instrument. And after the sample mixed solution is added into the hole and the reaction is finished, absorbing the sample mixed solution, cleaning the test hole by using deionized water, adding 100 mu l of deionized water into the test hole, and detecting a second absorbance value of the test hole under a specific wavelength.
Preferably, the SARS-COV-2 capture antibody is one of an S protein antibody capturing SARS-CoV-2 virus and ACE2 protein, and the SARS-COV-2 marker antibody is the other of an S protein antibody capturing SARS-CoV-2 virus and ACE2 protein.
The quantitative detection method of the novel coronavirus particle (SARS-CoV-2) based on the nano-plasma resonance sensing chip can solve the problem of higher detection cost of a commercial Biacore SPR biosensor system and can improve a virus particle detection mode.
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In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below. It should be apparent that the drawings in the following description are merely exemplary, and that other embodiments can be derived from the drawings provided by those of ordinary skill in the art without inventive effort.
Fig. 1 schematically shows a flow chart of the steps of the quantitative detection method of the novel coronavirus particles based on the nanoplasmon resonance sensor chip according to the preferred embodiment of the invention.
Fig. 2 schematically shows a schematic diagram of a quantitative detection method of novel coronavirus particles based on a nanoplasmon resonance sensor chip according to a preferred embodiment of the present invention.
Fig. 3 schematically shows a principle schematic diagram of a quantitative detection method of novel coronavirus particles based on a nanoplasmon resonance sensor chip according to a preferred embodiment of the invention.
Fig. 4 and 5 are spectral diagrams of samples of specific refractive indices provided by embodiments of the present invention.
FIG. 6 is a graph showing the results of detecting the difference in absorbance of a novel coronavirus particle (SARS-CoV-2) at different concentrations according to the present invention.
It should be noted that the structures, ratios, sizes, and the like shown in the specification are only used for matching with the contents disclosed in the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the technical content disclosed in the present invention without affecting the efficacy and the achievable purpose of the present invention.
Detailed Description
In order that the present disclosure may be more clearly and readily understood, reference will now be made in detail to the present disclosure as illustrated in the accompanying drawings.
Fig. 1 schematically shows a flow chart of steps of a quantitative detection method of novel coronavirus particles based on a nanoplasmon resonance sensing chip according to a preferred embodiment of the present invention, fig. 2 schematically shows a schematic diagram of a quantitative detection method of novel coronavirus particles based on a nanoplasmon resonance sensing chip according to a preferred embodiment of the present invention, and fig. 3 schematically shows a principle schematic diagram of a quantitative detection method of novel coronavirus particles based on a nanoplasmon resonance sensing chip according to a preferred embodiment of the present invention.
As shown in fig. 1, fig. 2 and fig. 3, the method for quantitatively detecting the novel coronavirus particles based on the nanoplasmon resonance sensor chip according to the preferred embodiment of the present invention comprises:
first step S1: preparing a pore plate and a nano plasma resonance sensing chip, and arranging the nano plasma resonance sensing chip in a pore of the pore plate to prepare a plasma photon resonance coupling effect device;
for example, the well plate is a 96 well plate.
Second step S2: preparing nano gold particles for amplifying detection signals; for example, the gold nanoparticles are gold nanoparticles having a particle size of 15 to 80nm, preferably 20 to 70 nm.
Specifically, for example, a method of preparing a gold nanoparticle for amplifying a detection signal includes: adding 1mL of 1% chloroauric acid solution into 100mL of deionized water, heating to boil, adding 0.6-4mL of 1% trisodium citrate solution, heating, stirring with a magnetic stirrer, changing the color of the solution from black to orange-red, and stopping heating after the solution is constant in color and heating for 15 min. And then standing and cooling to room temperature to obtain a clear and transparent Au nano-particle dispersion solution, namely the final nano-gold particles.
Third step S3: modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance sensing chip for specifically detecting the titer of SARS-CoV-2 virus particles; specifically, the SARS-COV-2 capture antibody is an S protein antibody or ACE2 protein that captures SARS-COV-2 virus.
For example, the method for modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance chip comprises the following steps: placing the nano plasma resonance chip in 1-10mM 11-mercaptoundecanoic acid solution for 1-12 hours at room temperature, cleaning with 70% ethanol, and blow-drying with nitrogen; adding 400 × 10-3M 1-ethyl-3-3-dimethylaminopropyl carbodiimide and 100 × 10-3M N-hydroxysuccinimide into a 1:1 mixed solution, and incubating for 30-60 minutes; immediately placing a monoclonal antibody SARS-COV-2 with the concentration of 50 mu g/ml into the culture medium after the incubation is finished, incubating the culture medium for 3 to 5 hours at room temperature, washing the culture medium twice by PBS, and drying the culture medium by nitrogen; adding 0.5-1.5% bovine serum albumin blocking solution, incubating at room temperature for 20-60 min, removing bovine serum albumin blocking solution, adding 10% ethanolamine solution, and incubating at room temperature for 30-60 min to cover unreacted NHS ester; after being washed twice by deionized water, the mixture is placed in a dry environment after being dried by nitrogen.
Fourth step S4: marking the nano gold particles by using a SARS-COV-2 marked antibody; specifically, the SARS-COV-2 marker antibody is S protein antibody of SARS-CoV-2 virus or ACE2 protein. In practice, for example, the SARS-COV-2 capture antibody and the labeled antibody may be interchanged.
For example, the method of labeling gold nanoparticles with SARS-COV-2 labeled antibody comprises: regulating the pH of the gold nanoparticle solution to the isoelectric point of SARS-COV-2 labeled antibody or ACE2 protein by using 0.1M K2CO3 solution, adding a proper amount of SARS-COV-2 labeled antibody or ACE2 protein antibody, mixing uniformly, incubating at room temperature for 30 minutes, adding bovine serum albumin blocking solution with the final concentration of 0.5-1.5%, and incubating at room temperature for 10 minutes. Setting the rotation speed of the centrifuge at 5000-.
Fifth step S5: adding SARS-CoV-2 virus particle samples with different titers into the marked nano gold particles to prepare a mixed solution; of course, this step includes preparing standard solutions of SARS-COV-2 virions at different titers for measuring the titer of SARS-COV-2 virions in the sample; for example, standard samples (1 x 10) of novel coronavirus particles (SARS-CoV-2) were prepared at different titer ranges3vp/ml～1*1010vp/ml); the dilution base solution is PBS buffer solution.
Sixth step S6: adding the mixed solution into a pore plate to be detected to complete the capture of the SARS-CoV-2 virus particle and marked nano gold particle compound;
specifically, for example, the capturing method of SARS-COV-2 virus S-protein antibody or ACE2 protein and labeled gold nanoparticle complex comprises: adding SARS-COV-2 virus particles into SARS-COV-2 virus S-protein antibody or ACE2 protein labeled nano-gold particle complex solution, mixing, adding the mixture into a chip orifice plate covered with SARS-COV-2 capture antibody or protein, and placing the orifice plate into an enzyme labeling instrument for shake reaction for 5-30 minutes.
Seventh step S7: the quantitative detection of SARS-CoV-2 virus particle is realized by detecting the variation curve of the absorbance difference value under specific wavelength before and after sample loading along with time.
For example, the specific wavelength is one or more of a wavelength of 580nm, a wavelength of 605nm, a wavelength of 620nm, and a wavelength of 640 nm.
Specifically, before sample addition, the chip pore plate covered with the SARS-COV-2 capture antibody or protein is taken out, 100. mu.l of deionized water is added into the hole to be tested, and a first absorbance value of the hole to be tested under a specific wavelength is detected by using an enzyme-labeling instrument. And after the sample mixed solution is added into the hole and the reaction is finished, absorbing the sample mixed solution, cleaning the test hole by using deionized water (2 times), adding 100 mu l of deionized water into the test hole, and detecting a second absorbance value of the test hole under a specific wavelength.
Or before sample adding, taking out the chip pore plate covered with the SARS-COV-2 capture antibody or protein, adding the sample mixed solution into the hole to be tested, and detecting the change value of the absorbance of the test hole along with the time under the specific wavelength.
The difference between the absorbance of the test hole before and after sample application can be used to characterize the conjugation amount between the SARS-COV-2 virus particles and the SARS-COV-2 antibody, thereby realizing the quantitative detection of the SARS-COV-2 virus particles.
The embodiment of the invention at least has the following advantages: the quantitative determination of virus particles is realized by using a common enzyme-labeling instrument and the nano-gold particles, and the device can reduce the system requirements of special instruments to the maximum extent. The method has the advantages of good selectivity, high sensitivity and the like, and because the gold nanoparticles marked by the specific antibody can be specifically combined with S-protein on the surface of SARS-COV-2 virus particles, the detection signals can be further amplified without additional detection marking substances, the detection process is simplified, and the detection cost is reduced; the chip has extremely high sensitivity to changes in the surface refractive index by transmitting light intensity changes in a specific wavelength range (580, 605, 620, 640 nm).
It should be noted that the terms "first", "second", "third", and the like in the description are used for distinguishing various components, elements, steps, and the like in the description, and are not used for indicating a logical relationship or a sequential relationship between the various components, elements, steps, and the like, unless otherwise specified.
It is to be understood that while the present invention has been described in conjunction with the preferred embodiments thereof, it is not intended to limit the invention to those embodiments. It will be apparent to those skilled in the art from this disclosure that many changes and modifications can be made, or equivalents modified, in the embodiments of the invention without departing from the scope of the invention. Therefore, any simple modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention are still within the scope of the protection of the technical solution of the present invention, unless the contents of the technical solution of the present invention are departed.
Claims (10)
1. A novel quantitative detection method of coronavirus particles based on nano-plasmon resonance is characterized by comprising the following steps:
the first step is as follows: preparing a pore plate and a nano plasma resonance sensing chip, and arranging the nano plasma resonance sensing chip in a pore of the pore plate to prepare a plasma photon resonance coupling effect device;
the second step is as follows: preparing nano gold particles for amplifying detection signals;
the third step: modifying SARS-COV-2 capture antibody on the surface of the nano plasma resonance sensing chip for specifically detecting the titer of SARS-CoV-2 virus particles;
the fourth step: marking the nano gold particles by using a SARS-COV-2 marked antibody;
the fifth step: adding SARS-CoV-2 virus particle samples with different titers into the marked nano gold particles to prepare a mixed solution;
a sixth step: adding the mixed solution into a pore plate to be detected to complete the capture of the SARS-CoV-2 virus particle and marked nano gold particle compound;
a seventh step of: the quantitative detection of SARS-CoV-2 virus particle is realized by detecting the variation curve of the absorbance difference value under specific wavelength before and after sample loading along with time.
2. The nanoplasmon resonance-based novel quantitative detection method for coronavirus particles according to claim 1, wherein the specific wavelength is one or more of 580nm wavelength, 605nm wavelength, 620nm wavelength and 640nm wavelength.
3. The quantitative detection method for the novel coronavirus particles based on nanoplasmon resonance as claimed in claim 1 or 2, wherein the gold nanoparticles are gold nanoparticles having a particle size of 15-80 nm.
4. The quantitative detection method of novel coronavirus particles based on nanoplasmon resonance as claimed in claim 1 or 2, wherein the gold nanoparticles are gold nanoparticles having a particle size of 20-70 nm.
5. The nanoplasmon resonance-based quantitative detection method for novel coronavirus particles according to claim 1 or 2, wherein the method for preparing gold nanoparticles for amplifying detection signals comprises: adding a chloroauric acid solution into deionized water, heating to boil, then adding a trisodium citrate solution, continuing to heat, stirring by using a magnetic stirrer, changing the color of the solution from black to orange red, heating for a preset time, stopping heating, then standing the solution, and cooling to room temperature to obtain an Au nanoparticle dispersed solution as a gold nanoparticle.
6. The method for quantitatively detecting the novel coronavirus particles based on nanoplasmon resonance as claimed in claim 1 or 2, wherein the method for modifying the surface of the nanoplasmon resonance chip with the SARS-COV-2 capture antibody comprises: placing the nano plasma resonance chip in 11-mercaptoundecanoic acid solution at room temperature, cleaning with ethanol, and drying with nitrogen; adding 400 × 10-3M 1-ethyl-3-3-dimethylaminopropyl carbodiimide and 100 × 10-3M N-hydroxysuccinimide into a 1:1 mixed solution for incubation; after incubation, putting monoclonal antibody SARS-COV-2 capture antibody into the cell, incubating at room temperature, cleaning, and drying; adding bovine serum albumin blocking solution for incubation at room temperature, removing the bovine serum albumin blocking solution, adding ethanolamine solution for incubation at room temperature to cover unreacted NHS ester; then the glass is cleaned by deionized water, dried by nitrogen and placed in a dry environment.
7. The nanoplasmon resonance-based novel quantitative detection method for coronavirus particles according to claim 1 or 2, wherein the method for labeling gold nanoparticles with SARS-COV-2 labeled antibody comprises: regulating the pH of the gold nanoparticle solution to the isoelectric point of SARS-COV-2 labeled antibody or ACE2 protein by using K2CO3 solution, adding the SARS-COV-2 labeled antibody or ACE2 protein antibody, mixing uniformly, incubating at room temperature, adding bovine serum albumin blocking solution with final concentration of 0.5-1.5%, and incubating at room temperature; setting the rotation speed of the centrifuge at 5000-.
8. The nanoplasmon resonance-based novel quantitative detection method for coronavirus particles according to claim 1 or 2, wherein the capturing of SARS-CoV-2 virus particles and labeled nanogold particle complexes comprises: adding SARS-COV-2 virus particles into SARS-COV-2 virus S-protein antibody or ACE2 protein labeled nano gold particle complex solution, mixing, adding the mixture into a chip orifice plate covered with SARS-COV-2 capture antibody or protein, and placing the orifice plate into an enzyme labeling instrument for oscillation reaction.
9. The method of claim 1 or 2, wherein the step of measuring the time-dependent variation of the absorbance difference at a specific wavelength before and after loading the sample comprises: before sample adding, taking out the chip pore plate covered with the SARS-COV-2 capture antibody or protein, adding 100 mul deionized water into the hole to be tested, and detecting the first absorbance value of the hole to be tested under the specific wavelength by using an enzyme-labeling instrument. And after the sample mixed solution is added into the hole and the reaction is finished, absorbing the sample mixed solution, cleaning the test hole by using deionized water, adding 100 mu l of deionized water into the test hole, and detecting a second absorbance value of the test hole under a specific wavelength.
10. The nanoplasmon resonance-based novel quantitative detection method for coronavirus particles according to claim 1 or 2, wherein the SARS-COV-2 capture antibody is one of an S protein antibody capturing SARS-CoV-2 virus and ACE2 protein, and the SARS-COV-2 marker antibody is the other of the S protein antibody capturing SARS-CoV-2 virus and ACE2 protein.
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|---|---|---|---|---|
| WO2021189058A1 (en) * | 2020-03-20 | 2021-09-23 | Victor Manneh | Coronavirus assay |
| WO2022007251A1 (en) * | 2020-07-06 | 2022-01-13 | 量准（上海）医疗器械有限公司 | Nano-plasmon resonance-based quantitative detection method for novel coronavirus particles |
| CN113484521A (en) * | 2021-05-19 | 2021-10-08 | 南京医科大学第二附属医院 | ACE2 protein biochip and preparation method and application thereof |
| CN113552041A (en) * | 2021-06-08 | 2021-10-26 | 上海交通大学 | Exosome subtype analysis method based on single particle imaging |
| CN113533252A (en) * | 2021-06-22 | 2021-10-22 | 中山大学 | Sensor based on strong coupling system, preparation method and application thereof |
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