CN115508331A - Raman sensor for rapidly detecting new coronavirus based on integrated optical flow control type microstructure optical fiber and SERS substrate - Google Patents
Raman sensor for rapidly detecting new coronavirus based on integrated optical flow control type microstructure optical fiber and SERS substrate Download PDFInfo
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- 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
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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Abstract
The invention provides 1. A Raman sensor for rapidly detecting new coronavirus based on integration of an optical flow control type microstructure optical fiber and an SERS substrate, which comprises the following steps: step one, preparing nano silver sol by a sol-gel method; connecting nano graphene oxide by a chemical bonding method; step three, growing the nano gold particles; step four, modifying the microstructure optical fiber; step five, modifying the microstructure optical fiber by the receptor domain; and step six, preparing a Raman detection device. The invention prepares an SERS substrate specifically combined with SARS-CoV-2 spike protein, and combines with a hollow optical fiber to form a light flow control Raman detector, thereby realizing the rapid detection of SARS-CoV-2 spike protein. The microfluidic Raman SARS-CoV-2 spike protein sensor has the advantages of high integration degree, high sensitivity, high response speed, low cost and simple operation.
Description
Technical Field
The invention relates to a Raman sensor for rapidly detecting a new coronavirus based on an integrated optical flow control type microstructure optical fiber and an SERS substrate, in particular to a Raman sensor for rapidly detecting SARS (severe acute respiratory syndrome) -CoV-2, which is based on a natural micro-fluidic optical fiber, integrates SERS (surface enhanced Raman scattering) integrated by taking nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 as a special substrate and has a flow sample introduction function.
Background
The current methods for detecting viruses include reverse transcription-polymerase chain reaction (RT-PCR), fluorescence detection, immunological detection, and the like. However, the above methods require long detection time and need to label the sample in advance, which are tedious and time-consuming to operate, and the detection result is susceptible to a large number of factors such as specimen quality, virus infection site and expression level. In contrast, the surface enhanced Raman scattering SERS detection method has the advantages of no marker, high speed, high sensitivity and the like, and has good application prospects in the aspects of quick and accurate diagnosis and disease propagation prevention.
In the process of detecting the new coronavirus, in order to avoid sample leakage in the detection process and large-area virus propagation, a small amount of detection samples are extracted during detection. Optical fiber sensors have received much attention due to their high sensitivity, anti-electromagnetic interference, and small size. In particular, some specially structured optical fibers, such as hollow fibers, have microstructured channels therein. Due to the special internal structure and flexible characteristics, the probe can be widely used for detecting biological and chemical trace samples.
At present, gold and silver nanoparticles are widely applied to Raman detection as common SERS substrates. Meanwhile, due to the fact that the novel two-dimensional nano materials such as graphene oxide can be combined with the nano particles to form a composite SERS substrate due to the special structure of the novel two-dimensional nano materials, the SERS detection effect can be greatly improved.
Disclosure of Invention
The purpose of the invention is: in order to solve the problems of high cost, complex operation, long time consumption and the like of a new coronavirus detection method, a Raman sensor for quickly detecting the new coronavirus based on an integrated optical flow control type microstructure optical fiber and an SERS substrate is provided, and particularly, a Raman sensor for quickly detecting SARS-CoV-2, which is based on the microstructure optical fiber, can be specifically combined with the SARS-CoV-2 by taking nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 as a special SERS substrate. The sensor has the advantages of both a Raman sensor and an optical fiber sensor, and has the advantages of high sensitivity, high response speed, small sample consumption, strong anti-interference capability, flowing sample introduction and the like.
The invention is realized by the following steps:
an integrated Raman sensor based on an optical flow control type microstructure optical fiber and an SERS substrate comprises: the device comprises a light path, an SERS substrate, a Raman detection system, a flow sample introduction system and the like. The optical path consists of a light source of the portable Raman spectrometer and a hollow optical fiber with a suspended core and a microstructure channel inside; the SERS substrate is constructed on the surface of a suspension core grown in the hollow optical fiber by a chemical bonding method; the Raman detection system comprises a spectrum detection device of the portable Raman spectrometer and Raman spectrum analysis software; the flow sample injection function is realized by a micro-injection pump.
The preparation process of the optofluidic Raman sensor for detecting the new coronavirus comprises the following steps of:
1. preparing nano silver sol by a sol-gel method: first, 30mg of silver nitrate was dissolved in 100mL of deionized water and heated. Next, 20mg of sodium citrate was dissolved in 2mL of deionized water to prepare a sodium citrate solution. When the silver nitrate solution is heated to boiling, the sodium citrate solution is slowly dropped. Finally, after the mixture is heated for 20 minutes by the residual temperature, the mixture is cooled to the room temperature, and the silver nano particles with the size of about 80nm are prepared.
Alternatively, the residual temperature heating time may be 10 to 30 minutes.
Alternatively, 20mg of sodium citrate was dissolved in 3mL of deionized water.
2. Connecting nano graphene oxide by a chemical bonding method: mixing the silver nano solution obtained in the first step with the graphene oxide solution according to the ratio of 1. And selecting single-layer nano Graphene Oxide (GO). First, a GO solution was prepared with deionized water at a concentration of 0.5 mg/mL. The GO solution needs to be pre-treated before the two solutions are mixed. That is, 96mg of polyvinylpyrrolidone (PVP) was added to 24mL of GO solution and dissolved in 12mL of deionized water after centrifugation. Then, adding 3mL of PVP modified GO solution into a mixed solution of 12mL of PDDA (30 wt%) solution and KCl to obtain a PDDA functionalized graphene oxide (GO/PDDA) solution, and dissolving the PDDA functionalized graphene oxide (GO/PDDA) solution in 6mL of deionized water after centrifugation. And finally, mixing the GO/PDDA solution and the Ag NPs and stirring for 3 minutes to obtain the GO/Ag NPs SERS substrate.
Optionally, the silver nano solution is mixed with the graphene oxide solution in a ratio of 1.
Alternatively, the GO/PDDA solution and the Ag NPs are mixed and stirred for a period of 2-5 minutes.
3. And (3) growing the nano gold particles: and mixing the nano-silver/nano-graphene oxide solution obtained in the second step with the gold nano-solution under the action of PVP according to the proportion of 1. 1mL of 1% PVP was mixed with 100. Mu.L of 100mM ascorbic acid and 100. Mu.L of 200mM NaOH solution. After stirring for 5 min, 50. Mu.L of nanosilver/nanographene oxide and 100. Mu.L of 2mM HAuCl 4 The solution was added to the above solution. And kept shaking for 3h in the dark. And finally, centrifuging the mixture and dissolving the precipitate in 50 mu L of ultrapure water to prepare the nano-silver/nano-graphene oxide/nano-gold SERS substrate.
Optionally, the nano silver/nano graphene oxide solution is mixed with the gold nanoparticle solution in a ratio of 1.
Optionally, the shaking is maintained in the dark for a period of 2 to 4 hours.
4. Modifying the microstructure optical fiber: and (4) growing the SERS substrate composed of the nano silver/nano graphene oxide/nano gold synthesized in the step three on the surface of the suspension core in the microstructure optical fiber. In order to construct the in-fiber Raman signal enhancement substrate around the suspension core, the nano-silver/nano-graphene oxide/nano-gold is fixed around the suspension core through a chemical bond. The hollow fiber was pretreated by first passing piranha solution (4 volumes H) 2 SO 4 And 1 volume H 2 O 2 ) Injecting into MHF; hydroxyl (-OH) groups on the surface of the hanging core are then exposed. Subsequently, 3-amino group is injectedPropyltriethoxysilane (APTES, volume ratio of APTES to ethanol 1 2 ). The amino and the exposed hydroxyl are combined to serve as a bridge for connecting nano silver/nano graphene oxide/nano gold. And finally, depositing the nano silver/nano graphene oxide/nano gold on the inner surface of the MHF through the reaction between the amino group and the nano silver/nano graphene oxide/nano gold. The last process is repeated several times in order to grow a large amount of particles to the surface of the suspended core. And then washing with deionized water to obtain the optical fiber optofluidic online SERS sensing optical fiber.
Optionally, piranha solution H 2 SO 4 And H 2 O 2 May be 2.
Alternatively, the volume ratio of APTES to ethanol may be 1.
5. Modification of microstructured optical fiber by acceptor domain: thoroughly washing the Raman sensor with the SERS substrate with the nano silver/nano graphene oxide/nano gold grown on the surface of the suspended core in the four-microstructure optical fiber by using deionized water and using N 2 And (5) drying. Subsequently, 1. Mu.L of ACE2 solution (30 pg) was passed into the microstructure channel and placed in an incubator at constant temperature and humidity (25 ℃;75%, w/w) for 4 hours. And preparing the novel Raman sensor for detecting the coronavirus based on the integration of the microstructure optical fiber and the SERS.
Alternatively, 0.5 μ L ACE2 solution (30 pg) was passed into the microstructure channel.
Alternatively, the incubation time in the incubator can be 2 to 5 hours.
6. Preparing a Raman detection device: taking the nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 in the fifth step as the surface of the microstructure optical fiber of the special SERS substrate capable of being specifically combined with SARS-CoV-2 for micropore etching, and utilizing CO 2 And etching micropores by the laser while not damaging the suspended core so that a detection sample solution can flow out, and preparing the microfluidic Raman sensor. Meanwhile, a gold film is plated at the tail end of the optical fiber by an optical fiber film plating machine to prepare a reflecting layer, so that the reflective microfluidic Raman sensor is constructed.
Alternatively, the surface of the optical fiber may be polished to form the micro-holes by an optical fiber polishing machine.
Alternatively, silver or aluminum film may be used as the reflecting layer at the tail end of the optical fiber.
The invention provides the micro-fluidic online Raman sensor based on the micro-structural optical fiber, which has the advantages of high sensitivity, high response speed, low cost and simple operation, and has the most remarkable advantages of integration and miniaturization of the device. The micro-type Raman sensor for the new coronavirus is expected to be applied to rapid and efficient detection of the new coronavirus.
Compared with the prior art, the invention has the beneficial effects that:
the invention prepares an SERS substrate specifically combined with SARS-CoV-2 spike protein, and combines with a hollow optical fiber to form a light flow control Raman detector, thereby realizing the rapid detection of SARS-CoV-2 spike protein. The microfluidic Raman SARS-CoV-2 spike protein sensor has the advantages of high integration degree, high sensitivity, high response speed, low cost and simple operation.
Drawings
FIG. 1 is a schematic diagram of a process for synthesizing nano-silver/nano-graphene oxide/nano-gold;
FIG. 2 is a schematic diagram of a process of using ACE2 to modify a microstructure hollow optical fiber with nano silver/nano graphene oxide/nano gold grown;
FIG. 3 is a connection diagram of each part of the micro-structured fiber microfluidic Raman sensor;
FIG. 4 is a schematic diagram of a micro-structured fiber microfluidic Raman sensor construction process;
FIG. 5 is a Raman spectrum of a micro-fluidic Raman sensor based on a microstructure optical fiber for detecting rhodamine solution.
Detailed Description
Please refer to fig. 1 and fig. 2 respectively for the process of constructing the micro-structured hollow optical fiber with ACE2 modification and nano silver/nano graphene oxide/nano gold growth. Referring to fig. 3, the connection mode of each part of the micro-structured fiber microfluidic raman sensor provided by the present invention refers to fig. 4, the left side of the drawing is a perspective view, and the right side is a cross-sectional view of the fiber end face. Each part is numbered separately: a micro-structure hollow optical fiber [1], a hollow optical fiber inner wall [1-1], a micro-structure channel [1-2] of the hollow optical fiber, a suspension core [1-3] of the hollow optical fiber, an ACE2 modified SERS substrate [1-4] growing on the surface of the suspension core, an ACE2 layer [1-5] in the SERS substrate, a graphene oxide layer [1-6] in the SERS substrate, a silver nanoparticle layer [1-7] in the SERS substrate, a gold nanoparticle layer [1-8] in the SERS substrate and a sample outlet [2].
The invention relates to a preparation method of a Raman sensor for rapidly detecting new coronavirus based on integrated optical flow control type microstructure optical fiber and SERS substrate, which comprises the following specific embodiments:
1. preparing nano silver sol by a sol-gel method: first, 30mg of silver nitrate was dissolved in 100mL of deionized water and heated by an electric furnace. Next, 20mg of sodium citrate was dissolved in 2mL of deionized water to make a sodium citrate solution. When the silver nitrate solution is heated to boiling, 0.1mL of sodium citrate solution is slowly dropped, after 2 minutes, the rest of the sodium citrate solution is slowly and gradually dropped into the silver nitrate solution, and the reaction is carried out for 10 minutes. And finally, turning off the power supply, heating for 20 minutes by using the residual temperature, putting the solution into a magnetic stirrer, stirring and cooling the solution to room temperature to obtain the silver nano particles with the size of about 80 nm.
Preferably, in this embodiment, the residual temperature heating time is 20 minutes.
Preferably, in this example, 20mg of sodium citrate is dissolved in 2mL of deionized water.
2. Connecting nano graphene oxide by a chemical bonding method: mixing the silver nano solution obtained in the first step with the graphene oxide solution according to the ratio of 1. Selecting single-layer nano Graphene Oxide (GO), and preparing a GO solution with the concentration of 0.5mg/mL by using deionized water. The GO solution needs to be pre-treated before the two solutions are mixed. That is, 96mg of polyvinylpyrrolidone (PVP) was added to 24mL of GO solution, stirred with a magnetic stirrer for 1.5h, centrifuged 3 times to remove impurities and dissolved in 12mL of deionized water. Then, 3mL of PVP modified GO solution is added into 12mL of a PDDA (30 wt%) solution and a KCl mixed solution, the mixture is stirred for 30 minutes to obtain a PDDA functionalized graphene oxide (GO/PDDA) solution, and after 2 times of centrifugation, redundant impurities are removed, and the PDDA functionalized graphene oxide solution is dissolved in 6mL of deionized water. And finally, mixing the GO/PDDA solution and the Ag NPs according to the proportion of 10.
Preferably, in the present embodiment, the silver nano solution and the graphene oxide solution are mixed in a ratio of 1.
Preferably, in this example, the GO/PDDA solution and the Ag NPs are mixed and stirred for 3 minutes.
3. And (3) growing the nano gold particles: and mixing the nano-silver/nano-graphene oxide solution obtained in the second step with the gold nano-solution under the action of PVP according to the proportion of 1. 1mL of 1% PVP was mixed with 100. Mu.L of 100mM ascorbic acid and 100. Mu.L of 200mM NaOH solution. After stirring with a magnetic stirrer for 5 minutes, 50. Mu.L of nano-silver/nano-graphene oxide and 100. Mu.L of 2mM HAuCl 4 The solution was added to the above solution. And kept shaking in the dark for 3h. And finally, centrifuging the mixture for 5 times to remove redundant impurities, and dissolving the concentrated precipitate in 50 mu L of deionized water to prepare the nano-silver/nano-graphene oxide/nano-gold SERS substrate.
Preferably, in the present embodiment, the nano silver/nano graphene oxide solution is mixed with the gold nanoparticle solution under the action of PVP in a ratio of 1.
Preferably, in the present embodiment, the time for keeping the shaking in the dark is 3h.
4. Modifying the microstructure optical fiber: and (3) growing the SERS substrate composed of the nano silver/nano graphene oxide/nano gold synthesized in the step three on the surface of the suspension core in the microstructure optical fiber. In order to construct the in-fiber Raman signal enhancement substrate around the suspension core, the nano-silver/nano-graphene oxide/nano-gold is fixed around the suspension core through a chemical bond. The hollow optical fiber is pretreated by first adding a piranha solution (4 volumes H) 2 SO 4 And 1 volume H 2 O 2 ) Injecting into MHF; repeatedly introducing the piranha solution into the hollow fiber for 3 times, wherein the solution stays in the hollow fiber for 30 min after each introduction. Subsequently, 3-aminopropyltriethoxysilane (APTES, 1 volume ratio of APTES to ethanol) was injected 4 times and internally held for 30 minutes to form an amino group (-NH) 2 ). The amino and the exposed hydroxyl are combined to be used as a bridge for connecting nano silver/nano graphene oxide/nano gold. And repeatedly washing and drying the processed microstructure hollow optical fiber by using deionized water. And finally, depositing the nano silver/nano graphene oxide/nano gold on the inner surface of the MHF through the reaction between the amino group and the nano silver/nano graphene oxide/nano gold. The last process is repeated several times in order to grow a large number of particles into the fiber. And then washing with deionized water to prepare the optofluidic online SERS sensing optical fiber in the optical fiber.
Preferably, in this embodiment, piranha solution H 2 SO 4 And H 2 O 2 May be 4.
Preferably, in this embodiment, the volume ratio of APTES to ethanol may be 1.
5. Modification of microstructured optical fiber by acceptor domain: thoroughly washing the Raman sensor with the SERS substrate with the nano silver/nano graphene oxide/nano gold grown on the surface of the suspended core in the four-microstructure optical fiber by using deionized water and using N 2 And (5) drying. Subsequently, 1 μ L of ACE2 solution (30 pg) was passed into a micro-structured channel, and in order to allow ACE2 to grow over the SERS substrate surface, after the first ACE2 pass for 1 hour, ACE2 was repeatedly passed, and finally placed in an incubator under constant temperature and humidity conditions (25 ℃;75%, w/w) for 4 hours. And preparing the novel Raman sensor for detecting the coronavirus based on the integration of the microstructure optical fiber and the SERS.
Preferably, in this example, 1 μ L of ACE2 solution (30 pg) is passed into the microstructure channel.
Preferably, in this embodiment, the cultivation time in the incubator may be 4 hours.
6. Preparing a Raman detection device: taking the nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 in the fifth step as the surface of the microstructure optical fiber of the special SERS substrate capable of being specifically combined with SARS-CoV-2 for micro-crystallizationEtching holes with CO 2 And etching micropores by a laser while not damaging the suspension core, wherein the distance between the micropores and the tail end of the optical fiber is 15cm, and a detection sample solution can enter from the tail end and flow out from the micropores to prepare the microfluidic Raman sensor. Meanwhile, a gold film is plated on the tail end of the optical fiber by an optical fiber film plating machine for 5 minutes, and the gold film is used for preparing a reflecting layer and constructing a reflecting microfluidic Raman sensor.
Preferably, in this embodiment, CO is used 2 The laser etches micro holes without damaging the suspended core.
Preferably, in this embodiment, the fiber end is coated with gold as a reflective layer.
Please refer to fig. 5 for a raman spectrum for rhodamine solution detection based on the micro-structured fiber microfluidic raman sensor. The rhodamine solution with the concentration of 10-14M is injected, and the Raman characteristic peak of the rhodamine can be obviously detected. The Raman sensor based on the integration of the optical flow control type microstructure optical fiber and the SERS substrate has good sensitivity and detection capability.
The working principle of the novel crown virus Raman sensor based on the integration of the optical flow control type microstructure optical fiber and the SERS substrate is as follows: the cell receptor angiotensin converting enzyme 2 (ACE 2) is modified on the surface of a suspended core of the micro-structured hollow optical fiber, and when a SARS-CoV-2 spike protein sample passes through a micro-structured channel in the hollow optical fiber, the sample can be combined with the ACE2 modified composite SERS substrate on the surface of the suspended core to form a recognition and Receptor Binding Domain (RBD) of SARS-CoV-2 spike protein. Meanwhile, the evanescent field can collect Raman signals into the suspension core. This binding domain successfully inhibits ACE2, which originally had a distinct raman characteristic peak. The SARS-CoV-2 spike protein RBD targets the short β 5 and β 6 chains, α 4 and α 5 helices and loops on ACE2, which are rich in phenylalanine and amide III for C-N stretching and N-H bending. The phenomenon of SERS signal intensity quenching occurs across the entire spectrum.
Gold and silver nanoparticles are widely used as common SERS enhancement substrates, the Raman enhancement principle of the gold and silver nanoparticles is based on the LSPR effect, and the electromagnetic field, namely the Raman enhancement effect, can be changed by changing the size of particles and the arrangement distance between the particles. The composite SERS substrate synthesized by the nano gold and the nano silver has a stronger Raman enhancement effect compared with the SERS substrate formed by a single particle. Meanwhile, the special structure in the two-dimensional material graphene oxide is utilized to embed nano silver and nano gold into the surface of the nano graphene oxide, which means that the distance between nano particles is greatly shortened, so that a stronger Raman enhancement effect can be obtained, and detection of a trace amount of detection sample with lower concentration is facilitated.
Because the hollow optical fiber is a purely natural optical flow control device and is internally provided with the suspension core and the microstructure channel, the detection sample can flow into the microstructure channel to be directly contacted with the suspension core. Meanwhile, the suspension core can grow special materials on the surface of the suspension core by a chemical method to realize a series of sensing effects. That is, when SARS-CoV-2 flows into the hollow optical fiber, it can contact with ACE2 modified nano-silver/nano-graphene oxide/nano-gold SERS substrate growing on the surface of the suspended core, and the excited raman signal is collected by the evanescent field of the suspended core and transmitted to the detector. Thereby obtaining a raman spectrum.
Claims (5)
1. The utility model provides a quick detect new coronavirus Raman sensor based on light stream accuse formula microstructure optic fibre and SERS base integrated form which characterized in that: the method comprises the following steps:
step one, preparing nano silver sol by a sol-gel method: dripping a sodium citrate solution into the silver nitrate solution which is boiled by utilizing an oxidation-reduction method until the two solutions are completely reacted and cooled to room temperature to prepare silver nano particles with the size of about 80 nm;
step two, connecting the nano graphene oxide by a chemical bonding method: mixing the silver nano solution obtained in the step one with a graphene oxide solution modified by polyvinylpyrrolidone (PVP) and PDDA (30 wt%) solution according to a ratio of 1;
step three, growing the nano gold particles: mixing the nano-silver/nano-graphene oxide solution obtained in the second step with the gold nano-solution under the action of PVP according to the proportion of 1;
step four, microstructure optical fiber modification: growing the SERS substrate composed of the nano silver/nano graphene oxide/nano gold synthesized in the third step on the surface of the suspension core in the microstructure optical fiber under the action of chemical bonding, and washing with deionized water to prepare the fiber internal optofluidic online SERS sensing optical fiber;
step five, modifying the microstructure optical fiber by the receptor domain: thoroughly washing a Raman sensor device of the SERS substrate with the nano silver/nano graphene oxide/nano gold grown on the surface of the suspended core in the four-step microstructure optical fiber by using deionized water, drying by using N2, and then introducing an ACE2 solution for standing for 3 hours to prepare a new coronavirus detection Raman sensor based on the integration of the microstructure optical fiber and the SERS;
step six, preparing a Raman detection device: taking the nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 in the fifth step as the surface of the microstructure optical fiber of the special SERS substrate capable of being specifically combined with SARS-CoV-2 for micropore etching, and utilizing CO 2 And etching micropores by the laser while not damaging the suspension core so that a detection sample solution can flow out, and preparing the microfluidic Raman sensor.
2. The integrated Raman sensor for rapidly detecting new coronavirus based on optical flow control type microstructure optical fiber and SERS substrate as claimed in claim 1, wherein: a natural micro-fluidic optical fiber device is internally provided with a suspension core and a micro-structural channel, and can realize online monitoring on trace samples; meanwhile, the optical fiber is beneficial to integration and miniaturization of the device.
3. The Raman sensor for rapidly detecting the new coronavirus based on the integration of the optical flow-controlled microstructure optical fiber and the SERS substrate as claimed in claim 1, wherein: in the microstructure optical fiber, the prepared nano silver/nano graphene oxide/nano gold is grown on the surface of the suspension core by a chemical bonding method to form the optical flow control Raman sensing device with the Raman enhancement effect.
4. The integrated Raman sensor for rapidly detecting new coronavirus based on optical flow control type microstructure optical fiber and SERS substrate as claimed in claim 1, wherein: the microstructure channel in the sensor can enable a sample solution to flow in from one end and flow out from the surface micropores, has a flowing sample introduction function, and realizes a photo-fluidic Raman sensing device.
5. The integrated Raman sensor for rapidly detecting new coronavirus based on optical flow control type microstructure optical fiber and SERS substrate as claimed in claim 1, wherein: the nano silver/nano graphene oxide/nano gold particles functionalized by ACE2 are constructed to be used as a special SERS substrate and can be specifically combined with SARS-CoV-2, and the nano silver/nano graphene oxide/nano gold particles have high sensitivity and rapid detection characteristics when being used for eliminating external interference.
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