CN108872194A - A kind of method of sandwich structure SERS detection pathogen - Google Patents
A kind of method of sandwich structure SERS detection pathogen Download PDFInfo
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- CN108872194A CN108872194A CN201810958582.2A CN201810958582A CN108872194A CN 108872194 A CN108872194 A CN 108872194A CN 201810958582 A CN201810958582 A CN 201810958582A CN 108872194 A CN108872194 A CN 108872194A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/65—Raman scattering
- G01N21/658—Raman scattering enhancement Raman, e.g. surface plasmons
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
Abstract
The invention belongs to technical field of biological, disclose a kind of method of sandwich structure SERS detection pathogen.Antibacterial peptide function magnetic nano-particle and bacterium are incubated for, the magnetic nano-particle for having adsorbed bacterium is separated under external magnetic field, obtain magnetic nano-particle-bacterial complex, then it is incubated for altogether with the graphene oxide-silver gold-covered nano compound SERS probe modified mercaptophenyl boronic acid, form sandwich structure composed by antibacterial peptide function magnetic nano-particle-bacterium-SERS probe, the sandwich structure is separated under the action of external magnetic field, detection and identification is carried out to different bacterium by SERS.
Description
Technical field
The invention belongs to technical field of biological, and in particular to a kind of sandwich structure SERS detection pathogen
Method.
Background technique
Bacteria Detection is a key areas of relationship public safety.The method for being conventionally used to Bacteria Detection has plate count
Method, polymerase chain reaction (PCR), Enzyme-linked Immunosorbent Assay (ELSA) etc., they are as sensitive, efficient microorganism quantitative approach
It is widely used.But there is the Bacteria Culture time for needing to grow very much, special sample pre-treatments, poor anti jamming capability in them
The disadvantages of, therefore, quick, high sensitivity, high specificity method of detecting bacterium is developed, is of great significance.
Surface enhanced Raman spectroscopy (SERS) has many advantages, such as that quick, efficient, specificity is high in Bacteria Detection, due to not
Commaterial has its distinctive finger-print, and SERS is also good qualitative tool simultaneously.
When using SERS detection bacterium, the unit for specificity crawl bacterium is the pass for carrying out bacterium and effectively detecting
Key.Unit currently used for grabbing bacterium includes antibody, aptamers, antibiotic, phenyl boric acid etc..Antibody has very high special
Property, but the price of antibody is high, stability is low；The high stability of aptamers, specificity energy that is also high, but screening at present
Aptamers type for specificity capture bacterium is less；The price of antibiotic is low, and stability is high, energy specific adsorption bacterium,
But antibiotic is small-molecule substance, it is less to the recognition site of bacterium.Antibacterial peptide has price low, the stable advantage of property,
Also there is specific adsorption ability to bacterium, compared to antibiotic, antibacterial peptide due to be long-chain polypeptide, grab bacterium when have
More recognition sites.But how the bacterium of crawl is separated and identified, it is still necessary to design new inspection policies.
Summary of the invention
In place of the above shortcoming and defect of the existing technology, the purpose of the present invention is to provide a kind of sandwich folders
The method of core structure SERS detection pathogen.The method of the present invention passes through antibacterial peptide function magnetic nano-particle and graphite oxide
Alkene-silver gold-covered nano compound collaboration composition sandwich structure is used for the identification and detection of pathogen.
The object of the invention is achieved through the following technical solutions：
A kind of method of sandwich structure SERS detection pathogen, includes the following steps：
(1) preparation of antibacterial peptide function magnetic nano-particle：
Magnetic nano-particle is subjected to condensation reaction with antibacterial peptide after surface carboxylation is modified, obtains antibacterial peptide function
Magnetic nano-particle；
(2) to graphene oxide-silver gold-covered nano compound SERS probe preparation of mercaptophenyl boronic acid modification：
Graphene oxide is obtained into graphene oxide-silver with silver-colored gold-covered nano particle hybrid reaction after sulfhydrylation is modified
Gold-covered nano compound obtains the graphene oxide-silver modified mercaptophenyl boronic acid then and to mercaptophenyl boronic acid hybrid reaction
Gold-covered nano compound SERS probe；
(3) sandwich structure SERS detects pathogen：
(a) antibacterial peptide function magnetic nano-particle and bacterium are incubated for, will have been adsorbed under external magnetic field thin
The magnetic nano-particle of bacterium is separated, and magnetic nano-particle-bacterial complex is obtained；
(b) by magnetic nano-particle-bacterial complex of step (a) and to the graphene oxide-of mercaptophenyl boronic acid modification
Silver-colored gold-covered nano compound SERS probe is incubated for altogether, forms antibacterial peptide function magnetic nano-particle-bacterium-SERS probe institute group
At sandwich structure, the sandwich structure is separated under the action of external magnetic field；
(c) the isolated sandwich structure of step (b) is subjected to detection and identification to different bacterium by SERS.
Further, magnetic nano-particle described in step (1) refers to the nanoparticle with superparamagnetic, including but unlimited
In Fe3O4、MnFe2O4Deng.
Further, the method for the modification of surface carboxylation described in step (1) is as follows：
By magnetic nano-particle and ethyl orthosilicate (TEOS) hybrid reaction, silanization magnetic nano-particle is obtained, then
With three sodium alkoxide hybrid reaction of carboxyethylsilane, the magnetic nano-particle of surface carboxylation modification is obtained.
Further, antibacterial peptide described in step (1) includes that bacitracin, bombesin, polymyxin e, Xenopus laevis element etc. have
The antibacterial peptide of bacterium recognition capability.
Further, the method for the modification of sulfhydrylation described in step (2) is as follows：
Graphene oxide is distributed in dehydrated alcohol, 1- ethyl -3- (3- dimethylaminopropyl) phosphinylidyne is then added
Diimine (EDC) condensing agent is added mercaptoethylmaine reaction, obtains sulfhydrylation graphene oxide after being uniformly mixed.
Further, silver gold-covered nano particle described in step (2) is prepared via a method which to obtain：
By HAuCl under stirring condition4It is added in the ultrapure water of boiling, sodium citrate is then added and is stirred to react, obtains
The dispersion liquid of gold nanoparticle is cooled to room temperature, and ascorbic acid solution is then added and is uniformly mixed, AgNO is added dropwise3Solution
It is stirred to react, obtains silver-colored gold-covered nano particle.
Further, the mass ratio of silver gold-covered nano particle described in step (2) and graphene oxide is (0.5~15):
1.More preferably 10:1.
Detection method of the invention has the following advantages that and beneficial effect：
(1) modified antimicrobial peptide has price low, property is stablized, the advantage more than binding site for capturing bacterium；
(2) magnetic nano-particle is combined, can be used for the enrichment and removal of impurities of sample, reach higher detection sensitivity；
(3) silver-colored gold-covered nano particle is integrated to above graphene oxide, plays the role of protection to silver-colored gold filled, prevents
Silver-colored gold filled is oxidized, and graphene oxide increases the adsorbance of 4-MPBA；
(4) after being integrated to 4-MPBA using variety classes bacterium, specific variations are had occurred in the Raman peaks of 4-MPBA, right
Variety classes bacterium realizes differentiation and identifies.
Detailed description of the invention
Fig. 1 is the synthetic method and characterization of bacitracin functionalized magnetic nano particle in the embodiment of the present invention：(A) bacitracin
Functional magnetic Fe3O4The synthesis step schematic diagram of NPs；(B) magnetic Fe3O4The TEM of NPs schemes；(C) silanization magnetic Fe3O4NPs
TEM figure；(D) magnetic Fe3O4NPs, silanization magnetic Fe3O4NPs and bacitracin functional magnetic Fe3O4The hysteresis loop of NPs
Figure；(E) magnetic Fe3O4NPs (a), silanization magnetic Fe3O4NPs (b), bacitracin functional magnetic Fe3O4The infrared light of NPs (c)
Spectrogram；(F) bacitracin functional magnetic Fe3O4The figure of NPs under an optical microscope；(G) bacitracin functional magnetic Fe3O4NPs
It is incubated for Escherichia coli, the figure after Magneto separate under an optical microscope.
Fig. 2 is the synthetic method and characterization of 4-MPAB-GO-Au@Ag SERS tags in the embodiment of the present invention：(A)4-
The synthesis step schematic diagram of MPBA-GO-Au@Ag SERS tags；(B) be respectively from left to right Au@AgNPs, Au@AgNPs and
The outside drawing of HS-GO mixing 5min, Au@AgNPs and HS-GO mixing 10min；(C~G) different proportion Au@AgNs and HS-GO knot
Close later TEM figure.
Fig. 3 is the test result figure of graphene oxide enhancing substrate stability in the embodiment of the present invention.
Fig. 4 is that three kinds of pathogenic bacteria (Pseudomonas aeruginosa Escherichia coli are detected and identified in the embodiment of the present invention
(p.aeruginosa), staphylococcus aureus (S.aureus), Escherichia coli (E.coil)) process principle and result figure.
The formation basic theory and result figure of sandwich structure in Fig. 5 embodiment of the present invention：(A) in the presence of there is no bacterium,
Sandwich structure cannot be formed, SERS tags cannot be come out by Magneto separate；(B) in the presence of there is no bacterium, Raman microscope
The magnetic Fe of black can only be observed down3O4Aggregate；(C) with the presence of bacterium when, sandwich structure is formd,
SERS tags is come out by Magneto separate；(D) with the presence of bacterium when, the substance of right metallic luster is observed under Raman microscope
Occur, because there is the introducing of SERS tags；(E) TEM of simple Escherichia coli；(F) Escherichia coli form sandwich structure
TEM afterwards.
SERS identifies the SERS figure of three kinds of variety classes pathogenic bacteria (a) 4-MPBA in Fig. 6 embodiment of the present invention；(b) green pus
Bacillus SERS figure；(c) staphylococcus aureus SERS schemes；(d) Escherichia coli SERS schemes；(e) bacitracin functionalization-Fe3O4SERS
Figure.
PCA is combined to identify the map and PCA result figure of three kinds of variety classes pathogenic bacteria in Fig. 7 embodiment of the present invention：(A)4-
MPBA；(B) staphylococcus aureus；(C) Pseudomonas aeruginosa；(D) Escherichia coli；(E) PCA result.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.
Embodiment
(1) preparation of antibacterial peptide function magnetic nano-particle：
(a) magnetic ferroferric oxide nano-particles (Fe3O4NPs synthesis)：By 1.35g FeCl3·6H2O is added to
In 40mL ethylene glycol, stirring forms uniform solution.Continuously add 1.0g polyethylene glycol (number-average molecular weight 10000) and
3.6g NaAc·6H2O, stirring is to being completely dissolved.Gained mixture is transferred in 100mL autoclave, at 200 DEG C
Lower heating 8h.It will be washed respectively 3 times after products therefrom Magneto separate with ethyl alcohol and deionized water, obtain magnetic Fe3O4NPs。
(b) magnetic Fe3O4NPs carries out silylation modification (package ethyl orthosilicate TEOS)：Take 3mL 7.5mg/mL
Fe3O4NPs solution (is scattered in 1.2wt.% ammonium hydroxide), and 200 μ L TEOS, ultrasonic 90min are added, continuously add 10 μ L TEOS, surpasses
Sound 90min will be washed respectively 3 times with ethyl alcohol and deionized water after products therefrom Magneto separate, obtain silanization magnetic Fe3O4NPs。
(c) silanization magnetic Fe3O4The carboxylated of NPs is modified：Take the silanization magnetic Fe of 1mL 10mg/mL3O4NPs is (molten
Solution is in 10mM phosphate buffer solution, pH=7.4), 80 μ L carboxyethylsilane, three sodium alkoxide is added, mixes 4h.Products therefrom exists
It is separated under magnetic field, is washed respectively 3 times with ethyl alcohol and deionized water, obtain carboxylated magnetic Fe3O4NPs。
(d) bacitracin functional magnetic Fe3O4The preparation of NPs：Take 10mg carboxylated magnetic Fe3O4NPs is added to 25mL
In PBS (pH=7.4) solution, 16.5mg EDC and 10g n-hydroxysuccinimide (NHS) are added, mixes 4h.Then add
Enter 20mg bacitracin (bacitracin), continuess to mix 3h.After products therefrom Magneto separate, distinguished with ethyl alcohol and deionized water
Washing 3 times, obtains bacitracin functional magnetic Fe3O4NPs。
Referring to attached drawing 1, A is bacitracin functional magnetic Fe3O4The synthesis step schematic diagram of NPs；B is naked obtained by step (a)
The magnetic Fe of dew3O4Transmission electron microscope (TEM) figure of NPs, it can be seen that prepared magnetic Fe3O4NPs diameter is about in 440nm；
C is silanization magnetic Fe obtained by step (b)3O4The TEM of NPs schemes, it can be seen that after modifying TEOS, magnetic
Fe3O4A thin layer of polymer has been wrapped up on the surface of NPs, forms core-shell structure；D is magnetic Fe3O4NPs(Fe3O4), silanization
Magnetic Fe3O4NPs(Fe3O4@SiO2) and bacitracin functional magnetic Fe3O4NPs(Fe3O4@SiO2@bacitracin) magnetic hysteresis
Loop line figure, it can be seen that exposed magnetic Fe3O4NPs has good superparamagnetism, after silylation modification, superparamagnetism
It decreases, continues to modify after bacitracin, and slightly lower, but generally, still there is preferable superparamagnetic
Property, it can be used for Magneto separate；E is exposed magnetic Fe3O4NPs (a), silanization magnetic Fe3O4What NPs (b) and bacitracin were modified
Magnetic Fe3O4The infrared spectrogram of NPs (c), it can be seen that magnetic Fe3O4NPs is infrared after silanization and modification bacitracin
Spectrogram all shows certain variation；F is exposed magnetic Fe3O4NPs and Escherichia coli are incubated for, and Magneto separate exists later
It is observed under optical microscopy, it can be seen that magnetic Fe3O4The aggregate of NPs is high-visible, but has not seen depositing for bacterium
?；G is bacitracin functional magnetic Fe3O4NPs and Escherichia coli are incubated for, and Magneto separate carries out under an optical microscope later
Observation has white point (Escherichia coli) occur, illustrates magnetic Fe compared to F, G3O4NPs is modified after bacitracin, can be fine
Ground grabs Escherichia coli.
(2) to graphene oxide-silver gold-covered nano compound SERS probe (4-MPBA-GO- of mercaptophenyl boronic acid modification
Au@Ag SERS tag) preparation：
(a) synthesis of silver-colored gold-covered nano particle (Au@AgNPs)：In the case where magnetic agitation, by HAuCl4(0.1M) adds
Enter in the ultrapure water to boil to 50mL, it is to be mixed uniformly after, sodium citrate (1% weight) solution of 0.75mL is added to above-mentioned
In boiling solution；Continue stirring 30 minutes, the dispersion liquid for obtaining a claret is the dispersion liquid of gold nanoparticle, then will
Dispersion liquid is cooled to room temperature, is filtered with 0.22 micron of miillpore filter, collects filtrate；The filtrate and anti-bad for taking 10mL to obtain
Hematic acid (0.1M) solution, is sufficiently mixed under magnetic stirring.Magnetic agitation is kept, at room temperature by 1mM AgNO3Dropwise, every
A drop is added dropwise within 30 seconds, is added in above-mentioned solution, continues stirring 30 minutes after dripping, obtains Au AgNPs solution.
(b) preparation of sulfhydrylation graphene oxide：100mg graphene oxide (GO) is taken to be added in 50mL ethyl alcohol, ultrasound
60min forms the GO alcohol dispersion liquid of 2mg/mL.Then, 1.9g EDC condensing agent is added, stirs 12h, makes the carboxyl on the surface GO
Sufficiently activation is added 50mL mercaptoethylmaine (1mM), stirs 4h.Products therefrom is centrifuged at 9000rpm, is washed with deionized water
Extra mercaptoethylmaine is removed, sulfhydrylation graphene oxide (HS-GO) has been obtained.
(c) preparation of graphene oxide-silver gold-covered nano compound (GO-Au@Ag NPs)：10mL Au@AgNPs is molten
Liquid is added in 2mL HS-GO (0.1mg/mL) dispersion liquid, stirs 2h, products therefrom is centrifuged at 9000rpm, uses deionization
Water washing 2 times, obtain GO-Au@Ag NPs.
(d) preparation of 4-MPBA-GO-Au@Ag SERS tag：Take 6mL to mercaptophenyl boronic acid (4-MPBA, 0.01mg/mL)
Solution and the obtained GO-Au@Ag NPs of step (c) carry out mixing 4h.Product is centrifuged at 9000rpm, uses deionized water
Washing 2 times, obtains 4-MPBA-GO-Au@Ag SERS tag.
Referring to attached drawing 2, A is the synthesis step schematic diagram of 4-MPBA-GO-Au@Ag；B is from left to right Au@Ag respectively
The outside drawing of NPs, Au@Au NPs and HS-GO mixing 5min, Au@Au NPs and HS-GO mixing 10min, it can be seen that Au@Au
NPs is added to after HS-GO, since the HS on GO has adsorbed Au@Ag NPs soon, so the color of Au@AgNPs is in short-term
It is interior to have changed a lot；C~G is that different quality ratio Au@AgNPs/GO (is 0.5 respectively:1,1:1,5:1,10:1,
15:1) TEM of the GO-Au@Ag NPs being prepared schemes, on figure as it can be seen that with Au@AgNPs amount increase, adsorb on GO lamella
Au@AgNPs gradually increase, but when ratio is more than 10:When 1, since Au@AgNPs largely exists, make on GO lamella
Au@AgNPs largely reunites, so 10:1 ratio is optimal.
Referring to attached drawing 3, the stability of Au@Ag NPs and GO-Au@Ag NPs compared, A and B are GO-Au@Ag respectively
NPs and Au@AgNPs stored in 4 DEG C 0 day, 7 days, 60 days after to the SERS reinforcing effect of R6G, as it can be seen that having existing for GO on figure
When, even if substrate reinforcing effect still has good SERS activity after storage 60 days, without GO there are when,
The reinforcing effect of substrate just significantly decreases after 7 days.C and D be respectively GO-Au@Ag NPs and Au@Ag NPs in the sun
To the SERS reinforcing effect of R6G after irradiation 0 hour, 12 hours, 72 hours, as it can be seen that when with the presence of GO on figure, substrate enhances
Effect has no significant effect under light illumination, and when depositing without GO, the SERS activity of substrate is significantly reduced.Illustrate GO in stabilization
Great role is played in substrate SERS activity.
(3) three kinds of pathogenic bacteria (Pseudomonas aeruginosa Escherichia coli of sandwich structure SERS detection and identification
(p.aeruginosa), staphylococcus aureus (S.aureus), Escherichia coli (E.coil))：
(a) by bacitracin functional magnetic Fe3O4NPs and the actual sample of bacterium are mixed, and 1h are incubated for, under magnetic field
By bacterium/magnetic Fe3O4NPs conjugate is separated, and unbonded bacterium is washed away with PBS, obtains magnetic nano-particle-bacterium
Compound；
(b) 4-MPBA-GO-Au@Ag SERS is added into step (a) gained magnetic nano-particle-bacterial complex
Tag is incubated for 1h, the SERS tag/ bacterium/magnetic Fe that will be obtained3O4NPs sandwich structure Magneto separate, is washed away more with PBS
Remaining SERS tag is detected for subsequent SERS.
(c) for different types of bacterium, the corresponding finger-print detected by SERS can be identified.
The process principle and result of three kinds of pathogenic bacteria of the detection of this step and identification are as shown in Fig. 4.
With reference to attached drawing 5, A and B indicate no bacterium there are when, since sandwich structure cannot be formed,
SERS tag cannot be come out by Magneto separate, and the Fe of black has been only seen under Raman microscope3O4Aggregate；C and D expression is having
Bacterium there are when, due to foring sandwich structure, SERS tags is come out by Magneto separate, under Raman microscope
There is the substance for having metallic luster, is caused by SERS tag；E is the TEM figure of simple Escherichia coli, and F is to form
The TEM of sandwich structure schemes, it can be seen that the adsorption of Escherichia coli has Fe3O4With GO-Au@Ag.
It is the result that sandwich structure strategy SERS identifies three kinds of pathogenic bacteria with reference to attached drawing 6.It is SERS respectively
Tags (a), Pseudomonas aeruginosa Escherichia coli (b), staphylococcus aureus (c), Escherichia coli (d), bacitracin functionalization Fe3O4NPs
(e) SERS map.As it can be seen that different types of bacterium presents different finger-print respectively on figure, can reflect easily
It does not come out.
With reference to attached drawing 7, it is to acquire the map of a large amount of bacterium using discriminant analysis (PCA), the result of identification is carried out
Statistical analysis, as it can be seen that being SERS tags (A), staphylococcus aureus (B), Pseudomonas aeruginosa Escherichia coli respectively on figure on figure
(C), Escherichia coli (D).E is PCA analysis as a result, figure is upper as it can be seen that three kinds of bacteriums can separate well, and illustrating can be very
Identify three kinds of different types of bacteriums well.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (8)
1. a kind of method of sandwich structure SERS detection pathogen, it is characterised in that include the following steps：
(1) preparation of antibacterial peptide function magnetic nano-particle：
Magnetic nano-particle is subjected to condensation reaction with antibacterial peptide after surface carboxylation is modified, obtains antibacterial peptide functionization magnetism
Nanoparticle；
(2) to graphene oxide-silver gold-covered nano compound SERS probe preparation of mercaptophenyl boronic acid modification：
It is covered with gold leaf to be obtained into graphene oxide-silver with silver-colored gold-covered nano particle hybrid reaction after sulfhydrylation is modified for graphene oxide
It is covered with gold leaf to obtain the graphene oxide-silver modified mercaptophenyl boronic acid then and to mercaptophenyl boronic acid hybrid reaction for nano-complex
Nano-complex SERS probe；
(3) sandwich structure SERS detects pathogen：
(a) antibacterial peptide function magnetic nano-particle and bacterium are incubated for, bacterium will have been adsorbed under external magnetic field
Magnetic nano-particle is separated, and magnetic nano-particle-bacterial complex is obtained；
(b) by magnetic nano-particle-bacterial complex of step (a) and to the graphene oxide-Yin Bao of mercaptophenyl boronic acid modification
Golden nano-complexes SERS probe is incubated for altogether, is formed composed by antibacterial peptide function magnetic nano-particle-bacterium-SERS probe
Sandwich structure separates the sandwich structure under the action of external magnetic field；
(c) the isolated sandwich structure of step (b) is subjected to detection and identification to different bacterium by SERS.
2. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that：Step
Suddenly magnetic nano-particle described in (1) refers to Fe3O4Or MnFe2O4。
3. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that step
Suddenly the method for the modification of surface carboxylation described in (1) is as follows：
By magnetic nano-particle and ethyl orthosilicate hybrid reaction, obtain silanization magnetic nano-particle, then with carboxy ethyl
Silantriol sodium salt hybrid reaction obtains the magnetic nano-particle of surface carboxylation modification.
4. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that：Step
Suddenly antibacterial peptide described in (1) refers to bacitracin, bombesin, polymyxin e or Xenopus laevis element.
5. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that step
Suddenly the method for the modification of sulfhydrylation described in (2) is as follows：
Graphene oxide is distributed in dehydrated alcohol, it is sub- that 1- ethyl -3- (3- dimethylaminopropyl) phosphinylidyne two is then added
Amine condensing agent is added mercaptoethylmaine reaction, obtains sulfhydrylation graphene oxide after being uniformly mixed.
6. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that step
Suddenly silver gold-covered nano particle described in (2) is prepared via a method which to obtain：
By HAuCl under stirring condition4It is added in the ultrapure water of boiling, sodium citrate is then added and is stirred to react, obtains gold nano
The dispersion liquid of particle is cooled to room temperature, and ascorbic acid solution is then added and is uniformly mixed, AgNO is added dropwise3Solution stirring is anti-
It answers, obtains silver-colored gold-covered nano particle.
7. a kind of method of sandwich structure SERS detection pathogen according to claim 1, it is characterised in that：Step
Suddenly the mass ratio of silver gold-covered nano particle and graphene oxide described in (2) is (0.5~15):1.
8. a kind of method of sandwich structure SERS detection pathogen according to claim 7, it is characterised in that：Institute
The mass ratio for stating silver-colored gold-covered nano particle and graphene oxide is 10:1.
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|---|---|---|---|---|
| CN110531075A (en) * | 2019-10-16 | 2019-12-03 | 军事科学院军事医学研究院军事兽医研究所 | Based on antibacterial peptide-phosphate nano compound pathogenic bacteria immunoassay biosensor and its preparation method and application |
| CN110927372A (en) * | 2019-11-26 | 2020-03-27 | 军事科学院军事医学研究院军事兽医研究所 | Colorimetric immunosensor based on double nano-composites and preparation method and application thereof |
| CN113092441A (en) * | 2021-04-08 | 2021-07-09 | 吉林大学 | Ultrasensitive biochip based on surface enhanced Raman scattering and preparation method thereof |
| CN113624736A (en) * | 2021-07-07 | 2021-11-09 | 张鑫 | Raman effect-based method for rapidly detecting concentration of microorganisms |
| CN114888276A (en) * | 2022-05-12 | 2022-08-12 | 集美大学 | Preparation method and application of chain-like high polymer modified nano particles |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102811943A (en) * | 2009-12-11 | 2012-12-05 | 韩国化学研究院 | Heterodimer Core-shell Nanoparticle In Which Raman-active Molecules Are Located At A Binding Portion Of A Nanoparticle Heterodimer, Use Thereof, And Method For Preparing Same |
| CN106442987A (en) * | 2016-09-30 | 2017-02-22 | 西南大学 | Staphylococcus-aureus fluorescence detection kit and application method thereof |
| CN106645090A (en) * | 2017-01-11 | 2017-05-10 | 华南师范大学 | Novel SERS substrate-based method for quantitatively testing pathogenic bacteria |
| CN106706910A (en) * | 2017-01-05 | 2017-05-24 | 江苏大学 | P-aminothiophenol modified gold nanorod-based food-borne pathogen detection method |
| KR20170131864A (en) * | 2016-05-23 | 2017-12-01 | 고려대학교 산학협력단 | Method for Preparing Porous SERS Substrate Using Cystein-rich Peptide Phage Display |
| CN107941779A (en) * | 2017-09-30 | 2018-04-20 | 中国人民解放军总医院 | The method of gold shell magnetic nanoparticles and nanometer label surface enhancing Raman scattering detection bacterium based on vancomycin modification |
-
2018
- 2018-08-22 CN CN201810958582.2A patent/CN108872194B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102811943A (en) * | 2009-12-11 | 2012-12-05 | 韩国化学研究院 | Heterodimer Core-shell Nanoparticle In Which Raman-active Molecules Are Located At A Binding Portion Of A Nanoparticle Heterodimer, Use Thereof, And Method For Preparing Same |
| KR20170131864A (en) * | 2016-05-23 | 2017-12-01 | 고려대학교 산학협력단 | Method for Preparing Porous SERS Substrate Using Cystein-rich Peptide Phage Display |
| CN106442987A (en) * | 2016-09-30 | 2017-02-22 | 西南大学 | Staphylococcus-aureus fluorescence detection kit and application method thereof |
| CN106706910A (en) * | 2017-01-05 | 2017-05-24 | 江苏大学 | P-aminothiophenol modified gold nanorod-based food-borne pathogen detection method |
| CN106645090A (en) * | 2017-01-11 | 2017-05-10 | 华南师范大学 | Novel SERS substrate-based method for quantitatively testing pathogenic bacteria |
| CN107941779A (en) * | 2017-09-30 | 2018-04-20 | 中国人民解放军总医院 | The method of gold shell magnetic nanoparticles and nanometer label surface enhancing Raman scattering detection bacterium based on vancomycin modification |
Non-Patent Citations (2)
| Title |
|---|
| JUNFENG WANG等: "Facile Synthesis of Au-Coated Magnetic Nanoparticles and Their Application in Bacteria Detection via a SERS Method", 《ACS APPL. MATER. INTERFACES》 * |
| 刘淑文等: "海水中致病菌检测研究进展", 《海洋科学》 * |
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|---|---|---|---|---|
| CN110531075A (en) * | 2019-10-16 | 2019-12-03 | 军事科学院军事医学研究院军事兽医研究所 | Based on antibacterial peptide-phosphate nano compound pathogenic bacteria immunoassay biosensor and its preparation method and application |
| CN110531075B (en) * | 2019-10-16 | 2022-06-14 | 军事科学院军事医学研究院军事兽医研究所 | Pathogenic bacteria immunoassay biosensor based on antibacterial peptide-phosphate nano compound and preparation method and application thereof |
| CN110927372A (en) * | 2019-11-26 | 2020-03-27 | 军事科学院军事医学研究院军事兽医研究所 | Colorimetric immunosensor based on double nano-composites and preparation method and application thereof |
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