CN109536571B - Nano biological probe for detecting pathogenic bacteria and preparation method thereof - Google Patents
Nano biological probe for detecting pathogenic bacteria and preparation method thereof Download PDFInfo
- Publication number
- CN109536571B CN109536571B CN201811217049.7A CN201811217049A CN109536571B CN 109536571 B CN109536571 B CN 109536571B CN 201811217049 A CN201811217049 A CN 201811217049A CN 109536571 B CN109536571 B CN 109536571B
- Authority
- CN
- China
- Prior art keywords
- ferroferric oxide
- silicon dioxide
- particles
- pathogenic bacteria
- biological probe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
-
- 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
- G01N33/535—Production of labelled immunochemicals with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56911—Bacteria
- G01N33/56938—Staphylococcus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/245—Escherichia (G)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/305—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F)
- G01N2333/31—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F) from Staphylococcus (G)
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a nano biological probe for detecting pathogenic bacteria and a preparation method thereof, wherein the nano biological probe is formed by coupling a magnetic core-shell nano structure and a specific recognition molecule aiming at the pathogenic bacteria, the magnetic core-shell nano structure takes ferroferric oxide particles as a core and a silicon dioxide layer embedded with platinum nano particles as a shell layer, the whole nano biological probe is negatively charged, and the nano biological probe has the activity of a simulated peroxidase. The nano biological probe provided by the invention has the function of simulating the activity of peroxidase, and can be used for quickly detecting whether pathogenic bacteria exist by using a colorimetric method. The preparation method provided by the invention is simple in process, strong in operability and environment-friendly, and the prepared nano biological probe can achieve the purpose and effect of rapidly detecting pathogenic bacteria and has a good application prospect.
Description
Technical Field
The invention relates to the field of novel nano material preparation, in particular to a nano biological probe for detecting pathogenic bacteria and a preparation method thereof.
Background
Pathogenic bacteria are microorganisms that cause diseases, also called pathogenic microorganisms, and include bacteria, viruses, spirochetes, rickettsiae, chlamydia, mycoplasma, fungi, actinomycetes, and the like.
Pathogenic bacteria not only seriously damage human health but also have a great impact on economy, and have become one of the most prominent public health problems internationally. The traditional method for detecting pathogenic bacteria (such as escherichia coli, staphylococcus aureus and the like) firstly collects a sample for enrichment culture, so that the number of detected bacteria reaches a detectable level in the sample, and then observation and a series of physiological and biochemical characteristic identification can be carried out according to morphological characteristics, the process is long in time consumption and generally needs 3-5 days. Therefore, there is an urgent need for a rapid, specific and sensitive method to detect pathogens, particularly for online, real-time monitoring.
Currently, methods for rapidly detecting pathogenic bacteria include small biochemical tests, immunological tests, nucleic acid probe-based methods, polymerase chain reaction, and the like. However, the above method also has problems of complicated equipment, false positive, etc., and requires operations such as early enrichment of bacteria, etc., and thus cannot fully satisfy the market demand.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a nano biological probe for detecting pathogenic bacteria and a preparation method thereof.
The invention provides a nano biological probe for detecting pathogenic bacteria, which is formed by coupling a magnetic core-shell nano structure and a specific recognition molecule aiming at the pathogenic bacteria, wherein the magnetic core-shell nano structure takes ferroferric oxide particles as a core and a silicon dioxide layer embedded with platinum particles as a shell, and the whole nano biological probe is negatively charged and has the activity of simulated peroxidase.
In the technical scheme, the magnetic core-shell nanostructure in the nano biological probe has superparamagnetism and simulated peroxidase activity, the nano biological probe firstly enriches pathogenic bacteria to be detected through the superparamagnetism, then the nano biological probe is used for simulating the peroxidase activity, hydrogen peroxide and TMB can be added into a system to be detected, qualitative or quantitative analysis of the pathogenic bacteria is carried out according to the existence of the final color based on an ELISA method, and the nano biological probe has the advantages of rapidness and high sensitivity, is particularly suitable for online real-time monitoring of the pathogenic bacteria, and has a good application prospect.
Preferably, the size of the magnetic core-shell nano structure is 30-50 nm, wherein the size of ferroferric oxide particles is 10-20 nm, the thickness of a silicon dioxide layer is 5-10 nm, and the size of platinum particles is 2-3 nm.
Preferably, the specific recognition molecule is one or more of an antibody, a targeting polypeptide, a nucleic acid aptamer and an antibiotic.
The invention also provides a preparation method of the nano biological probe, which comprises the following steps:
(1) Coating silicon dioxide on the ferroferric oxide in the organic phase by using a reverse microemulsion method to prepare ferroferric oxide-silicon dioxide particles;
(2) Performing amination modification on the ferroferric oxide-silicon dioxide particles to ensure that the surfaces of the ferroferric oxide-silicon dioxide particles are positively charged;
(3) Loading platinum particles on the surface of the positively charged ferroferric oxide-silicon dioxide particles through electrostatic adsorption to prepare a magnetic core-shell nano structure;
(4) And coupling the magnetic core-shell nano structure with a specific recognition molecule aiming at pathogenic bacteria.
The preparation method has the advantages of simple process, strong operability and environmental friendliness, and the prepared nano biological probe can achieve the purpose and effect of quickly detecting pathogenic bacteria.
Preferably, the step (1) specifically comprises: and dispersing ferroferric oxide dispersed in an organic phase in a polyethylene glycol octyl phenyl ether/n-hexanol/water mixed solution to form a reverse microemulsion system, and then adding ammonia water and tetraethoxysilane to hydrolyze the tetraethoxysilane in an alkaline environment to form a silicon dioxide coating layer.
Preferably, the ferroferric oxide is prepared by a solvothermal method, and the concentration of the ferroferric oxide dispersed in cyclohexane is 1-10 mg/mL, more preferably 1mg/mL.
Preferably, the molar ratio of the polyethylene glycol octyl phenyl ether to the n-hexanol to the water in the polyethylene glycol octyl phenyl ether/n-hexanol/water mixed solution is 1.
Preferably, the mass ratio of the ethyl orthosilicate, the ferroferric oxide and the ammonia water is 1.
Preferably, the step (2) specifically comprises: mixing the ferroferric oxide-silicon dioxide particles and 3-aminopropyltrimethoxy siloxane, and performing amination modification, wherein the mass ratio of the ferroferric oxide-silicon dioxide particles to the 3-aminopropyltrimethoxy siloxane is 1.
Preferably, the mass ratio of the positively charged ferroferric oxide-silica particles to the platinum particles in the step (3) is (1-1), preferably (10).
Preferably, the coupling method in step (4) is a glutaraldehyde spacer method or an EDC/NHS method, and more preferably a glutaraldehyde spacer method.
As a preferred embodiment, the preparation method comprises the following steps:
(1) Dispersing ferroferric oxide dispersed in an organic phase in a polyethylene glycol octyl phenyl ether/n-hexanol/water mixed solution to form a reverse microemulsion system, adding ammonia water and tetraethoxysilane to hydrolyze the tetraethoxysilane in an alkaline environment to form a silicon dioxide coating layer, and preparing ferroferric oxide-silicon dioxide particles;
(2) Mixing the ferroferric oxide-silicon dioxide particles and 3-aminopropyltrimethoxy siloxane, and performing amination modification to ensure that the surface of the mixture is positively charged;
(3) Loading platinum particles on the surface of the positively charged ferroferric oxide-silicon dioxide particles through electrostatic adsorption, wherein the mass ratio of the platinum particles to the positively charged ferroferric oxide-silicon dioxide particles is (1-1);
(4) And coupling the magnetic core-shell nano structure with a specificity recognition molecule aiming at pathogenic bacteria by adopting a glutaraldehyde spacer arm method.
The invention also aims to provide the application of the nano biological probe in the preparation of a pathogenic bacterium detection kit.
The nano biological probe provided by the invention has the function of simulating the activity of peroxidase, and can be used for quickly detecting whether pathogenic bacteria exist by using a colorimetric method. The preparation method provided by the invention is simple in process, strong in operability and environment-friendly, can achieve the purpose and effect of rapidly detecting pathogenic bacteria, and has a good application prospect.
Drawings
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 introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 shows Fe prepared in example 1 3 O 4 TEM image of @ Si-Pt;
FIG. 2 shows Fe prepared in example 1 3 O 4 Results of a simulated peroxidase activity assay of @ Si-Pt;
FIG. 3 shows Fe obtained in example 2 3 O 4 TEM image of @ Si-Pt;
FIG. 4 shows Fe obtained in example 3 3 O 4 TEM image of @ Si-Pt;
FIG. 5 shows Fe obtained in example 4 3 O 4 TEM image of @ Si;
FIG. 6 is a TMB absorption spectrum of Staphylococcus aureus of application example 1 at different concentrations;
FIG. 7 is a plot of the linear relationship between TMB coloration and Staphylococcus aureus counts in application example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.
Example 1
The embodiment provides a nano biological probe for detecting staphylococcus aureus, which is formed by coupling a magnetic core-shell nano structure and a specific antibody aiming at staphylococcus aureus, wherein the magnetic core-shell nano structure takes ferroferric oxide particles as a core and a silicon dioxide layer embedded with platinum particles as a shell, and the whole nano biological probe is negatively charged and has the activity of simulated peroxidase. The preparation method comprises the following steps:
(1) Dispersing ferroferric oxide nanoparticles (1 mg/mL,10 mL) dispersed in cyclohexane in 10mL of polyethylene glycol octyl phenyl ether (Triton X-100)/n-hexanol/water mixed solution (Triton X-100, n-hexanol, water molar ratio of 1;
(2) Mixing ferroferric oxide-silicon dioxide particles (1 mg/mL,5 mL) and 3-aminopropyl trimethoxy siloxane (APTMS, 200 mu L), and performing amination modification for 14h to ensure that the surface of the mixture is positively charged;
(3) Mixing and reacting positively charged ferroferric oxide-silicon dioxide particles (1 mg/mL,5 mL) and negatively charged platinum nanoparticles (1 mg/mL,0.5 mL) for 14h, and loading the platinum nanoparticles on the surfaces of the ferroferric oxide-silicon dioxide particles through electrostatic adsorption to obtain a magnetic core-shell nanostructure which takes the ferroferric oxide particles as cores and a silicon dioxide layer embedded with the platinum particles as a shell and is marked as Fe 3 O 4 @Si-Pt；
(4) Using glutaraldehyde spacer method to mix Fe 3 O 4 And (3) coupling the @ Si-Pt with a specific antibody aiming at the staphylococcus aureus to obtain the target product.
FIG. 1 shows Fe prepared in this example 3 O 4 TEM image of @ Si-Pt. As can be seen from the figure, fe is present in this example 3 O 4 The grain size of the @ Si-Pt grains is 30-50 nm, wherein the size of the ferroferric oxide grains is 10-20 nm, the thickness of the silicon dioxide layer is 5-10 nm, the size of the platinum nano-particles is 2-3nm, fe 3 O 4 Particles of @ Si-PtThe particles are of a core-shell structure taking ferroferric oxide particles as a single core, and have uniform size and good dispersibility.
The Fe prepared in this example was analyzed 3 O 4 The simulated peroxidase activity of @ Si-Pt was performed as follows: selecting sodium acetate buffer (pH = 4) to prepare 1mL of reaction system, wherein 940. Mu.L of buffer, 20. Mu.L of hydrogen peroxide (initial concentration 50 mM), 20. Mu.L of TMB (initial concentration 50 mM), and Fe 3 O 4 @ Si-Pt 20. Mu.L (initial concentration 1 mg/mL), reacted for 30 minutes, and 200. Mu.L each was placed in a 96-well plate, and the absorption spectrum at 652nm was measured with a microplate reader.
As shown in FIG. 2, it can be seen that the amount of Fe in the solution is larger than that of hydrogen peroxide 3 O 4 @Si，Fe 3 O 4 The mimic peroxidase activity of @ Si-Pt is significantly enhanced, TMB has significant absorption at 652nm, and its mimic peroxidase activity is not significantly affected after modification of specific antibodies.
Example 2
This example provides a nanobioprobe for detecting staphylococcus aureus, which is different from example 1 in that: in the step (3), the positively charged ferroferric oxide-silicon dioxide particles (1 mg/mL,2 mL) and the negatively charged platinum nanoparticles (1 mg/mL,2 mL) are mixed and reacted for 14h.
FIG. 3 shows Fe prepared in this example 3 O 4 TEM image of @ Si-Pt. As can be seen from the figure, fe prepared in this example 3 O 4 The size of the @ Si-Pt particles is uniform, the platinum particles on the surface are more, but the aggregation phenomenon partially occurs, so that the overall particle morphology becomes non-uniform, and the later application in a biological system is not facilitated.
Example 3
This example provides a nano-biological probe for detecting staphylococcus aureus, which is different from example 1 in that: in the step (1), the molar ratio of Triton X-100, n-hexanol and water in the polyethylene glycol octyl phenyl ether (Triton X-100)/n-hexanol/water mixed solution is 1.
FIG. 4 shows Fe prepared in this example 3 O 4 TEM image of @ Si-Pt. As can be seen from the figure, the reverse microemulsion system formed in this example is not ideal, resulting in poor silica coating effect and formation of multi-core.
Example 4
This example provides a ferroferric oxide-silica particle (Fe) 3 O 4 @ Si) comprising: ferroferric oxide nanoparticles (1 mg/mL,10 mL) dispersed in cyclohexane are dispersed in 10mL of polyethylene glycol octyl phenyl ether (Triton X-100)/n-hexanol/water mixed solution (Triton X-100, n-hexanol, water molar ratio of 1.
FIG. 5 shows Fe prepared in this example 3 O 4 TEM image of @ Si. It can be seen from the figure that an excess of tetraethylorthosilicate results in a thickening of the silica coating, a relatively smaller ferroferric oxide core and the presence of empty silica spheres.
Example 5
The embodiment provides a nano biological probe for detecting escherichia coli, which is formed by coupling a magnetic core-shell nano structure and a specific antibody aiming at the escherichia coli, wherein the magnetic core-shell nano structure takes ferroferric oxide particles as a core and a silicon dioxide layer embedded with platinum particles as a shell layer, and the whole nano biological probe is negatively charged and has the activity of simulating peroxidase. The preparation method comprises the following steps:
(1) To (3) the same as in example 1;
(4) Using glutaraldehyde spacer method to mix Fe 3 O 4 Coupling the @ Si-Pt with a specific antibody aiming at the escherichia coli to obtain a target product.
Application example 1
The nano biological probe provided by the embodiment 1 is applied to the rapid detection of staphylococcus aureus based on an ELISA method, and comprises the following steps:
(1) Coupling 2% BSA and 50mg/mL vancomycin by an EDC/NHS method to serve as capture probes, and coating the capture probes at the bottom of a 96-well plate;
(2) Golden yellow colourStaphylococcal concentration gradient was designed as negative control, 10 2 、10 3 、10 4 And 10 5 cfu/mL；
(3) Adding 150 mu L of staphylococcus aureus into each hole, and shaking the mixture for 1 hour at 37 ℃ in a shaking table;
(4) Sucking out the bacterial liquid, and washing with PBS for three times;
(5) Adding 100 mu L of PBS and 20 mu L of the nano biological probe provided in the example 1 with the concentration of 1mg/mL into each hole, and incubating for 1 hour;
(6) The supernatant was aspirated and washed three times with PBS;
(7) mu.L of acetic acid buffer, 2. Mu.L of hydrogen peroxide and TMB (50 mM) were added to each well, and the absorbance at 652nm was measured after 1 hour.
The results are shown in FIG. 6, which is the TMB absorption spectrum of Staphylococcus aureus at different concentrations. A fitting graph of the linear relation between the TMB coloration and the number of staphylococcus aureus is obtained according to FIG. 6, and as shown in FIG. 7, it can be seen that the TMB coloration and the number of bacteria have a good linear relation, and the coloration becomes more obvious as the number of staphylococcus aureus increases.
Application example 2
The nano biological probe provided in application example 3 was used to detect staphylococcus aureus by the same detection method as in application example 1, and as a result, the linear relationship between TMB color development and staphylococcus aureus number was inferior to application example 1.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (2)
1. A nano biological probe for detecting pathogenic bacteria is characterized by being formed by coupling a magnetic core-shell nano structure and a specific recognition molecule aiming at the pathogenic bacteria, wherein the magnetic core-shell nano structure takes ferroferric oxide particles as a core and a silicon dioxide layer embedded with platinum particles as a shell, is integrally negatively charged and has the activity of simulated peroxidase; the specific recognition molecule is one or more of an antibody, a targeting polypeptide, a nucleic acid aptamer and an antibiotic;
the size of the magnetic core-shell nano structure is 30-50 nm, wherein the size of ferroferric oxide particles is 10-20 nm, the thickness of a silicon dioxide layer is 5-10 nm, and the size of platinum particles is 2-3 nm;
the preparation method of the nano biological probe comprises the following steps:
(1) Dispersing ferroferric oxide dispersed in an organic phase in a polyethylene glycol octyl phenyl ether/n-hexanol/water mixed solution to form a reverse microemulsion system, adding ammonia water and tetraethoxysilane to hydrolyze the tetraethoxysilane in an alkaline environment to form a silicon dioxide coating layer, and preparing ferroferric oxide-silicon dioxide particles; wherein the concentration of ferroferric oxide dispersed in the organic phase is 1-10 mg/mL; the molar ratio of the polyethylene glycol octyl phenyl ether to the n-hexanol to the water in the polyethylene glycol octyl phenyl ether/n-hexanol/water mixed solution is 1; the mass ratio of ethyl orthosilicate, ferroferric oxide and ammonia water is 1;
(2) Mixing the ferroferric oxide-silicon dioxide particles and 3-aminopropyltrimethoxy siloxane, and performing amination modification, wherein the mass ratio of the ferroferric oxide-silicon dioxide particles to the 3-aminopropyltrimethoxy siloxane is (1);
(3) Loading platinum particles on the surface of the positively charged ferroferric oxide-silicon dioxide particles through electrostatic adsorption, wherein the mass ratio of the platinum particles to the positively charged ferroferric oxide-silicon dioxide particles is 1;
(4) And coupling the magnetic core-shell nano structure with a specificity recognition molecule aiming at pathogenic bacteria by adopting a glutaraldehyde spacer arm method.
2. The use of the nanobody probe of claim 1 in the preparation of a pathogen detection kit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811217049.7A CN109536571B (en) | 2018-10-18 | 2018-10-18 | Nano biological probe for detecting pathogenic bacteria and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811217049.7A CN109536571B (en) | 2018-10-18 | 2018-10-18 | Nano biological probe for detecting pathogenic bacteria and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109536571A CN109536571A (en) | 2019-03-29 |
| CN109536571B true CN109536571B (en) | 2022-11-04 |
Family
ID=65844405
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811217049.7A Active CN109536571B (en) | 2018-10-18 | 2018-10-18 | Nano biological probe for detecting pathogenic bacteria and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109536571B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102537287B1 (en) * | 2021-04-23 | 2023-05-26 | 재단법인대구경북과학기술원 | Microstructure for active collection of microorganisms and active collection method of microorganisms using the same |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0599926A (en) * | 1991-10-11 | 1993-04-23 | Tdk Corp | Method for measuring bacterium, cell and virus |
| CN101672771A (en) * | 2009-09-23 | 2010-03-17 | 东南大学 | Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection |
| CN102645536A (en) * | 2012-04-19 | 2012-08-22 | 沈鹤柏 | Method for detecting staphylococcus aureus |
| CN104749365A (en) * | 2013-12-31 | 2015-07-01 | 中国科学院上海微系统与信息技术研究所 | Difunctional composite nanosphere and method for rapidly detecting food-borne pathogenic bacteria |
| CN106248933A (en) * | 2016-08-19 | 2016-12-21 | 浙江工商大学 | The method quickly detecting the rugged Cronobacter sakazakii of slope based on PGM and magnetic Nano microsphere |
| CN108187741A (en) * | 2018-01-09 | 2018-06-22 | 天津大学 | More carboxyl magnetic Nano silicon ball solid-borne platinum catalysts and its preparation method and application |
-
2018
- 2018-10-18 CN CN201811217049.7A patent/CN109536571B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0599926A (en) * | 1991-10-11 | 1993-04-23 | Tdk Corp | Method for measuring bacterium, cell and virus |
| CN101672771A (en) * | 2009-09-23 | 2010-03-17 | 东南大学 | Method for applying magnetic gamma-Fe2O3 nano-particle mimetic enzyme to biological detection |
| CN102645536A (en) * | 2012-04-19 | 2012-08-22 | 沈鹤柏 | Method for detecting staphylococcus aureus |
| CN104749365A (en) * | 2013-12-31 | 2015-07-01 | 中国科学院上海微系统与信息技术研究所 | Difunctional composite nanosphere and method for rapidly detecting food-borne pathogenic bacteria |
| CN106248933A (en) * | 2016-08-19 | 2016-12-21 | 浙江工商大学 | The method quickly detecting the rugged Cronobacter sakazakii of slope based on PGM and magnetic Nano microsphere |
| CN108187741A (en) * | 2018-01-09 | 2018-06-22 | 天津大学 | More carboxyl magnetic Nano silicon ball solid-borne platinum catalysts and its preparation method and application |
Non-Patent Citations (2)
| Title |
|---|
| A novel hierachically-nanostructured Pt/SiO2/Fe3O4 catalyst with high activity and recyclability towards hydrosilylation;Huachao Zai et,al.;《RSC Advances》;20161010;第6卷(第100期);8522页 右栏第二段、第98523页 右栏第一段 * |
| Construction of surface charge-controlled reduced graphene oxide-loaded Fe3O4 and Pt nanohybrid for peroxidase mimic with enhanced catalytic activity;LanLiu et,al.;《Analytica Chimica Acta》;20180731;第1014卷(第9期);摘要、第78页 左栏第3段、结论 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109536571A (en) | 2019-03-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Li et al. | Rapid identification and antibiotic susceptibility test of pathogens in blood based on magnetic separation and surface-enhanced Raman scattering | |
| Zhao et al. | Rapid detection of Listeria monocytogenes in food by biofunctionalized magnetic nanoparticle based on nuclear magnetic resonance | |
| Su et al. | Gold nanoparticles as colorimetric sensor: A case study on E. coli O157: H7 as a model for Gram-negative bacteria | |
| Wang et al. | Development of colloidal gold immunochromatographic signal-amplifying system for ultrasensitive detection of Escherichia coli O157: H7 in milk | |
| CN101907556B (en) | Method for detecting the colon bacillus by combining magnetic nanoparticle enrichment with bi-color flow cytometry | |
| ES2566478T3 (en) | Rapid procedure to detect microorganisms with magnetic particles | |
| CN102928590A (en) | Kit adopting fluorescent quantum dots to quickly screen, separate and detect salmonella | |
| Fei et al. | A sandwich electrochemical immunoassay for Salmonella pullorum and Salmonella gallinarum based on a AuNPs/SiO 2/Fe 3 O 4 adsorbing antibody and 4 channel screen printed carbon electrode electrodeposited gold nanoparticles | |
| WO2011144012A1 (en) | Magnetic kit for rapid detection of microorganisms and its preparation and usage | |
| CN106366197B (en) | HER2, EGFR, EpCAM and MUC1 multiple antibody immunomagnetic beads and preparation method thereof | |
| US9766237B2 (en) | Method of capturing bacteria on polylysine-coated microspheres | |
| WO2011081075A1 (en) | Method for extracting staphylococcus aureus antigen, reagent for extracting staphylococcus aureus antigen, and method for testing staphylococcus aureus | |
| Ling et al. | Rapid method for detection of Staphylococcus aureus in feces | |
| CN107389919A (en) | A kind of label-free fluorescence aptamer sensor and its preparation method and application | |
| CN113866408A (en) | Detecting food-borne intestinal pathogenic bacteria O157 based on the aptamer, the nanoparticle and the quantum dot label: h7 method | |
| CN109536571B (en) | Nano biological probe for detecting pathogenic bacteria and preparation method thereof | |
| Sun et al. | A nonenzymatic optical immunoassay strategy for detection of Salmonella infection based on blue silica nanoparticles | |
| CN105527427B (en) | A kind of method of quick detection Listeria monocytogenes | |
| Yu et al. | Simultaneous detection of pathogenic bacteria using agglutination test based on colored silica nanoparticles | |
| Fan et al. | Rapid and simultaneous detection of Salmonella spp., Escherichia coli O157: H7, and Listeria monocytogenes in meat using multiplex immunomagnetic separation and multiplex real-time PCR | |
| Ge et al. | Lateral flow immunoassay for visible detection of human brucellosis based on blue silica nanoparticles | |
| Fang et al. | Identification of Salmonella using colony-print and detection with antibody-coated gold nanoparticles | |
| CN105527428B (en) | A kind of quick detection Escherichia coli O 157：H7 method | |
| Ye et al. | An ultrasensitive sandwich immunoassay with a glucometer readout for portable and quantitative detection of Cronobacter sakazakii | |
| CN109507258B (en) | Electrochemical immunosensor and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |