KR101277696B1 - Methods for Separating, Concentrating and Detecting Microorganisms in One-Step Using Targeting-Gold Magnetic Nanoparticles - Google Patents

Abstract

The present invention relates to targeting-gold magnetic nanoparticles and uses thereof. Nanoparticles of the present invention comprises (i) a core comprising a magnetic material, (ii) a shell comprising gold and (iii) a targeting moiety coupled to the shell, wherein the targeting is coupled to the shell. Moieties (eg, antibodies to surface antigens of target microorganisms) can be used to effectively isolate, concentrate, and detect target microorganisms in one step. Thus, the magnetic nanoparticle-based method of the present invention not only makes the isolation and concentration of microorganisms very simple compared to conventional complicated and time-consuming methods, but also enables efficient detection at the same time.

Description

Targeting-Gold Magnetic Nanoparticles-Methods for Separating, Concentrating and Detecting Microorganisms in One-Step Using Targeting-Gold Magnetic Nanoparticles}

The present invention relates to a one-step separation, concentration and detection method of targeting-gold magnetic nanoparticle-based microorganisms.

In order to detect harmful microorganisms contained in the medium in the food, medical and environmental fields, only microorganisms contained in the medium are selectively separated and then detected through a microorganism culture process for a certain time to increase the detection signal according to the microorganism concentration. In general, the microbial incubation time is about 12 hours for E. coli hemorrhagic E. coli, depending on the microorganism takes 12 to 48 hours. In order to shorten the detection time, a technique for rapidly separating and concentrating microorganisms in a sample medium is being developed based on immunomagnetism.

 Immunomagnetism is a technique of immobilizing an antibody that selectively reacts with a microorganism on the surface of a nano or micro-sized particle having magnetic properties to induce selective reaction with a microorganism in a sample substrate through an antigen-antibody reaction, and then separating only the microorganism with a magnet. to be. Conventional immunomagnetism is a technique for selectively separating and concentrating microorganisms in a sample substrate and using another detection method (platelet smearing, colorimetric method, real-time PCR, SPR and ELISA, etc.) to identify the separated microorganisms.

For example, Korean Patent No. 0962286 discloses a method for improving sensitivity by increasing surface plasmon resonance using magnetic core gold nanoparticles in detecting food poisoning bacteria by surface plasmon resonance (SPR). This method has a complicated problem that requires the use of expensive SPR equipment and separate separation, concentration and detection process.

Therefore, there is an urgent need in the art for a new method capable of simultaneously separating, harvesting and detecting microorganisms to improve the complex and time-consuming conventional methods.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The inventors have sought to develop nanoparticle-based methods that can effectively separate target microorganisms in a sample. As a result, (i) a core comprising a magnetic material, (ii) a shell comprising gold and (iii) a targeting moiety that is bound to the surface of the shell and binds to the surface antigen of the microorganism. The present invention has been completed by confirming that targeting-gold magnetic nanoparticles can selectively separate / concentrate and detect one-step target microorganisms in a sample at the same time without special treatment.

Accordingly, it is an object of the present invention to provide a one-step separation, concentration and detection method of microorganisms using gold magnetic nanoparticles.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a method for one-step separation, concentration and detection of microorganisms using gold magnetic nanoparticles comprising the following steps:

(a) a targeting comprising a (i) core comprising magnetic material, (ii) a shell comprising gold and (iii) a targeting moiety bound to the surface of the shell and binding to the surface antigen of the microorganism Contacting the sample with gold magnetic nanoparticles; And

(b) applying the result of step (a) to a magnetically applied column to simultaneously separate, concentrate and detect the microorganisms, wherein the magnetic application is performed at a portion of the column and the magnetic application The visible light is irradiated to the portion of the column where the microorganism is separated to measure the absorbance.

The inventors have sought to develop nanoparticle-based methods that can effectively separate target microorganisms in a sample. As a result, (i) a core comprising a magnetic material, (ii) a shell comprising gold and (iii) a targeting moiety that is bound to the surface of the shell and binds to the surface antigen of the microorganism. Using targeting-gold magnetic nanoparticles, it was confirmed that the target microorganisms in the sample can be selectively separated / concentrated and detected in one step without any special treatment at the same time.

As used herein, the term “one-step” refers to the separation, concentration and detection of microorganisms in a sample through a single process of the sample to be analyzed and the target-gold magnetic nanoparticles. More specifically, it means that the microorganisms in the sample are separated, concentrated and detected by one step of loading the reactant of the sample and the targeting-gold magnetic nanoparticles into the column used in the present invention.

One-step methods of microbial isolation, concentration, and detection of the present invention are magnetic nanoparticle-based methods.

According to the invention, step (a) of the invention is carried out by contacting the sample with the targeting-gold magnetic nanoparticles.

The targeting-gold magnetic nanoparticles of the present invention comprise (a) a core comprising a magnetic material, (b) a shell comprising gold and (c) a targeting moiety coupled to the shell.

According to a preferred embodiment of the present invention, the core comprising the magnetic material of the present invention means a material having a size of 1-100 nm preferably. The core including the magnetic material used in the present invention is not particularly limited in shape and shape as long as it has a nano-size material (for example, particles, tubes, rods, or tetrahedrons).

According to a preferred embodiment of the present invention, the core comprising the magnetic material used in the present invention is a nanoparticle. The term “nanoparticle” refers to particles of various materials having a diameter in nano units, preferably 5-100 nm, more preferably 8-50 nm, and most preferably 10-30 nm. The nanoparticles are not particularly limited as long as they are particles having a nano size. For example, metal nanoparticle, metal oxide particle, etc. are mentioned. Specific examples of the metal particles include Au, Ag, Pd, Pt, Cu, Ni, Co, Fe, Mn, Ru, Rh, Os, Ir and the like. Metal oxide particles refer to compounds represented by the chemical formula M _x O _y (wherein M represents a metal, O represents oxygen, and x and y represent an integer), such as Fe ₂ O ₃ , Fe ₃ O ₄ , Ag ₂ O , TiO ₂ , SiO ₂ , and the like, but are not limited thereto.

More preferably, the core including the magnetic material used in the present invention means a metal oxide nanoparticle. As used herein, the term “metal oxide nanoparticles” refers to metal particles having a diameter in nano units, preferably 1-100 nm, more preferably 1-50 nm, most preferably 5-30 nm (eg, Iron, nickel, platinum, manganese) and oxides thereof, including, but not limited to, all nano-sized particles having a magnetic property. This small particle size allows the nanoparticles of the invention to easily bind to the cell of interest (eg, bacteria).

Even more preferably, in the targeting-gold magnetic nanoparticles used in the present invention, the magnetic core is a metal oxide represented by M _x O _y (wherein M is a metal, O is oxygen and x and y are integers). , Most preferably Fe ₂ O ₃ or Fe ₃ O ₄ .

According to the most preferred embodiment of the present invention, in the targeting-gold magnetic nanoparticles used in the present invention, the shell comprises gold. Gold is easy to manufacture in the form of stable particles, easy to control the size, unlike the heavy metals such as manganese, aluminum, cadmium, lead, mercury, cobalt, nickel, beryllium, harmless to the human body has a high bio-compatibility.

The gold nanoparticles used in the present invention can be prepared, for example, as follows: HAuCl ₄ is used as a gold source, and sodium citrate is used as a reducing agent to reduce HAuCl ₄ to prepare gold nanoparticles. In this case, the size of the gold nanoparticles can be adjusted by varying the citrate added. That is, the size of the gold nanoparticles decreases as the amount of citrate increases, so that nucleation increases.

When the gold nanoparticles become larger than 100 nm in diameter, their properties as nanoparticles are not only lost, but the binding of the gold surface having no properties of nanomaterials to functional groups such as thiol groups is weak. Particles are difficult to manufacture. Therefore, the gold nanoparticles of the present invention preferably have a diameter of 5-100 nm, more preferably 8-50 nm, and most preferably 10-30 nm.

Preferably, the surface of the gold magnetic nanoparticles of the present invention is coated with a material that can be used for binding to the targeting moiety. For example, the substance may be mercaptohexadecanoic acid (MHDA), cystamine, polyethylene glycol (PEG), dextran, polyvinylpyrrolidone, albumin, polyvinyl alcohol, polyethyleneimine, chitosan, Hyaluronic Acid, Polylactic Glycolic Acid (PLGA), Polylactic Acid, Polylactide, Polyglycolic Acid, Polyamino Acid, Polyacetal, Polytherabolate, Polycarbonate, Polycarbolactone Polyacrylic Acid, Polymethacrylic Acid , Polysaccharides, polyketals, polyethers, polyamides, polymaleic anhydrides, polymethylvinyl ethers and copolymers of the above polymers, but are not limited thereto. Most preferably, mercapdohexadecanoic acid.

As used herein, the term “targeting moiety” means any substance capable of binding to an antigen present on the surface of a target microorganism. According to a preferred embodiment of the invention, the targeting moiety of the invention comprises a protein, peptide, antibody or fragment thereof, more preferably a peptide, antibody or fragment thereof, most preferably an antibody or fragment thereof It includes.

Targeting moieties of the present invention comprises an F _ab and F _c region of antibodies, and is non-covalently bonded to the surface antigen of a target microbe through the F _ab regions. As used herein, the term “F _ab site” refers to a site of an antibody that interacts with a surface antigen (eg, a receptor) of a target microorganism. In general, cleavage of full-length antibodies with papain yields two sites, the F _ab region and the Fc region. The F _ab regions of all antibodies in one class have nearly identical characteristics.

According to a preferred embodiment, F _ab and F _ab portion means a portion of the IgG.

On the other hand, in the range having the property of binding to the surface antigens of the target microbe F _ab sites it is conventional may be a part sequence of the F _ab regions.

According to a preferred embodiment of the invention, the targeting moiety is an antibody, aptamer, polypeptide, peptide, ligand, lectin, sugar, lipid, glycolipid or nucleic acid comprising the F _ab region of the antibody, more preferably the antibody An antibody, aptamer or ligand comprising the F _ab region of, most preferably an antibody.

If the targeting moiety is an antibody comprising a F _ab site, a full length antibody may be used as the targeting moiety. Full length antibodies include polyclonal or monoclonal antibodies. Antibodies that bind to specific surface molecules of a cell can be prepared by methods commonly practiced in the art, such as fusion methods (Kohler and Milstein, European Journal of Immunology , 6: 511-519 (1976)), recombinant DNA methods ( US Pat. No. 4,816,56) or phage antibody library method (Clackson et al, Nature , 352: 624-628 (1991) and Marks et al, J. Mol. Biol. , 222: 58, 1-597 (1991) It can be prepared by). General procedures for antibody preparation are described in Harlow, E. and Lane, D., Using Antibodies: A Laboratory Manual , Cold Spring Harbor Press, New York, 1999; Zola, H., Monoclonal Antibodies: A Manual of Techniques , CRC Press, Inc., Boca Raton, Florida, 1984; And Coligan, CURRENT PROTOCOLS IN IMMUNOLOGY , Wiley / Greene, NY, 1991, which are incorporated herein by reference. For example, the preparation of hybridoma cells producing monoclonal antibodies is accomplished by fusing an immortalized cell line with an antibody-producing lymphocyte, and the techniques necessary for this process are well known and readily practicable by those skilled in the art. Polyclonal antibodies can be obtained by injecting a cell surface molecule as an antigen into a suitable animal, collecting antisera from the animal and then isolating the antibody from the antisera using known affinity techniques.

Next, step (b) of the present invention is to apply the result of step (a) to the magnetically applied column to simultaneously isolate, concentrate and detect microorganisms in one step. The magnetic application is performed at a predetermined portion of the column, and the magnetic light is applied to the portion of the column where the microorganisms are separated, and thus the absorbance is measured.

According to a preferred embodiment of the present invention, the column used in the present invention includes a membrane filter thereon and the targeting-gold magnetic nanoparticles are bound in the result of step (a) applied to the column by the membrane filter. Delay the flow of microorganisms.

According to a preferred embodiment of the invention, the membrane filter which can be used in the present invention has a size of 50-1,000 nm, more preferably has a size of 70-900 nm, even more preferably 90-850 nm in size, most preferably 100-800 nm.

Target microorganisms to which the gold magnetic nanoparticles of the present invention are bound may be separated under magnetic (eg, under permanent magnets). Thus, the method of the present invention can utilize size and magnetism to isolate and concentrate the target microorganism more specifically and effectively.

According to a preferred embodiment of the present invention, the magnetic application is made by a permanent magnet or an electromagnet.

Furthermore, step (b) of the present invention measures the absorbance of visible region to the result of step (a) described above, so that the method of the present invention can quantitatively measure the amount of the target microorganism isolated.

According to a preferred embodiment of the present invention, the measurement of the absorbance is performed using a spectrophotometer connected to the column, through which not only the detection of the target microorganism but also the amount of the microorganism can be detected quantitatively.

As used herein, the term “quantitative” means that it is possible to numerically detect the amount of the target microorganism bound to the gold nanoparticles of the present invention. More specifically, by irradiating light in the visible region with respect to the target microorganism-gold magnetic nanoparticle complex captured under the magnetic (eg permanent magnet) by measuring the change in absorbance and converting the change in absorbance by the number of target microorganisms Is done.

According to a preferred embodiment of the present invention, the binding of the captured target microorganism-gold magnetic nanoparticle complex and the target microorganism has a linear correlation with the absorbance change of the gold magnetic nanoparticles (see FIG. 7).

According to a preferred embodiment of the present invention, the visible light region used in the present invention has a range of 500-600 nm, more preferably in the range of 520-560 nm, most preferably in the range of 530-540 nm. Has

According to a preferred embodiment of the invention, the quantitative detection range of the target microorganism (eg E. coli) detected by the method of the invention is 10 ^{1-10 9} cells, more preferably 10 ^{1-10 8} cells and most preferably Is 10 ¹ -10 ⁶ cells.

According to a preferred embodiment of the invention, the target microorganism which can be isolated, concentrated and detected using the method of the invention is determined by the targeting moiety described above.

Target microorganisms that can be isolated, concentrated and detected in the present invention are preferably harmful microorganisms, more preferably harmful eukaryotes and harmful bacteria, for example E. coli (e.g. E. coli 0157: H7), Salmonella (Salmonella; e. g., Salmonella enteritidis, Salmonella typhi), Shh Kella (Shigella; e. g., Shigella dysenteriae), Enterobacter bacteria three (Enterobacteriaceae; for example, Klebsiella pneumoniae), Pseudomonas (Pseudomonas; e.g., Pseudomonas aeruginosa), morak Cellar (Moraxella ; e.g., Moraxella catarrhalis), Helicobacter pylori (Helicobacter; e.g., Helicobacter pylori), Pomona's (Stenotrophomonas by stacking notes; and the like e.g., Stenotrophomonas maltophilia), but the embodiment is not limited thereto. For example, using targeting-gold magnetic nanoparticles coupled with targeting moieties specific for antigens located on the surface of food poisoning-inducing microorganisms (eg, Salmonella enteritidis), the presence of food poisoning-inducing microorganisms in a sample can be detected. Not only can it be done, but it can also be separated (/ enriched).

As described above, the present invention is a novel method with improved convenience, sensitivity, and accuracy for separating, concentrating, and detecting microorganisms in one-step using targeting-gold magnetic nanoparticles.

The present inventors have already suggested through the Republic of Korea Patent No. 0962286 that microorganisms can be detected using magnetic core gold nanoparticles. However, the method described in the patent document is basically based on SPR, and this method has to use an expensive SPR device and has a complicated process. The inventors have devised and completed the present invention in order to overcome the problems of the prior art, and through the present invention a method for separating, concentrating and detecting microorganisms in one-step using a simpler and less expensive device. present.

The features and advantages of the present invention are summarized as follows:

(a) The present invention enables targeting, gold magnetic nanoparticles to separate, concentrate and detect target microorganisms in a sample simultaneously and in one-step.

(b) The gold magnetic nanoparticle-based method of the present invention not only makes the isolation and concentration of microorganisms very simple compared to conventional complicated and time-consuming methods, but also enables efficient detection at the same time.

1 is a result of observing the gold magnetic nanoparticles of the present invention that can selectively react with microorganisms.
Figure 2 is a schematic representation of the process of producing the magnetic gold nanoparticles of the present invention.
3 is a view illustrating a process of separating and reacting the gold magnetic nanoparticles of the present invention with a sample.
4 is a diagram illustrating a process of separating target microorganisms using a membrane filter.
Figure 5 is a schematic diagram showing the process of separating and concentrating the microorganism to which the gold magnetic nanoparticles of the present invention are bound.
6 is a view showing a process (a) and microscopic results (b) of measuring the amount of separated microorganisms.
7 is an absorbance result of measuring the microorganism to which the gold magnetic nanoparticles of the present invention are bound in the visible region.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Fabrication of Au-magnetic Particles that Can React with Microorganisms selectively

25 ml of a mixed aqueous solution of 0.5M NaOH and 0.1M FeCl ₂ 4H ₂ O, 0.2M Fe ₃ Cl ₆ 4H ₂ O, and 0.4M HCl was stirred at 80 rpm at a speed of 2,000 rpm for 30 minutes using a mechanical stirrer. Then, the supernatant was separated from the stirring solution using a permanent magnet. After neutralization with 0.04 M HCl, washed with distilled water and ethanol and dispersed in ethanol to the final 200 ml to prepare a nanoparticle-size Fe ₂ O ₃ or Fe ₃ O ₄ magnetic core. As described above, in order to modify the prepared magnetic core surface with an amine functional group, 1.0 M aminopropyltriethoxysilane dissolved in 50 ml of ethanol and the dispersed nanoparticles were reacted for 24 hours. After centrifugation or permanent magnet washing with ethanol three to five times and finally dispersed in 100 ml distilled water.

Subsequently, gold nanoparticles having a magnetic core were added with 70 ml of distilled water in which 0.08 g of HAuCl ₄ 3H ₂ O was dissolved in 10 ml of the dispersion of the nanoparticles surface-modified with the amine group, and stirred under reflux, followed by 1% sodium site. The reaction was carried out for 2 hours after the addition of sodium citrate. Purification using a permanent magnet to produce gold magnetic nanoparticles (Fig. 1).

Sample preparation technology using immunomagnetic-based gold magnetic nanoparticles

After reacting the prepared gold magnetic nanoparticles with ethanol in which 1 mM mercaptohexadecanoic acid (MHDA) was dissolved for 12 hours, MHDA-fixed gold magnetic nanoparticles were separated using a permanent magnet. And purified. The separated and purified gold magnetic nanoparticles were reacted with 0.4 M EDC (N- (3-Dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride) and 0.1 M NHS (N-Hydroxysulfosuccinimide) for 5 minutes, followed by antibody (10 The antibody was immobilized on the surface of the magnetic gold nanoparticles by reacting with a solution of PBS (pH 7.4) dissolved in μg / ml of anti-0157) for 2 hours. Thereafter, the reaction was coated with BSA for 30 minutes to eliminate the nonspecific reaction, and then the antibody-fixed gold magnetic nanoparticles were purified using permanent magnets (FIG. 2).

PBS solution contaminated with various concentrations of Escherichia coli ( Esherichia coli 0157: H7) was used as a model sample to verify the performance of the prepared antibody-fixed gold magnetic nanoparticles. To a 250 mL PBS model sample containing various concentrations of microorganisms (0, 10, 10 ³ , 10 ⁵ and 10 ⁶ cells / ml) was added 200 μl of the gold magnetic nanoparticles prepared above and stirred for 5 minutes. The solution was filtered using a nylon filter having a pore size of 450 nm to separate only microorganisms having a size of several micrometers and gold magnetic nanoparticles adsorbed on the surface of the microorganisms (FIGS. 3 and 4).

The microorganisms on the filter were redispersed using 5 ml distilled water and the dispersed microorganisms were separated and concentrated using permanent magnets (FIG. 5). In addition, a UV-Vis spectrometer was installed opposite the permanent magnet and the permanent magnet was used as a reflector to confirm the visible absorption spectrum change of the gold nanoparticles adsorbed on the surface of the microorganism in the reflection mode at the same time (concentration). 6a).

Figure 6b is measured by reacting E. coli O157: H7 with gold magnetic nanoparticles (pictured left) and by reacting E. coli O157: H7 with gold magnetic nanoparticles, separating them using a filter, and then redispersing them. (Picture on the right) A picture of an electron microscope. That it is not substantially present in the ambient H7: As can be seen in 6b, magnetic gold nanoparticles E. coli O157: is adsorbed selectively respond well to the surface of the H7 and Poultry magnetic nanoparticles unreacted E. coli O157 I could confirm it.

7 shows the results of confirming the concentration of E. coli O157: H7 using a visible absorption spectrum in the process of separating / concentrating E. coli O157: H7 reacted with gold magnetic nanoparticles using permanent magnets. The result was measured 10 minutes after the start of the concentration, and is the result of the visible spectrum versus the concentration of E. coli O157: H7 contaminated with 250 ml of the initial model sample PBS (FIG. 8, left panel). As the concentration of E. coli O157: H7 (0, 10, 10 ³ , 10 ⁵ and 10 ⁶ cells / ml) increased, the absorption intensity increased at 530-540 nm, which is the intrinsic absorption wavelength of the magnetic nanoparticles. Could know. In addition, it can be seen that the linear increase in the wavelength intensity for each concentration of E. coli O157: H7 at the maximum absorption wavelength (540 nm) indicated by the arrow in the left panel of FIG. 7 (right panel of FIG. 7). An absorption intensity of about 0.01 in a sample contaminated with E. coli O157: H7 (concentration 0 cells / ml) is a signal of unreacted gold magnetic nanoparticles that are not completely isolated from the filter. This corresponds to a negligibly small signal of less than 10% compared to the 10 cell / ml concentration. Therefore, it can be seen that simultaneous detection of sample pretreatment using gold magnetic nanoparticles is a highly sensitive detection method capable of measuring up to 10 cells / ml in a model sample.

As a result, the gold magnetic nanoparticles are nanometer-sized multifunctional particles having both magnetic properties affected by permanent magnets and gold nanoparticles absorbing certain visible region (530-540 nm). Existing immunomagnetic and detection methods could be applied simultaneously in sample preparation. The present invention provides a method for real-time monitoring of the separation / concentration process of a specific microorganism in a sample, and by simplifying the whole process by unifying the existing microbial sample pretreatment and detection process, the accuracy, reproducibility and reliability of sample pretreatment are improved and trace amount is reduced. A method for detecting microorganisms at high speed has been proposed.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (14)

One-step separation, concentration and detection of microorganisms using gold magnetic nanoparticles comprising the following steps:
(a) a targeting comprising a (i) core comprising magnetic material, (ii) a shell comprising gold and (iii) a targeting moiety bound to the surface of the shell and binding to the surface antigen of the microorganism Contacting the sample with gold magnetic nanoparticles; And
(b) applying the result of step (a) to a magnetically applied column to simultaneously separate, concentrate and detect the microorganisms, wherein the magnetic application is performed at a portion of the column and the magnetic application The visible light is irradiated to the portion of the column where the microorganism is separated to measure the absorbance.
The method of claim 1, wherein the magnetic application is performed by a permanent magnet or an electromagnet.
The method of claim 1, wherein the absorbance measurement is performed using a spectrophotometer connected to the column.
The method of claim 1, wherein the absorbance measurement enables detection of the microorganism and the detection of the microorganism is quantitative detection.
The method of claim 1, wherein the magnetic material is M _x O _y (wherein M is Fe, Au, Pt, Ni or Mn, O is oxygen, and x and y represent an integer). .
The method of claim 5, wherein the M _x O _y is Fe ₂ O ₃ or Fe ₃ O ₄ .
The method of claim 1, wherein the targeting moiety is an antibody or fragment thereof.
The flow of microorganisms of claim 1, wherein the column includes a membrane filter thereon and the targeting-gold magnetic nanoparticles are combined in the result of step (a) applied to the column by the membrane filter. Delaying the process.
The method of claim 1, wherein the visible region is in the range of 500-600 nm.
The method of claim 1, wherein the gold magnetic nanoparticles are nanoparticles having a size of 5-100 nm.
The method of claim 1, wherein the surface of the magnetic gold nanoparticles is coated with a material that can be used for binding to the targeting moiety.
The method of claim 11, wherein the surface of the gold magnetic nanoparticles are mercaptohexadecanoic acid (MHDA), cystamine, polyethylene glycol (PEG), dextran, polyvinylpyrrolidone, albumin, polyvinyl Alcohol, Polyethylenimine, Chitosan, Hyaluronic Acid, Polylactic Glycolic Acid (PLGA), Polylactic Acid, Polylactide, Polyglycolic Acid, Polyamino Acid, Polyacetal, Polytherabolic Acid, Polycarbonate Tonic polyacrylic acid, polymethacrylic acid, polysaccharide, polyketal, polyether, polyamide, polymaleic anhydride, polymethylvinyl ether or a copolymer of the above polymer.
13. The method of claim 12, wherein the surface of the gold magnetic nanoparticles is coated with mercapdohexadecanoic acid.
The method of claim 1, wherein the microorganism is a food poisoning-inducing microorganism.

Images