CN108088994A - A kind of hollow core shell nanoparticles and preparation method, test strips and test method - Google Patents
A kind of hollow core shell nanoparticles and preparation method, test strips and test method Download PDFInfo
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Abstract
The invention discloses a kind of hollow core shell nanoparticles and preparation method, test strips and test methods, can quantitatively detect the concentration of determinand in sample solution, improve detection sensitivity.The hollow core shell nanoparticles, including nucleome and housing, the housing is located on the outside of nucleome, has spacing between housing and nucleome, the outer surface of the housing and inner surface are marked with Raman molecular respectively, and the outer surface of the nucleome is marked with Raman molecular；The housing is made of electrum, and the nucleome is made of gold.
Description
Technical field
The present invention relates to field of biological medicine, it particularly relates to a kind of hollow core shell nanoparticles and preparation side
Method, test strips and test method.
Background technology
Immuno-chromatographic assay technology based on gold nano grain (Au NPs) is in monoclonal antibody technique and novel nano
The vitro detection diagnostic method based on film to grow up on material technology obtains extensive use, bag in various fields at present
Include allergy, early pregnancy, heart infarction marker, infectious disease, environmental contaminants, drugs etc..Only by taking early pregnancy detects as an example, required for annual
Kit/test strips dosage have reached 107It is a.Immunochromatography testing principle based on Au NPs is as follows：Specificity is caught
It obtains antibody to be fixed on nitrocellulose (Nitro Cellulose, NC) film, forms band；The Au of identification antibody will be combined
NPs compounds are supported on the fiberglass packing at NC membrane samples end, and gold-labelled pad is made；Using the capillarity of NC membrane micropores, make
Sample solution is moved from test strips one end to the other end；It is added dropwise when solution to be measured is flowed through at gold-labelled pad in sample pad, solution
Middle determinand is combined with the identification antibody on Au NPs surfaces, forms " Au NPs- identify antibody-determinand " compound, when this is multiple
Close logistics after tested area when, the capture antibody being fixed on nitrocellulose filter is captured, formation double antibodies sandwich structure.With
The structure is built up test section, is just present with red stripes at the detection line T on film, and nature controlling line C is then examining
Whether active survey coated albumen on immunological probe Au NPs.Immuno-chromatographic assay technology based on Au NPs is easy to be quick,
Only with the naked eye can judging result, in past 10 years increasingly established it detects (point-of-care by bed
Testing, POCT) in status.But the technology still remains the challenge of two aspects at present：It is how to realize first
The methods of high sensitivity detection, immunochromatography detection sensitivity is much smaller than enzyme linked immunological and immunofluorescence at present；Secondly, meat is passed through
It is a qualitative results to observe and survey band colour developing, and the eyes of different people are larger for the sensitivity differences of color, sensitive
It spends low.
Exemplified by being diagnosed with acute myocardial injury (acute myocardial infarction, AMI), ECG examination must
Peripheral blood Myocardial Troponin I (cardiac troponin I, cTnI) content detection, which must be combined, can just provide finally clearly
Diagnosis.CTnI is one of three subunits for forming troponin complex, has the Cardiac-specific of height, blood level content
From the influence of the other diseases such as Skeletal muscle injury, strenuous exercise, nephrosis.The content of cTnI is generally low in healthy population peripheral blood
In 0.06ng/mL, and for the patient of acute myocardial infarction AMI, after myocardial cell damage, different types of egg in cardiac muscle cell
It is discharged into vain in blood, after AMI occurs within 2.2~6.8h, the concentration rapid increase of peripheral blood cTnI reaches after 11.2h
Peak 195.9ng/mL, and sustainable 4~10 days.It is detection cardiac muscle stalk this period that cTnI concentration rises to peak by normal value
Dead Best Times.In patient for myocardial infarction occurs in clinic, there is about 25% patient in myocardial infarction early stage disease
Shape unobvious, the patient of nearly half are not in the feature of myocardial necrosis in Electrocardiography.Therefore, after myocardial necrosis, the heart
The detection of flesh marker has vital meaning to myocardial infarction early diagnosis.At present, traditional enzyme-linked immunization (ELISA)
Although highly sensitive detection can be realized, detecting step is comparatively laborious, can not realize quick testing requirements.And based on gold nano
The immunochromatographic method of particle can realize that rapid in-vitro diagnosis has been widely used in cTnI detections, detection range 5ng/mL
~1 μ g/mL, detectable minimum concentration are 5ng/mL.On the basis of immunochromatographic method, Xu et al. is reported to be received with superparamagnetism
Rice grain replace gold nano grain quickly detected for cTnI immunochromatographies, using magnetic analytical meter to magnetic signal in detection line into
It has gone detection, has realized cTnI and quantitatively detect, minimal detectable concentration can reach 0.01ng/mL.Cui groups select GoldMag
Gold-magnetic particles equally realize the fast qualitative of cTnI or quantitative detection as carrier.But existed based on magnetic immunochromatographic method
It is easily disturbed during detection be subject to the earth and ambient enviroment magnetic field.
The content of the invention
Technical problem：The technical problems to be solved by the invention are：It is proposed a kind of hollow core shell nanoparticles and preparation side
Method, test strips and test method can quantitatively detect the concentration of determinand in sample solution, improve detection sensitivity.
Technical solution：In order to solve the above technical problems, the embodiment of the present invention adopts the following technical scheme that：
A kind of hollow core shell nanoparticles, including nucleome and housing, the housing is located on the outside of nucleome, housing and nucleome it
Between have spacing, the outer surface of the housing and inner surface are marked with Raman molecular respectively, and the outer surface of the nucleome is marked with drawing
Graceful molecule；The housing is made of electrum, and the nucleome is made of gold.
As preference, the hollow core shell nanoparticles are used for Surface enhanced Raman scattering immuno-chromatographic test paper strip.
A kind of preparation method of hollow core shell nanoparticles, the described method includes：
Step 10) prepares gold seeds using hydro-thermal method：
Gold grain is prepared by seed growth-curing method in step 20)：
Step 30) prepares Au@Ag gold and silver core shell nanoparticles by seed mediated growth method：
Step 40) is prepared using Au Ag gold and silver core shell nanoparticles prepared by step 30) as template is sacrificed in Au Au-Ag
Empty core shell nanoparticles.
As preference, the step 10) includes：After heating water to boiling, chlorauric acid solution and lemon are sequentially added
Acid sodium solution prepares gold seeds；The size of gold seeds is 10~15nm.
As preference, the step 20) includes：After heating water to boiling, sodium citrate solution, chlorine gold are sequentially added
Gold seeds prepared by acid solution, quinol solution and step 10), is prepared gold grain；The size of gold grain for 30~
100nm；Nile blue solution is added in into solution, by electrostatic absorption principle, in one layer of Raman molecular of gold grain surface modification.
As preference, the step 30) includes：Gold grain prepared by step 20) is seed, under stirring at normal temperature, according to
Secondary addition ascorbic acid solution and silver nitrate solution, grow into silver-colored shell on gold grain surface, are added with stirring NBA solution, pass through
Electrostatic Absorption Raman molecular in silver surface modification, is made Au Ag gold and silver core shell nanoparticles.
As preference, the step 40) includes：Au@Ag gold and silver core shell nanoparticles solution prepared by step 30) is added
AuCl is added dropwise to after seething with excitement in heat2 -Solution after reaction, is obtained by filtration Au@Au-Ag nanoparticles solutions, adds
NBA solution modifies one layer of Raman molecular in particle surface, the hollow core shell nanoparticles of Au Au-Ag is made.
A kind of test strips containing the hollow core shell nanoparticles, the test strips include bottom liner, sample pad, nanometer
Particle bonding pad, chromatographic film and water absorption pad；Chromatographic film is equipped with the detection line being mutually parallel and nature controlling line；Sample pad, nanometer
Grain bonding pad, chromatographic film, water absorption pad is closely coupled successively and is attached on bottom liner；Wherein, adhere on nano particle bonding pad anti-
The hollow core shell nanoparticles of body mark；Attachment capture antibody in detection line.
As preference, the hollow core shell nanoparticles include nucleome and housing, and the housing is located on the outside of nucleome, shell
There is spacing between body and nucleome, the outer surface of the housing and inner surface are marked with Raman molecular, the appearance of the nucleome respectively
Face is marked with Raman molecular；The housing is made of electrum, and the nucleome is made of gold.
A kind of detection method that clinical marker is carried out using the test strips, this method are included：Determinand will be contained
Sample solution is added dropwise by well in sample pad, after standing, utilizes Raman spectrometer acquisition ELISA test strip line position
Raman spectrum according to the relative intensity of Raman spectrum, obtains testing concentration.
Advantageous effect：It is received with existing SERS immunochromatography techniques using spherical gold nano grain or Au@Ag gold and silver nucleocapsids
Rice grain is compared, and the present invention is had the advantages that using the hollow core shell nanoparticles of Au Au-Ag：
1. compared with spherical shape Au Au@Ag gold and silver core shell nanoparticles used in traditional test strip, in Au@Au-Ag
Empty core shell nanoparticles have stronger SERS activity, substantially increase ELISA test strip sensitivity.
The hollow core shell nanoparticles nucleocapsid spacing of 2.Au Au-Ag is adjustable, possesses stronger SERS activity.As Au@Au-Ag
Nucleocapsid spacing reduces, and the SERS activity of particle then exponentially increases, and when less than 2nm, increases especially pronounced.Pass through change
Au@Ag nano particle Ag shell thicknesses, Au@Au-Ag nucleocapsid spacing is adjustable in the range of 2nm, is conducive to obtain SERS activity most
Excellent immunochromatography detection probe.
3. for spherical gold nano grain, Raman molecular is modified in particle surface, during subsequently test strips are prepared
Easily come off from particle surface.In the present embodiment, the nucleocapsid surfaces externally and internally and its spacing of the hollow core shell nanoparticles of Au Au-Ag
Raman molecular can be modified, and Raman molecular is subject to shell to protect, and from external interference, begins in follow-up test strips preparation process
It keeps stablizing eventually.Raman molecular keeps stablizing, and the preparation and detection that can ensure test strips have very high repeatability.
4.Au Ag core shell nanoparticles and the hollow core shell nanoparticles of Au Au-Ag because shell contain silver element all have compared with
Strong SERS activity, but Au@Ag shells are made of fine silver, need to be carried on polyester fiber film simultaneously in test strips step is prepared
It is dry, it is easy to oxidation deactivation in follow-up preserve, and the shell of Au@Au-Ag is electrum shell, it is severe in oxidation corrosion etc.
Still it can keep stable in environment, the test strips of making are easy to preserve.
5. previous preparation method can only obtain surfactant (cetyl trimethylammonium bromide) or polymer molecule
The Au-Ag alloy nanoparticles of claddings such as (polyvinylpyrrolidones), in subsequent applications, particle surface is difficult to modify antibody etc.
Other molecules.Au@Au-Ag nano particles prepared by the present invention are stablized by sodium citrate, are easy to surface modification, follow-up to try
The preparation process of paper slip is consistent as the Traditional immunochromatographic test strips of probe with the Au nano particles stablized using sodium citrate, possesses
Ready-made preparation process without doing additional improvement, reduces production cost.
It is more enough to prepare more stable, the stronger surface increasing of signal using the hollow core shell nanoparticles of Au Au-Ag as material
Strong Raman scattering immuno-chromatographic test paper strip, realizes the quick, highly sensitive of clinical marker, half-quantitative detection.
Description of the drawings
Fig. 1 is the structure diagram of the hollow core shell nanoparticles of Au Au-Ag of the embodiment of the present invention；
Fig. 2 is the structure diagram of the test strips of the embodiment of the present invention；
Fig. 3 is gold nano grain (Au), gold and silver core shell nanoparticles (Au Ag) and the hollow core shell nanoparticles of Au Au-Ag
SERS performance comparison figures；
Fig. 4 is the optical photograph obtained after ELISA test strip various concentration cTnI in the embodiment of the present invention；
Fig. 5 is the Raman spectrogram that ELISA test strip various concentration cTnI is obtained in the embodiment of the present invention；
Fig. 6 is in the embodiment of the present invention, and test strips are with Raman spectrum main peak 592cm in Fig. 5-1The standard that intensity is set up
Graph (logarithmic transformed).
Have in figure：Hollow core shell nanoparticles 1, identification antibody 2, capture antibody 4, bottom liner 5, sample pad 6, are received determinand 3
Rice grain bonding pad 7, chromatographic film 8, detection line 9, nature controlling line 10, water absorption pad 11, nucleome 101, housing 102, Raman molecular 103.
Specific embodiment
With reference to specific embodiment, the present invention is further explained.It is to be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In addition, it should also be understood that, after the content that the present invention is told about has been read, people in the art
Member makes various changes or modifications the present invention, and such equivalent forms equally fall within the application model as defined in the appended claims
It encloses.
As shown in Figure 1, the embodiment of the present invention provides a kind of hollow core shell nanoparticles, including nucleome and housing.Housing position
On the outside of nucleome, there is spacing between housing and nucleome.The outer surface of housing and inner surface are marked with Raman molecular respectively.Nucleome
Outer surface is marked with Raman molecular.Housing is made of electrum, and nucleome is made of gold.The hollow core shell nanoparticles are used for table
Face enhances Raman scattering immuno-chromatographic test paper strip.
Hollow core shell nanoparticles in the embodiment, there is spacing between nucleome and housing, and adjustable, a little between nucleocapsid
The irregular electrum beam column bridge joint of small size, therefore, nucleocapsid can keep being relatively fixed.
It is marked with Raman molecular respectively in the outer surface of housing and inner surface, the outer surface of nucleome is marked with Raman molecular.
Individual particle modifies Raman molecular as much as possible, so as to improve detection signal strength.Housing is made of electrum, and nucleome is
Gold is made, and chemical property is stablized, and is not easy to be aoxidized, is conducive to the preservation of test strips.
The preparation method of the hollow core shell nanoparticles of above-described embodiment, including：
Step 10) prepares gold seeds using hydro-thermal method：After heating water to boiling, chlorauric acid solution and lemon are sequentially added
Lemon acid sodium solution, prepares gold seeds.
Gold grain is prepared by seed growth-curing method in step 20)：After heating water to boiling, lemon is sequentially added
Gold seeds prepared by lemon acid sodium solution, chlorauric acid solution, quinol solution and step 10), is prepared gold grain；To solution
Middle addition Nile blue (should be full name：NileBlue A, text in referred to as：NBA) solution, by electrostatic absorption principle, in gold grain
One layer of Raman molecular of surface modification.
Step 30) prepares Au@Ag gold and silver core shell nanoparticles by seed mediated growth method：Gold grain prepared by step 20)
For seed, under stirring at normal temperature, ascorbic acid solution and silver nitrate solution are sequentially added, silver-colored shell is grown on gold grain surface, stirs
Mix it is lower addition NBA solution, by Electrostatic Absorption silver surface modification on Raman molecular, Au Ag gold and silver core shell nanoparticles are made.
Step 40) is prepared using Au Ag gold and silver core shell nanoparticles prepared by step 30) as template is sacrificed in Au Au-Ag
Empty core shell nanoparticles：After Au@Ag gold and silver core shell nanoparticles solution prepared by step 30) is heated to boiling, it is added dropwise
AuCl2 -Solution after reaction, is obtained by filtration Au@Au-Ag nanoparticles solutions, adds NBA solution, repaiied in particle surface
One layer of Raman molecular is adornd, the hollow core shell nanoparticles of Au Au-Ag are made.
In above-described embodiment, in the step 10), the size of gold seeds is 10~15nm；In the step 20), gold
The size of grain is 30~100nm.
As shown in Fig. 2, the embodiment of the present invention also provides a kind of test strips containing above-mentioned hollow core shell nanoparticles, including
Bottom liner 5, sample pad 6, nano particle bonding pad 7, chromatographic film 8 and water absorption pad 11.Chromatographic film 8 is equipped with the detection line being mutually parallel
9 and nature controlling line 10；Sample pad 6, nano particle bonding pad 7, chromatographic film 8, water absorption pad 11 are closely coupled successively and are attached to bottom liner 5
On；Wherein, antibody and hollow core shell nanoparticles are adhered on nano particle bonding pad 7；Attachment capture antibody in detection line 9.
In the embodiment, hollow core shell nanoparticles 1 include nucleome 101 and housing 102.Housing 102 is located at outside nucleome 101
There is spacing in side between housing 102 and nucleome 101.The outer surface of housing 102 and inner surface are marked with Raman molecular 103 respectively.Core
The outer surface of body 101 is marked with Raman molecular 103.Housing 102 is made of electrum, and nucleome 101 is made of gold.
Above-mentioned test strips carry out the detection method of clinical marker, including：By the sample solution containing determinand 3 by adding
Sample hole is added dropwise in sample pad 6, and after standing, the Raman spectrum of 9 position of ELISA test strip line is gathered using Raman spectrometer, according to
The relative intensity of Raman spectrum obtains the concentration of determinand 3.Time of repose can be 10-20min, such as 15min.
Nano particle bonding pad 7 is equipped with the identification antibody 2 of nanoparticle label.Nano particle bonding pad 7 is fine by polyester
Dimension film is made.Sample pad 6 is made of glass fibre membrane.Chromatographic film 8 is made of nitrocellulose filter.9 antibody of detection line is capture
Antibody 4, the antibody of nanoparticle label is identification antibody.Capture antibody 4 mutually can match to form folder with identification antibody 2 for 2 plants
The monoclonal antibody of core structure.
The embodiment of the present invention prepares immuno-chromatographic test paper strip with the hollow core shell nanoparticles of Au Au-Ag, passes through Raman spectrum
Detection, realizes the quick, highly sensitive of clinical marker, half-quantitative detection.Sample is detected using Raman spectroscopy and
Discriminating have many advantages, such as it is non-contact, without destruction, without particular sample preparation process, and since influence of the water to Raman spectrum is non-
It is often small, so Raman spectrum can be widely used in the detection for possessing bioactivity sample in biosystem.Raman scattering
The intensity of spectrum is only the 10 of incident intensity-10, signal is too weak so that being difficult to collect detection.Surface Enhanced Raman Scattering Spectrum skill
The appearance of art (Surface Enhanced Raman Scattering, SERS) greatly improves the inspection of Raman spectroscopy
Survey sensitivity.When the molecule with Raman active is contacted with metal Nano structure, pass through the Electromagnetic enhancement of metal Nano structure
It is acted on Chemical enhancement effect, can be greatly enhanced the Raman spectrum of molecule, enhance ability up to 104~107.To be checked
In the case that survey object concentration is relatively low, highly sensitive detection is realized, and gold, silver nano particle can make the letter of its surface Raman molecule
Number intensity obtains greatly enhancing, and the intensity of Raman spectrum and the quantity of gold, silver nano particle have positive correlation, thus will
The SERS characteristics of gold, silver nano particle are detected applied to immunochromatography, you can realize that quick, highly sensitive, quantification detection will
It asks.
A specific example is enumerated below：
Quick detection for cTnI prepares the SERS immune chromatography test papers of the hollow core shell nanoparticles marks of Au Au-Ag
The step of and interpretation of result：
(1) the hollow core shell nanoparticles of Au Au-Ag of Raman molecular Nile blue (NBA) mark are prepared：A) first with biography
System hydro-thermal method prepares the gold seeds of 15nm：By 96mL water be heated to boiling after sequentially add 1mL concentration be 25mM gold chloride it is molten
Liquid and 3mL concentration are 1% sodium citrate solution；B) uniform 50nm gold grains are prepared by seed growth-curing method：It will
After 93mL water is heated to boiling, sequentially add 1mL concentration be 15mM sodium citrate solutions, 1mL concentration be 25mM chlorauric acid solutions,
1mL concentration is 25mM quinol solutions and the above-mentioned 15nm gold seedses of 4mL, 50nm gold grain solution is prepared, to gold grain
It is 10 that 0.5mL concentration is added in solution-4The NBA solution of M, by electrostatic absorption principle in one layer of Raman molecular of surface modification；c)
Au@Ag gold and silver core shell nanoparticles are prepared again by seed mediated growth method：Using the 50nm gold grains of the above-mentioned preparations of 50mL as seed,
Under stirring at normal temperature, sequentially add the ascorbic acid solution that 0.25mL concentration is 0.1M and the silver nitrate that 1.25mL concentration is 10mM is molten
Liquid grows into silver-colored shell on gold nano grain surface, is added with stirring 0.5mL concentration as 10-4The NBA solution of M, passes through Electrostatic Absorption
The Raman molecular in silver surface modification；D) the hollow core shell nanoparticles of Au Au-Ag are prepared using Au Ag as template is sacrificed：By 50mL
After above-mentioned Au@Ag nanoparticles solutions are heated to boiling, the AuCl that 24mL concentration is 1mM is added in2 -Solution filters after reaction
Au@Au-Ag nanoparticles solutions are obtained, add 0.5mL concentration as 10-4The NBA solution of M modifies one layer of drawing in particle surface
Graceful molecule.
(2) product of involved three different phases is in the hollow core shell nanoparticles preparation process of above-mentioned Au Au-Ag
50nm gold nano grains, Au Ag core shell nanoparticles and the hollow core shell nanoparticles of Au Au-Ag, because each self-structure is modified respectively
One layer, two layers, three layers of NBA Raman moleculars, by Raman spectrometer detect the identical lower three kinds of nanoparticles solutions of granule density
Raman spectrum, the results are shown in Figure 3.As can be seen from Figure 3：The hollow core shell nanoparticles of Au Au-Ag possess most strong SERS
Activity, much larger than two times and three times of gold nano grain and Au Ag core shell nanoparticles intensity.
(3) the above-mentioned hollow core shell nanoparticles pH value of solution of Au Au-Ag for having modified NBA molecules is adjusted to 8.5, is added in and is known
Other antibody (one plant of anti-human H-cTnI monoclonal antibody A) and the hollow core shell nanoparticles solution reactions of Au Au-Ag, centrifuging must sink
It forms sediment, precipitation is resuspended, must identify the hollow core shell nanoparticles solution of Au Au-Ag of antibody mark.
(4) it is polyester fiber film is (pure for 1% ox blood containing mass fraction with the Tris-HCl buffer solutions of 0.05M, pH9.0
Albumen, mass fraction 0.02%NaN3, mass fraction 0.1%PVA, mass fraction 0.9%NaCl, mass fraction be
10% sucrose and mass fraction are 0.1%TritonX-100) impregnate after, drying, the Au@Au- that above-mentioned identification antibody is used to mark
The hollow core shell nanoparticles solution spray printings of Ag are positioned in 37 DEG C of drying boxes dry standby.
(5) with capture antibody (one plant of anti-human H-cTnI monoclonal antibody B), detection is drawn in spray on nitrocellulose membrane has been activated
Line, vacuum drying；Then sprayed with sheep anti-mouse igg on nitrocellulose membrane has been activated and draw nature controlling line, vacuum drying；
(6) it is pasted onto according to the order of sample pad, Au@Au-Ag nanoparticle labels pad, nitrocellulose filter, water absorption pad
On the plastic plate of single side pressure sensitive adhesive, cutting is packed into plastic clip；
(7) test strips use：In the sample pad that sample solution is added dropwise by well, 15min is stood；
(8) testing result is read：A) use visually observes detection line color, and detection line can develop the color, then can qualitatively judge
Contain determinand in sample solution；B) Raman signal is detected in ELISA test strip line position using Raman spectrometer, according to Raman
Concentration of the signal strength quantitative analysis determinand in sample solution.
Fig. 4 is the optical photograph obtained after ELISA test strip various concentration cTnI.It is consistent with conventional method, it detects by an unaided eye
Various concentration cTnI immunochromatography testing results are merely able to tell the detection line in 5ng/mL and concentrations above test strips.
Fig. 5 is the Raman spectrum obtained using Raman spectrometer test strip detection line, it can be seen that with cTnI
The increase of concentration, the intensity of Raman spectrum gradually increase.With the main peak 592cm of Raman molecular NBA spectrum-1Intensity is ordinate,
The concentration of cTnI is abscissa, makes standard curve after carrying out Logarithm conversion, as shown in Figure 6.As can be seen from Figure 6：NBA light
The intensity of spectrum and cTnI concentration are in a linear relationship, and detection limit can reach 0.09ng/mL, and 50 are improved compared with visual results
Times.
The present invention replaces currently used spherical gold nano grain and gold and silver using the hollow core shell nanoparticles of Au Au-Ag
Core shell nanoparticles improve the detection sensitivity and storage stability of Surface enhanced Raman scattering immuno-chromatographic test paper strip, real
Show that human serum marker is simple and quick, highly sensitive, half-quantitative detection.
The basic principles, main features and advantages of the invention have been shown and described above.Those skilled in the art should
Understand, the present invention from above-mentioned specific embodiment limitation, the description in above-mentioned specific embodiment and specification be intended merely into
One step illustrates the principle of the present invention, without departing from the spirit and scope of the present invention, the present invention also have various change and
It improves, these changes and improvements all fall within the protetion scope of the claimed invention.The scope of protection of present invention will by right
Ask book and its equivalent thereof.
Claims (10)
1. a kind of hollow core shell nanoparticles, which is characterized in that including nucleome (101) and housing (102), the housing (102)
On the outside of nucleome (101), there are spacing, the outer surface of the housing (102) and Nei Biao between housing (102) and nucleome (101)
Face is marked with Raman molecular (103) respectively, and the outer surface of the nucleome (101) is marked with Raman molecular (103)；The housing
(102) it is made of electrum, the nucleome (101) is made of gold.
2. hollow core shell nanoparticles described in accordance with the claim 1, which is characterized in that the hollow core shell nanoparticles are used for
Surface enhanced Raman scattering immuno-chromatographic test paper strip.
3. a kind of preparation method of hollow core shell nanoparticles described in claim 1, which is characterized in that the described method includes：
Step 10) prepares gold seeds using hydro-thermal method；
Gold grain is prepared by seed growth-curing method in step 20)；
Step 30) prepares Au@Ag gold and silver core shell nanoparticles by seed mediated growth method；
Step 40) prepares Au Au-Ag hollow cores using Au Ag gold and silver core shell nanoparticles prepared by step 30) as template is sacrificed
Core-shell nanoparticles.
4. the preparation method of hollow core shell nanoparticles described in accordance with the claim 3, which is characterized in that the step 10) bag
It includes：After heating water to boiling, chlorauric acid solution and sodium citrate solution are sequentially added, prepares gold seeds；The step 10)
In, the size of gold seeds is 10~15nm.
5. the preparation method of hollow core shell nanoparticles described in accordance with the claim 3, which is characterized in that the step 20) bag
It includes：After heating water to boiling, sequentially add sodium citrate solution, chlorauric acid solution, quinol solution and step 10) and prepare
Gold seeds, gold grain is prepared；In the step 20), the size of gold grain is 30~100nm；Into gold grain solution
Nile blue solution is added in, by electrostatic absorption principle, in one layer of Raman molecular of gold grain surface modification.
6. the preparation method of hollow core shell nanoparticles described in accordance with the claim 3, which is characterized in that the step 30) bag
It includes：Gold grain prepared by step 20) under stirring at normal temperature, sequentially adds ascorbic acid solution and silver nitrate solution for seed,
Gold grain surface grows into silver-colored shell, is added with stirring NBA solution, by Electrostatic Absorption silver surface modification on Raman molecular, system
Into Au@Ag gold and silver core shell nanoparticles.
7. the preparation method of hollow core shell nanoparticles described in accordance with the claim 3, which is characterized in that the step 40) bag
It includes：After Au@Ag gold and silver core shell nanoparticles solution prepared by step 30) is heated to boiling, AuCl is added dropwise2 -Solution, instead
After answering, Au@Au-Ag nanoparticles solutions are obtained by filtration, add NBA solution, one layer of Raman point is modified in particle surface
The hollow core shell nanoparticles of Au Au-Ag are made in son.
A kind of 8. test strips containing hollow core shell nanoparticles described in claim 1, which is characterized in that the test strips bag
Include bottom liner (5), sample pad (6), nano particle bonding pad (7), chromatographic film (8) and water absorption pad (11)；Chromatographic film (8) is equipped with phase
Mutually parallel detection line (9) and nature controlling line (10)；Sample pad (6), nano particle bonding pad (7), chromatographic film (8), water absorption pad
(11) it is closely coupled successively and be attached on bottom liner (5)；Wherein, the hollow of antibody mark is adhered on nano particle bonding pad (7)
Core shell nanoparticles；Attachment capture antibody in detection line (9).
9. according to the test strips described in claim 8, which is characterized in that the hollow core shell nanoparticles include nucleome and shell
Body, the housing are located on the outside of nucleome, have spacing between housing and nucleome, the outer surface of the housing and inner surface mark respectively
There is Raman molecular, the outer surface of the nucleome is marked with Raman molecular；The housing is made of electrum, and the nucleome is gold
It is made.
A kind of 10. detection method that clinical marker is carried out using test strips described in claim 8, which is characterized in that this method
Including：
The sample solution that determinand (3) will be contained is added dropwise by well in sample pad (6), after standing, utilizes Raman spectrum
Instrument gathers the Raman spectrum of ELISA test strip line (9) position, according to the relative intensity of Raman spectrum, obtains the dense of determinand (3)
Degree.
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