CN104819967A - Novel gold nanorod composite material and method thereof for detecting concentration of calf thymus DNA (Deoxyribonucleic Acid) - Google Patents
Novel gold nanorod composite material and method thereof for detecting concentration of calf thymus DNA (Deoxyribonucleic Acid) Download PDFInfo
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- CN104819967A CN104819967A CN201510206233.1A CN201510206233A CN104819967A CN 104819967 A CN104819967 A CN 104819967A CN 201510206233 A CN201510206233 A CN 201510206233A CN 104819967 A CN104819967 A CN 104819967A
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Abstract
The invention discloses a novel gold nanorod composite material wrapped by a fluorescent dye and with a core shell structure and a method thereof for detecting concentration of calf thymus DNA (Deoxyribonucleic Acid). On the basis that the experimental conditions of optimal detection comprise concentration of the composite material (Au NRs@SiO2-AO), pH value of reaction, different adding sequences and reaction time and under the optimum condition, the fluorescence emission spectrum after action of the calf thymus DNAs of different concentrations and the composite material is detected by using a fluorescence spectrophotometer, and finally the linear relation between the fluorescence emission spectrum and the ctDNA concentration is analyzed. The application method is a detection means enhanced by using fluorescence, and has a stable signal and good linearity and realizes rapid and easy detection during detection of the calf thymus DNA.
Description
Technical field
The present invention relates to analytical chemistry field, particularly a kind of novel gold nanorods compound substance and for detecting calf thymus DNA.
Background technology
Gold nanorods (Au NRs) is a kind of nano material of anisotropic one dimension, and it has a wide range of applications on biology and biomedicine as a kind of novel substitute.Because gold nanorods has superpower radiativity and non-radiation type, it can be used in the following aspects, the biology transmission etc. of such as biology sensor, bio-imaging, gene and medicine.[see 1.Nusz G.J., Marinakos S.M., Curry A.C., et al. Label-free plasmonic detection of biomolecular binding by a single gold nanorod. Anal. Chem., 2008, 80, 2. 984-989. Huang X.H., Neretina S., El-Sayed M.A., Gold nanorods:from synthesis and properties to biological and biomedical applications. Adv. Mater., 2009, 21, 4880-4910. 3. Takahashi H., Niidome Y., Yamada S., Gold nanorod-sensitized cell death:microscopic observation of single living cells irradiated by pulsed near-infrared laser light in the presence of gold nanorods. Chem. Commun., 2006, 35, 500-501.] nanometer rods has very high specific surface area, it is often chosen as nano-carrier and carrys out the various material of load to build multifunctional nano probe.[see Xiao Z.Y., Ji C.W., Shi J.J., et al. DNA self-assembly of targeted near-infrared-responsive gold nanoparticles for cancer thermo-chemotherapy. Angew. Chem. Int. Edit., 2012,124,12023-12027.] at present, the biology sensor based on gold nanorods mainly depends on surface plasma body resonant vibration.[see Yu C.X., Nakshatri H., Irudayaraj J., Identity profiling of cell surface markers by multiplex gold nanorod probes. Nano. Lett., 2007,7,2300-2306.] therefore, in the field of fluorescence probe, the biology sensor built based on gold nanorods is very important.
Summary of the invention
The object of this invention is to provide a kind of coated fluorescent dye, there is the compound substance of nucleocapsid structure and for detecting calf thymus DNA.
To achieve these goals, the present invention is by the following technical solutions:
A kind of compound substance of novel gold nanorods, coating mesoporous silicon doping acridine orange and there is the gold nanorods of nucleocapsid structure, the concentration of this compound substance is 0.6489 nM, the major axis of the gold nanorods used is 100 ± 4 nm, and minor axis is 28 ± 3 nm, and the TEM image of the gold nanorods used as shown in Figure 1, gold nanorods in building-up process, tetraethyl orthosilicate, the methanol solution of 20 %), and compound concentration is the acridine orange of 10 mg/mL, volume ratio be 3000:12:5.
Synthesized compound substance adopts the method for Fluorescence Increasing to detect calf thymus DNA;
Following steps are adopted to detect calf thymus DNA:
Step one: detect variable concentrations, the ctDNA of 0-18 μ g/mL is to the response of fluorescence probe corresponding to compound substance;
Step 2: make fluorescence intensity linearity curve corresponding to variable concentrations ctDNA according to the experimental result of step one;
Step 3: add testing sample, tests its response to fluorescence probe corresponding to compound substance, brings the linearity curve that step 2 obtains into, calculate its concentration.
In described step one, the experiment condition of first optimum detection, comprises compound substance (Au NRs@SiO
2-AO) concentration, the pH value of reaction, different addition sequences and reaction time, then at optimum conditions, fluorescence emission spectrum after utilizing fluorescence spectrophotometer to detect variable concentrations calf thymus DNA and this compound substance effect, ultimate analysis goes out the linear relationship between emission spectrum and concentration.
(1) the Au NRs@SiO of different concentration is chosen
2-AO (0 nM, 0.1625 nM, 0.1083 nM, 0.0813 nM, 0.065 nM, 0.0433 nM, 0.0325 nM), detects its fluorescence intensity, and the concentration analyzing best compound substance is 0.1083 nM;
(2) select B-R damping fluid as the correctives of pH in reaction environment.We have selected pH from 1.8 to 8.7, and then after adding finite concentration ctDNA, we detect Au NRs@SiO respectively
2the fluorescence spectrum of-AO, analyzing best pH value in reaction is 5.7;
(3) following three kinds of materials are detected: Au NRs@SiO
2-AO, BR damping fluid and ctDNA, the different addition sequence of three is on the impact of experimental fluorescence intensities, and having carried out analysis discovery to the result of experiment ought (Au NRs@SiO in the following sequence
2-AO, BR damping fluid, ctDNA) fluorescence signal that obtains when detecting is the strongest, illustrates that now Fluorescence Increasing value is maximum, so show that the addition sequence of the best is followed successively by: Au NRs SiO
2-AO, BR damping fluid, ctDNA;
(4) duration of contact that detection fluorescence probe is different from ctDNA, on the impact of fluorescence signal stability, show that the best reaction time is 1 min;
(5) under above top condition, the ctDNA of variable concentrations is detected on the impact of fluorescence probe corresponding to this compound substance.
A kind of compound substance based on gold nanorods is for detecting the application of calf thymus DNA.
Introducing due to mesoporous silicon can increase the adsorbance of acridine orange, specific surface area and its stability of gold nanorods can be improved further after being coated on gold nanorods surface simultaneously, we devise a kind of novel coated gold nanorods compound substance of the doped meso-porous silicon of fluorescence probe-acridine orange, and for detecting the method for calf thymus DNA
Beneficial effect of the present invention:
(1) application process of the present invention is applicable to other nanocomposite load fluorescent dyes or medicine for fields such as coherent detections.
(2) detection means described in application process of the present invention is the method for the Fluorescence Increasing adopted, and has potential value to field of nucleic acid detection.
(3) application process of the present invention, when detecting calf thymus DNA, signal stabilization, linearly well, detects quick, simple.
Accompanying drawing explanation
The TEM image of the gold nanorods that the core-shell material that Fig. 1 synthesizes uses
The core-shell composite material of this gold nanorods of Fig. 2 detects the fluorescence response curve corresponding with variable concentrations ctDNA effect, and wherein the concentration of the ctDNA that a-o is corresponding is followed successively by 0, and 0.5,1.5,2.5,3.5,4.5,5.5,6.5,7.5,8.5,9.5,10.5,13.5,15.5,18 μ g/mL.
The fluorescence intensity linearity curve that Fig. 3 variable concentrations ctDNA is corresponding.Insertion figure is (F-F
0)/F
0with the linear correlation curve of ctDNA concentration.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment 1:
A kind of compound substance of novel gold nanorods, coating mesoporous silicon doping acridine orange and there is the gold nanorods of nucleocapsid structure, the concentration of this compound substance is 0.6489 nM, the major axis of the gold nanorods used is 100 ± 4 nm minor axises is 28 ± 3 nm, as shown in Figure 1, the volume ratio of the gold nanorods in building-up process, tetraethyl orthosilicate (methanol solutions of 20 %) and acridine orange (compound concentration is 10 mg/mL) is 3000:12:5 to the TEM image of the gold nanorods used.
Synthesized compound substance adopts the method for Fluorescence Increasing to detect calf thymus DNA;
Following steps are adopted to detect calf thymus DNA:
Step one: detect variable concentrations, the ctDNA of 0-18 μ g/mL is to the response of fluorescence probe corresponding to compound substance;
Step 2: make fluorescence intensity linearity curve corresponding to variable concentrations ctDNA according to the experimental result of step one;
Step 3: add testing sample, tests its response to fluorescence probe corresponding to compound substance, brings the linearity curve that step 2 obtains into, calculate its concentration.
The optimization of experiment condition:
Au NRs@SiO
2the concentration optimization of-AO: the Au NRs@SiO choosing different concentration
2-AO (0 nM, 0.1625 nM, 0.1083 nM, 0.0813 nM, 0.065 nM, 0.0433 nM, 0.0325 nM), detects its fluorescence intensity, analyzes the concentration of best compound substance;
Reaction pH optimizes: select B-R damping fluid as the correctives of pH in reaction environment.We have selected pH from 1.8 to 8.7, and then after adding finite concentration ctDNA, we detect Au NRs@SiO respectively
2the fluorescence spectrum of-AO, analyzes best pH value in reaction;
Addition sequence is optimized: detect following three kinds of materials: Au NRs@SiO
2-AO, BR damping fluid and ctDNA, the different addition sequence of three is on the impact of experimental fluorescence intensities;
Reaction time is optimized: detect fluorescence probe duration of contact different from ctDNA to the impact of fluorescence signal stability, draw the reaction time of the best;
Detect ctDNA: under above top condition, detect the ctDNA of variable concentrations (0-18 μ g/mL) to the response of fluorescence probe corresponding to this compound substance, corresponding response curve is for shown in Fig. 2.Wherein sensing range can from 0.5 μ g/mL to 10 μ g/mL, and detect and be limited to 0.1667 μ g/mL, linearly dependent coefficient is 0.9985, and corresponding linearity curve is for shown in Fig. 3.Therefore, this kind of compound substance can be used for quantitatively detecting calf thymus DNA, and the rapid simple and stable of testing process.
More than show and describe ultimate principle of the present invention and principal character and advantage of the present invention.The technician of the industry should be appreciated that; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and instructions just illustrates principle of the present invention; without departing from the spirit and scope of the present invention; the present invention also has various changes and modifications; these changes and improvements all fall in the claimed scope of the invention, and application claims protection domain is defined by its equivalent of appending claims.
Claims (4)
1. a novel gold nanorods compound substance, it is characterized in that coating mesoporous silicon doping acridine orange and have the gold nanorods of nucleocapsid structure, the gold nanorods in this compound substance building-up process, tetraethyl orthosilicate (methanol solutions of 20 %) and compound concentration are the volume ratio of the acridine orange of 10 mg/mL is 3000:12:5.
2. novel gold nanorods compound substance according to claim 1 is for detecting a method for the concentration of calf thymus DNA, specifically comprises the steps:
Step one: detect variable concentrations, the ctDNA of 0-18 μ g/mL is to the response of fluorescence probe corresponding to novel gold nanorods compound substance;
Step 2: make fluorescence intensity linearity curve corresponding to variable concentrations ctDNA according to the experimental result of step one;
Step 3: add testing sample, tests its response to fluorescence probe corresponding to novel gold nanorods compound substance, brings the linearity curve that step 2 obtains into, calculate its concentration.
3. the compound substance of novel gold nanorods according to claim 2 is for detecting the method for the concentration of calf thymus DNA, it is characterized in that the concentration of novel gold nanorods compound substance in step one is 0.1083 nM, use B-R damping fluid to regulate reaction system PH to be 5.7, addition sequence is followed successively by: Au NRs@SiO
2-AO, BR damping fluid, ctDNA; Detecting fluorescence probe duration of contact different from ctDNA to the impact of fluorescence signal stability, is 1 min at once.
4. the compound substance of novel gold nanorods according to claim 2 is for detecting the method for the concentration of calf thymus DNA, it is characterized in that in step 2, the slit width exciting and launch of fluorescence spectrophotometer is respectively 3 nm and 5 nm.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102346148A (en) * | 2011-08-18 | 2012-02-08 | 江南大学 | Method for detecting surface enhanced Raman scattering of cancer cells based on self-assembled material |
| US20120308444A1 (en) * | 2011-05-30 | 2012-12-06 | Jimin Zhu | Lateral Flow Immunoassay for Detecting Cardiac Troponin I and Myoglobin |
| CN103076327A (en) * | 2012-10-25 | 2013-05-01 | 宁波大学 | Method and kit for quantificationally detecting lead ions by using gold label silver staining technology |
| CN103308462A (en) * | 2013-06-26 | 2013-09-18 | 南京邮电大学 | Surface plasma resonance probe with silver-gold core-satellite structure and preparation method thereof |
| WO2014035039A1 (en) * | 2012-08-27 | 2014-03-06 | 고려대학교산학협력단 | Gene transcription analysis method using surface plasmon resonance sensor |
| CN103776772A (en) * | 2014-02-14 | 2014-05-07 | 国家纳米科学中心 | Method for detecting DNA (deoxyribonucleic acid) on basis of gold nanocone accumulated circular dichroism signal |
| CN103884693A (en) * | 2012-12-20 | 2014-06-25 | 江南大学 | Preparation method for monodispersed and low-biotoxicity gold nanorods, and use for detection of allergen |
-
2015
- 2015-04-28 CN CN201510206233.1A patent/CN104819967A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120308444A1 (en) * | 2011-05-30 | 2012-12-06 | Jimin Zhu | Lateral Flow Immunoassay for Detecting Cardiac Troponin I and Myoglobin |
| CN102346148A (en) * | 2011-08-18 | 2012-02-08 | 江南大学 | Method for detecting surface enhanced Raman scattering of cancer cells based on self-assembled material |
| WO2014035039A1 (en) * | 2012-08-27 | 2014-03-06 | 고려대학교산학협력단 | Gene transcription analysis method using surface plasmon resonance sensor |
| CN103076327A (en) * | 2012-10-25 | 2013-05-01 | 宁波大学 | Method and kit for quantificationally detecting lead ions by using gold label silver staining technology |
| CN103884693A (en) * | 2012-12-20 | 2014-06-25 | 江南大学 | Preparation method for monodispersed and low-biotoxicity gold nanorods, and use for detection of allergen |
| CN103308462A (en) * | 2013-06-26 | 2013-09-18 | 南京邮电大学 | Surface plasma resonance probe with silver-gold core-satellite structure and preparation method thereof |
| CN103776772A (en) * | 2014-02-14 | 2014-05-07 | 国家纳米科学中心 | Method for detecting DNA (deoxyribonucleic acid) on basis of gold nanocone accumulated circular dichroism signal |
Non-Patent Citations (4)
| Title |
|---|
| FLÁVIO LAURETTI ET AL: "Use of acridine orange staining for the detection of rotavirus RNA in polyacrylamide gels", 《JOURNAL OF VIROLOGICAL METHODS》 * |
| TIAN MING ET AL: "Strong Polarization Dependence of Plasmon-Enhanced Fluorescence on Single Gold Nanorods", 《NANO LETTERS》 * |
| XIANG-QUN GUO ET AL: "DNA-Dye Fluorescence Enhancement Based on Shifting the Dimer-Monomer Equilibrium of Fluorescent Dye", 《APPLIED SPECTROSCOPY》 * |
| 徐良敏 等: "纳米金胶体对吖啶橙水溶液的荧光增强效应", 《宁夏师范学院学报（自然科学）》 * |
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Application publication date: 20150805 |