WO2019088527A2 - Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc - Google Patents

Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc Download PDF

Info

Publication number
WO2019088527A2
WO2019088527A2 PCT/KR2018/012339 KR2018012339W WO2019088527A2 WO 2019088527 A2 WO2019088527 A2 WO 2019088527A2 KR 2018012339 W KR2018012339 W KR 2018012339W WO 2019088527 A2 WO2019088527 A2 WO 2019088527A2
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
zinc oxide
nanostar
pathogen
mixture
Prior art date
Application number
PCT/KR2018/012339
Other languages
English (en)
Korean (ko)
Other versions
WO2019088527A3 (fr
Inventor
신용
리우후이팡
Original Assignee
울산대학교 산학협력단
재단법인 아산사회복지재단
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 울산대학교 산학협력단, 재단법인 아산사회복지재단 filed Critical 울산대학교 산학협력단
Priority to US16/759,366 priority Critical patent/US11603526B2/en
Priority to EP18874738.0A priority patent/EP3705581A4/fr
Priority claimed from KR1020180124184A external-priority patent/KR102090877B1/ko
Publication of WO2019088527A2 publication Critical patent/WO2019088527A2/fr
Publication of WO2019088527A3 publication Critical patent/WO2019088527A3/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
    • C12N15/1006—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor by means of a solid support carrier, e.g. particles, polymers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1003—Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor

Definitions

  • the present invention relates to a method for extracting pathogenic bacteria and nucleic acids using zinc oxide nanostars.
  • Nucleic acid is an important analytical tool for identifying disease states, and DNA biomarkers, such as single nucleotide polymorphism (SNP), mutation or DNA methylation, And provides an important clue to providing a great opportunity for prognosis and surveillance as well as diagnosing and monitoring the condition of the disease during the early stages of the disease.
  • SNP single nucleotide polymorphism
  • nucleic acids such as DNA are present at very low physiological concentrations compared to other components such as proteins (e.g., tens of nanograms of DNA versus a few tens of micrograms of protein per microliter of whole blood), DNA is effectively extracted from clinical samples Preconcentration is very important for subsequent processes such as amplification and detection.
  • nucleic acids have been increasing in various fields such as biotechnology, diagnostic medicine, pharmacology, and metabolism. Therefore, efforts are being made to separate nucleic acids more rapidly and neatly from various biological samples.
  • nucleic acid adsorbing materials are a carrier that specifically adsorbs only nucleic acid from various kinds of substances contained in the cell lysis solution such as genomic DNA, plasmid DNA, messenger RNA, protein.
  • An object of the present invention is to provide a gentle composition for a pathogen including a zinc oxide nanostar, a pathogen ligation method and kit using the same, A composition for extracting nucleic acid including zinc oxide nanostar, a nucleic acid extraction method and kit using the same, and a kit.
  • the present invention provides a germicidal composition for a pathogen comprising zinc oxide nanostar.
  • the present invention also provides a kit for a pathogen comprising the composition.
  • the present invention provides a pathogen method comprising contacting a zinc oxide nanostar with a sample containing a pathogen.
  • the present invention also provides a composition for extracting nucleic acid comprising zinc oxide nanostar.
  • the present invention also provides a kit for nucleic acid extraction comprising the above composition.
  • the present invention provides a method for producing a zinc nanostructure comprising: a first step of adding a zinc oxide nanostar to a nucleic acid sample and reacting the mixture to prepare a mixture; And a second step of extracting the nucleic acid from the mixture.
  • the present invention provides a method for producing a zinc nanostructure comprising: a first step of adding a zinc oxide nanostar to a nucleic acid sample and reacting the mixture to prepare a mixture; Adding diatomite modified with a silane compound to the mixture and adding the mixture to a mixture of dimethyl adipimidate (DMA), dimethylpimelimidate (DMP), dimethyl suberimidate (DMS) and dimethyl
  • DMA dimethyl adipimidate
  • DMP dimethylpimelimidate
  • DMS dimethyl suberimidate
  • a second step of preparing a reaction mixture by adding at least one selected from the group consisting of 3,3'-dithiobispropionimidate (DTBP), 3,3'-dithiobispropionimidate (DTBP);
  • DTBP 3,3'-dithiobispropionimidate
  • DTBP 3,3'-dithiobispropionimidate
  • the nucleic acid extraction method using the zinc oxide nanostructure of the present invention can extract the nucleic acid by dissolving the cells of the pathogen without using a lysis buffer and is capable of extracting nucleic acid from a variety of substances including a high concentration of salts contained in the lysis buffer Thereby preventing nucleic acid degradation and fragmentation caused by the nucleic acid degradation.
  • the zinc oxide nanostar (200 to 900 nm) of the present invention is superior to conventional zinc oxide nanoparticles (20 to 50 nm) in cell solubility to increase nucleic acid extraction efficiency and can be extracted at room temperature without heating It can be used as a field diagnostic method.
  • FIG. 3 compares the solubility and nucleic acid extraction efficiency of zinc oxide nanostars using brucella bacteria with zinc oxide nanoparticles and a conventional nucleic acid extraction kit (Qiagen).
  • FIG. 4 compares the solubility and nucleic acid extraction efficiency of zinc oxide nanostars using Brucella, Escherichia coli, Staphylococcus aureus, and Bacillus cereus with a conventional nucleic acid extraction kit (Qiagen).
  • FIG. 5 confirms that nucleic acid extraction is possible by combining zinc oxide nanostar with a column of a conventional nucleic acid extraction kit (Qiagen).
  • FIG. 6 shows that the zinc oxide nanostar is combined with diatomaceous earth or the same type 2 functional imidoester to extract nucleic acid.
  • FIG. 8 is a graph comparing (A) DNA extraction and (B) RNA extraction efficiency of a conventional nucleic acid extraction kit (Qiagen), zinc oxide nanostar, and zinc oxide nanoparticles using HCT116 as a colon cancer cell line.
  • the inventors of the present invention have developed an extraction method capable of dissolving a pathogen and extracting a nucleic acid, and the ZnO nanostar (ZnO NS) of the present invention is superior to conventional zinc oxide nanoparticles in dissolving a cell membrane and nucleus of a pathogen
  • the nucleic acid extracting method using the zinc oxide nanostar of the present invention is capable of extracting nucleic acid of high concentration and high concentration without lysis buffer and heating step,
  • the present invention has been accomplished on the basis of the finding that nucleic acid can be extracted and an immediate on-the-spot diagnosis is possible.
  • the present invention provides a mycelial composition for pathogen comprising zinc oxide nanostar.
  • the zinc oxide nanostructure refers to a nanoparticle in which a plurality of sharp projections are annularly arranged.
  • the zinc oxide nanostars have an average particle diameter of 200 to 900 nm, preferably an average particle diameter of 350 to 900 nm, more preferably an average particle diameter of 500 nm, but are not limited thereto Specify.
  • the pathogen is a microorganism and the microorganism may be, but is not limited to, a virus, a bacterium, a fungus, a protozoa, a ricketta or a spirotherte.
  • the present invention also provides a kit for a pathogen comprising the composition.
  • the present invention provides a pathogen method comprising contacting a zinc oxide nanostar with a sample containing a pathogen.
  • the sample containing the pathogenic agent may be used as a sample of a subject suspected of being infected with a pathogen such as feces, urine, tears, saliva, external secretion of skin, external secretion of the respiratory tract, external secretion of the intestinal tract, external secretion of the digestive tract, Spinal fluid, lymph fluid, body fluids, and tissue, but is not limited thereto.
  • a pathogen such as feces, urine, tears, saliva, external secretion of skin, external secretion of the respiratory tract, external secretion of the intestinal tract, external secretion of the digestive tract, Spinal fluid, lymph fluid, body fluids, and tissue, but is not limited thereto.
  • the pathogen is a microorganism and the microorganism may be, but is not limited to, a virus, a bacterium, a fungus, a protozoa, a ricketta or a spirotherte.
  • the present invention also provides a composition for extracting nucleic acid comprising zinc oxide nanostar.
  • the zinc oxide nanostructure refers to a nanoparticle in which a plurality of sharp projections are annularly arranged.
  • the zinc oxide nanostars have an average particle diameter of 200 to 900 nm, preferably an average particle diameter of 350 to 900 nm, more preferably an average particle diameter of 500 nm, but are not limited thereto Specify.
  • the nucleic acid may be DNA or RNA.
  • the present invention also provides a kit for nucleic acid extraction comprising the above composition.
  • the present invention provides a method for producing a zinc nanostructure comprising: a first step of adding a zinc oxide nanostar to a nucleic acid sample and reacting the mixture to prepare a mixture; And a second step of extracting the nucleic acid from the mixture.
  • the present invention provides a method for producing a zinc nanostructure comprising: a first step of adding a zinc oxide nanostar to a nucleic acid sample and reacting the mixture to prepare a mixture; Adding diatomite modified with a silane compound to the mixture and adding the mixture to a mixture of dimethyl adipimidate (DMA), dimethylpimelimidate (DMP), dimethyl suberimidate (DMS) and dimethyl
  • DMA dimethyl adipimidate
  • DMP dimethylpimelimidate
  • DMS dimethyl suberimidate
  • a second step of preparing a reaction mixture by adding at least one selected from the group consisting of 3,3'-dithiobispropionimidate (DTBP), 3,3'-dithiobispropionimidate (DTBP);
  • DTBP 3,3'-dithiobispropionimidate
  • DTBP 3,3'-dithiobispropionimidate
  • the silane compound may be a compound represented by the following general formula (1), but is not limited thereto.
  • R 1 to R 3 may be the same or different and are any one of C 1 to C 4 alkyl or C 1 to C 4 alkoxy
  • R 4 is amino (C 1 to C 10) alkyl, 3- (2 amino (C1 to C4) alkylamino] (C1 to C4) alkyl or 3- [2- (2-amino (C1 to C4) alkylamino) (C1 to C4) alkylamino] (C1 to C4) alkyl.
  • the silane compound may be at least one selected from the group consisting of (3-aminopropyl) triethoxysilane (APTES), (3-aminopropyl) trimethoxysilane, (1-aminomethyl) triethoxysilane, (2-aminoethyl) triethoxysilane, (4-aminobutyl) triethoxysilane, (5-aminopentyl) triethoxysilane, (6-aminohexyl) triethoxysilane, 3-aminopropyl (diethoxy) methylsilane 3-aminopropyl (diethoxy) methylsilane; APDMS), N- [3- (trimethoxysilyl) propyl] ethylenediamine, N- [3- (trimethoxysilyl ) Propyl] diethylenetriamine, [3- (2-aminoethylamino) propyl] trimethoxysilane ([3
  • the present invention also relates to a process for preparing a mixture by adding zinc nitrate hexahydrate and hexadecyltrimethylammonium bromide to water; A second step of heating the mixture at 85 to 95 ⁇ for 30 to 80 minutes to prepare a reaction mixture; And a third step of adding a solution of ammonium hydroxide to the reaction mixture to prepare a colloidal solution.
  • Zinc nitrate hexahydrate Zn NO 3 * 6H 2 O, 98%), ammonium hydroxide solution (28% NH 3 in H 2 O, 99.99% trace metals), (3-aminopropyl (3-aminopropyl) triethoxysilane (APTES, 98%) was purchased from Sigma-Aldrich (St Louis, MO, USA).
  • Hexadecyltrimethylammonium bromide (CTAB) C 19 H 42 BrN,> 98%) was purchased from Tokyo Chemical Industry.
  • Commercial zinc oxide nanoparticles (dispersion, 20 wt% in size less than 40 nm in H 2 O) were used as controls.
  • biomaterials of diatomaceous earth were purchased from Sigma-Aldrich.
  • Milli-Q water, 99% ethyl alcohol, phosphate buffered saline (PBS, 10x, pH 7.4) and streptavidin conjugated magnetic beads (Dynabeads® MyOne TM Streptavidin C1) Respectively.
  • FE-SEM field-emission scanning electron microscopy
  • XRD X-ray diffraction
  • FT-IR Fastier transform infrared
  • a centrifuge (CF-5, 100 to 240 Vas, 50/60 Hz, 8 W), Vortex Mixer (T5AL, 60 Hz, 30 W, 250 V) and MSH- .
  • the Ariamx real-time PCR system (Agilent technologies), the Gene Amp PCR system 9700 (LSK), the Submerge-Mini, the gel documentation system and the Nanodrop 2000 (PeqLab) Respectively.
  • the cells used in the experiment were HCT116 (ATCC CCL-247), a colon cancer cell line.
  • Brucella ovis ATCC 25840
  • Escherichia coli E coli, ATCC 25922
  • Staphylococcus aureus Staphylococcus aureus S. aureus
  • Bacillus cereus B cereus
  • Zno nanostar (Zno NS) crystals were synthesized by hydrothermal method in alkaline medium.
  • reaction temperature, reaction time and stirring speed were controlled for uniform production of zinc oxide nanostars.
  • the reaction flask of the ice box was immediately taken out and centrifuged, dried at room temperature and washed with Milli-Q water. All syntheses were carried out without any special treatment, and finally the samples were stored in ethanol (99%).
  • zinc oxide nanostar has a particle size of 200 to 600 nm on average and shows a particle shape in which a plurality of sharp projections are annularly arranged
  • zinc oxide nanoparticles have an average of 20 ≪ / RTI > to 60 nm and exhibits an irregular spherical particle morphology.
  • the structures of zinc oxide nanostars and zinc oxide nanoparticles are different from each other by the Raman technique.
  • the zinc oxide nanostructure of the present invention has a particle size larger than that of the conventional zinc oxide nanoparticles and has a particle shape in which a plurality of sharp projections are annularly arranged to more easily dissolve cell membranes and nuclei of pathogenic bacteria Respectively.
  • the commercialization kit was used for comparative analysis of dissolution characteristics of zinc oxide nanostars.
  • the amount and purity of the extracted nucleic acid was measured with Ariamx real-time PCR system, Gene Amp PCR system 9700, electrophoresis apparatus, electrophoresis gel recording apparatus and Nanodrop 2000.
  • FIG. 3 and the following Table 1 show the results of using the conventional nucleic acid extraction kit (Qiagen) + lysis buffer, zinc oxide nanoparticles + dissolution buffer unused, zinc oxide nanostar + dissolution buffer unused, conventional nucleic acid extraction kit
  • Qiagen nucleic acid extraction kit
  • FIG. 3 and the following Table 1 show the results of using the conventional nucleic acid extraction kit (Qiagen) + lysis buffer, zinc oxide nanoparticles + dissolution buffer unused, zinc oxide nanostar + dissolution buffer unused, conventional nucleic acid extraction kit
  • dissolution characteristics of zinc oxide nanostar were comparatively analyzed using Brucella, Escherichia coli, Staphylococcus aureus and Bacillus cereus as in the above method.
  • the zinc oxide nanostar of the present invention was superior to the conventional nucleic acid extraction kit in solubility and nucleic acid extraction in all four kinds of bacteria.
  • FIG. 5 is an analysis of whether the zinc oxide nanostar of the present invention is capable of nucleic acid extraction by grafting into a conventional nucleic acid extraction kit (Qiagen) column. As a result of comparing nucleic acid extraction efficiencies with different conditions as shown in Table 2 below, it was confirmed that zinc oxide nanostar could efficiently extract nucleic acid even at room temperature without heating.
  • Qiagen nucleic acid extraction kit
  • A1 A2 B1 B2 C1 C2 Nucleic acid extract composition Qiagen Qiagen Qiagen + zinc oxide nanostar Qiagen + zinc oxide nanostar Qiagen Qiagen Dissolution buffer use use unused unused unused unused Temperature (°C) 56 Room temperature 56 Room temperature 56 Room temperature Time (minutes) 10 10 10 10 10 10 10
  • the diatomite pore structure can accommodate a variety of molecules, and the high density silanol groups on the walls are beneficial in linking with functional amine groups, so the washed diatomite is functionalized with APTES.
  • the detailed experimental procedure is as follows.
  • diatomite functionalized with APTES was dispersed in distilled water at a concentration of 50 mg / ml.
  • the characteristics of APTES - functionalized diatomite and pure diatomite were analyzed by field emission scanning electron microscopy, Fourier transform infrared spectroscopy and Raman microscopy system.
  • nucleic acid was extracted by adding diatomaceous earth functionalized with APTES and an imidoester crosslinking agent (Dimethyl suberimidate. 2 HCl, DMS) to a 5 ml tube containing the prepared biological sample.
  • an imidoester crosslinking agent Dimethyl suberimidate. 2 HCl, DMS
  • RNA extraction 100 ⁇ l of the sample solution was added to a 1.5 ml tube containing 10 ⁇ l of protease K and 10 ⁇ l of a lysis buffer (M-SDS lysis buffer or zinc oxide nanostar solution), mixed using a pipette, Min. ≪ / RTI > 10 [mu] l of DNase was added for RNA extraction.
  • M-SDS lysis buffer or zinc oxide nanostar solution a pipette, Min. ≪ / RTI &gt
  • 10 [mu] l of DNase was added for RNA extraction.
  • the cells were incubated at 56 ° C for 10 minutes for M-SDS lysis or mixed with a pipette for zinc oxide nanostar dissolution and cultured at room temperature for 2 minutes.
  • the amount and purity of the extracted nucleic acid was measured with Ariamx real-time PCR system, Gene Amp PCR system 9700, electrophoresis apparatus, electrophoresis gel recording apparatus and Nanodrop 2000.
  • the zinc oxide nanostar of the present invention is a diatomaceous earth functionalized with APTES and a homobifunctional imidoester [DMA (Dimethyl adipimidate), Dimethyl pimelimidate (DMP) (DMS) or dimethyl 3,3'-dithiobispropionimidate (DTBP)], as shown in Table 1 below.
  • DMA Dimethyl adipimidate
  • DMP Dimethyl pimelimidate
  • DTBP dimethyl 3,3'-dithiobispropionimidate
  • the zinc oxide nanostar of the present invention was grafted together with diatomite and DMS functionalized with APTES and the nucleic acid extraction efficiency was compared according to the conditions as shown in Table 3. As a result, Extraction was possible. In addition, since it does not use a lysis buffer, it can be applied to the detection technique through the final PCR by giving less damage to the nucleic acid.
  • the extraction efficiency of the nucleic acid according to the particle size was compared, and the extraction efficiency of the nucleic acid of the zinc oxide nanostar having the particle size of 350 to 900 nm was the best, Zinc oxide nanostar with particle size of.
  • Example 7 Nucleic acid extraction method using zinc oxide nanostar
  • Zinc oxide nanostar was synthesized by hydrothermal method on alkaline medium.
  • reaction temperature, reaction time and stirring speed were controlled for uniform production of zinc oxide nanostars.
  • the zinc oxide nanostar solution was added to a 1.5 ml tube containing 100 ⁇ ⁇ of brucellosis (CFU: 10 ⁇ 5 / ml), and gently mixed with a pipette within 10 minutes at room temperature.
  • CFU brucellosis
  • the prepared sample was transferred to a Qiagen column, washed, and then the DNA was eluted with 60 Q of Qiagen elution buffer.
  • diatomite-APTES 2 mg / ml 50 mg / ml, 40 [mu] l was added and then 100 [mu] l of a 100 mg / ml DMS solution was added. After mixing, the cells were incubated at 56 ° C for 10 minutes for M-SDS lysis, or mixed with a pipette for zinc oxide nanostar dissolution and cultured at room temperature for 2 minutes.
  • optical strip lid was placed in the well, being careful not to cross-contaminate and stain the surface of the lid.
  • Step 1 95 ° C, 10 min
  • Step 2 95 ⁇ , 10 seconds; 60 ⁇ , 20 seconds; 72 ° C, 20 seconds (40 to 45 cycles)
  • Step 3 72 ° C, 10 min
  • Example 8 Solubility test of zinc oxide nanostar using eukaryotic cells
  • HCT116 cells In order to verify the usefulness of zinc oxide nanostars in eukaryotic cells, HCT116 cells, a colon cancer cell line, were serially diluted to a concentration of 1 to 10 4 cells / 100 ⁇ l, and then zinc oxide nanostar, zinc oxide nano- And extraction efficiency of DNA and RNA was analyzed using a conventional nucleic acid extraction kit (Qiagen).
  • zinc oxide nanostars were able to detect DNA and RNA 1 to 1.5 cycles earlier than zinc oxide nanoparticles (20 nm) .
  • zinc oxide nanostar extracts nucleic acids at a higher concentration and purity than conventional nucleic acid extraction kits, and even in a small number of cells, high concentrations and high purity (3.5 ⁇ 2.3 ng / ⁇ l and 1.73 ⁇ 0.26 at 10 1 cells / ).
  • Zinc oxide nanostars enable rapid cell lysis at room temperature without the use of lysis buffers in various types of cells, making them useful as diagnostic systems in clinical settings.

Abstract

La présente invention concerne un procédé de lyse de pathogènes et d'extraction d'acides nucléiques utilisant une nanoétoile d'oxyde de zinc. Un procédé d'extraction d'acides nucléiques utilisant une nanoétoile d'oxyde de zinc selon la présente invention permet l'extraction d'acides nucléiques par lyse cellulaire d'agents pathogènes sans utiliser de tampon de lyse et permet l'extraction d'acides nucléiques à une pureté élevée et à une concentration élevée tout en empêchant la dégradation et la fragmentation des acides nucléiques attribuées à divers matériaux dans un tampon de lyse, notamment une concentration élevée de sels. En outre, les nanoétoiles d'oxyde de zinc (200 à 900 nm) selon la présente invention, qui sont des nanoparticules d'oxyde de zinc classiques supérieures (20 à 50 nm) en termes de potentiel de lyse cellulaire, améliorent l'efficacité d'extraction d'acides nucléiques grâce à leur potentiel de lyse cellulaire et permettent l'extraction à température ambiante sans étape de chauffage, ce qui permet de trouver une application en tant que procédé de diagnostic dans le domaine.
PCT/KR2018/012339 2017-10-30 2018-10-18 Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc WO2019088527A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US16/759,366 US11603526B2 (en) 2017-10-30 2018-10-18 Pathogen lysis and nucleic acid extraction method using zinc oxide nanostar
EP18874738.0A EP3705581A4 (fr) 2017-10-30 2018-10-18 Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
KR20170142590 2017-10-30
KR10-2017-0142590 2017-10-30
KR10-2018-0124184 2018-10-18
KR1020180124184A KR102090877B1 (ko) 2017-10-30 2018-10-18 산화아연 나노스타를 이용한 병원체 용균 및 핵산 추출 방법

Publications (2)

Publication Number Publication Date
WO2019088527A2 true WO2019088527A2 (fr) 2019-05-09
WO2019088527A3 WO2019088527A3 (fr) 2019-07-04

Family

ID=66331463

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2018/012339 WO2019088527A2 (fr) 2017-10-30 2018-10-18 Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc

Country Status (1)

Country Link
WO (1) WO2019088527A2 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114988456A (zh) * 2022-06-21 2022-09-02 深圳市尚维高科有限公司 氧化锌复合颗粒及制备方法及其在核酸检测中的病毒裂解应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2010078399A2 (fr) * 2008-12-31 2010-07-08 3M Innovative Properties Company Dispositifs d'échantillonnage et procédés de concentration de microorganismes
ES2706761T3 (es) * 2012-12-13 2019-04-01 One Biomed Pte Ltd Métodos sin marcadores para el aislamiento y análisis de ácidos nucleicos en dispositivos en fase sólida
KR20150142492A (ko) * 2014-06-12 2015-12-22 호서대학교 산학협력단 산화아연 나노입자를 포함하는 알레르기 염증 질환 치료용 조성물

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BIOSENSORS AND BIOELECTRONICS, vol. 99, 2018, pages 443 - 449
See also references of EP3705581A4

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114988456A (zh) * 2022-06-21 2022-09-02 深圳市尚维高科有限公司 氧化锌复合颗粒及制备方法及其在核酸检测中的病毒裂解应用
CN114988456B (zh) * 2022-06-21 2023-10-03 深圳市尚维高科有限公司 氧化锌复合颗粒及制备方法及其在核酸检测中的病毒裂解应用

Also Published As

Publication number Publication date
WO2019088527A3 (fr) 2019-07-04

Similar Documents

Publication Publication Date Title
KR102090877B1 (ko) 산화아연 나노스타를 이용한 병원체 용균 및 핵산 추출 방법
JP5651011B2 (ja) ポリヌクレオチド捕捉材料およびその使用方法
KR20080005213A (ko) 생물학 샘플로부터의 ｒｎａ 격리 강화 방법
JP2008513007A (ja) 生物学材料および細胞材料からの核酸の単離方法
WO2019088527A2 (fr) Procédé de lyse de pathogènes et d'extraction d'acides nucléiques à l'aide de nanoétoiles d'oxyde de zinc
KR20090003205A (ko) Ｒｎａ를 추출하는 방법
WO2010137903A2 (fr) Procédé de criblage d'une substance candidate médicamenteuse qui inhibe l'interaction protéine-protéine cible pour développer un nouveau concept de médicament
KR20090003219A (ko) 핵산를 추출하는 방법
WO2015199423A1 (fr) Procédé d'isolement d'acide nucléique à l'aide de particule magnétique
WO2018131826A1 (fr) Procédé et composition servant à isoler de l'adn plasmidique à l'aide d'une nanoparticule magnétique revêtue d'amine
JPWO2016152763A1 (ja) 核酸の回収方法
WO2012002597A1 (fr) Amorce destinée à diagnostiquer le virus de l'hépatite b, sonde, kit comprenant celles-ci, et procédé de diagnostic du virus de l'hépatite b utilisant le kit
WO2009102171A2 (fr) Particules magnétiques de silice de forme sphérique et processus de préparation de celles-ci
CN1043941A (zh) 34-脱(n-乙酰葡萄糖胺基)-34-脱氧-太可普劳宁的c63酰胺衍生物
WO2022102885A1 (fr) Technique de diagnostic du stress associé à la dermatite atopique utilisant des miarn dérivés d'exosomes
WO2022097984A1 (fr) Procédé de production de vésicules extracellulaires dérivées d'un agrégat cellulaire tridimensionnel de type sphéroïde
WO2018169307A1 (fr) Procédé de concentration microbienne au moyen de diatomite et procédé d'extraction d'acides nucléiques
WO2021141369A1 (fr) Procédé de détection d'arn à base de sonde d'adn simple brin
WO2018048194A1 (fr) Composition et procédé pour améliorer la sensibilité et la spécificité de la détection d'acides nucléiques à l'aide d'une protéine dcas9 et de la liaison d'arng à une séquence d'acide nucléique cible
WO2021221368A1 (fr) Procédé de filtration de vésicule dérivée de cellules
WO2019088429A1 (fr) Procédé d'extraction d'un biomatériau issu d'une biopsie liquide à l'aide d'imidoester homobifonctionnel
WO2011142646A2 (fr) Procédé de détection de hpv (papillomavirus humain) et de son génotype
EP3368690A1 (fr) Procédé de diagnostic prénatal à l'aide de la pcr numérique.
JP3788999B2 (ja) 抄紙斑点の原因となる微生物の検知方法及びそれに用いるプライマー
WO2017051939A1 (fr) Procédé pour extraire et amplifier un acide nucléique à l'aide de particules magnétiques

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 18874738

Country of ref document: EP

Kind code of ref document: A2

ENP Entry into the national phase

Ref document number: 2018874738

Country of ref document: EP

Effective date: 20200602