WO2021015452A1 - Kit de détection de matériaux cibles et procédé de détection de matériaux cibles faisant appel à celui-ci - Google Patents

Kit de détection de matériaux cibles et procédé de détection de matériaux cibles faisant appel à celui-ci Download PDF

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WO2021015452A1
WO2021015452A1 PCT/KR2020/008894 KR2020008894W WO2021015452A1 WO 2021015452 A1 WO2021015452 A1 WO 2021015452A1 KR 2020008894 W KR2020008894 W KR 2020008894W WO 2021015452 A1 WO2021015452 A1 WO 2021015452A1
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nucleic acid
isothermal amplification
kit
hydrogel
inner primer
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Korean (ko)
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이민영
정병호
최동일
우아영
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사회복지법인 삼성생명공익재단
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
    • C12Q1/701—Specific hybridization probes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/531—Production of immunochemical test materials
    • G01N33/532—Production of labelled immunochemicals
    • G01N33/533—Production of labelled immunochemicals with fluorescent label
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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    • C12Q2527/00—Reactions demanding special reaction conditions
    • C12Q2527/101—Temperature
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    • C12Q2537/00—Reactions characterised by the reaction format or use of a specific feature
    • C12Q2537/10—Reactions characterised by the reaction format or use of a specific feature the purpose or use of
    • C12Q2537/143—Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
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    • C12Q2563/00—Nucleic acid detection characterized by the use of physical, structural and functional properties
    • C12Q2563/107—Nucleic acid detection characterized by the use of physical, structural and functional properties fluorescence
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    • C12Q2565/00—Nucleic acid analysis characterised by mode or means of detection
    • C12Q2565/50—Detection characterised by immobilisation to a surface
    • C12Q2565/537—Detection characterised by immobilisation to a surface characterised by the capture oligonucleotide acting as a primer
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • It relates to a target substance detection kit and a method of detecting a target substance using the same.
  • Conventional methods for identifying pathogens of acute respiratory infectious diseases include traditional cell culture, shell vial culture, immunofluorescence staining, rapid antigen detection, and nucleic acid amplification.
  • the traditional cell culture method has the advantage that it is possible to diagnose and isolate various viruses and/or bacteria, but it takes a long period of 3 to 10 days for diagnosis, and an additional method of immunofluorescence staining is required, and is performed by a skilled person.
  • diagnosis due to the presence of viruses or bacteria that cannot be cultured.
  • the rapid culture method is useful for diagnosing a virus that is not well cultured, and even an unskilled person can easily confirm the cytopathic effect, but it takes 24 to 72 hours to diagnose, and there are viruses or bacteria that cannot be cultured, so the diagnosis is limited. have. In addition, there is a risk that the examiner will become infected.
  • Recently, methods to identify respiratory pathogens have been developed using a molecular biological method, multiplex PCR. Multiple PCR is a method of simultaneously amplifying several genes by using a combination of more than one pair of primers.
  • there is a limit to the number of discrimination and diagnosis and since it takes about 5 hours or more for diagnosis, there is a limitation to apply to on-site diagnosis.
  • One aspect relates to a kit for multiple detection of a target substance including a substrate on which a nucleic acid probe or antibody capable of specifically binding to an isothermal amplification product of a target substance is immobilized on or inside a hydrogel.
  • Another aspect comprises obtaining a nucleic acid sample from a biological sample of the subject; And contacting the sample to a substrate on which a nucleic acid probe or antibody capable of specifically binding to an isothermal amplification product of a target material is immobilized on the surface or inside of the hydrogel, comprising the step of contacting the sample.
  • One aspect is a substrate on which a nucleic acid probe or antibody capable of specifically binding to an isothermal amplification product of a target nucleic acid is immobilized on the surface or inside of a hydrogel; And it provides a kit for multiple detection of a target substance comprising a set of primers capable of specifically binding to a target nucleic acid.
  • the kit can detect a target substance more quickly and conveniently because it can detect a specific gene in the field even if it is not in a laboratory environment.
  • the antibody may be, for example, anti-hapten.
  • the target substance detection kit may be in the form of a miniaturized diagnostic chip, for example, a lab on a chip.
  • the term "isothermal amplification” refers to a method of amplifying a target nucleic acid under constant reaction temperature conditions. While the conventional PCR method has to change the temperature while going through the steps of denaturation, annealing, and extension, the isothermal amplification method has the advantage that conjugation and extension are possible at a constant temperature. This is the Taq. Instead of DNA polymerase, Bst. This was made possible by using a DNA polymerase. Bst. DNA polymerase is Taq.
  • the primer may be an inner primer set, an outer primer set, and/or a loop primer set.
  • Isothermal amplification methods include HAD (Helicase-Dependent Amplification), RPA (Recombinase Polymerase Amplification), RCA (Rolling Circle Amplification), LAMP (Loop mediated isothermal amplification), NASBA (Nucleic Acid-Sequence-Based Amplification), TMA (Transcription Mediated Amplification). ), SMART (Signal Mediated Amplification of RNA Technology), SDA (Strand Displacement Amplification), IMDA (Isothermal Multiple Displacement Amplification), SPIA (Single Primer Isothermal Amplification), HDA (Helicase-Dependent Amplification), or a combination thereof. .
  • a nucleic acid probe or antibody capable of specifically binding to the isothermal amplification product of the target nucleic acid may be three-dimensionally immobilized on the surface or inside of the hydrogel. Since the nucleic acid probe or antibody is immobilized on the surface or inside of the hydrogel, cross-reaction or interference reaction does not occur between the probe or antibody, and thus, there is an advantage in that it is easy for multiple diagnosis.
  • the probe may be a set of forward inner primers (FIP) or a set of backward inner primers (BIP) of the target nucleic acid.
  • the outer primer set may include a sequence complementary to a sequence closer to both ends of the target nucleic acid than the inner primer set, and the inner primer set may include a sequence closer to the center of the target nucleic acid than the outer primer set.
  • the nucleic acid probe may be a linker capable of binding to the surface or the interior of the hydrogel is bonded to the 5'end.
  • a linker capable of binding to the surface or the interior of the hydrogel may be bonded to the 5'end of the forward inner primer and/or the reverse inner primer.
  • the linker may be, for example, a compound including an acrylicdite, a thiol group, or a derivative thereof.
  • the linker may be bonded to the surface or the interior of the hydrogel by covalent bonding with the hydrogel.
  • the nucleic acid forward inner primer and/or the reverse inner primer may have a labeling material bound to the 5'end, and the labeling material may be a reporter or a quencher.
  • the reporter is, for example, fluorescein, fluorescein chlorotriazinyl, rhodamine green, rhodamine red, tetramethylrhodamine, FITC, Oregon green, Alexa Fluor, FAM, JOE, ROX, HEX, Texas Red, TET, TRITC, TAMRA, Cyanine series dyes and siadicarbocyanin (thiadicarbocyanine) It may be one or more selected from the group consisting of dyes.
  • the quencher is, for example, Dabcyl, TAMRA, Eclipse, DDQ, QSY, Blackberry Quencher, Black Hole Quencher, Qxl, Iowa black FQ, Iowa Black It may be one or more selected from the group RQ and IRDye QC-1.
  • the kit may further include a double-stranded fluorescent dye, and the dye may be chelated to DNA.
  • the fluorescent dye may be, for example, SYBR green.
  • the fluorescent dye hardly shows a fluorescent signal before being chelated to the double-stranded DNA, but after chelating to the double-stranded DNA, the fluorescent signal is greatly increased, so that a target substance can be detected.
  • the isothermal amplification product of the target nucleic acid to the antibody when the antibody is immobilized on the surface or inside of the hydrogel, the isothermal amplification product of the target nucleic acid to the antibody; And an antibody capable of specifically binding to the isothermal amplification product of the target nucleic acid may form a complex.
  • One or more antibodies may be immobilized on the surface or inside of the hydrogel.
  • the antibody may be, for example, streptavidin, anti-digoxigenin, anti-tamra, anti-texasred, anti-hapten, or the like. Specifically, it may be to form an antigen-antibody complex using an antigen capable of specifically binding to the antibody.
  • the hapten capable of specifically binding to the anti-hapten is bound to the 5'end of the primer, and the anti-hapten and the hapten-primer It may be to form an antigen-antibody complex of.
  • the 5'hapten-primer participates in the isothermal amplification reaction, and the isothermal amplification product including the 5'hapten-primer may specifically bind to the anti-hapten fixed on the surface or inside of the hydrogel (Fig. 2).
  • the streptavidin and biotin capable of specifically binding thereto may form a streptavidin-biotin complex.
  • a forward inner primer or a reverse inner primer in which biotin is bound to the 5'end; And a product isothermal amplified by the external primer and streptavidin may form a complex.
  • nucleic acid probe when a nucleic acid probe is immobilized on the surface or inside of a hydrogel, an isothermal amplification product of a target nucleic acid to the probe; And a probe capable of specifically binding to the isothermal amplification product of the target nucleic acid may form a complex.
  • One or more nucleic acid probes may be immobilized on the surface or inside of the hydrogel.
  • an isothermal amplification product of a target nucleic acid may be complementarily bound to the nucleic acid probe.
  • FIG. 3A is a concrete embodiment of Figure 3a, a forward internal primer or probe fixed on the surface or inside of a hydrogel and a linker is bonded to the end, and a reverse internal primer included in the composition for isothermal amplification and a fluorescent material is bonded to the end. The detection method used is shown.
  • a linker capable of binding to the surface or inside of the hydrogel is bonded to the end of the forward inner primer or probe, and the fluorescent labeling material is bonded to the reverse inner primer. Therefore, the isothermal amplification product of the target substance can be specifically detected by binding with the primer.
  • the reverse internal primer is included in the composition for amplification of the lamp, and the isothermal amplification product participating in the isothermal amplification reaction is complementarily bound to the forward internal primer or probe fixed on the hydrogel, thereby specifically detecting the target material. have.
  • the isothermal amplification product is bound to one or more antibodies or nucleic acid probes immobilized on the surface or inside of the hydrogel, so that several target substances can be simultaneously detected.
  • a nucleic acid primer, probe, or antibody that specifically binds to at least one bacterial or viral nucleic acid is immobilized on the surface or inside of the hydrogel, so that the isothermal amplification product binds to the primer, probe or antibody, thereby positioning the hydrogel.
  • the hydrogel may be PEG (polyehthylene glycol), PEGDA (polyethylene glycol diacrylate), PEGDMA (polyethylene glycol dimetacrylate), photoinitiator, or a combination thereof.
  • the photoinitiator is, for example, 2-hydroxy-2-methylpropiophenone (2-Hydroxy-2-methylpropiophenone) or lithium phenyl-2,4,6-trimethylbenzol phosphate (Lithium phenyl-2,4,6- trimethylbenzoylphosphate).
  • the hydrogel may further contain one or more selected from the group consisting of streptavidin, anti-digoxigenin, anti-TAMRA, anti-texathread, and graphene.
  • the streptavidin may be itself or may be coated on beads.
  • the hydrogel contains streptavidin-coated beads, the pore size of the gel may be increased.
  • the isothermal amplification method has a concentration of primers 10 to 1,000 times higher during reaction than the conventional PCR method. Fluorescent labeling materials for detecting amplification products exhibit fluorescence after chelating DNA double strands, but in general, fluorescence when the concentration of single strands is high. Therefore, the graphene may be added to reduce false positives in fluorescence detection of isothermal amplification products.
  • the graphene may be added, for example, to a hydrogel or may be additionally added to a hydrogel to which streptavidin, anti-digoxigenin, anti-TAMRA or anti-texathread has been added.
  • the graphene is bound to a single strand and is difficult to chelate with a fluorescent labeling material, and the fluorescent labeling material bound to a single strand may be quenched fluorescence by graphene.
  • the double strand since the double strand does not bind to graphene, it can be chelated with a fluorescent dye, so that the fluorescent background by the primer can be removed.
  • the target material is Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Metanumovirus, Metanumovirus, Coronavirus 229E (Coronavirus 229E), Coronavirus NL63 (Coronavirus NL63), and Coronavirus OC43 (Coronavirus OC43) may be one or more selected from the group consisting of.
  • the nucleic acid probe or antibody is immobilized on the surface or inside of the hydrogel, it is possible to detect several target substances simultaneously by specifically binding to the isothermal amplification product.
  • a fluorescent signal can be generated by the forward internal primer or the reverse internal primer is bound to a fluorescent dye or a fluorescent dye capable of binding to double-stranded DNA is displaced, and the isothermal amplification product can be detected by monitoring the emitted fluorescence.
  • Another aspect comprises obtaining a nucleic acid sample from a biological sample of the subject; Contacting the sample with a substrate on which a nucleic acid probe or antibody capable of specifically binding to an isothermal amplification product of a target nucleic acid is immobilized on the surface or inside of the hydrogel; And it provides a method for multiple detection of a target substance comprising the step of performing an isothermal amplification reaction by adding a composition for isothermal amplification. By simultaneously detecting a target substance by the above method multiplexedly, it is possible to quickly diagnose whether an individual is infected with viruses and/or bacteria.
  • the method comprises obtaining a nucleic acid sample from a biological sample of the subject.
  • the individual is a target for detecting acute respiratory infectious diseases, for example, a target for predicting the likelihood of an acute respiratory infectious disease, a target for diagnosing the state of an acute respiratory infectious disease, and a target for determining the prognosis of an acute respiratory infectious disease , It means a target for determining the dosage of a drug for preventing or treating acute respiratory infectious diseases, a target for determining a treatment method according to the progression of acute respiratory infectious diseases, etc.
  • the individual may be a vertebrate animal, specifically mammals, amphibians, reptiles, birds, etc., more specifically, may be a mammal, for example, a human (Homo sapiens), a Korean I can.
  • the sample may include a sample such as tissue, cells, whole blood, serum, plasma, saliva, sputum, cerebrospinal fluid or urine isolated from an individual.
  • the method includes contacting the sample with a substrate on which a probe or antibody capable of specifically binding to an isothermal amplification product of a target nucleic acid is immobilized on the surface or inside of the hydrogel. Details of the substrate are as described above.
  • the method includes performing an isothermal amplification reaction by adding a composition for isothermal amplification.
  • the composition for isothermal amplification includes an inner primer set and an outer primer set. Details of the inner primer set and the outer primer set are as described above.
  • the composition may further include a DNA polymerase, dNTPs, and a buffer for performing an isothermal amplification reaction.
  • the isothermal amplification reaction may be performed at 30 to 75°C for 20 to 120 minutes.
  • the isothermal amplification reaction may be performed at, for example, 30 to 75°C, 30 to 70°C, 30 to 65°C, 50 to 75°C, or 55 to 70°C.
  • the isothermal amplification reaction may be performed for, for example, 20 to 120 minutes, 20 to 100 minutes, 30 to 80 minutes, 30 to 60 minutes, 50 to 120 minutes, 50 to 100 minutes, or 50 to 80 minutes.
  • the reaction temperature is less than the above range, there is a problem that not only the Bst polymerase is not sufficiently activated, but also the probe and the primer are abnormally bound to the nucleic acid. If the reaction temperature exceeds the above range, the Bst polymerase is not sufficiently activated, and There is a problem in that the primer cannot bind to the nucleic acid.
  • the reaction time is less than the above range, there is a problem in that the amplification reaction does not sufficiently occur, resulting in a false negative result, and when the reaction time exceeds the above range, a non-specific amplification reaction may occur, resulting in a false positive result.
  • the probe fixed to the hydrogel coupled with the isothermal amplification product amplified through the isothermal amplification reaction as described above can be complementarily combined with the inner primer set and the outer primer set to which the fluorescent labeling material in the composition for isothermal amplification is bound. At this time, the target material may be detected by measuring the fluorescence signal emitted from the fluorescent labeling material.
  • an isothermal amplification reaction was performed by combining a fluorescent dye with an inner primer to measure fluorescence, and after the reaction, it was possible to confirm whether or not fluorescence was detected through a fluorescence microscope under UV. Therefore, since several target substances can be simultaneously detected according to the above method, it is possible to quickly diagnose whether a virus and/or bacteria are infected in the field.
  • a probe or antibody capable of specifically binding to the isothermal amplification product of the target nucleic acid is immobilized on the substrate, on the surface or inside of the hydrogel, without cross-reaction or interference reaction between the probes or antibodies. Multiple target substances can be detected simultaneously.
  • the isothermal amplification method it is possible not only to quickly diagnose in the field, but also to construct a diagnostic test system having excellent sensitivity and specificity.
  • FIG. 1 is a schematic diagram of a kit for detecting a target substance using a three-dimensional gel containing a nucleic acid specifically binding to an isothermal amplification product.
  • FIG. 2 is a schematic diagram showing the operating principle of the target substance detection kit, and shows a detection method using a hydrogel immobilized with anti-heptene.
  • 3A is a schematic diagram showing the operating principle of a target substance detection kit, and shows a detection method using a nucleic acid primer or a hydrogel to which a probe is bound.
  • Figure 3b is a concrete embodiment of Figure 3a, a forward internal primer or probe fixed on the surface or inside of a hydrogel and a linker is bonded to the end, and a reverse internal primer included in the composition for isothermal amplification and a fluorescent material is bonded to the end. The detection method used is shown.
  • 4A is a diagram showing a substrate on which streptavidin, anti-digoxigenin, anti-texathread and anti-TAMRA are fixed, respectively.
  • 4B is a diagram showing a substrate to which an acrylic dite forward inner primer is bonded to a hydrogel.
  • 5 is a result of isothermal amplification and fluorescence analysis using a biotin-forward inner primer, a FAM-reverse inner primer, and an outer primer in a hydrogel fixed with streptavidin.
  • FIG. 6 is a result of isothermal amplification and fluorescence analysis using a forward inner primer, a FAM-reverse inner primer, and an outer primer in a hydrogel fixed with an acrylicdite-nucleic acid primer.
  • Primer sets for isothermal amplification reactions for each bacterial and viral genome were prepared using Primer Explore (http://primerexplorer.jp/e/index.html), and the base sequences of the primers are shown in Table 1 below.
  • polyethylene glycol diacrylate Poly(ethyleneglycol) diacrylate
  • 2-hydroxy-2-methylpropiophenone 2-Hydroxy-2-methylpropiophenone
  • a solution in which 10 ⁇ M of a nucleic acid probe that specifically binds an isothermal amplification product is dissolved in distilled water is mixed with the first gel composition in a ratio of 1:10, and then vortexed for about 5 seconds to prepare a second gel composition solution.
  • the probe an acrylate-forward inner primer for streptavidin or Chlamydophila pneumonia was used.
  • 0.5 ⁇ l of the second gel composition solution was dispensed into each well of a 96-well plate at 1 ⁇ m intervals and gelled using UV.
  • a target substance detection kit with a gel spot to which a nucleic acid that specifically binds to an isothermal amplification product for a target substance is fixed or bound was prepared.
  • Chlamydophila pneumonia Omp A gene DNA was synthesized by Bioneer Co., Ltd. and used as a template.
  • An isothermal amplification composition for Chlamydophila pneumonia was added to the target substance detection kit prepared in Example 1 and reacted at 60° C. for 1 hour. Thereafter, fluorescence was analyzed by washing with a washing buffer.
  • the forward outer primer, biotin-forward inner primer, reverse outer primer, FITC-reverse outer primer, reverse inner primer, dNTP, Bst polymerase and buffer are isothermal. It was used as an amplification composition.
  • the forward outer primer, the forward inner primer, the reverse outer primer, the FITC-reverse outer primer, the reverse inner primer, dNTP, Bst polymerase and The buffer was used as an isothermal amplification composition.
  • FIG. 4 is a result of fluorescence analysis by isothermal amplification after fixing streptavidin to a gel
  • FIG. 5 is a result of fluorescence analysis by isothermal amplification after binding an Acrydite-nucleic acid probe to the gel.
  • Positive is a group containing a target substance
  • Negative is a group that does not contain a target substance
  • w/o dntp is a group that does not contain nucleoside triphosphate
  • a non-target probe is a group in which a nucleic acid probe for a substance other than the target substance is immobilized on the hydrogel

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Abstract

La présente invention concerne un kit pour la détection multiple de matériaux cibles, comprenant un substrat sur lequel sont immobilisés des sondes nucléiques ou des anticorps, capables de se lier spécifiquement à des produits d'amplification isotherme de matériaux cibles sur une surface ou à l'intérieur d'un hydrogel. En faisant appel au kit, un diagnostic rapide sur site est permis et un système de test de diagnostic de maladie ayant une excellente sensibilité et une excellente spécificité peut être construit.
PCT/KR2020/008894 2019-07-25 2020-07-08 Kit de détection de matériaux cibles et procédé de détection de matériaux cibles faisant appel à celui-ci WO2021015452A1 (fr)

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KR1020190090481A KR20210012553A (ko) 2019-07-25 2019-07-25 표적 물질의 검출용 키트 및 이를 이용하여 표적 물질을 검출하는 방법

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