KR930700682A - Elements and Methods for Amplifying and Detecting Nucleic Acids by Bound Probes - Google Patents

Abstract

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Description

Elements and Methods for Amplifying and Detecting Nucleic Acids by Bound Probes

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The figure is a graph showing the results of a comparative analysis performed as described in Example 2 later.

Claims (15)

(a) a nucleic acid reagent comprising a first polymer having a glass transition temperature of 70 ° C. or higher and having an average diameter of 0.1 to 3 μm, and an oligonucleotide covalently bound to the particle, and (b) the first A nucleic acid reagent composition comprising a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the polymer, wherein the nucleic acid reagent composition comprises a mixture of water-insoluble adhesives present in the composition at 1-20% by weight An element comprising a sealable support. The element according to claim 1, wherein the support has a hydrophilic surface group provided by corona discharge treatment, chromic acid treatment, or high frequency electromagnetic field treatment in the presence of a reactive gas, or by having a hydrophilic support layer in which the composition is distributed. The element of claim 1 wherein said polymer particles have an average diameter of 0.5 to 2 μm. The element of claim 1, wherein the glass transition temperature of the first polymer is 70 to 175 ° C. and the glass transition temperature of the second polymer is 30 to 120 ° C. lower than the glass transition temperature of the first polymer. The element of claim 1 wherein said adhesive is present in said composition at 2 intrinsic 10% by weight. The process of claim 1 wherein the first polymer is (a) active halogen, carboxy, amidine, activated 2-substituted ethyl sulfonyl, activated 2-substituted ethylcarbonyl, active ester, vinylsulfonyl, vinyl 0.1 to 60% by weight of repeat units derived from at least one ethylenically unsaturated polymerizable monomer having a reactive group selected from the group consisting of carbonyl, aldehyde, hydrazine, epoxy, amino and sulfhydryl, (b) one providing hydrophobicity 60 to 99.9% by weight of repeating units derived from at least one ethylenically unsaturated polymerizable monomer, and (c) repetitions derived from at least one ethylenically unsaturated polymerizable monomer other than the monomers defined in (a) and (b) above. An element comprising from 0 to 15% by weight. The method of claim 1, wherein the second polymer (d) 55 to 100% by weight repeating units derived from at least one ethylenically unsaturated polymerizable monomer providing hydrophobicity, (e) to form a water-soluble homopolymer or hydrophilic 0 to 45% by weight of repeating units derived from at least one ethylenically unsaturated polymerizable monomer providing, and (f) repeating units derived from at least one ethylenically unsaturated polymerizable monomer providing crosslinking to the second polymer. An element comprising 0 to 15% by weight. The element of claim 1, wherein said composition is distributed in a defined region of said support. (a) a nucleic acid reagent comprising a first polymer having a glass transition temperature of 70 ° C. or higher and having an average diameter of 0.1 to 3 μl, and an oligonucleotide covalently bonded to the particle, and (b) the first A sealable composition comprising a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the polymer, the composition comprising a mixture of water-insoluble adhesives present in the composition at 1 to 20% by weight; A self-contained test device comprising an element comprising a support. (A) a nucleic acid reagent comprising (a) a first polymer having a glass transition temperature of 70 ° C. or more and having an average diameter of 0.1 to 3 μm, and an oligonucleotide covalently bound to the particle, and (b) Sealing a nucleic acid reagent composition comprising a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the first polymer, the mixture comprising a mixture of water-insoluble adhesives present in the composition at 1 to 20% by weight. (B) heating the composition at a temperature sufficient for and for such a time to deposit the deposited nucleic acid reagent composition to the support, if possible. (1) (a) a nucleic acid comprising a first polymer having a glass transition temperature of 70 ° C. or higher and having an average diameter of 0.1 to 3 μm, and an oligonucleotide complementary to the target nucleic acid covalently bound to the particle A reagent, and (b) a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the first polymer, the mixture comprising a water-insoluble adhesive present in the composition at 1 to 20% by weight. And an element comprising a sealable support having a composition distributed thereon, and (2) a detection reagent for detecting a hybridization product of the nucleic acid reagent oligonucleotide and the target nucleic acid in the element. The kit of claim 11, wherein the detection reagent is a biotinylated primer complementary to the target nucleic acid. (A) (a) a nucleic acid comprising a first polymer having a glass transition temperature of 70 ° C. or higher and having an average diameter of 0.1 to 3 μm, and an oligonucleotide complementary to the target nucleic acid covalently bound to the particle A reagent, and (b) a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the first polymer, the mixture comprising a water-insoluble adhesive present in the composition at 1 to 20% by weight. Contacting a sample comprising the sealable support with a nucleic acid reagent composition distributed thereon and a sample believed to contain the target nucleic acid to form a hybridization product of the target nucleic acid and the nucleic acid reagent on the element, B) detecting the presence of the hybridization product as an indicator of the presence of the target nucleic acid in the sample. Detection method. (A) amplifying a target nucleic acid in a test sample, (B) (a) a particle composed of a first polymer having a glass transition temperature of 70 ° C. or higher and having an average diameter of 0.1 to 3 μl, and covalently bound to the particle A nucleic acid reagent comprising an oligonucleotide complementary to the target nucleic acid, and (b) a second polymer having a glass transition temperature at least 30 ° C. lower than the glass transition temperature of the first polymer, wherein the composition comprises 1 to Contacting the amplified target nucleic acid with an amplified target nucleic acid and an element comprising a sealable support having a nucleic acid reagent composition comprising a mixture of a water-insoluble adhesive present at 20% by weight. Forming a hybridization product of a nucleic acid reagent, and (C) the presence of the hybridization product as a stock indicating the presence of the target nucleic acid in the sample. Method for amplifying and detecting a target nucleic acid, comprising detecting. ※ Note: The disclosure is based on the initial application.