# US 2025/0066844 A1 · Composition and method for amplifying nucleic acid sequences > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2025/0066844 A1 - Document ID: US20250066844A1 - Status: Published application - Kind: A1 - Application number: 18/722,223 - Filed: 2022-12-13 - Published: 2025-02-27 - Earliest claimed: 2021-12-22 - Assignees: Bforcure - Applicants: Bforcure - Inventors: Maël LE BERRE; Christophe PANNETIER; Francesco VILLA; Maryna MURLYKINA; Julien CROZZOLO - Web page: https://patentgrep.com/patent/US20250066844A1 ## CPC - C12Q 1/6844: Nucleic acid amplification reactions - C12Q 1/686: Polymerase chain reaction [PCR] - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES ## Abstract A composition is disclosed for the amplification of nucleic acid sequences, which is intended to be mixed with a biological sample containing one or more target nucleic acid sequences and with primers specific for the target nucleic acid sequence(s), and fluorescent probes for revealing an amplification of the target nucleic acid sequence(s). The composition includes reagents and enzymes for nucleic acid application. The composition also includes a polymerizable reagent, which polymerizes under the influence of a stimulation chosen from a heat stimulation and a light stimulation, so as to form a hydrogel which locally encapsulates each target nucleic acid sequence when the composition is exposed to the stimulation. A method using said composition is also disclosed. ## Claims ### Claim 1 A masterbatch composition for the amplification of nucleic acid sequences, intended to be mixed with a biological sample containing one or more target nucleic acid sequence(s) and specific primers of the target nucleic acid sequence(s), as well as fluorescent probes for revealing an amplification of the target nucleic acid sequence(s), the composition comprising: reagents and nucleic acid amplification enzymes, wherein the composition further comprises a polymerisable reagent, which polymerises under the influence of a stimulation selected from among a thermal stimulation and a light stimulation, so as to form a hydrogel locally encapsulating each target nucleic acid sequence when said composition is exposed to said stimulation. ### Claim 2 (depends on 1) The composition according to claim 1, wherein the polymerisable reagent is selected from among a photosensitive reagent that polymerises when exposed to light, and a heat-sensitive reagent that polymerises when exposed to a threshold temperature. ### Claim 3 (depends on 1) The composition according to claim 1, wherein the polymerisable reagent is selected from among the class of PEGs and other water-soluble polymers compatible with PCR. ### Claim 4 (depends on 1) The composition according to claim 1, wherein the polymerisable reagent is selected from among the class of PEGs and other water-soluble polymers compatible with PCR, capable of photosensitive or heat-sensitive reactions such as radical, ionic, dimerisation or Diels-Aider reactions. ### Claim 5 (depends on 1) The composition according to claim 1, wherein the polymerisable reagent is present in a content from 2% to 25%, preferably from 5% to 15%, by weight per unit volume (w/V). ### Claim 6 (depends on 1) The composition according to claim 1, further comprising an initiator agent selected from among a photo-initiator and a thermo-initiator, in a content from 0.02% to 1% by weight per unit volume (w/V). ### Claim 7 (depends on 1) The composition according to claim 1, comprising the specific primers of the target nucleic acid sequence(s), as well as fluorescent probes for revealing an amplification of the target nucleic acid sequence(s). ### Claim 8 (depends on 1) The composition according to claim 1, wherein the composition remains liquid before application of said stimulation for a time period comprised between several hours and several months, and said composition forms the hydrogel after application of said stimulation within a time period shorter than or equal to 10 minutes. ### Claim 9 (depends on 1) A method for amplifying nucleic acid sequences comprising the following steps: (i) providing a composition according to claim 1; (ii) mixing said composition with a biological sample to be analysed; (iii) applying a stimulation selected from among a thermal stimulation and a light stimulation so as to polymerise the mixture of step (ii) to obtaining a hydrogel; (iv) exposing the hydrogel to an amplification reaction of acid sequences. ### Claim 10 (depends on 9) The method according to claim 9, wherein the provision step (i) is maintained for a time period comprised between several hours and several months. ### Claim 11 (depends on 9) The method according to claim 9, wherein said reaction is a polymerase chain reaction. ### Claim 12 (depends on 9) The amplification method according to claim 9, wherein step (iii) of applying a stimulation includes exposing said composition to a UV light or a threshold temperature. ### Claim 13 (depends on 9) The amplification method according to claim 9, wherein the hydrogel is obtained within a time period shorter than or equal to 10 minutes. ### Claim 14 (depends on 9) The amplification method according to claim 9, wherein step (iii) includes a reaction selected from among the group formed by a bio-orthogonal thiol-ene reaction and a dimerisation reaction. ### Claim 15 (depends on 6) The composition according claim 6, wherein the photo-initiator is lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP), and the thermo-initiator is 2,2′-azobis [2-(2-imidazolin-2-yl) propane] dihydrochloride.