• 专利标题:   Photoresponsive gel microsphere useful for detecting nucleic acid, preferably Proteus mirabilis nucleic acid is prepared by synthesizing ferrosoferric oxide-graphene oxide-silicon dioxide complex material, mixing with microdroplet solution, PCR reagent and sample and cutting into microdroplets.
  • 专利号:   CN113856573-A
  • 发明人:   WANG Y, YU J, ZHAO Y, ZHANG L, YE F
  • 专利权人:   UNIV CHINESE ACAD SCI WENZHOU INST
  • 国际专利分类:   B01J013/00, B01J013/02, C12Q001/06, C12Q001/6851, C12Q001/689
  • 专利详细信息:   CN113856573-A 31 Dec 2021 B01J-013/00 202224 Chinese
  • 申请详细信息:   CN113856573-A CN11312020 08 Nov 2021
  • 优先权号:   CN11312020

▎ 摘  要

NOVELTY - Photoresponsive gel microsphere is prepared by (a) synthesizing ferrosoferric oxide nanospheres by hydrothermal method, mixing the ferric oxide nanospheres, dopamine, hydrochloric acid, and tris(hydroxymethyl)aminomethane base, replacing the solution with a single-layer graphene oxide dispersion liquid, repeatedly cleaning, mixing the ferrosoferric oxide nanoparticles with ethyl orthosilicon dioxidete under alkaline conditions to generate ferrosoferric oxide-graphene oxide-silicon dioxide complex, washing and dispersing in water and (b) uniformly mixing the ferrosoferric oxide-graphene oxide-silicon dioxide complex material, microdroplet raw material solution, PCR reagent and sample to be tested to obtain inner phase, taking HFE7500 oil phase containing fluorine-containing surfactant as the outer phase, injecting the inner phase and outer phase respectively into the chip through a syringe pump, and cutting the inner phase by the outer phase inside the chip to obtain microdroplets. USE - The photoresponsive gel microsphere is useful for detecting nucleic acid, preferably P.mirabilis nucleic acid by dPCR (claimed) and in fields such as health and public safety. ADVANTAGE - The photoresponsive gel microsphere has efficient near-infrared light heat generation and magnetic response, can solidify into spheres that can store the information of single molecules and is cost-effective. The detection method is effective. DETAILED DESCRIPTION - Photoresponsive gel microsphere is prepared by (a) synthesizing ferrosoferric oxide nanospheres by hydrothermal method, uniformly mixing the ferric oxide nanospheres, dopamine, hydrochloric acid, and tris(hydroxymethyl)aminomethane base for 6 hours until the solution turns brown, and the surface of the ferric oxide nanospheres is loaded with a polydopamine layer, replacing the solution with a single-layer graphene oxide dispersion liquid, uniformly mixing and stirring the mixture at room temperature for 6 hours to generate ferrosoferric oxide nanoparticles with few layers of graphene oxide on the surface, repeatedly cleaning with ethanol and deionized water, uniformly mixing the ferrosoferric oxide nanoparticles with graphene oxide on the surface with ethyl orthosilicon dioxidete at 60degreesC under alkaline conditions for 1 hour to generate ferrosoferric oxide-graphene oxide-silicon dioxide complex, repeatedly washing with ethanol and deionized water and dispersing in water and (b) uniformly mixing the ferrosoferric oxide-graphene oxide-silicon dioxide complex material obtained in step (a), microdroplet raw material solution, PCR reagent and sample to be tested to obtain inner phase, taking HFE7500 oil phase containing fluorine-containing surfactant as the outer phase, injecting the inner phase and outer phase respectively into the chip through a syringe pump, and cutting the inner phase by the outer phase inside the chip to obtain microdroplets, which are light-responsive variable temperature gel microspheres. An INDEPENDENT CLAIM is included for use of the photoresponsive gel microsphere for detecting nucleic acid, preferably Proteus mirabilis nucleic acid by digital PCR (dPCR), where the detection method involves subjecting the microspheres to programmed near-infrared light to adjust the temperature of the gel microspheres, reading the fluorescence signal of the gel microspheres by a fluorescence microscope, counting the fluorescence ratio and estimating the content of the initial P.mirabilis nucleic acid molecules.