• 专利标题:   Electro chemiluminescence sensor based on carbon dots and gold-silver sulfide resonance energy transfer to detect promyelocytic leukemia/retinoic acid receptor alpha fusion gene in acute and early young particle, comprises carbon dot as donor, gold-silver sulfide core-shell nanoparticle as acceptor.
  • 专利号:   CN116067948-A
  • 发明人:   ZHANG Y, HAN S, ZHANG Z, LIN M, LIU A, LEI Y
  • 专利权人:   UNIV FUJIAN MEDICAL
  • 国际专利分类:   C12Q001/6818, C12Q001/6825, C12Q001/6886, G01N021/76, G01N027/30, G01N027/327
  • 专利详细信息:   CN116067948-A 05 May 2023 G01N-021/76 202356 Chinese
  • 申请详细信息:   CN116067948-A CN10154064 23 Feb 2023
  • 优先权号:   CN10154064

▎ 摘  要

NOVELTY - Electro chemiluminescence sensor based on carbon dots and Au@Ag2S (gold-silver sulfide) resonance energy transfer comprises carbon dots(CDs)/Poly(amidoamine)(PAMAM) as the donor in the electrochemiluminescence resonance energy transfer /reduced graphene CDs/PAMAM/rGO(reduced graphene oxide), gold-silver sulfide core-shell nanoparticles (Au@Ag2S NPs) in resonance energy transfer as acceptor which efficiently quench carbon dot Electrochemiluminescence (ECL), binding capture DNA probe and target DNA, Au @ Ag2S NPs marks auxiliary DNA probe to hybridize and forms a sandwich structure, establishs a high sensitivity and selective electrochemiluminescence sensor, which can be used for quantitative detection of acute promyelocytic leukemia PML/RAR fusion gene. USE - Electro chemiluminescence sensor based on carbon dots and gold-silver sulfide resonance energy transfer to detect promyelocytic leukemia/retinoic acid receptor alpha fusion gene in acute and early young particle. ADVANTAGE - The ECL biosensor has simple and convenient, low cost, high sensitivity and it is expected to be a tool for acute promyelocytic leukemia (Activity: Cytostatic)diagnosis and treatment. DETAILED DESCRIPTION - An INDEPENDENT CLAIM is included for preparing electro chemiluminescence sensor, which involves (1) polishing glassy carbon electrode (GCE) in 1.0, 0.3 and 0.05 μm aluminum oxide (Al2O3) powder, placing in 1:1 nitric acid (HNO3) for ultrasonication in absolute ethanol and ultrapure water and dried with nitrogen (N2), then uniformly dripping the 5 L of dispersed CDs/PAMAM/rGO solution on the GCE surface, drying in an oven at 45°C to form a film to obtain a CDs/PAMAM/rGO modified electrode, (2) immersing the CDs/PAMAM/rGO modified electrode in 1 mL of a mixed solution containing 20 mg ethylene dichloride (EDC) and 10 mg (N-hydroxysuccinimide) (NHS) and activating for 0.5 hours, using ultrapure water to wash and clean, and drying with nitrogen, adding dropwise the CDs/PAMAM/rGO 8 µL of capture DNA probes on the surface of reduced graphene oxide (rGO) modified electrode, after selfassembling for 2 hours at room temperature, using tris(hydroxymethyl)aminomethane (TrisHCl) buffer solution to wash, drying with nitrogen, assembling the capture DNA probe on the CDs/PAMAM/rGO modified electrode through amide bonds to prepare a ssDNA/CDs/PAMAM/rGO modified electrode, placing the prepared ssDNA/CDs/PAMAM/rGO modified electrode into 100 µL hybridization solution containing Au@Ag2S NPs-labeled auxiliary DNA probe DNA-Au@Ag2S NPs and target DNA, and hybridize at 37 °C for 1 hours, at this time, capturing DNA probe and target DNA, Au@Ag2S NPs mark auxiliary DNA probe hybridization, forming a sandwich structure, taking out the electrode of the structure and using Tris-HCl buffer solution with pH 7.40 to wash, drying with nitrogen to obtain dsDNA/CDs/PAMAM/rGO modified electrode, then performing electrochemical luminescence detection, Au @ Ag2S NPs is close to CDs immobilized on surface of dsDNA/CDs/PAMAM/rGO modified electrode which can generate quenching effect on carbon point electro-chemiluminescence signal, according to the sandwich structure to form the change value of the front and back ECL signal, realizing the high sensitivity quantitative detection of acute promyelocytic leukemia PML/RAR fusion gene target DNA. The linear concentration range is 0.005-500 pM; regression equation delta IECL/a.u. =3511.52 + 829.19 * lgC (pM), the correlation coefficient (r)=0.996, the detection limit reaches 0.72 fM.