Method for detecting thrombin through nanogold
A detection method, thrombin technology, applied in the field of biomarker detection
Inactive Publication Date: 2022-07-29
CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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[0004] In view of this, in order to solve the existing detection problem of relying on nano-gold aggregation, the present invention provides a thrombin detection method that relies on nano-gold from agglomeration to depolymerization, which can detect thrombin conveniently, economically and with high sensitivity
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[0024] like figure 1 As shown, the experimental centrifuge tubes were cleaned;
[0025] Hybridization stock solution preparation: TB-cApt1@AuNPs:TB-Apt1@AuNPs=2.5 (molar ratio), after mixing evenly, put it in a refrigerator at 4°C and let it stand for >=3h;
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Abstract
The invention discloses a method for detecting thrombin by using nanogold. According to the method, two oligonucleotide sequences are designed and are respectively marked as TB-Apt1 and TB-cApt1, the TB-Apt1 is a thrombin nucleic acid aptamer, and the TB-cApt1 is a complementary chain of the thrombin nucleic acid aptamer. The oligonucleotides are anchored on the surface of the nanogold through a plurality of adenine fragments respectively. And the TB-Apt1 and the TB-cApt1 are hybridized with each other, so that the nanogold is agglomerated. After the thrombin is added, the TB-Apt1 is combined with the thrombin, a hybrid chain formed by the TB-Apt1 and the TB-cApt1 is separated, and the nanogold is changed into a dispersed state from an agglomerated state. The degree of the dispersion state of the nanogold in the solution is in positive correlation with the thrombin concentration. And the thrombin concentration can be calculated by observing the absorption light spectrum intensity of the nanogold solution. According to the method, the surface of the nanogold is modified with oligonucleotide by using an adenine fragment, so that the cost of nucleic acid functionalized nanogold is reduced, low-concentration detection of thrombin is realized in the process of inducing the nanogold from agglomeration to depolymerization by using thrombin, and the sensitivity of the detection method is improved.
Description
technical field [0001] The invention relates to the technical field of biomarker detection, in particular to the design of protein detection, in particular to a method for detecting thrombin with nano-gold. Background technique [0002] Thrombin, as a major serine protease in the coagulation cascade, plays a key role in both physiological and pathological coagulation. Thrombin is often used as a biomarker to provide a basis for the early detection and treatment of diseases. The highly sensitive detection of thrombin is of great significance in clinical diagnosis. [0003] Gold nanoparticles are often used in combination with nucleic acid aptamers to develop colorimetric detection methods due to their sensitive localized surface plasmon resonance sensing and easy synthesis and modification. In this process, the agglomeration of gold nanoparticles is usually induced by the target, and the detection is performed by observing the changes in the absorption light spectrum of the ...
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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/573G01N33/543G01N33/532
CPCG01N33/573G01N33/54346G01N33/532G01N2333/974
Inventor 吴江黄昱
Owner CHONGQING INST OF GREEN & INTELLIGENT TECH CHINESE ACADEMY OF SCI
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