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A nanocomposite probe, composition and fluorescence quantitative kit for highly sensitive fluorescent quantitative detection of serum tumor markers

A technology for fluorescent quantitative detection and tumor markers, applied in the field of fluorescent quantitative detection, can solve the problems of complex operation, low content of tumor markers, short storage time of reagents, etc., achieve high catalytic activity, improve sensitivity and specificity, and improve sensitivity Effect

Active Publication Date: 2020-09-01
LINYI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Because the content of tumor markers in the body is generally very low in the early stage of cancer, it is difficult for conventional detection methods to perform accurate quantitative and specific analysis, and it is easy to lead to false positive and false negative results
For example, enzyme-linked immunoassay is widely used at present, but the target patients can only be detected after the tumor has developed to a certain stage; radioimmunoassay is also the main method for tumor marker detection, but this method has complex operations. , Unstable measurement results, short storage time of reagents, radioactive contamination, expensive instruments, etc.

Method used

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  • A nanocomposite probe, composition and fluorescence quantitative kit for highly sensitive fluorescent quantitative detection of serum tumor markers
  • A nanocomposite probe, composition and fluorescence quantitative kit for highly sensitive fluorescent quantitative detection of serum tumor markers
  • A nanocomposite probe, composition and fluorescence quantitative kit for highly sensitive fluorescent quantitative detection of serum tumor markers

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preparation example Construction

[0042] The nanocomposite probe containing a large amount of DNase chains and a small amount of serum tumor marker secondary antibody to be detected can be obtained by adopting the preparation sequence described in the present invention, so that the probe is rich in a large amount of DNase chains, shortening the reaction time and rapidly amplifying Fluorescent signal for high sensitivity detection.

[0043] Specifically, the nanocomposite probe of the present invention is prepared according to the following steps:

[0044] S1, modifying gold nanoparticles by asymmetric modification with DNA enzyme chains to obtain gold nanoparticles-enzyme chains;

[0045] S2. Mix the secondary antibody of the serum tumor marker to be detected with the gold nanoparticle-enzyme chain obtained in step S1 for asymmetric modification to obtain a nanocomposite probe.

[0046] In the present invention, the DNA enzyme chain is modified on the surface of the gold nanoparticle by means of asymmetric mo...

Embodiment 1

[0064] The preparation of embodiment 1 nanocomposite probe

[0065] 0.1 mL of tannic acid-coated AuNPs (30nm, 50nM) was drop-coated on a positively charged clean glass slide for 3 hours at room temperature, and the tannic acid-coated AuNPs were adsorbed on the positively charged glass slide by electrostatic adsorption. On the slide, the glass slide appeared lavender, washed with a large amount of ultrapure water, and 100 μL of DNase chain (10 -6 M) Drop coating on a glass slide to asymmetrically modify the AuNPs adsorbed on the glass slide. After reacting for 12 hours, rinse with a large amount of ultrapure water, blow dry with nitrogen, and place the slide in 1.0 mL of ultrapure water for 5 minutes to remove the adsorption of AuNPs and the slide. The solution obtained by sonication was centrifuged to obtain a precipitate (8000r / min, 10min), washed with ultrapure water, and redispersed in 0.1mL of ultrapure water to obtain asymmetrically modified AuNPs-enzyme chains.

[0066...

Embodiment 2

[0068] The preparation of embodiment 2 nanocomposite probes

[0069] 0.1mL of tannic acid-coated AuNPs (30nm, 50nM) was drop-coated on a positively charged clean glass slide for 2.5 hours at room temperature, and the tannic acid-coated AuNPs were adsorbed on the positively charged glass slide by electrostatic adsorption. On the slide, the glass slide appeared lavender, washed with a large amount of ultrapure water, and 100 μL enzyme chain (10 -6 M) Drop coating on a glass slide to asymmetrically modify the AuNPs adsorbed on the glass slide. After reacting for 14 hours, rinse with a large amount of ultrapure water, blow dry with nitrogen, and place the glass slide in 1.0 mL of ultrapure water for 8 minutes to remove the adsorption of AuNPs and the glass slide. The solution obtained by ultrasonication was centrifuged to obtain a precipitate (9000r / min, 8min), which was washed with ultrapure water and redispersed in 0.1mL of ultrapure water to obtain asymmetrically modified AuNP...

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Abstract

The invention relates to the field of fluorescent quantitative detection and provides a high-sensitivity nano composite probe for the fluorescent quantitative detection of a blood serum tumor marker.The nano composite probe comprises gold nanoparticles loaded with a blood serum tumor marker secondary antibody to be detected and a DNA (Deoxyribonucleic Acid) enzyme chain. The nano composite probeprovided by the invention takes the gold nanoparticles as carriers and a compound formed by loading deoxyribozyme with a high catalytic property keeps the high catalytic property and recognition capability of the deoxyribozyme, and also introduces a signal transduction function of a nano-material, so that recognition and signal transduction functions are integrated. The sensitivity and specificityof the fluorescent quantitative detection are remarkably improved by combining an efficient specific recognition function of the tumor marker, high recognition and catalytic properties of the deoxyribozyme, and signal transduction and fluorescent amplification functions of the gold nanoparticles; the detections sensitivity can reach 0.01ng / mL.

Description

technical field [0001] The invention relates to the field of fluorescence quantitative detection, in particular to a nano-composite probe for high-sensitivity fluorescence quantitative detection of serum tumor markers, a composition comprising the nano-composite probe and a fluorescence quantitative kit. Background technique [0002] In cancer research and clinical practice, tumor markers have great practical value in tumor screening, diagnosis, prognosis, evaluation of treatment efficacy, and follow-up observation of high-risk groups. Tumor markers were first proposed in 1978 by Herberman at the "Human Immunity and Tumor Immunology Diagnosis" meeting held by the National Cancer Institute of the United States. The following year it was recognized by participants at the 7th UK Conference on Oncogenesis Biology and Medicine and began to be publicly cited. [0003] Tumor markers refer to a class of substances that are abnormally secreted or shed by tumor cells during the occur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/533G01N33/577G01N33/543
CPCG01N33/533G01N33/54346G01N33/57484G01N33/577
Inventor 周宏刘静张宁波张书圣
Owner LINYI UNIVERSITY
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