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A combined detection instrument for multiple breast cancer tumor markers based on a nucleic acid aptamer

A nucleic acid aptamer and breast cancer tumor technology, which is applied in the fields of heavy metal detection and biomedical detection, can solve problems such as reducing the detection environment requirements, and achieve the effects of increasing storage time, reducing detection costs, and reducing requirements

Inactive Publication Date: 2018-09-11
HANGZHOU DIANZI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of combined detection of multiple tumor markers in breast cancer diagnosis and reduce the detection environment requirements, the present invention provides a detection idea based on multiple heavy metal ion-labeled nucleic acid aptamers instead of traditional antibodies, which can realize the detection of multiple breast cancer Simultaneous detection of cancer tumor markers, with the characteristics of simple operation and rapid detection

Method used

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  • A combined detection instrument for multiple breast cancer tumor markers based on a nucleic acid aptamer
  • A combined detection instrument for multiple breast cancer tumor markers based on a nucleic acid aptamer
  • A combined detection instrument for multiple breast cancer tumor markers based on a nucleic acid aptamer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: The preparation process of the marker connected with the MUC1 nucleic acid aptamer is as follows:

[0040] (1) Add 10uL of apoferritin to 4mL of acetic acid-sodium acetate buffer solution, and then add 50uL respectively at a concentration of (1-1.5) g·L -1 Heavy metal ions Pb 2+ , to get apoferritin / heavy metal ion Pb 2+ Mixture; add acetic acid-sodium acetate buffer solution in above-mentioned mixture, centrifuge, ultrafiltration obtains and contains heavy metal ion Pb 2+ of apoferritin. The protein will release its wrapped heavy metal ion Pb under acidic conditions 2+ .

[0041] (2) Prepare (0.5-1) mg·mL by dissolving graphene oxide in deionized water -1 Dispersion solution; Add chloroauric acid (5mM) and sodium hydroxide (1mg·mL -1 ) each (0.2-0.5) mL, sonicate (2-2.5) hours, and centrifuge for 10-15 minutes to obtain a graphene-nano-gold composite; finally use the dithiol alkane chain to modify the graphene-nano-gold composite for later use.

[004...

Embodiment 2

[0044] Embodiment 2: The preparation process of the marker connected with HER-2 nucleic acid aptamer is as follows:

[0045] (1) Add 10uL of apoferritin to 4mL of acetic acid-sodium acetate buffer solution, and then add 50uL respectively at a concentration of (1-1.5) g·L -1 heavy metal ion Cd 2+ , to get apoferritin / heavy metal ion Cd 2+ Mixture; add acetic acid-sodium acetate buffer solution in above-mentioned mixture, centrifuge, ultrafiltration obtains and contains heavy metal ion Cd 2+ of apoferritin. The protein will release its wrapped heavy metal ions under acidic conditions.

[0046] (2) Prepare (0.5-1) mg·mL by dissolving graphene oxide in deionized water -1 Dispersion solution; Add chloroauric acid (5mM) and sodium hydroxide (1mg·mL -1 ) each (0.2-0.5) mL, sonicate (2-2.5) hours, and centrifuge for 10-15 minutes to obtain a graphene-nano-gold composite; finally use the dithiol alkane chain to modify the graphene-nano-gold composite for later use.

[0047] (3) D...

Embodiment 3

[0049] Example 3: Preparation of magnetic bead-aptamer composite material: magnetic beads coated with a silicon layer on the outer surface, and the magnetic beads are modified with nucleic acid aptamers of MUC1 and HER-2, which can be purchased.

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Abstract

The invention discloses a combined detection instrument for multiple breast cancer tumor markers based on a nucleic acid aptamer. The combined detection instrument includes signal markers, magnetic bead-aptamer composite material, an electrolytic cell, a three-electrode measuring device and a measurement and analysis system based on a single-chip microcomputer STM8L. The signal markers include themarkers connected with the MUC 1 nucleic acid aptamer and HER-2 nucleic acid aptamer. The tradition protein antibody is replaced by adopting the multiple metal ion labels to mark the nucleic acid aptamer of different breast cancer tumor markers, the simultaneous combined detection for the multiple breast cancer tumor markers is realized, and the detection cost is reduced. The protein antibody isreplaced by adopting the nucleic acid aptamer, the requirements for the detection environment are reduced, and the storage time of the signal markers is improved.

Description

technical field [0001] The invention belongs to the technical field of heavy metal detection and biomedical detection, and relates to a simultaneous detection of multiple breast cancer tumor markers based on multiple metal ion tag-labeled nucleic acid aptamers, which is used to provide assistance for early diagnosis of breast cancer. Background technique [0002] Cancer has always been a killer that threatens human life. Studies have shown that the cure rate of early cancer is as high as 80%. However, there are as many as 200 types of cancer, most of which have no obvious symptoms in the early stage, and most cancer patients often miss the best time for treatment when they are discovered. For women, breast cancer is the most common cancer and the second leading cause of death. And more than 90% of them are due to the metastasis of cancer cells. Therefore, the detection of early breast cancer is particularly important. However, like other cancers, early detection of breas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N27/48
CPCG01N27/48G01N33/57415G01N33/57484G01N2800/50
Inventor 樊凯李杜娟徐枫厉力华刘红英苏畅姚睿曹佳斐
Owner HANGZHOU DIANZI UNIV
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