Biosensor for ctDNA detection and preparation method thereof

A biosensor and nano-gold particle technology, which is applied in the field of DNA tumor analysis, can solve problems such as limited use and reduced practicability of ctDNA biosensors, and achieve the effect of achieving a dynamic range

Active Publication Date: 2021-07-30
山东雷泽生物科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, current biosensors have a limitation: the physics of single-site binding produces a hyperbolic dose-response curve with a useful dynamic range spanning a fixed change i...

Method used

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  • Biosensor for ctDNA detection and preparation method thereof
  • Biosensor for ctDNA detection and preparation method thereof
  • Biosensor for ctDNA detection and preparation method thereof

Examples

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Embodiment 1

[0022] Embodiment 1 of the present invention provides a method for preparing a biosensor for ctDNA detection, comprising the following steps:

[0023] Step S1, preparing nano-gold particles: Add chloroauric acid and ultrapure water to the two-necked flask, set the temperature of the oil bath to 300°C, set the sensor to 130°C, and heat the oil bath to 600rpm 120°C, adjust the rotation speed to 1100rpm, then put the two-necked flask containing chloroauric acid and ultrapure water into the oil bath, and start heating the oil bath. After 10 minutes, the solution in the two-necked flask starts to boil, and then add lemon Sodium acid solution, after boiling, 3 minutes, the magneton has purple, indicating that crystal nuclei have begun to form; 5 minutes, the whole solution has obvious purple; 8 minutes, the solution is purple, the crystal nucleus grows, and the solution begins to turn red; 9 minutes, wine red , as the number of particles increases, the red color deepens; after 12 mi...

Embodiment 2

[0027] The steps of preparing the biosensor in Example 2 differ from Example 1 only in that the nucleotide sequence of the DNA used in Example 2 is shown in SEQ ID No.2, and the rest are basically the same as in Example 1.

Embodiment 3

[0029] The steps of preparing the biosensor in Example 3 differ from Example 1 only in that the nucleotide sequence of the DNA used in Example 3 is shown in SEQ ID No.3, and the rest are basically the same as in Example 1.

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Abstract

The invention provides a preparation method of a biosensor for ctDNA detection, which comprises the following steps: preparing gold nanoparticles: mixing and heating chloroauric acid and ultrapure water, then adding a sodium citrate solution, continuously conducting heating and reacting, conducting cooling to room temperature after the reaction is finished, conducting filtering to obtain the gold nanoparticles, and conducting preserving at low temperature for later use; and conducting DNA modification: carrying out annealing treatment on DNA, then uniformly mixing the DNA subjected to annealing treatment with the prepared gold nanoparticles, carrying out standing treatment, then adding a sodium citrate solution, then adding a phosphate buffer solution, carrying out standing, finally adding water, and carrying out centrifugal washing, thus obtaining the biosensor for ctDNA detection.

Description

technical field [0001] The invention relates to the technical field of DNA tumor analysis, in particular to a biosensor for ctDNA detection and a preparation method thereof. Background technique [0002] Tumor is a serious disease that threatens human health. Currently, the main body of cancer diagnosis, staging, and treatment decisions is tissue biopsy. Although tissue biopsy is very useful, the invasiveness and inherent selection preference of biopsy limit its usefulness as a real-time monitoring tool, making it difficult to use for early diagnosis, and Invasive tissue biopsies often cause great pain to patients. In order to overcome this shortcoming, people began to look for various tumor biomarkers, such as protein markers, circulating tumor cells and circulating tumor DNA (circulating tumor DNA, ctDNA) and so on. In recent years, studies have shown that the tumor mutation information carried by ctDNA of tumor patients has a good consistency with tumor tissue. Therefo...

Claims

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Application Information

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IPC IPC(8): C12Q1/6825
CPCC12Q1/6825C12Q2563/137C12Q2565/607C12Q2563/155C12Q2525/307
Inventor 顾梦寒林美华夏帆张健
Owner 山东雷泽生物科技有限责任公司
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