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PSA detection method based on magnetic core-gold satellite assembly body

A PSA detection and assembly technology, applied in measurement devices, instruments, material analysis by optical means, etc., can solve the problems of limited use of antibody stability, harsh antibody conditions, and long time required to reach the detection limit. Low, high selectivity, easy to operate effect

Active Publication Date: 2017-03-22
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

As we all know, the conditions for antibody preparation are extremely harsh and expensive, and it takes a long time; in addition, the stability of antibodies further limits their widespread use

Method used

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  • PSA detection method based on magnetic core-gold satellite assembly body
  • PSA detection method based on magnetic core-gold satellite assembly body
  • PSA detection method based on magnetic core-gold satellite assembly body

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Such as figure 1 As shown, the quantitative detection method of the prostate-specific antigen PSA provided in this embodiment comprises the following steps:

[0048] (1) Preparation of capture probes

[0049] Take 100 μL of Fe surface-modified with carboxyl groups 3 o 4 , purchased from Aladdin Reagent (Shanghai) Co., Ltd., washed twice with Tris-HCl buffer, and finally adjusted to 500 μL. Then prepare EDC (2 mg / mL) and NHS solution (0.5 mg / mL), first add 30 μL of EDC solution to the above solution, and after 20 min, add the same volume of NHS solution, and the mixture is continuously stirred for 1.5 h. Subsequently, 50 μL (10 μM) of PSA aptamer was added, and after reacting for 3 hours, the previous magnetic beads were washed twice with Tris-HCl buffer solution, and resuspended for use to obtain capture probes.

[0050] (2) Preparation of gold nanoparticles

[0051] Under constant stirring, 100 mL of 1 mM chloroauric acid (HAuCl 4 ) solution was heated to boiling, ...

Embodiment 2

[0062] Get the water body of the Guangzhou section of the Pearl River, the operation method is the same as in Example 1, and its Raman spectrum is recorded as Figure 5 As shown (three experiments), the signal strength is about 226 (a.u.), which is relatively close to the background signal strength, and it can be considered that the water source does not contain PSA.

Embodiment 3

[0064] In order to illustrate the specificity of the present invention, respectively prepare bovine serum albumin BSA and carcinoembryonic antigen CEA standard solution (4ng / mL), the specific implementation steps are with reference to example 1, the Raman signal intensity that obtains can refer to Figure 6 , indicating that the method has strong specificity.

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Abstract

The invention discloses a PSA detection method based on a magnetic core-gold satellite assembly body and belongs to the field of biological detection. The method includes the step that magnetic nanoparticles modified with PSA aptamer and gold nanoparticles modified with a sequence PSA CS partly complementary to the aptamer are assembled to form the core-satellite assembly body. On the basis that when a to-be-detected object PSA exists, structure change of the assembly body causes change of Raman signals of supernate in a system, detection of the PSA content can be achieved. The invention provides a surface enhanced Raman spectrum method based on the core-satellite assembly body of capture probe-Raman signal particles is provided so as to achieve PSA ultra-sensitive detection. The magnetic enrichment function of the magnetic capture probe is combined with the method, the Raman signals are obtained by detecting the supernate, and the method has the advantages of being high in sensitivity, specificity and selectivity, low in detection limit, easy to operate and convenient and quick to use and has very good application prospects.

Description

technical field [0001] The invention belongs to the field of biological detection and relates to a new method for detecting prostate specific antigen, in particular to a PSA detection method based on a magnetic nucleus-golden satellite assembly. Background technique [0002] Prostate Specific Antigen (PSA) is a single-chain glycoprotein of 237 amino acid residues produced by the endoplasmic vesicles of prostate epithelial cells, with a molecular weight of about 34kD, and is functionally kallikrein-like a serine protease. It is recognized as a better tumor marker for the diagnosis of prostate cancer. [0003] The standard method of assessing the risk of prostate cancer in the state of the art is to measure the level of PSA in the blood. For example, Chinese patent CN101070345B discloses a detection method for detecting prostate-specific antigen (PSA), which utilizes the monoclonal antibody against prostate-specific antigen invented by the patent, and the PSA detection range...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658
Inventor 胡勇军杨康朱挺锋余萌江宁静张文建
Owner SOUTH CHINA NORMAL UNIVERSITY