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Electrochemical method for detecting androgen receptors based on androgen receptor recognition element and G-quadruplex hybridization chain amplification reaction

A technology of androgen receptor and recognition element, which is applied in the field of analytical chemistry and clinical diagnosis, can solve the problems of inaccurate determination of receptor content, uneconomical, time-consuming, etc., and achieve improved detection sensitivity, high sensitivity, and high-sensitivity detection Effect

Active Publication Date: 2019-02-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Western blot and immunohistochemistry are typical semi-quantitative detection modes, which cannot accurately determine the receptor content
Although enzyme-linked immunosorbent assay can be used for quantitative detection, it requires specific labeled antibodies, which is time-consuming and uneconomical

Method used

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  • Electrochemical method for detecting androgen receptors based on androgen receptor recognition element and G-quadruplex hybridization chain amplification reaction
  • Electrochemical method for detecting androgen receptors based on androgen receptor recognition element and G-quadruplex hybridization chain amplification reaction

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Drawing of peak current and androgen receptor concentration standard curve in embodiment 1 DPV curve

[0038] ARBP was immobilized on the electrode, and the electrode obtained after incubation with different concentrations of AR samples, NspI cleavage, G-quadruplex hybridization chain amplification reaction, and G-quadruplex-heme complex formation was used as the working electrode, and the differential pulse Measured by voltammetry (DPV).

[0039] Specific steps are as follows:

[0040] (1) Formation of AR-binding probe ARBP: 0.1 μmol L -1 Thiol-modified P1 and 0.1 μmol L -1 Probe P2 that is partially complementary to it was prepared at 10mmol·L -1 In Tris-HCl buffer, undergo denaturation at 95°C for 5 minutes, and renaturation at room temperature for 2 hours to form partially complementary double-stranded DNA ARBP for future use;

[0041] (2) Pretreatment of G-quadruplex formation probe: use 10mmol·L -1 Dissolve Hp1, Hp2, Hp3 and Hp4 in PB buffer and dilute to ...

Embodiment 2

[0051] Example 2 Specific Analysis of AR Detection

[0052] The signal responses in the presence of three different proteins, androgen receptor (AR), immunoglobulin G (IgG) and human serum albumin (HSA), were compared. Compared with the increased signal generated by AR protein, the signal intensity of IgG and HSA is much lower, thus verifying that the constructed electrochemical sensor can well distinguish AR and other proteins.

Embodiment 3

[0053] Example 3 Detection of AR in Human Serum

[0054] Add a certain concentration of AR to the human serum sample, and react with the same operation and reagents as in Example 1 (1)-(9), measure the DPV curve, and calculate from the standard curve according to the peak current value in the DPV curve Calculate the concentration of the AR concentration and evaluate the recovery and relative standard deviation of the assay accordingly.

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Abstract

The invention relates to an electrochemical method for detecting androgen receptors based on an androgen receptor recognition element and a G-quadruplex hybridization chain amplification reaction, andbelongs to the technical field of analytical chemistry and clinical diagnosis. According to the method, by using the protective effect of an androgen receptor (AR) to an androgen receptor recognitionelement (ARE), the AR is bound to double-stranded DNA having a specific sequence (ARE) in the presence of the AR so as to protect from restriction endonuclease NspI cleavage, and a large number of G-quadruplex structures are aggregated on a DNA probe immobilized on the surface of an electrode and containing the ARE through sequence complementation by using a hybridization chain amplification reaction; and the G-quadruplex-heme complex having redox activity and bound on the surface of the electrode is positively correlated with the AR concentration, the peak current signal is determined by using a differential pulse voltammetry (DPV) method, and the quantitative analysis of the AR is achieved through the linear relationship between the current value and the AR concentration. According to the present invention, the method has advantages of high sensitivity, good specificity and the like, and is suitable for the quantitative analysis of the AR concentration in clinical samples such as human serum and the like.

Description

technical field [0001] The invention relates to an electrochemical method for detecting androgen receptor based on an androgen receptor recognition element and a G-quadruplex hybridization chain amplification reaction, belonging to the technical fields of analytical chemistry and clinical diagnosis. Background technique [0002] Androgen is a very important steroid hormone in the body. It is mainly testosterone and a small amount of dehydroepiandrosterone and androstenedione in the body. It plays an important physiological role in the male reproductive system, such as: affecting the development of male reproductive organs and the production of sperm Occurs and matures, promotes the appearance of secondary sex characteristics and maintains normal sexual function in men. The androgen effect must rely on its receptor—androgen receptor (AR). Without AR, androgen has no stimulating response to tissues. AR is an intermediary substance of androgen action. Studies have shown that A...

Claims

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Application Information

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IPC IPC(8): C12Q1/6816
CPCC12Q1/6816C12Q2521/301
Inventor 周楠迪姚斌斌田亚平
Owner JIANGNAN UNIV
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