Carboxyl ligand induced annihilation type ratio electrochemiluminescence aptamer sensing method for detecting Pb < 2 + >

A sensing method, electrochemical technology, applied in the direction of chemiluminescence/bioluminescence, analysis by making materials undergo chemical reactions, measurement devices, etc., can solve problems that affect the stability of the ECL system, interfere with the detection environment, and measure errors. Achieve excellent stability, improve sensitivity, and improve reliability

Pending Publication Date: 2022-06-17
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in some applications, the introduction of co-reactants may interfere with the detection environment and affect the stability of the ECL system, resulting in measurement errors

Method used

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  • Carboxyl ligand induced annihilation type ratio electrochemiluminescence aptamer sensing method for detecting Pb &lt; 2 + &gt;
  • Carboxyl ligand induced annihilation type ratio electrochemiluminescence aptamer sensing method for detecting Pb &lt; 2 + &gt;
  • Carboxyl ligand induced annihilation type ratio electrochemiluminescence aptamer sensing method for detecting Pb &lt; 2 + &gt;

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] according to figure 1 Described preparation process:

[0049] (1) Preparation of AuNPs and AuNPs-cDNA

[0050] 0.2mL 0.1M HAuCl 4 ·3H 2 O solution was dissolved in 25mL ultrapure water, stirred and dissolved to obtain a mixed solution. Then heat the mixed solution to boiling at 150°C; when the solution boils completely, add 0.25mL of 100mg·mL -1 Sodium citrate solution to obtain a red solution; then, the solution continued to react for 15 minutes and then cooled to room temperature to obtain an AuNPs solution; the obtained AuNPs solution was stored in a refrigerator at 4°C in the dark for later use;

[0051] Preparation of AuNPs-cDNA: First, mix 50 μL of 100 μM cDNA solution with a sulfhydryl group at the end and 100 μL of 10 mM tris(2-carbonylethyl)phosphate hydrochloride (TCEP) solution, and after standing for 1 h, an activated cDNA solution was obtained; then 150 μL of activated cDNA solution and 1850 μL of AuNPs solution were mixed, shaken and incubated for 12 ...

Embodiment 2

[0063] The selectivity of the annihilation-type ratiometric electrochemiluminescence aptasensor was investigated using the sensor prepared in Example 1;

[0064] When placing the ECL aptasensor in aqueous solution, the obtained I 阴 / I 阳 is a blank signal; when different ions (K + , Na + , Ca 2+ ,Mg 2+ , Fe 2+ , Fe 3+ ,Cd 2+ ,Cu 2+ ,Hg 2+ , Ni 2+ ,Mn 2+ ,Zn 2+ , Mix, Pb 2+ and Pb 2+ +Mix) to replace the blank sample, the obtained I 阴 / I 阳 is the test signal; using the absolute value of the difference between the test signal and the blank signal, it is recorded as |ΔI 阴 / I 阳 |, to examine its selection performance.

[0065] Figure 5 is the selectivity of the sensor: where blank refers to an aqueous solution, defined as a blank sample; K + , Na + , Ca 2+ ,Mg 2+ , Fe 2+ , Fe 3+ ,Cd 2+ ,Cu 2+ ,Hg 2+ , Ni 2+ ,Mn 2+ ,Zn 2+ , Mix is ​​used as an interference substance, and Mix is ​​a mixed solution of the above-mentioned 12 kinds of interference ions. ...

Embodiment 3

[0067] Using the sensor prepared in Example 1 to investigate the reproducibility of the annihilation-type ratiometric electrochemiluminescence aptasensor;

[0068] Five ECL aptasensors prepared in parallel in the same batch were respectively placed in a 1.0×10 -9 5 Pb of M 2 + When in solution, record the I within the batch 阴 / I 阳 ; 5 ECL aptasensors prepared in different batches were placed in a 1.0×10 -9 5 Pb of M 2+ When in solution, record the batch-to-batch I 阴 / I 阳 .

[0069] from Image 6 , it can be seen that the relative standard deviation (RSD) of the ECL signals of the five parallel aptasensors within a batch was 2.86%, and 2.62% between batches, proving that the sensor has good reproducibility.

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Abstract

The invention belongs to the field of biosensing detection, and relates to a sensing method for detecting an annihilation type ratio electrochemical luminescence aptamer induced by a carboxyl ligand of Pb < 2 + >. The invention develops an electrochemiluminescence aptamer detection method for detecting the annihilation type ratio of Pb < 2 + > by taking carboxyl terpyridyl ruthenium with strong cathode-anode signal intensity as a luminous body and fixing resonance energy transfer receptor gold nanoparticles of the carboxyl terpyridyl ruthenium by utilizing the hybridization effect of a Pb < 2 + > aptamer and complementary DNA (Deoxyribose Nucleic Acid) of the Pb < 2 + > aptamer. The reliability of the annihilation ratio electrochemical luminescence method is improved by combining the carboxyl terpyridyl ruthenium with excellent annihilation performance and the resonance energy transfer receptor gold nanoparticles which can be distinguished at the cathode and the anode; due to the introduction of the aptamer of the specific recognition element Pb < 2 + >, the selectivity of the electrochemical luminescence method is improved, and the specific analysis of Pb < 2 + > is realized; the method is high in sensitivity, good in selectivity and high in reliability.

Description

technical field [0001] The invention belongs to the field of biosensing detection, in particular to a method for detecting Pb 2+ A Carboxyl Ligand-Induced Annihilation-Type Ratiometric Electrochemiluminescence Aptamer Sensing Method. Background technique [0002] The problem of heavy metal pollution in soil has attracted great attention from governments and relevant international organizations. Among them, Pb 2+ As a common heavy metal ion in farmland soil, it is difficult to decompose, highly toxic, and easy to accumulate in organisms. They can pollute farmland soil, reduce the fertility of arable soil, and lead to a reduction in crop yields; in turn, they can affect the quality and safety of agricultural products and threaten human health through absorption and accumulation in plants. Currently, Pb 2+ The detection and analysis methods mainly include atomic absorption spectrometry, flame atomic absorption spectrometry, inductively coupled plasma-mass spectrometry, indu...

Claims

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Application Information

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IPC IPC(8): G01N27/327G01N27/416G01N21/76
CPCG01N27/308G01N27/3276G01N27/416G01N21/76
Inventor 李丽波刘晓红由天艳罗莉君毕晓雅赵万林
Owner JIANGSU UNIV
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