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Quick label-free detection method for lead ions

A lead ion, label-free technology, applied in the direction of material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of unfavorable rapid detection, troublesome operation, time-consuming and labor-consuming, etc., and achieve the effect of low detection limit, simple operation and high detection accuracy

Inactive Publication Date: 2012-06-27
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

However, it is often necessary to label fluorescent groups and quenching groups, which is cumbersome, time-consuming and labor-intensive, and is not conducive to rapid detection

Method used

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  • Quick label-free detection method for lead ions
  • Quick label-free detection method for lead ions
  • Quick label-free detection method for lead ions

Examples

Experimental program
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Effect test

Embodiment 1

[0034] The steps for label-free detection of lead ions using fluorescent copper nanoparticles formed based on random double-stranded DNA as a template are as follows:

[0035] 1) Dissolve random complementary double-stranded DNA with 3-(N-morpholine) propanesulfonate sodium salt (MOPS) buffer solution (10 mM, pH=7.5) containing 150 mM sodium acetate. The sequence of the randomly adopted DNA is : 5'-TACTCATACGCTCATACGTTCATCACGACTACACA-3' (SEQ ID NO: 1) (synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.), 500 nM two random complementary DNAs were mixed evenly, reacted in a 90°C water bath for 10 minutes, and then slowly Cool down to room temperature to obtain 500 nM random complementary double-stranded DNA solution;

[0036] 2) Add 1 mM sodium ascorbate to 500 mL of 500 nM complementary double-stranded DNA, mix well, then add 100 mM copper acetate, mix well, and react at room temperature for about 20 minutes in the dark to obtain random double-stranded DNA Fluorescent c...

Embodiment 2

[0042] In order to investigate the specificity of fluorescent copper nanoparticles synthesized based on random double-stranded DNA as a template for label-free detection of lead ions, a selectivity experiment was carried out as follows:

[0043] 1) Dissolve random complementary double-stranded DNA with 3-(N-morpholine) propanesulfonate sodium salt (MOPS) buffer solution (10 mM, pH=7.5) containing 150 mM sodium acetate. The sequence of the randomly adopted DNA is : 5'-ATGAACGTATGAGCG-3' (SEQ ID NO: 2) (synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.). Two random complementary DNAs of 500 nM were mixed evenly, reacted in a 90°C water bath for 10 minutes, and then slowly lowered to room temperature to obtain a 500 nM random complementary double-stranded DNA solution;

[0044] 2) Add 1 mM sodium ascorbate to 500 mL of 500 nM complementary double-stranded DNA, mix well, then add 100 mM copper sulfate, mix well, and react at room temperature for about 20 minutes in the da...

Embodiment 3

[0048] In order to investigate the use of fluorescent copper nanoparticles synthesized based on random double-stranded DNA as a template for label-free detection of lead ions in actual samples, the recovery rate experiment was carried out as follows:

[0049] 1) Dissolve random complementary double-stranded DNA with 3-(N-morpholine) propanesulfonate sodium salt (MOPS) buffer solution (10mM, pH=7.5) containing 150 mM sodium acetate. The sequence of the randomly used DNA is: 5'-AGTTGCAAGAAGATGACAGAGAAGT-3' (SEQ ID NO: 3) (synthesized by Sangon Bioengineering (Shanghai) Co., Ltd.). Two random complementary DNAs of 500 nM were mixed evenly, reacted in a 90°C water bath for 10 minutes, and then slowly lowered to room temperature to obtain 500 nM random complementary double-stranded DNA;

[0050] 2) Add 1 mM sodium ascorbate to 500 mL of 500 nM complementary double-stranded DNA, mix well, then add 100 mM copper nitrate, mix well, and react at room temperature for about 20 minutes in...

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Abstract

The invention discloses a quick label-free detection method for lead ions. In the presence of a reducing agent, bivalent copper ions form nano-copper particles randomly in a deoxyribonucleic acid (DNA) double-chain; after the lead ions are added, fluorescence of the fluorescent nano-copper particles is quenched; and concentration of the lead ions and fluorescence strength have good correlation so that the purpose for detecting the lead ions is achieved. The detection method has good sensitivity, detection limit of 5nM and good specificity, does not influence the detection on other ions, and can be used for detecting real samples. In addition, the recovery rate is 86%-108%. Furthermore, the quick label-free detection method for the lead ions is simple in operation, convenient in use, high in sensitivity and wide in selectivity, does not need special nucleotide sequence and complex instruments, and provides a novel detection means for quickly detecting lead ion content in the real samples.

Description

Technical field [0001] The present invention involves a test method of a lead ion, which specializes in a method of fast marking to detect lead ions. Background technique [0002] Lead ions are seriously harmful to the human body and are an important indicator of environmental monitoring.Its main toxic effect is anemia, neurological disorders and renal damage, and reproductive system damage.The US Environmental Protection Agency stipulates that the maximum allowable amount of lead ions in drinking water must not exceed 72nm.At present, the conventional detection methods of marks in the environment are atomic absorption method, atomic fluorescent spectrum, inductive plasma launch spectrum method, and electrochemical methods.However, these methods are cumbersome, and the troubled forward processing, special analysis technical personnel, and expensive instruments are not conducive to fast analysis and testing on the spot.In recent years, the method of using the GNA enzyme (17E DNAZY...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
Inventor 曾令文陈俊华
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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