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Dual-signal-amplification fluorescence sensor for detecting mercury ions and preparation method thereof

A fluorescent sensor and dual-signal technology, applied in the field of biosensors, can solve the problems of limiting the scope of use of traditional methods and real-time operability, and achieve the effects of fast response, mild reaction conditions, and improved sensitivity

Pending Publication Date: 2020-05-19
MINJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Long time pretreatment and expensive equipment limit the range of use and real-time operability of traditional methods

Method used

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  • Dual-signal-amplification fluorescence sensor for detecting mercury ions and preparation method thereof
  • Dual-signal-amplification fluorescence sensor for detecting mercury ions and preparation method thereof
  • Dual-signal-amplification fluorescence sensor for detecting mercury ions and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Template DNA probe, mercury ion recognition probe DNA probe, fuel probe, hairpin probe and fluorescent dye BHQ-2 and FAM modified hairpin probe sequence design are shown in Table 1;

[0046] Table 1. Complete sequence of sensors.

[0047]

Embodiment 2

[0048] Embodiment 2 Preparation of each component

[0049] (1) Synthesis of component a

[0050] Use 1×HEPES buffer to prepare probe S, F1 and F2 solutions with a concentration of 100 μM, take a 200 μL centrifuge tube, add 1 μL probe S (100 μM), 1 μL F1 (100 μM) and 1 μL F2 (100 μM), Supplement with 97 μL of 1×HEPES buffer (10 mM HEPES, 20 mM magnesium acetate, 150 mM sodium acetate, pH 7.4). Keep the centrifuge tube in a constant temperature metal bath at 30°C for 3 hours to fully hybridize and form the template DNA hybrid S / F1 / F2.

[0051] (2) Use 1×HEPES buffer to prepare a solution with a probe IP concentration of 1 μM, that is, component b.

[0052] (3) Use 1×HEPES buffer to prepare a solution with a concentration of F of 1 μM, that is, component c.

Embodiment 3

[0053] Embodiment 3 Annealing treatment of the hairpin probe

[0054] (1) Annealing of hairpin probe H1

[0055] Use 1×HEPES buffer to prepare probe H1 solution with a concentration of 2 μM, take a 200 μL centrifuge tube, add 1 μL H1 (100 μM) and 49 μL 1×HEPES buffer (10 mM HEPES, 20 mM magnesium acetate, 150 mM acetic acid sodium, pH 7.4). Keep the centrifuge tube in a constant temperature metal bath at 95°C for 5 minutes, and then quickly place it in ice water for 30 minutes to form a stable secondary structure.

[0056] (2) Annealing of fluorescent dye BHQ-2 and FAM-modified hairpin probe H2

[0057] Use 1×HEPES buffer to prepare a probe H2 solution with a concentration of 2 μM, take a 200 μL centrifuge tube, add 1 μL H2 (100 μM) and 49 μL 1×HEPES buffer (10 mM HEPES, 20 mM magnesium acetate, 150 mM acetic acid sodium, pH 7.4). Keep the centrifuge tube in a constant temperature metal bath at 95°C for 5 minutes, and then quickly place it in ice water for 30 minutes to fo...

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Abstract

The invention discloses a dual-signal-amplification fluorescence sensor for detecting mercury ions and a preparation method thereof. The fluorescence sensor comprises seven probes, namely a, b, c andd, namely a probe S, a probe F, a probe F1, a probe F2, a probe IP, a probe H1 and a probe H2; fuel stimulation isothermal amplification (FSP) primary circulation is realized based on specific recognition (T- Hg<2+> -T specific base mismatch) of the mercury ions and nucleic acid, an F2 chain released by the FSP can achieve chain displacement catalytic card assembly reaction (CHA) with H1 and H2, secondary circulation is achieved, and lastly the mercury ions are detected through fluorescence signal enhancement. The fluorescence sensor is advantaged in that the sensor has low detection limit, high sensitivity, high reaction speed, strong anti-interference capability, mild reaction conditions and the like, can make up for defects of a mercury ion detection method in the prior art, and realizes rapid and accurate quantitative detection of the mercury ions.

Description

technical field [0001] The invention belongs to the field of biosensors, and more specifically relates to a fluorescent sensor with dual signal amplification for detecting mercury ions and a preparation method thereof, in particular to a fuel-stimulated isothermal amplification (FSP) based on chain displacement regulation and catalytic hairpin assembly reaction ( CHA) double isothermal signal amplification circuit and fluorescent biosensor for fluorescent detection of mercury ions. Background technique [0002] Mercury pollution is one of the most common heavy metal pollution in water bodies, and the monitoring of heavy metal mercury ions in water bodies has attracted extensive attention. Mercury can be absorbed and enriched through human skin, respiratory tract, and food, and can combine with various proteins containing sulfhydryl groups, thereby causing damage to the human digestive tract, kidneys, liver, etc., and can also cause neurasthenia and other symptoms. cause gre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06C12Q1/682C12Q1/6825
CPCG01N21/643C09K11/06C12Q1/682C12Q1/6825G01N2021/6432C09K2211/1088
Inventor 邢超王军
Owner MINJIANG UNIV
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