Method for simultaneously detecting As < 3 + >, Pb < 2 + > and Hg < 2 + >

An as3 and nucleic acid technology, applied in measuring devices, biological testing, fluorescence/phosphorescence, etc., can solve problems that have not been reported in the literature, and achieve the effect of enhanced sensitivity and high selectivity detection

Pending Publication Date: 2021-08-24
QINGDAO UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] So far, based on enzyme cycle amplification technology, using hollow porous nanomaterials, using base recognition to construct nanocapsule-nucleic acid biomolecular complexes for the simultaneous detection of As 3+ , Hg 2+ and Pb 2+ technology has not yet been reported in the literature

Method used

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  • Method for simultaneously detecting As &lt; 3 + &gt;, Pb &lt; 2 + &gt; and Hg &lt; 2 + &gt;

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Embodiment 1

[0037] A preparation method proposed by the present invention that can detect As simultaneously 3+ , Pb 2+ and Hg 2+ The method for the nanocapsule-nucleic acid biomolecule complex based on exonuclease cycle amplification technology and biomolecular recognition technology comprises the following steps:

[0038] (1) Separately screen, design and synthesize three kinds of nucleic acid biorecognition molecules to ensure that they are respectively and only for the target metal ion As 3+ , Pb 2+ and Hg 2+ Have a specific response, but no response to any other coexisting ions and interference ions;

[0039] (2) Wash 20 μL of magnetic beads with Tris-HCl buffer solution with pH=7.4, add 400 μL of nano-gold carrier solution with hollow and porous structure, and add 200 μL of 11.664 mg / mL polydiallyldimethylammonium chloride solution , magnetically separated after 10 h at 37°C, and washed with a Tris-HCl buffer solution with pH = 7.4;

[0040] (3) Add 2 μL of fluorescent molecule...

Embodiment 2

[0049] A method proposed by the present invention that can simultaneously detect As 3+ , Pb 2+ and Hg 2+ method, including the following steps:

[0050] (1) Dispense 30 μL containing As 3+ , Pb 2+ and Hg 2+ The solution was added to the nanocapsule-nucleic acid biomolecule complex proposed by the present invention, 2.25 μL EXOⅢ shearing enzyme (45U) was added, diluted to 100 μL with a Tris-HCl buffer solution of pH=7.4, mixed evenly, and placed in a shaker The bed was reacted for 2 hours, and the reaction temperature was 37°C. Due to the target As 3+ , Pb 2+ and Hg 2+In the presence of the nucleic acid biomolecules assembled on the nanocapsule surface, the As 3+ , Pb 2+ and Hg 2+ A specific response occurs, and the three ions are specifically combined with their corresponding nucleic acid biomolecules, causing their conformation to change, falling off the surface of the nanocapsules, and opening the blocked pores, resulting in the fluorescent substances Cy5, RhB and ...

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Abstract

Environmental and water pollution caused by high-toxicity heavy metals is always a serious problem faced by human beings. As < 3 + >, Pb < 2 + > and Hg < 2 + > are environmental pollutants which have serious toxicity and are widely distributed, so that the environment and human beings can be seriously threatened even at low concentration. The standard of drinking water is strictly limited by the World Health Organization. Therefore, establishment of an efficient and sensitive analysis method capable of simultaneously detecting As < 3 + >, Pb < 2 + > and Hg < 2 + > is of great importance to the fields of environmental monitoring, food safety and the like. The invention provides a method capable of simultaneously detecting As < 3 + >, Pb < 2 + > and Hg < 2 + >, the method adopts a nanocapsule-nucleic acid biomolecule compound based on an exonuclease cyclic amplification technology, so that the As < 3 + >, Pb < 2 + > and Hg < 2 + > can be simultaneously detected, the detection sensitivity is remarkably improved on the premise of ensuring high specificity, and meanwhile, a wider linear detection range is obtained.

Description

technical field [0001] The present invention relates to As 3+ , Pb 2+ , Hg 2+ The simultaneous detection of, specifically relates to a simultaneous detection of As 3+ , Pb 2+ and Hg 2+ The nanocapsule-nucleic acid biomolecule complex based on exonuclease cycle amplification technology and biomolecular recognition technology and its preparation method and application belong to the field of environmental monitoring and nanobiotechnology. Background technique [0002] Environmental and water pollution caused by highly toxic heavy metals has always been a major concern of people, and they can enter the food chain through air, water and soil and seriously endanger the ecosystem. [0003] Arsenic is an element that exists widely in nature and has metal-like properties. Arsenic and its compounds have been recognized as carcinogens by many international authorities such as the International Agency for Research on Cancer under the World Health Organization (WHO) and the US Envi...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N21/64
CPCG01N33/5308G01N21/6486
Inventor 王卫董传诚侯晓珊罗细亮
Owner QINGDAO UNIV OF SCI & TECH
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