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Lead ion visual detection method

A detection method and technology for lead ions are applied in the field of rapid visual detection of lead ions, which can solve the problems of interfering lead ion detection and low detection sensitivity, and achieve the effects of high sensitivity and rapid detection, high detection sensitivity, and improved use efficiency.

Inactive Publication Date: 2017-03-29
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the above defects or improvement needs of the prior art, the present invention provides a rapid visual detection method for lead ions, the purpose of which is to treat DNase with graphene oxide, absorb unreacted free single-stranded nucleic acid, and combine double gold standard Probe detection technology improves the use efficiency of fragments produced after enzyme digestion, thereby improving detection sensitivity, thus solving the problem of false positive results in the detection of lead ion chromatography test paper based on DNase in the prior art that interferes with the detection and detection sensitivity of lead ions Low and other technical problems that impurity ions interfere with the detection results

Method used

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preparation example Construction

[0046] 1) Preparation of gold-labeled probe:

[0047] Preparation of the first gold-labeled probe: Add the sulfhydryl-modified first single-stranded DNA to the nano-gold solution, react for 24 hours, then add phosphate buffer and sodium dodecylsulfonate, mix at room temperature for 30 minutes, add chloride Aging at room temperature with sodium for 2 days, centrifuging at 4°C for 25 min, discarding the supernatant, and resuspending the precipitate with resuspension buffer to obtain the prepared first gold-labeled probe;

[0048] Preparation of the second gold-labeled probe: Add the sulfhydryl-modified second single-stranded DNA to the nano-gold solution, react for 24 hours, then add phosphate buffer and sodium dodecylsulfonate, mix at room temperature for 30 minutes, add chloride Aging with sodium at room temperature for 2 days, centrifuging at 4°C for 25 min, discarding the supernatant, and resuspending the precipitate with resuspension buffer to obtain the prepared second gol...

Embodiment 1

[0067] A rapid visual detection method for lead ions, comprising the steps of:

[0068] (1) Preparation of nano gold solution:

[0069] 100mL HAuCl 4 The aqueous solution (0.01%) was poured into a round bottom flask equipped with a reflux condenser and heated to boiling, then 2 mL of trisodium citrate solution (1%) was added to the flask. Continue to heat and reflux under strong magnetic stirring. After the color of the solution gradually changes from colorless and black to deep red, continue heating for 10 minutes, stop heating, cool to room temperature, filter with a 0.22 μm nylon membrane to remove large particles, and store it at temperature Set aside in the refrigerator at 4 °C.

[0070] (2) Preparation of gold standard probe:

[0071] Add 45 μL of thiol-modified first single-stranded DNA and second single-stranded DNA (100 μM, 1OD) to 14 mL of nano-gold solution, react for 24 h, add 100 mmol / L PB (phosphate buffer) and 10% SDS ( sodium dodecyl sulfate) to make the fi...

Embodiment 2

[0085] A kind of lead ion gold label chromatography test strip provided by the invention, such as figure 2 As shown, including the sample pad, gold standard pad, nitrocellulose membrane and absorbent paper carried on the PVC bottom plate in sequence, the gold standard pad is fixed with the first gold standard probe ( figure 2 The gold-labeled probe 1) and the second gold-labeled probe ( figure 2 The gold standard probe 2) in the medium; There are detection line and quality control line on the nitrocellulose membrane, and the third biotin probe is immobilized on the detection line ( figure 2 The biotin probe 3) and the fourth biotin probe ( figure 2 The biotin probe 4) in the quality control line is immobilized with the fifth biotin probe ( figure 2 The biotin probe 5) and the sixth biotin probe ( figure 2 Biotin probes in 6). The base chain GR-5S and the enzyme chain GR-5E are complementary paired, the excess free single chain is adsorbed by graphene oxide, and lead...

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Abstract

The invention discloses a lead ion fast visual detection method. The method comprises: performing hybrid hybridization of fundamental chains and enzyme chains to form a GR-5 DNA enzyme; adding oxidized graphene for treatment to remove unreacted free single-stranded nucleic acid to eliminate the false positive effect; spraying two kinds of gold label probes on a gold label mat; and adding two chains which are obtained from the GR-5 DNA enzyme which is a pretreated to-be-detected sample through cutting at the rA position, wherein the two chains together with the gold label probes and nucleotide sequences located on a detection line form a sandwich structure through chromatography, so that the chains are fixed at the detection line for color development, and excessive free nucleic acid is fixed at a quality control line for color development. According to the intensity of color development on the detection line, the lead ion concentration is obtained. Lead ions can be detected in 15 min at the room temperature, and the minimum limit of detection of the lead ions can reach 0.05 nM. The method is high in sensitivity and good in specificity, is not affected by other divalent metal ions, is simple and easy to operate, and can be directly applied to detection of lead ions in the environment.

Description

technical field [0001] The invention belongs to the field of metal ion detection, and more specifically relates to a rapid visual detection method for lead ions. Background technique [0002] Lead, as a heavy metal element, can cause environmental pollution and endanger human health. The damage of lead to the human body is multi-system and multi-organ, and it will cause toxic effects on the bone marrow hematopoietic system, cardiovascular system, immune system, nervous system and digestive system; as a central nervous system poison, it is harmful to children. Nervous system damage is more serious. After lead is discharged into the environment, most of it will enter the soil, and then enter the human body through grain and vegetables. Many countries have regulations on lead content in the environment and food. For example, China's "Soil Environmental Quality Standard" limits the maximum concentration of lead ions in farmland II soil to 50nM, and the U.S. Environmental Prot...

Claims

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Application Information

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IPC IPC(8): G01N33/558
CPCG01N33/558
Inventor 赵元弟王海波
Owner HUAZHONG UNIV OF SCI & TECH
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