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 prod

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Example Embodiment

[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 dodecyl sulfonate, mix at room temperature for 30 minutes, and add chlorination Sodium aging for 2 days at room temperature, centrifugation at 4°C for 25 min, discard the supernatant, and resuspend the pellet with resuspension buffer to obtain the first gold-labeled probe prepared;

[0048] Preparation of the second gold-labeled probe: Add the second single-stranded DNA modified with sulfhydryl group to the nano gold solution, react for 24 hours, then add phosphate buffer and sodium dodecyl sulfonate, mix at room temperature for 30 minutes, add chlorination Sodium aging for 2 days at room temperature, centrifugation at 4°C for 25 min, discard the supernatant, and resuspend the pellet with resuspension buffer to obtain the second gold-labeled probe p...

Example Embodiment

[0066] Example 1

[0067] A rapid visual detection method for lead ions includes the following steps:

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

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

[0070] (2) Preparation of gold-labeled probe:

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

Example Embodiment

[0084] Example 2

[0085] The invention provides a lead ion gold standard chromatography test strip, such as figure 2 As shown, it includes a sample pad, a gold label pad, a nitrocellulose membrane and absorbent paper which are sequentially mounted on a PVC bottom plate. The gold label pad is fixed with the first gold label probe ( figure 2 The gold-labeled probe 1) and the second gold-labeled probe ( figure 2 The gold standard probe 2); there are detection lines and quality control lines on the nitrocellulose membrane, and the third biotin probe ( figure 2 Biotin probe 3) and the fourth biotin probe ( figure 2 The biotin probe 4), the fifth biotin probe ( figure 2 Biotin probe 5) and the sixth biotin probe ( figure 2 The biotin probe in 6). The base chain GR-5S and the enzyme chain GR-5E undergo complementary pairing, the excess free single chains are adsorbed by graphene oxide, and lead ions are added to the sample pad as a loading solution.

[0086] The basal strand of GR-...

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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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