2019-nCoVs detection method and kit based on surface enhanced Raman spectroscopy

A surface-enhanced Raman and detection method technology, applied in reagent containers, Raman scattering, chemical instruments and methods, etc., can solve the problems of inability to obtain virus fingerprints, inability to reliably detect and identify, inability to adapt to hot spots, etc. The effect of reproducibility, high signal-to-noise ratio, and high sensitivity

Pending Publication Date: 2021-11-26
海澳华(黑龙江)生物医药技术有限公司
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Problems solved by technology

However, since the "hot spots" of SERS technology are usually less than 10nm space between gold or silver nanostructures, while biomacromolecules are larger than 10nm, for example, the size of viruses is usually about 100nm, which makes viruses and other biomacromolecules unable to adapt to SERS. "Hot spots" cannot be reliably detected and identified simply by using traditional SERS technology. In view of the above, the following improvements have been made in the prior art:
However, the method used cannot obtain the fingerprint of the virus itself, but indirectly detects it through the protein expressed after it infects the cell, and the fingerprint signal of the protein is easily affected by the inherent protein on the cell surface, resulting in poor accuracy.

Method used

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  • 2019-nCoVs detection method and kit based on surface enhanced Raman spectroscopy
  • 2019-nCoVs detection method and kit based on surface enhanced Raman spectroscopy
  • 2019-nCoVs detection method and kit based on surface enhanced Raman spectroscopy

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Experimental program
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Effect test

Embodiment 1

[0061] Silver sols with citrate ions on the surface were prepared according to Lee's method, where sodium citrate solution (1%, 4 mL) was added to slightly boiling silver nitrate solution (0.034 g, 200 mL) to reduce silver nanoparticles. Centrifuge 5 mL of sodium citrate-reduced silver sol (6500 rpm, 20 min, 20°C), remove the supernatant, and incubate 20 μL of centrifuged silver sol with 20 μL of potassium iodide solution (1 Mm) for more than 60 min at room temperature to obtain iodide ion-modified silver nanoparticles Sol;

Embodiment 2

[0063] (1) Add 3 μL of acetonitrile to the mixture of 20 μL of centrifuged silver sol and 20 μL of potassium iodide solution (1 Mm);

[0064] (2) Add 20 μL of influenza D virus sample (100 copies / mL) and 2 μL of Ca 2+ (0.01M CaCl 2 2H 2 O) mixing;

[0065] (3) The mixture is fully mixed and placed on the support for SERS detection. The laser wavelength is 633nm, the scanning time is 35s, and the laser energy is 28mW. Each test is accumulated once.

Embodiment 3

[0067] The difference between this embodiment and embodiment 2 is that the virus sample is influenza type C virus.

[0068] Such as figure 2 As shown, above-mentioned embodiment 2 and 3 have all obtained the SERS signal of virus constituent material through experimental verification, and all can obtain the fingerprint spectrum of different types of influenza virus at very low (100 copy number / mL),

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Abstract

The invention relates to a 2019-nCoV detection method and kit based on surface-enhanced Raman spectroscopy, and provides a biomacromolecule detection method and kit based on surface-enhanced Raman spectroscopy. The method comprises the following steps: firstly, preparing iodide ion modified silver nanoparticle sol as a mixture 1; adding acetonitrile into the mixture 1 to obtain a mixture 2; adding a sample to be detected into the mixture 2 to obtain a mixture 3; adding a substance capable of providing calcium ions into the mixture 3 to obtain a mixture 4; and carrying out SERS detection on the mixture 4. The method can be applied to actual clinical detection, can promote the application of SERS in the field of virus detection, and improves the handling capacity of sudden and new viral infectious diseases.

Description

technical field [0001] The invention belongs to the technical field of biomacromolecule detection, and in particular relates to a biomacromolecule detection method based on surface-enhanced Raman spectroscopy, and also relates to a kit. Background technique [0002] Biomacromolecules refer to macromolecules such as proteins, nucleic acids, and polysaccharides that exist in living cells. The structures are very complex, but their basic structural units are not complicated. Protein molecules are long chains of amino acid molecules arranged in a certain order. Amino acid molecules are the constituent materials of most living matter, and there are dozens of different amino acid molecules. The vast majority of enzymes in organisms belong to proteins, which are indispensable for organisms to maintain normal metabolic functions. Raman spectroscopy is a powerful means to study biomacromolecules. It can study the structure and changes of biomacromolecules in close to natural state ...

Claims

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Application Information

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IPC IPC(8): G01N21/65B01L3/00
CPCG01N21/658B01L3/52
Inventor 李洋孙建平高欣王晓童
Owner 海澳华(黑龙江)生物医药技术有限公司
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