Mass spectrum database establishing method based on virus identification

A technology for establishing a method and database, which is applied in the field of mass spectrometry database establishment based on virus identification, and achieves the effects of high sensitivity, high throughput and simple operation.

Pending Publication Date: 2020-06-02
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But there is no article report about the establishment of the virus mass spectrum database at present, so the purpose and significance of the present invention are to establish a mass spectrum database for rapid identification and distinction of different viruses, and use it in the fields of biosafety and public health

Method used

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  • Mass spectrum database establishing method based on virus identification
  • Mass spectrum database establishing method based on virus identification
  • Mass spectrum database establishing method based on virus identification

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Virus sample preprocessing and database establishment

[0027] Virus sample pretreatment process:

[0028] Take 5 μL of the virus sample, centrifuge at 10,000 rpm at 0°C for 10 min, and then remove the supernatant. Add 2 μL of 10% trifluoroacetic acid solution to the pellet to fully dissolve the virus particles, then add 2 μL of acetonitrile and vortex to mix well. The obtained virus samples are pre-treated samples.

[0029] Virus mass spectrum acquisition:

[0030] First weigh 5 mg of CHCA and dissolve it in 1 mL of acetonitrile / water (3:2 volume ratio, containing 1% trifluoroacetic acid by volume) mixed solvent to obtain a 7 mg / mL matrix stock solution.

[0031] Add 1 μL of sample solution dropwise on the MALDI target plate, after drying, add 1 μL of CHCA matrix dropwise, and then spot 8 groups of samples in parallel. After all the solutions were dried, 24 (8×3) virus mass spectra were collected. The MALDI-TOF MS parameters during acquisition are set as follows: ...

Embodiment 2

[0036] MALDI-TOF MS detection of virus samples

[0037] Virus sample pretreatment process:

[0038] Take 8 μL of virus samples, centrifuge at 20,000 rpm at 4°C for 30 min, and remove the supernatant. Add 5 μL of 30% trifluoroacetic acid solution to the pellet to fully dissolve the virus particles, then add 5 μL of acetonitrile, and vortex to mix well. The obtained virus samples are pre-treated samples.

[0039] Virus mass spectrum collection:

[0040] First weigh 10 mg DHB and dissolve it in 2 mL of acetonitrile / water (3:2 volume ratio, containing 1% trifluoroacetic acid by volume) mixed solvent to obtain a 5 mg / mL matrix stock solution.

[0041] Add 1 μL of sample solution dropwise on the MALDI target plate, after drying, add 1 μL of LDHB matrix dropwise, and then spot 8 groups of samples in parallel. After all the solutions were dried, 24 (8×3) virus mass spectra were collected. The MALDI-TOF MS parameters during acquisition are set as follows: laser energy (60%), frequ...

Embodiment 3

[0046] MALDI-TOF MS detection of virus samples

[0047] Virus sample pretreatment process:

[0048] Take 10 μL of the virus sample, centrifuge at 10° C. at 20,000 rpm for 45 minutes, and remove the supernatant. Add 5 μL of 50% trifluoroacetic acid solution to the precipitate to fully dissolve the virus particles, then add 5 μL of acetonitrile, and vortex well. The obtained virus samples are pre-treated samples.

[0049] Virus mass spectrum collection:

[0050] First weigh 20 mg of SA and dissolve it in 1 mL of acetonitrile / water (3:2 volume ratio, containing 0.1% trifluoroacetic acid by volume) mixed solvent to obtain a 20 mg / mL matrix stock solution.

[0051] Add 1 μL of sample solution dropwise on the MALDI target plate, after drying, add 1 μL of SA matrix dropwise, and then spot 8 groups of samples in parallel. After all the solutions were dried, 24 (8×3) virus mass spectra were collected. The MALDI-TOF MS parameters during acquisition are set as follows: laser energy ...

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Abstract

The invention relates to a mass spectrum database establishing method based on virus identification and application thereof. Establishment of the database mainly comprises two parts: firstly, providing a processing method aiming at a sample to obtain an improved primary mass spectrogram of a sample to be detected; secondly, establishing a database that is mainly established for a primary mass spectrogram of a sample to be detected and characteristic spectra of a low molecular weight end and a high molecular weight end. The method has a remarkable effect on virus identification, has the advantages of high-throughput analysis, detection and identification, and can be applied to the fields of biosafety and public health.

Description

technical field [0001] The invention relates to a method for establishing a mass spectrum database based on virus identification and its application. [0002] technical background [0003] As we all know, a virus is a kind of non-cellular organism with small individual, no complete cell structure, only one kind of genetic material (DNA or RNA), and must parasitize in living cells and multiply by replication. Each virus has its own unique genetic material and protein composition, and has a variety of shapes, mainly spherical, rod-shaped and tadpole-shaped. There is a genome composed of one or more nucleic acid molecules (DNA or RNA) inside, and a protective shell of protein or lipoprotein outside. The prevalence and outbreak of viruses will have a huge impact on human production and life, and seriously threaten public health. Therefore, the rapid and accurate identification and detection of viruses, as well as the prevention and control of viral diseases are very important a...

Claims

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Application Information

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IPC IPC(8): G01N27/62G01N33/569
CPCG01N27/622G01N33/56983
Inventor 张丽华韩国斌刘路宽杨开广梁振张玉奎
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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