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Identification method of sea snake venom and application thereof

An identification method and snake venom technology are applied in the identification of sea snake venom, and the snake venom protein analysis technology is used to identify the species of snake venom. The problem is to overcome the complex and cumbersome operation process, obvious advantages and high detection sensitivity.

Pending Publication Date: 2021-12-17
中国人民解放军海军特色医学中心 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the agglutination test method has the problems of low detection sensitivity, low correct detection rate and obvious cross-reaction
Some scholars have revealed that there are serious cross-reactions in the identification of snake venom proteins by ELISA, which is very unfavorable for the identification of snake venom
Aptamer technology has the disadvantage that the operation process is very complicated and tedious, such as "A method for detecting and identifying snake venom species using aptamer technology" (Chinese invention patent application CN102323400A)
Polymerase chain reaction is to identify from the perspective of DNA, but there are few reports on related research
[0008] Invention patent application CN 110426522 A discloses an identification method and its application of Viper venom venom. SDS-PAGE electrophoresis combined with mass spectrometry is used to identify Viper venom. The identification method can clearly identify the type of protein , but the equipment used in mass spectrometry is expensive, the test time is long, and the manual operation is heavy

Method used

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  • Identification method of sea snake venom and application thereof
  • Identification method of sea snake venom and application thereof
  • Identification method of sea snake venom and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1 A kind of identification method of sea snake venom

[0054] Proceed as follows:

[0055] (1) Sea snake venom (including sea snake venom and sea snake venom) was dissolved in solution A and prepared to 30mg / mL;

[0056] (2) Ultraviolet Spectrum Scanning Analysis

[0057] Dilute the sea snake venom to 1mg / ml, set the spectral scanning wavelength to 200-400nm, scan at a medium speed, and obtain the spectrum of the ultraviolet spectral scanning (see figure 1 ), with the highest absorption peaks at 210-220nm and 280nm.

[0058] (3) Carry out SDS-PAGE electrophoresis, the protein gel concentration is 17.5%, and the formula is as follows in Table 2:

[0059] Table 2 Protein glue concentration formula

[0060]

[0061] (4) Coomassie brilliant blue staining; SDS-PAGE electrophoresis see figure 1 , 2 main protein bands (15kD, 6-9kD respectively) can be distinguished (see figure 2 Main Band 1 and Main Band 2 in );

[0062] (5) LC-ESI-MS / MS mass spectrometry...

Embodiment 2

[0074] Example 2 Application of a sea snake venom identification method in the preparation of anti-sea snake venom serum for the treatment of sea snake venom poisoning

[0075] Proceed as follows:

[0076] (1) Sea snake venom (including sea snake venom and sea snake venom) freeze-dried powder, solution A (see Example 1) was dissolved, and then scanned by ultraviolet spectrum (see image 3 ), with the existing spectrogram ( figure 1 ) for comparison;

[0077] (2) Carry out SDS-PAGE electrophoresis and LC-ESI-MS / MS mass spectrometry identification, and the SDS-PAGE electrophoresis results ( Figure 4 ) and the existing electrophoretic pattern ( figure 2 ) comparison, confirming that the protein has two main bands, which are the main protein corresponding to sea snake venom;

[0078] (3) Immunize the horses after detoxifying formaldehyde from the identified sea snake venom;

[0079] (4) Horses immune to sea snake venom are transferred to the production department for the pr...

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Abstract

The invention discloses an identification method of sea snake venom, which comprises the following steps of: (1) selecting freeze-dried powder of sea snake venom (containing Lapemis curtus venom and Hydrophis cyanocinctus venom) from the origin of China, dissolving the freeze-dried powder and preparing into a protein solution; (2) carrying out analysis and preliminary identification on the sea snake venom by adopting ultraviolet spectrum scanning; and (3) carrying out clear identification by adopting SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and LC-ESI-MS / MS (Liquid Chromatography-Mass Spectrometry). In addition, the invention further discloses application of the identification method of the sea snake venom in preparation of anti-sea snake venom serum for treating sea snake venom poisoning. Laboratory identification can be carried out on the sea snake venom, quality control can be carried out on the sea snake venom more accurately, and then the specific medicine anti-sea snake venom serum for treating sea snake venom poisoning is produced. The identification method has the advantages of high detection sensitivity, high correct detection rate, no cross reaction, effective shortening of the test time, and great reduction of the manual operation cost.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a method for identifying sea snake venom, in particular to a method for identifying the type of snake venom using snake venom protein analysis technology; in addition, the invention also relates to the application of the identification method, and the identified snake venom can be used for Production of antivenom. Background technique [0002] Venomous snakebites are common in subtropical and tropical regions. In the rural areas of Asia, Africa, Latin America and Oceania, venomous snakebite is a particularly prominent public health problem, and the incidence of snakebite in my country shows a trend of increasing year by year. [0003] The venom is mostly light yellow or milky white translucent viscous liquid with more than 100 components. Each snake venom contains a variety of different toxic components, and the content of various toxic components in different snake venoms is qu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33G01N21/31G01N27/447G01N30/02G01N30/72G01N30/88
CPCG01N21/33G01N21/31G01N27/447G01N30/02G01N30/72G01N30/88G01N2030/8822G01N2030/8831
Inventor 张黎明杨智柳国艳范铁炯王博史小月王倩倩陈则
Owner 中国人民解放军海军特色医学中心