Method for identifying vipera russelli venom and application of method

An identification method for Viper rotundum, which is applied to the identification of types of snake venom by using snake venom protein analysis technology. The application of this identification method, in the field of identification of Viper rotundus venom, can solve the complicated and cumbersome operation process and low detection sensitivity , low correct detection rate and other issues, to achieve the effect of simple operation process, high detection sensitivity and high correct detection rate

Active Publication Date: 2019-11-08
SHANGHAI SERUM BIOTECH
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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 is a serious cross-reaction in the identification of snake venom protein snake venom by ELISA, but it 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 related research reports

Method used

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  • Method for identifying vipera russelli venom and application of method
  • Method for identifying vipera russelli venom and application of method
  • Method for identifying vipera russelli venom and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1 A kind of identification method of the viper venom

[0044] Proceed as follows:

[0045] (1) Select a batch of viperin toxin, dissolve it with solution A and make it 40mg / mL;

[0046] (2) Carry out SDS-PAGE electrophoresis, the protein separation gel concentration is 15%, and the formula is as follows in Table 2:

[0047] Table 2 SDS-PAGE protein separation gel and stacking gel formula

[0048]

[0049] (3) Coomassie brilliant blue staining; see SDS-PAGE electrophoresis figure 1 , 13 protein bands could be distinguished (see figure 1 Middle 1-13);

[0050] (4) Cut out the corresponding protein band, add a small amount of solution H to cover the protein band;

[0051] (5) LC-MS / MS mass spectrometry identification: the sample is separated from the mobile phase in the liquid mass spectrometer, and after being ionized, the mass analyzer of the mass spectrometer separates the ion fragments according to the mass number, and the mass spectrogram is obtaine...

Embodiment 2

[0064] Example 2 Application of an identification method of viper venom in the preparation of anti-viper venom for the treatment of viper venom poisoning

[0065] Proceed as follows:

[0066] (1) Viper venom purchased from Yiwu Chunfeng Snake Industry Co., Ltd., dissolved in solution A, and subjected to SDS-PAGE electrophoresis;

[0067] (2) SDS-PAGE electrophoresis results (see figure 2 ) and the existing electrophoretic pattern ( figure 1 ) comparison, it was confirmed that the protein was 13 major bands, which were the corresponding viper venom protein;

[0068] (3) Immunize the horses after detoxification of the identified viper venom formaldehyde;

[0069] (4) Horses immune to viper venom, after blood collection by apheresis, are transferred to the production department for the production of anti-viper venom, and anti-viper venom stock solution and products are obtained;

[0070] The production process of the above anti-viper venom is to identify the type of snake ve...

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Abstract

The invention discloses a method for identifying a vipera russelli venom. The method comprises the following steps of (1) dissolving vipera russelli venom toxin lyophilized powder to prepare a proteinsolution; (2) performing SDS-PAGE electrophoresis analysis; (3) conducting LC-MS/MS identification. In addition, the invention discloses application of the method for identifying the vipera russellivenom in preparing anti-viper-venom serum for treating viper venom poisoning. By means of the method, laboratory identification can be conducted on the viper venom to more accurately control the quality of the viper venom; it can be ensured that the special-effect anti-viper-venom serum for treating viper venom poisoning is safe, effective and controllable in quality, and the method has very important significance in the aspects of viper species identification, viper wound diagnosis and treatment, application of other viper venom and the like. The method is high in detection sensitivity and correct detection rate and simple in operation process and has no cross reaction.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals, and relates to a method for identifying the venom of Viper viper, 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, in the anti-venom The production of serum, the identification of venomous snake species, the diagnosis and treatment of snakebite and the application of other snake venoms are of great significance. 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] Snake venom is mostly light yellow or milky white translucent viscous liquid, with more than 100 components. Eac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/6848G01N2550/00G01N2570/00
Inventor 陈则杨智王德慧罗敏李鑫卞聪华范铁炯
Owner SHANGHAI SERUM BIOTECH
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