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A method for detecting virus antigen content of Japanese encephalitis vaccine

A technology for the content of Japanese encephalitis virus and antigen, which is applied to measuring devices, biological tests, material inspection products, etc., can solve the problems of not being able to find reports from foreign companies, and the method of detecting the content of Japanese encephalitis virus antigen is not disclosed, and achieve The effect of shortening the experiment time and saving experiment funds

Active Publication Date: 2015-11-18
LIAONING CHENGDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, there is no domestic method for detecting JE virus antigen content, and no relevant reports from foreign companies can be found.

Method used

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  • A method for detecting virus antigen content of Japanese encephalitis vaccine
  • A method for detecting virus antigen content of Japanese encephalitis vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] This method for detecting the content of Japanese encephalitis virus antigen has the following steps:

[0054] Preparation of a Rabbit Anti-Japanese Encephalitis Polyclonal Antibody

[0055] 1. Inject antigen preparation (emulsification) and shake CFA (complete Freund's adjuvant) or IFA (incomplete Freund's adjuvant). Mix vigorously with the same volume of Japanese encephalitis virus purified stock solution in a glass test tube at 4°C. until a cream is formed. Squeeze out a drop onto the surface of cold water, if it forms a drop on the surface of the water, it is ready for use. If the droplets disperse, continue mixing. The reconstituted emulsion is transferred to a 2.5ml syringe ready for injection.

[0056] 2 Immunization program

[0057] Before injection, blood was taken from the ear of the rabbit as a blank control for the experimental serum

[0058] primary immunization

[0059] The injection site is the back of the shoulders and the vicinity of the two hips...

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Abstract

The invention relates to a method for detecting the content of a virus antigen in an encephalitis B vaccine. The method is characterized in that the method comprises the following steps: coating a 96-aperture enzyme label plate with an encephalitis B virus polyclonal antibody, respectively adding encephalitis B positive and negative reference substances and a gradient-diluted sample for detection after the coating, incubating at 37DEG C, cleaning the enzyme label plate, adding an encephalitis B virus E protein monoclonal antibody, incubating at 37DEG C, cleaning the enzyme label plate, adding a horseradish peroxidase conjugate, incubating at 37DEG C, cleaning the enzyme label plate, adding a substrate coloring solution for coloring, reading the OD value by using ELIASA after the coloring terminates, and calculating the P / N value to determine the antigen content. The method realizes the rapid detection of the antigen content of encephalitis B and the discrimination of the authenticity of the encephalitis B vaccine, and provides favorable data support for the production of the encephalitis B vaccine.

Description

technical field [0001] The present invention relates to a detection method of biological preparations, in particular to a method for detecting the content of Japanese encephalitis virus antigen by using the principle of double-antibody sandwich ELISA to quickly detect the content of Japanese encephalitis virus antigen and to identify the authenticity of the Japanese encephalitis vaccine. Belongs to the field of biotechnology. It is especially suitable for use in the quality control link of Japanese encephalitis vaccine production enterprises. Background technique [0002] Japanese encephalitis is an acute infectious disease caused by Japanese encephalitis virus and transmitted by mosquitoes. JE has a high mortality rate and disability rate, and is one of the main infectious diseases that threaten the health of the population, especially children. my country is a high-endemic area of ​​JE. In the 1960s and early 1970s, a large-scale epidemic occurred across the country. Aft...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68G01N33/569G01N33/543
CPCG01N33/54373G01N33/56983G01N2333/005
Inventor 高旭哲战辛刘畅张玥
Owner LIAONING CHENGDA BIOTECH