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Detection method of influenza virus split vaccine monovalent stock solution hemagglutinin

A technology of influenza virus and detection method, which is applied in the biological field, can solve the problems of low economy, low test accuracy, and low detection efficiency, and achieve the effects of accelerating production progress, alleviating high prices, and improving stability

Pending Publication Date: 2022-03-15
LIAONING CHENGDA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the detection of influenza virus hemagglutinin by the conventional one-way immunodiffusion method has the following deficiencies: 1) factors such as the concentration of agarose, the toughness of the gel plate, the staining time, the decolorization time, the shape and the depth of the precipitation ring are used for the detection of the hemagglutination test. The results are affected, resulting in low test accuracy; 2) The test time often takes 2 days, the test cycle is long, and the test efficiency is low; 3) The amount of antigen standard products is large, and there may be a shortage of antigen standard products, which is economical. Low

Method used

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  • Detection method of influenza virus split vaccine monovalent stock solution hemagglutinin
  • Detection method of influenza virus split vaccine monovalent stock solution hemagglutinin
  • Detection method of influenza virus split vaccine monovalent stock solution hemagglutinin

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0071] Preparation example 1 provides a kind of H1N1 type influenza virus antigen reference product, and its preparation steps are:

[0072] S1. Add 30 mL of water to the container, heat to boil, add 10 g of maltose, stir until completely dissolved, add water and phosphate to prepare a 0.1 g / mL solution with a pH of 7.4 to obtain a lyoprotectant;

[0073] S2. Take the monovalent stock solution of the H1N1 influenza virus split vaccine (taken from the company's production workshop), and dilute it with 0.02mol / LPBS buffer solution to diluent A with a hemagglutinin content of 42 μg / mL;

[0074] S3. Take 200 mL of the diluent A obtained in the step S2 and add the lyoprotectant obtained in the step S1 to obtain a reference solution with a hemagglutinin content of 28 μg / mL;

[0075] S4. Divide the reference product solution obtained in step S3 into freeze-dried bottles, and put it into a -40°C low-temperature refrigerator for pre-freezing for 4 hours;

[0076] S5. When the freezing...

preparation example 2

[0104] Preparation Example 2 provides a H3N2 influenza virus antigen reference product. The difference between the preparation steps and Preparation Example 1 is that the monovalent stock solution of H1N1 influenza virus split vaccine is replaced by the monovalent stock solution of H3N2 influenza virus split vaccine. Virus standard antiserum was replaced with H3N2 influenza virus standard antiserum (batch number 19 / 316, purchased from NIBSC), and H1N1 influenza virus standard antigen was replaced with H3N2 influenza virus standard antigen (batch number 20 / 108, purchased from NIBSC) , and the different steps are as follows:

[0105] S2. Take the monovalent stock solution of the H3N2 influenza virus split vaccine (taken from the company's production workshop), and dilute it with 0.02mol / LPBS buffer solution to diluent A with a hemagglutinin content of 36 μg / mL;

[0106] S3. Take 200 mL of the diluent A obtained in the step S2, and add the lyoprotectant obtained in the step S1 to...

preparation example 3

[0110] Preparation Example 3 provides a BV type influenza virus antigen reference product. The difference between the preparation steps and Preparation Example 1 is that the monovalent stock solution of H1N1 influenza virus split vaccine is replaced by the monovalent stock solution of BV type influenza virus split vaccine. Virus standard antiserum was replaced with BV influenza virus standard antiserum (batch number 19 / 218, purchased from NIBSC), H1N1 influenza virus standard antigen was replaced with BV influenza virus standard antigen (batch number 19 / 208, purchased from NIBSC) , and the different steps are as follows:

[0111] S2. Take the monovalent stock solution of the BV-type influenza virus split vaccine (taken from the production workshop of our company), and dilute it to a dilution A with a hemagglutinin content of 42 μg / mL with 0.02mol / L PBS buffer solution;

[0112] S3. Take 200 mL of the diluent A obtained in the step S2 and add the lyoprotectant obtained in the s...

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Abstract

The invention discloses a method for detecting hemagglutinin of an influenza virus split vaccine monovalent stock solution. The method comprises the following steps: S1, respectively adding a splitting agent into the influenza virus split vaccine monovalent stock solution and an influenza virus antigen reference for splitting; s2, respectively detecting the hemagglutination titer of the split influenza virus split vaccine monovalent stock solution and the hemagglutination titer of the split influenza virus antigen reference; s3, calculating a hemagglutination titer standard value of the influenza virus split vaccine monovalent stock solution according to the hemagglutination titer detection value of the influenza virus split vaccine monovalent stock solution; and S4, calculating the hemagglutinin standard value of the influenza virus split vaccine monovalent stock solution according to the hemagglutinin titer standard value of the influenza virus split vaccine monovalent stock solution. According to the detection method provided by the invention, the stability, the accuracy and the sensitivity of a detection result are improved, the detection time is shortened from 2 days to 2 hours or less, and the production progress of influenza vaccines is accelerated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a detection method of biological preparations, in particular to a detection method of hemagglutinin in monovalent stock solution of influenza virus split vaccine. Background technique [0002] Hemagglutinin is a glycoprotein on the surface of influenza virus. Hemagglutinin can induce protective immune responses in a variety of hosts and is currently the only indicator reflecting the effectiveness of influenza vaccines. Therefore, how to quickly and accurately measure the hemagglutinin of influenza vaccine can evaluate the efficacy of influenza vaccine, and also has a positive effect on preventing influenza virus infection. [0003] The principle of the conventional one-way immunodiffusion method for detecting influenza virus hemagglutinin is as follows: the specific binding of antigen and antibody forms a precipitation ring, and the diameter of the precipitation ring is measured after ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/86G01N33/569
CPCG01N33/86G01N33/56983G01N2333/11
Inventor 苏文全闫秀琦任克达于艳孙非非吴琼廖辉周荔葆杨俊伟毛昱高军张庶民
Owner LIAONING CHENGDA BIOTECH
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