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A rapid qualitative and quantitative detection method for oil adjuvant vaccines

A quantitative detection method and quantitative detection technology, which are applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of no universal and accurate method for quantitative detection, and can not respond well to the effective antigen content. Simple, powerful demulsification effect

Active Publication Date: 2019-04-23
SHANGHAI SHEN LIAN BIOMEDICAL CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Can not reflect the effective antigen content in synthetic peptide vaccine against foot-and-mouth disease
[0006] At present, there is no universal and accurate method for the quantitative detection of antigen content in synthetic peptide vaccines for FMD

Method used

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  • A rapid qualitative and quantitative detection method for oil adjuvant vaccines
  • A rapid qualitative and quantitative detection method for oil adjuvant vaccines
  • A rapid qualitative and quantitative detection method for oil adjuvant vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1) select the commercially available porcine foot-and-mouth disease vaccine (vaccine concentration is 50 μ g / mL) and process according to the following method of demulsification:

[0041] The vaccine to be tested is mixed with a demulsifier, the organic solvent in the demulsifier is acetonitrile, the acid is trifluoroacetic acid, and the volume ratio of acetonitrile to trifluoroacetic acid is 100:0.05. Take 8ml of the vaccine to be tested and put it in a centrifuge tube. Mix the vaccine and demulsifier at a volume ratio of 8:2, shake and mix well, and centrifuge at 3000G for 15 minutes at 4°C. After centrifugation, carefully extract the lower aqueous phase with a 10ml syringe. After liquid chromatography analysis, it was compared with the theoretical antigen concentration standard in the vaccine, wherein, the integral information of the peak position of the HPLC detection spectrum sample of the theoretical antigen concentration in the vaccine is shown in Table 1, and the...

Embodiment 2

[0060] This embodiment provides a rapid qualitative and quantitative detection method for oil adjuvant vaccines, specifically using the following steps:

[0061] 1) select the commercially available porcine foot-and-mouth disease vaccine (vaccine concentration is 50 μ g / mL) and process according to the following method of demulsification:

[0062] The vaccine to be tested is mixed with a demulsifier, the organic solvent in the demulsifier is acetonitrile, the acid is trifluoroacetic acid, and the volume ratio of acetonitrile to trifluoroacetic acid is 100:0.05. Take 5ml of the vaccine to be tested and mix it with the demulsifier at a volume ratio of 5:5, shake and mix well, and centrifuge at 3000G for 15 minutes at 4°C to obtain the aqueous phase antigen sample.

[0063] The foot-and-mouth disease vaccine was demulsified by the method of this example, and the demulsification efficiency was 90.3%, and the dilution effect was 33.33%.

[0064] 2) Competitive ELISA detection

[...

Embodiment 3

[0078] This embodiment provides a rapid qualitative and quantitative detection method for oil adjuvant vaccines, specifically using the following steps:

[0079] 1) select the commercially available porcine foot-and-mouth disease vaccine (vaccine concentration is 50 μ g / mL) and process according to the following method of demulsification:

[0080] The vaccine to be tested is mixed with the demulsifier, the organic solvent in the demulsifier is ethanol, the acid is hydrochloric acid, and the volume ratio of ethanol to hydrochloric acid is 100:0.33, 7ml of the vaccine to be tested is mixed with the demulsifier at a volume ratio of 7:3, Shake and mix well, and centrifuge at 3000G for 15 minutes at 4°C to obtain an aqueous phase antigen sample.

[0081] The foot-and-mouth disease vaccine was demulsified by the method of this example, the demulsification efficiency was 93.5%, and the dilution effect was 53.85%.

[0082] 2) Competitive ELISA detection

[0083] The competition ELIS...

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Abstract

The present invention provides a rapid qualitative and quantitative detection method for an oil adjuvant vaccine. The rapid qualitative and quantitative detection method comprises: demulsifying a foot-and-mouth disease synthetic peptide vaccine, and carrying out qualitative and quantitative detection on the demulsified antigen sample by using a competitive ELISA method, wherein the demulsifying method comprises: mixing a foot-and-mouth disease synthetic vaccine and a demulsifying agent, and layering to obtain a water phase antigen sample, the demulsifying agent comprises the mixture of an organic solvent and an acid, the organic solvent is one selected from acetonitrile, n-butanol, benzyl alcohol, butyl diethylene glycol and ethanol, and the acid is one selected from hydrochloric acid, acetic acid, formic acid and trifluoroacetic acid. According to the present invention, the demulsifying ability of the demulsifying method is strong, wherein the sample can be determined without the purification treatment so as to shorten the detection time; and the antigen concentration is subjected to qualitative and quantitative detection by using the competitive ELISA method, such that the slope of the obtained standard curve can be greater than the slope of the standard curve obtained through the conventional indirect competition method so as to substantially improve the detection sensitivity.

Description

technical field [0001] The invention relates to the technical field of oil adjuvant vaccine detection, in particular to a rapid qualitative and quantitative detection method for oil adjuvant vaccines. Background technique [0002] The existing vaccine quality standards stipulate that the efficacy test must use this animal for testing. Since the country implements a 100% enhanced immunization policy, it is difficult to select susceptible animals for testing, and animal challenge has high requirements for experimental facilities (BSL3 level laboratory), time-consuming (more than one month), and large capital expenditure. If the serum neutralization test is used to select susceptible animals, it is technically difficult to exclude non-susceptible animals with cellular immunity, and the problem of irregular test data often occurs in practice, which affects the accuracy of the test. Therefore, the quality inspection of foot-and-mouth disease vaccines, especially vaccines for cat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543
CPCG01N33/543
Inventor 刘自立马贵军石海芳俞爱敏姬明放
Owner SHANGHAI SHEN LIAN BIOMEDICAL CORP
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