Method for measuring free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate

A technology of Streptococcus pneumoniae and capsular polysaccharide, applied in the field of vaccine detection, can solve problems such as result interference, and achieve the effects of reducing analysis cost, low analysis cost and simple operation

Inactive Publication Date: 2018-05-04
华兰生物疫苗股份有限公司 +2
View PDF3 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the literature method, the addition of sodium deoxycholate has some interference on the results of the anthrone method.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for measuring free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate
  • Method for measuring free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate
  • Method for measuring free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0056] The detection method of the present invention is carried out repetitive experimental verification, and the sample to be tested is selected pneumococcus 19A type, and concrete experimental procedure is as follows:

[0057] (1) Take Streptococcus pneumoniae capsular polysaccharide type 19A as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide type 19A as sample 2 (take 3 different concentrations of free polysaccharide samples for 3 parallel detections, label be sample a1, sample a2, sample a3, sample b1, sample b2, sample b3, sample c1, sample c2, sample c3);

[0058] (2) Prepare Streptococcus pneumoniae type 19A capsular polysaccharide combined sugar standard solution;

[0059] (3) Prepare standard substance gradient solution, prepare 5%, 10%, 20%, 40%, 60%, 80%, 100% standard substance solution in colorimetric tube, each tube is supplemented with ultrapure water respectively;

[0060] (4) Take 1.5 parts by volume of the c...

experiment example 2

[0077] The detection method of the present invention is subjected to a sample addition recovery rate experiment to detect the conjugate corresponding to the pneumococcal type 19A capsular polysaccharide.

[0078] The specific experimental steps are as follows:

[0079] (1) Take Streptococcus pneumoniae capsular polysaccharide type 19A as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide type 19A as sample 2 (take 3 different concentrations of free polysaccharide samples for 3 parallel detections, label be sample a1, sample a2, sample a3, sample b1, sample b2, sample b3, sample c1, sample c2, sample c3);

[0080] (2) Prepare Streptococcus pneumoniae type 19A capsular polysaccharide combined sugar standard solution;

[0081] (3) Prepare standard substance gradient solution, prepare 5%, 10%, 20%, 40%, 60%, 80%, 100% standard substance solution in colorimetric tube, each tube is supplemented with ultrapure water respectively;

[0...

Embodiment 3

[0097] In this experimental example, it is tested whether sodium deoxycholate interferes with the experimental results in the method of the present invention.

[0098] The experimental method is a detection method provided by the present invention, the sample to be tested is pneumococcus type 2, and the steps are as follows:

[0099] (1) Take Streptococcus pneumoniae capsular polysaccharide as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide as sample 2;

[0100] (2) According to the difference of different glycoforms of Streptococcus pneumoniae capsular polysaccharide conjugates, prepare a Streptococcus pneumoniae capsular polysaccharide combined sugar standard solution;

[0101] (3) Prepare standard substance gradient solution, prepare 10%, 20%, 40%, 60%, 80%, 100% standard substance solution in colorimetric tube, each tube is supplemented with ultrapure water respectively;

[0102] (4) Take 1.5 parts by volume of the capsul...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a method for measuring free polysaccharide in a streptococcus pneumoniae capsular polysaccharide conjugate. The method is characterized by comprising the following steps: respectively adding a sodium deoxycholate solution into streptococcus pneumoniae capsular polysaccharide and the conjugate corresponding to the streptococcus pneumoniae capsular polysaccharide, and adjusting the pH value to 2-6, centrifuging and collecting supernatant; adding a chromogenic reagent into the supernatant of the streptococcus pneumoniae capsular polysaccharide for carrying out a chromogenicreaction; adding the chromogenic reagent into the supernatant of the conjugate corresponding to the streptococcus pneumoniae capsular polysaccharide for carrying out a chromogenic reaction; measuringabsorbance, and calculating the content of the free polysaccharide. The invention develops the method, which is low in analysis cost and simple to operate, for quantitative detection of the free polysaccharide in the streptococcus pneumoniae capsular polysaccharide conjugate.

Description

Technical field: [0001] The invention belongs to the field of vaccine detection, and in particular relates to a method for determining the content of free polysaccharides in capsular polysaccharide conjugates of Streptococcus pneumoniae. Background technique: [0002] Streptococcus pneumoniae is the main pathogen of bacterial lobar pneumonia, meningitis, and bronchitis, and 5% to 10% of normal people carry such bacteria in the upper respiratory tract. People who are susceptible to this type of bacteria are infants and the elderly with poor resistance. About 800,000 to 1,000,000 children under the age of 5 die each year from various diseases caused by Streptococcus pneumoniae. At the same time, bacterial pneumonia induced by pneumococcus is also a major cause of death in the elderly over 60 years old. [0003] Since the capsular polysaccharide, the main virulence factor of Streptococcus pneumoniae, is a T-cell-independent antigen, and the immune system of newborns is not yet...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/78G01N21/33
CPCG01N21/33G01N21/78
Inventor 安文琪李青鹿泽华李辛欣常聪郝一楠
Owner 华兰生物疫苗股份有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap