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Method for determining free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate

A technology of Streptococcus pneumoniae and capsular polysaccharide, which is applied in the field of vaccine detection, can solve the problems of result interference and other problems, and achieve the effect of reducing analysis cost, low analysis cost and good detection effect

Inactive Publication Date: 2018-08-10
华兰生物疫苗股份有限公司 +2
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  • 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

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  • Method for determining free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate
  • Method for determining free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate
  • Method for determining free polysaccharide in streptococcus pneumoniae capsular polysaccharide conjugate

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0044] The detection method of the present invention is carried out repetitive experimental verification, and the sample to be tested is selected pneumococcus type 1, and concrete experimental procedure is as follows:

[0045] (1) Take Streptococcus pneumoniae capsular polysaccharide type 1 as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide type 1 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);

[0046] (2) According to the difference of different glycoforms of Streptococcus pneumoniae capsular polysaccharide type 1 conjugates, prepare a Streptococcus pneumoniae capsular polysaccharide combined sugar standard solution;

[0047] (3) Prepare standard substance gradient solution, prepare 5%, 10%, 20%, 40%, 60%, 80%, 100% standard substance solution in colorimetric tube...

experiment example 2

[0066] The detection method of the present invention is subjected to a sample addition recovery rate experiment, and the conjugate corresponding to the pneumococcal type 1 capsular polysaccharide is detected.

[0067] The experimental method is a detection method provided by the present invention, the sample to be tested is pneumococcus type 1, and the steps are as follows:

[0068] (1) Take Streptococcus pneumoniae capsular polysaccharide type 1 as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide type 1 as sample 2 (take the same batch of conjugate samples with known free polysaccharide concentration and add a variety of different content of the combined sugar solution sample is detected, marked as sample a, sample b, sample c);

[0069] (2) Prepare Streptococcus pneumoniae type 1 capsular polysaccharide combined sugar standard solution;

[0070] (3) Prepare standard substance gradient solution, prepare 5%, 10%, 20%, 40%, 60%...

Embodiment 3

[0086] In this experimental example, it is tested whether sodium deoxycholate interferes with the experimental results in the method of the present invention.

[0087] The experimental method is a detection method provided by the present invention, the sample to be tested is pneumococcus type 1, and the steps are as follows:

[0088] (1) Take Streptococcus pneumoniae capsular polysaccharide as sample 1, and take the conjugate corresponding to Streptococcus pneumoniae capsular polysaccharide as sample 2;

[0089] (2) Prepare Streptococcus pneumoniae type 1 capsular polysaccharide combined sugar standard solution;

[0090] (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;

[0091] (4) Take 1.5 parts by volume of the capsular polysaccharide of Streptococcus pneumoniae and place it in a centrifuge tube of 2.5 parts by volume, and take ...

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Abstract

The invention provides a method for determining the content of free polysaccharide in a streptococcus pneumoniae capsular polysaccharide conjugate. The scheme comprises: adding a sodium deoxycholate solution to streptococcus pneumoniae capsular polysaccharide and a conjugate corresponding to the streptococcus pneumoniae capsular polysaccharide, adjusting the pH to 2-6, and performing centrifugation to collect the supernatant; adding a color reagent to the streptococcus pneumoniae capsular polysaccharide supernatant to carry out a color development reaction; adding a color reagent to the supernatant of the conjugate corresponding to the streptococcus pneumoniae capsular polysaccharide to carry out a color development reaction; and determining the absorbance and calculating the content of free polysaccharide. The method for quantitative determination of free polysaccharide in a streptococcus pneumoniae capsular polysaccharide conjugate is low in analytical cost and simple to operate.

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

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Application Information

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IPC IPC(8): G01N21/33
CPCG01N21/33
Inventor 安文琪李青许来芳杨艳李会云王海旗
Owner 华兰生物疫苗股份有限公司
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