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Method for determining dissociation amylase content of pneumonia streptococcus capsular polysaccharide combo

A technology of Streptococcus pneumoniae and capsular polysaccharide, applied in the biological field, can solve the problem of inability to accurately measure the content of free polysaccharides in the capsular polysaccharide conjugate of Streptococcus pneumoniae

Active Publication Date: 2010-05-12
云南沃森生物技术股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The method of the present invention overcomes the deficiency that the existing methods cannot accurately measure the free polysaccharide content of the Streptococcus pneumoniae capsular polysaccharide conjugate, and the method can accurately and simply measure the content of the free polysaccharide in the Streptococcus pneumoniae capsular polysaccharide conjugate

Method used

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  • Method for determining dissociation amylase content of pneumonia streptococcus capsular polysaccharide combo
  • Method for determining dissociation amylase content of pneumonia streptococcus capsular polysaccharide combo
  • Method for determining dissociation amylase content of pneumonia streptococcus capsular polysaccharide combo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Take Streptococcus pneumoniae type 1 capsular polysaccharide conjugate as sample 1; take the derivative solution corresponding to the test Streptococcus pneumoniae type 1 capsular polysaccharide conjugate as sample 3.

[0021] Take another 4.5 ml of Streptococcus pneumoniae type 1 capsular polysaccharide conjugate and place it in a 50 ml centrifuge tube; take 4.5 ml of the derivative solution corresponding to the Streptococcus pneumoniae type 1 capsular polysaccharide conjugate and place it in another 50 ml centrifuge tube. Add 1.125ml of 5mol / L sodium chloride solution to each centrifuge tube, shake and mix, then add 22.5ml of absolute ethanol, shake and mix, set at -20℃ for 70 hours; centrifuge at 5℃, 4500rpm for 60 minutes, discard the supernatant liquid.

[0022] Add 0.75ml water for injection (that is, 0% ethanol) to each centrifuge tube, shake and mix, and let stand at room temperature for 1 hour; then add 2.25ml of water for injection, shake and mix, and stand at room...

Embodiment 2

[0030] Take Streptococcus pneumoniae type 2 capsular polysaccharide conjugate as sample 1, and take the derivative solution corresponding to the test Streptococcus pneumoniae type 2 capsular polysaccharide conjugate as sample 3.

[0031] Take another 4.5ml of Streptococcus pneumoniae type 2 capsular polysaccharide conjugate and place it in a 50ml centrifuge tube; take 4.5ml of the derivative solution corresponding to the Streptococcus pneumoniae type 2 capsular polysaccharide conjugate and place it in another 50ml centrifuge tube. Add 1.125ml of 5mol / L sodium chloride solution to each centrifuge tube, shake and mix, then add 22.5ml of absolute ethanol, shake and mix, set at -20°C for 90 hours; centrifuge at 5°C, 6500rpm for 90 minutes, discard the supernatant liquid.

[0032] Add 0.75ml 60% ethanol solution to each centrifuge tube, shake and mix, and let stand at room temperature for 1 hour; then add 2.25ml 60% ethanol solution, shake and mix, and stand at room temperature for 2 ho...

Embodiment 3

[0040] Take Streptococcus pneumoniae type 3 capsular polysaccharide conjugate as sample 1, and take the derivative solution corresponding to the test Streptococcus pneumoniae type 3 capsular polysaccharide conjugate as sample 3.

[0041] Take another 4.5ml of Streptococcus pneumoniae type 3 capsular polysaccharide conjugate and place it in a 50ml centrifuge tube; take 4.5ml of the derivative solution corresponding to Streptococcus pneumoniae type 3 capsular polysaccharide conjugate and place it in another 50ml centrifuge tube. Add 1.125ml of 5mol / L sodium chloride solution to each centrifuge tube, shake and mix, then add 22.5ml of absolute ethanol, shake and mix, set at -20℃ for 90 hours; centrifuge at 5℃, 5500rpm for 70 minutes, discard the supernatant liquid.

[0042] Add 0.75ml 40% ethanol solution to each centrifuge tube, shake and mix, and stand at room temperature for 1 hour; then add 2.25ml 40% ethanol solution, shake and mix, and stand at room temperature for 2 hours; centr...

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Abstract

This invention relates to one test method for free polysaccharide content of pneumococcal peritonitis compound in biological technique field, which comprises the following steps: a, fetching same content of the compound and adding ethanol without water and common salt for 70 to 90 hours under -20 degrees then removing upper liquid by anticentripetal and using proper agent to clear deposition to collect upper liquid through filter salt and ethanol; b, using anthranone to test sample one and two to compute free polysaccharide content.

Description

Technical field: [0001] The present invention belongs to the field of biotechnology, and more specifically, relates to a method for determining the content of free polysaccharides in a Streptococcus pneumoniae capsular polysaccharide conjugate. Background technique: [0002] Streptococcus pneumoniae is an important pathogen causing bacterial pneumonia, meningitis, pleurisy, endocarditis, and otitis media. It is the most common cause of bacterial pneumonia in children in developing countries. 1 to 2 million people die from pneumococcal infections worldwide every year. The incidence of pneumonia in China has always been high. In the past 50 years, the mortality rate of bacteremia among patients with pneumococcal infection has been hovering between 25-29%. In addition, pneumonia is also one of the main causes of death for people over 60. Since the drug-resistant strains of Streptococcus pneumoniae are as high as 96%, it has become a worldwide public problem. [0003] In 1983, the U...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N31/00G01N1/34G01N33/00C12Q1/00G06F19/00
Inventor 黄镇吴凯
Owner 云南沃森生物技术股份有限公司
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