A kind of preparation method of Streptococcus agalactiae type-specific antiserum

A Streptococcus lactis and antiserum technology, applied in the preparation methods of peptides, chemical instruments and methods, from serum immunoglobulins, etc., can solve the problems of low antibody titers, multiple injections of viable bacteria, lack of methods, etc. The effect of large serum volume, relatively low cost, and outstanding large-scale application prospects

Active Publication Date: 2020-11-10
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The present invention aims at the technical defects of the prior art, and provides a preparation method for adding a new type of adjuvant to prepare Streptococcus agalactiae type-specific antiserum, so as to solve the technical problem of lack of relevant methods in the prior art
[0007] Another technical problem to be solved by the present invention is that the preparation of Streptococcus agalactiae type-specific antiserum has a low antibody titer and requires multiple injections of live bacteria

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 (preparation of streptococcus agalactiae specific antiserum)

[0035] 1 Antigen preparation

[0036] 1.1 Preparation of inactivated vaccine

[0037] 1.1.1 Preparation of bacteria solution After culturing the freeze-dried strains of the standard strain of Streptococcus agalactiae in nutrient broth for 18 hours, inoculate a blood agar plate containing 7% sterile sheep defibrinated blood for streaking for purification and culture, then pick out the typical A single colony was inoculated in THB broth medium supplemented with 2% newborn bovine serum and 8g / L glucose and subcultured 5 times, and the time of each culture was 6 hours.

[0038] 1.1.2 Preparation of inactivated vaccines The bacteria solution prepared in section 1.1.1 was concentrated by centrifugation at a speed of 4000rpm, centrifugation time was 30min, and the concentration factor was 5 times. Add formaldehyde solution to the concentrated bacteria solution of Streptococcus agalactiae at a ratio o...

Embodiment 2

[0054] Embodiment 2 (preparation of streptococcus agalactiae specific antiserum)

[0055] 1 Antigen preparation

[0056] 1.1 Preparation of inactivated vaccine

[0057] 1.1.1 Preparation of bacteria solution After culturing the freeze-dried strains of Streptococcus agalactiae standard strains in nutrient broth for 12 hours, inoculate a blood agar plate containing 7% sterile sheep defibrinated blood for streaking for purification and culture, and then pick out typical strains. A single colony was inoculated in THB broth medium supplemented with 3% neonatal bovine serum and 4g / L glucose and subcultured 5 times, and the time of each culture was 5 hours.

[0058] 1.1.2 Preparation of inactivated vaccine Preparation 1.1.1 The prepared bacterial solution was concentrated by centrifugation, wherein the centrifugal speed was 6000rpm, the centrifugation time was 20min, and the concentration factor was 7 times. Add formaldehyde solution to the concentrated Streptococcus agalactiae bac...

Embodiment 3

[0074] Embodiment 3 (preparation of streptococcus agalactiae specific antiserum)

[0075] 1 Antigen preparation

[0076] 1.1 Preparation of inactivated vaccine

[0077] 1.1.1 Preparation of bacteria solution After culturing the freeze-dried strains of Streptococcus agalactiae standard strains in nutrient broth for 12 hours, inoculate a blood agar plate containing 7% sterile sheep defibrinated blood for streaking for purification and culture, and then pick out typical strains. The single bacterium colony was inoculated in THB broth medium supplemented with 5% newborn bovine serum and 10g / L glucose and subcultured 5 times, and the time of each culture was 8 hours.

[0078] 1.1.2 Preparation of inactivated vaccine Preparation 1.1.1 The prepared bacterial liquid was concentrated by centrifugation, wherein the centrifugal speed was 8000rpm, the centrifugation time was 15min, and the concentration ratio was 10 times. Add formaldehyde solution to the concentrated bacteria solution ...

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Abstract

The invention provides a preparation method of streptococcus agalactiae type specific antiserum. The method includes: preparing antigens, screening domestic rabbits, performing immunization and harvesting and identifying serum. The preparation method has the advantages that a streptococcus agalactiae inactivated vaccine is used to perform basic immunization on the domestic rabbits, and using the inactivated vaccine and inactivated antigens to perform booster immunization to prepare the streptococcus agalactiae type specific antiserum. The preparation method has the advantages that the prepared streptococcus agalactiae type specific antiserum is high in antibody value as compared with adjuvant-free streptococcus agalactiae antigens; the streptococcus agalactiae type specific antiserum is cheap and high in safety as compared with immunization using a Freund's adjuvant; the streptococcus agalactiae type specific antiserum is simple and high in safety as compared with booster immunization using live bacterium challenge, the domestic rabbits are used to prepare positive serum, and the streptococcus agalactiae type specific antiserum is low in cost and has an outstanding large-scale application prospect.

Description

technical field [0001] The invention relates to the technical field of biological products, in particular to a preparation method of Streptococcus agalactiae type-specific antiserum. Background technique [0002] Streptococcus agalactiae (S. agalactiae) is one of the main pathogens causing mastitis in cows. It is highly contagious and often exists in the skin, teats and udders of cows. The mammary glands are carried and spread mechanically, attaching to the mammary duct wall and destroying it, causing chronic mastitis, the incubation period can reach several weeks or months, and it is not easy to find out. At the same time, Streptococcus agalactiae is also an important cause of puerperal sepsis in pregnant women and neonatal meningitis. It parasitizes in the maternal reproductive tract and can cause infection in babies. It can also cause postpartum infection, bacteremia, endocarditis, skin and soft tissue infection, and osteomyelitis. [0003] Streptococcus agalactiae is ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/12C07K16/06C07K1/34
CPCC07K16/06C07K16/1275
Inventor 丁旭娜吕茂杰杨保收付旭彬梁武
Owner TIANJIN RINGPU BIO TECH
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