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New mycoplasma hyopneumoniae strain and vaccine composite of new mycoplasma hyopneumoniae

A technology of mycoplasma hyopneumoniae and vaccine composition, applied in vaccines, multivalent vaccines, bacteria, etc., can solve the problems of poor clinical prevention and treatment effects, and achieve the effect of not affecting weight gain and reducing the loss of lung lesions

Active Publication Date: 2015-03-25
PU LIKE BIO ENG +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The above literature shows that the clinical prevention and treatment effect of Mycoplasma hyopneumoniae J strain used in the vaccine is not good for the clinical prevention and treatment of Mycoplasma hyopneumoniae

Method used

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  • New mycoplasma hyopneumoniae strain and vaccine composite of new mycoplasma hyopneumoniae
  • New mycoplasma hyopneumoniae strain and vaccine composite of new mycoplasma hyopneumoniae
  • New mycoplasma hyopneumoniae strain and vaccine composite of new mycoplasma hyopneumoniae

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1, isolation and identification of Mycoplasma hyopneumoniae HN0613 strain

[0042] 1. Materials and methods

[0043] 1.1 Materials

[0044] 1.1.1 Source of disease materials In 2006, 16 lungs suspected of being caused by mycoplasma swine pneumonia were collected from various places in Henan Province.

[0045] 1.1.2 Medium Mycoplasma hyopneumoniae Friis medium was purchased from BD Company; pig serum was purchased from GIBCO Company; phenol red indicator was purchased from American AMRESCO Company.

[0046] 1.1.3 Biochemical reagents Chemical reagents were purchased from Sinopharm Chemical Reagent Co., Ltd. Bacterial Genomic DNA Extraction Kit was purchased from TIANGEN Company, and Dieter's Staining Kit was purchased from Chongqing Pangtong Medical Instrument Co., Ltd. Biochemical identification reagents were purchased from Hangzhou Tianhe Microbiological Reagent Co., Ltd.

[0047] 1.1.4 Negative and positive serum Rabbit anti-Mycoplasma hyopneumoniae lip...

Embodiment 2

[0084] The preparation of the combination vaccine antigen of embodiment 2 mycoplasma pneumonia vaccine composition and porcine circovirus type 2, mycoplasma hyopneumoniae

[0085] 1.1 Preparation of bacteria (poison) species for production

[0086] The preparation of porcine circovirus type 2 SH: the virus seed PCV2SH strain was diluted 1:9 with MEM liquid medium (prepared according to the instruction manual with the MEM dry powder medium purchased from Invitrogen Company of the United States), and then diluted by 5% of the volume of the cell culture medium. % inoculated on PK15 (ATCC, preservation number is CCL-33) monolayer cells, adsorbed at 37°C for 30 minutes, added cell maintenance solution (add 4% calf serum and 2mmol / L D-glucosamine hydrochloride in MEM liquid medium) ), cultured at 37°C for 4 days, frozen and thawed 2 to 3 times, and the virus was harvested, and the virus titer was 10 6.5 TCID 50 / ml.

[0087] Preparation of Mycoplasma hyopneumoniae strains: After ...

Embodiment 3

[0124] Embodiment 3, the preparation of composition vaccine:

[0125] In order to compare the immune effect of the vaccine, Montanide is selected when preparing the composition vaccine TM Gel adjuvant (produced by SEPPIC, France) was used to prepare porcine circovirus disease (whole virus antigen or ORF2 protein), dual inactivated vaccine against mycoplasma pneumoniae and single vaccine against mycoplasma pneumoniae. The specific configuration is as follows:

[0126] Preparation method: formula is as table 5. Get the PCV2 antigen and Mycoplasma hyopneumoniae antigen prepared in Example 1. The concentrated antigen solution was prepared into a mixed antigen solution or directly prepared into an antigen solution according to the final antigen content in the joint vaccine and the single vaccine, and then the antigen solution was mixed with Gel01 adjuvant (produced by SEPPIC, France) at a ratio of 90:10 (V / V) mixing, replenish volume with PBS solution with pH 7.2 and stir at 30...

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Abstract

The invention provides a mycoplasma hyopneumoniae strain HN0613 which has better immunogenicity proved through isolation and identification, also provides a mycoplasma hyopneumoniae antigen prepared by using the mycoplasma hyopneumoniae strain HN0613 and a mycoplasma hyopneumoniae vaccine containing the mycoplasma hyopneumoniae antigen. The invention further provides a porcine circovirus type 2 / mycoplasma hyopneumoniae bicombinant vaccine, containing a PCV2 antigen (inactivated PCV2 antigen or PCV2ORF2 protein), an inactivated mycoplasma hyopneumoniae and vaccine adjuvants. Through injection of the bicombinant vaccine, the aim of preventing PCV and mycoplasma hyopneumoniae can be achieved and the effect of prevention and protection from mycoplasma hyopneumoniae infection can be achieved further.

Description

[0001] This case is a divisional application of invention patent application 201210224389.9, and the filing date of the original application is June 29, 2012. technical field [0002] The invention relates to a novel mycoplasma hyopneumoniae and a vaccine composition, belonging to the field of veterinary vaccines. Background technique [0003] Mycoplasma hyopneumoniae is the causative agent of mycoplasma pneumonia in swine. Mycoplasma swine pneumonia is a chronic consumptive respiratory disease of pigs with a high clinical infection rate. The disease causes a persistent cough that lasts for several weeks, loss of luster in the coat, growth retardation, and a stocky appearance. Once the pig herd is infected with Mycoplasma hyopneumoniae, it is difficult to remove it, which not only seriously affects the growth and development of the pig herd, but also increases the dosage of drugs, and is prone to secondary infection of PRRSV (porcine blue ear disease virus), PCV (porcine ci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61K39/12A61K2039/54A61K2039/545A61K2039/70C12N2750/10034C12N1/205C12R2001/35
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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