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Novel mycoplasma hyopneumoniae bacterial strain and vaccine composition thereof

A technology of mycoplasma hyopneumoniae and vaccine composition, which is applied in the direction of bacteria, microorganisms, antibacterial drugs, etc., can solve the problems of poor clinical prevention and treatment effects, and achieve the effects of improving immune efficacy, good protection effect, and small loss of lung lesions

Active Publication Date: 2014-12-17
PU LIKE BIO ENG
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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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  • Novel mycoplasma hyopneumoniae bacterial strain and vaccine composition thereof
  • Novel mycoplasma hyopneumoniae bacterial strain and vaccine composition thereof
  • Novel mycoplasma hyopneumoniae bacterial strain and vaccine composition thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1, isolation and identification of Mycoplasma hyopneumoniae HN0613 strain

[0041] 1. Materials and methods

[0042] 1.1 Materials

[0043] 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.

[0044] 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.

[0045] 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.

[0046] 1.1.4 Negative and positive serum Rabbit anti-Mycoplasma hyopneumoniae lip...

Embodiment 2

[0083] The preparation of the combination vaccine antigen of embodiment 2 mycoplasma pneumonia vaccine composition and porcine circovirus type 2, mycoplasma hyopneumoniae

[0084] 1.1 Preparation of bacteria (virus) species for production

[0085] Preparation of porcine circovirus type 2 SH: the virus seed PCV2SH strain was diluted 1:9 with MEM liquid medium (prepared with MEM dry powder medium purchased from Invitrogen Company of the United States according to the instructions), and then divided into 5% of the volume of cell culture medium. % inoculated in PK15 (ATCC, preservation number CCL-33) monolayer cells, adsorbed at 37°C for 30 minutes, added cell maintenance solution (4% calf serum and 2mmol / L D-glucosamine hydrochloride were added to MEM liquid medium) ), cultivated at 37°C for 4 days, freeze-thawed 2-3 times, harvested the virus, and the virus titer was 10 6.5 TCID 50 / ml.

[0086] Preparation of Mycoplasma hyopneumoniae strains: After unsealing the freeze-dried...

Embodiment 3

[0123] Embodiment 3, the preparation of composition vaccine:

[0124] In order to compare the immune effect of the vaccine, Montanide is selected when preparing the composition vaccine TM Gel adjuvant (produced by Sepic 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:

[0125] 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 of the combined vaccine and the single vaccine, and then the antigen solution was mixed with Gel 01 adjuvant (produced by SEPPIC, France) at 90: 10 (V / V) mix, supplement the volume with PBS solution with pH 7.2 and stir at ...

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Abstract

The invention provides a mycoplasma hyopneumoniae strain HN0613 which is isolated and identified to have better immunogenicity, and further provides a mycoplasma hyopneumoniae antigen prepared from the mycoplasma hyopneumoniae strain HN0613 and mycoplasma pneumonia pneumovax containing the mycoplasma hyopneumoniae antigen. The invention further provides a bivalent combined vaccine of porcine circovirus II and mycoplasma hyopneumoniae. The duplex combination vaccine comprises PCV (Porcine Circovirus) antigen II (inactivated porcine circovirus antigen II or PCV20 RF2 protein), inactivated mycoplasma hyopneumoniae and vaccine adjuvant. The combination vaccine can achieve the purpose of preventing the porcine circovirus disease and the mycoplasma pneumonia swine by injection once, and has a protective effect of preventing mycoplasma hyopneumoniae infection.

Description

technical field [0001] The invention relates to a novel mycoplasma hyopneumoniae and a vaccine composition, belonging to the field of veterinary vaccines. Background technique [0002] 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, 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 circovirus), etc., resulting in Multiple vaccinations have failed. Mycoplasma hyopneumoniae is one of the important pathogens of porcine respiratory syndrome, and t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61K39/116A61K39/295A61P31/04A61P31/14A61P31/16A61P31/20A61P31/22C12R1/35A61K39/10A61K39/102A61K39/12A61K39/145A61K39/245
Inventor 张许科孙进忠白朝勇
Owner PU LIKE BIO ENG
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