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Mycoplasma hyopneumoniae, vaccine composition and application thereof

A technology of Mycoplasma hyopneumoniae and a vaccine composition, applied in the biological field, can solve the problems of increased vaccine production cost, slow growth, extremely high requirements, etc., and achieves the effects of good immunogenicity, high culture titer, and reduced usage

Active Publication Date: 2021-11-09
PULIKE BIOLOGICAL ENG INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mycoplasma hyopneumoniae has extremely high requirements on the medium and grows slowly, which greatly increases the production cost of the vaccine

Method used

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  • Mycoplasma hyopneumoniae, vaccine composition and application thereof
  • Mycoplasma hyopneumoniae, vaccine composition and application thereof
  • Mycoplasma hyopneumoniae, vaccine composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0015] As an embodiment of the present invention, the culture of the Mycoplasma hyopneumoniae GZ strain of the present invention is a culture of passage 1 to 42.

[0016] The term "vaccine composition" used in the present invention refers to a pharmaceutical composition containing the immunogenicity of Mycoplasma hyopneumoniae, which can induce, stimulate or enhance the immune response of pigs against Mycoplasma pneumoniae. The vaccine composition comprises immune dose of attenuated live vaccine, inactivated vaccine, subunit vaccine or synthetic peptide vaccine of Mycoplasma hyopneumoniae strain.

[0017] As used herein, the term "inactivated vaccine", also called inactivated vaccine, refers to a suspension of inactivated virus used as an antigen to generate immunity. Examples of inactivated vaccines include whole virus vaccines and split vaccines. Inactivated vaccines can be readily produced using known methods. For example, whole virus inactivated vaccines can be obtained ...

Embodiment 1

[0050] Example 1 Isolation and Identification of Mycoplasma Hyopneumoniae GZ Strain

[0051] 1. Source of disease material

[0052] The disease materials came from 20 lungs suspected to be caused by mycoplasma swine pneumonia collected from a pig farm in Henan Province in 2016.

[0053] 2. Multiplex PCR identification of disease materials

[0054] 2.1 Primer design and synthesis

[0055] Multiplex PCR (Mycoplasma hyopneumoniae, Mycoplasma suis and Mycoplasma hyorhina) primer reference (T.stakenbory et al. A multiplex PCR to Identify Porcine Mycoplasma Present in Broth Cultures) was synthesized:

[0056] mhp-f: 5'-TTCAAAGGAGCCTTCAAGCTTC-3';

[0057] mhr-f: 5'-GGGAAGAAAAAAATTAGGTAGGG-3';

[0058] mfl-f: 5'-CGGGATGTAGCAATACATTCAG-3';

[0059] m-r: 5'-AGAGGCATGATGATTTGACGTC-3';

[0060] It is estimated that the lengths of the amplified fragments of Mycoplasma hyopneumoniae, Mycoplasma hyorhinosus and Mycoplasma suis are 1000bp, 1129bp and 754bp respectively.

[0061] 2.2 DN...

Embodiment 2

[0084] The preparation of embodiment 2 mycoplasma pneumoniae GZ strain inactivated vaccine of swine

[0085] 1. Preparation of strains for production

[0086] After the freeze-dried strain of Mycoplasma hyopneumoniae GZ strain is unsealed, inoculate the liquid medium with 10% inoculum amount, cultivate at 37°C for 3-5 days, harvest when the pH value drops to 6.8-7.0, and use it as a first-class production seed after pure inspection . Take the first-grade seeds and inoculate the liquid medium with 5% inoculum amount, cultivate them at 37°C for 3-5 days, harvest when the pH value drops to 6.8-7.0, pass the pure inspection, and use them as the second-grade production seeds.

[0087] Liquid medium formula: beef heart extract (BD company) 300ml, double distilled water 360ml, correct the pH value to 7.4, sterilize at 121°C for 15 minutes, then add the following filter sterilization components: Hank's balanced salt solution (10×) 40ml, 0.25 (W / V) phenol red 10ml, horse serum 200ml,...

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Abstract

The invention provides a Mycoplasma hyopneumoniae GZ strain, which has better immunogenicity, and the prepared vaccine composition can completely protect pigs, and the GZ strain has high titer during amplification preparation, is easy to cultivate, and can produce a large amount of antigen. The antigen prepared by it can be used with other antigens at the same time, and the immune effects produced by each will not be affected by each other.

Description

technical field [0001] The invention relates to a mycoplasma hyopneumoniae, a vaccine composition prepared by the same and an application thereof, belonging to the field of biotechnology. Background technique [0002] Mycoplasma pneumonia (Mycoplasma Pneumoniae of swine, MPS), also known as swine asthma, is a chronic respiratory infectious disease caused by Mycoplasma hyopneumoniae (Mhp), characterized by high infectivity, high morbidity and low mortality. Features, mainly manifested as cough, dyspnea, fleshy or marble-like lesions in lung tissue, mainly lung lesions, especially pancreatoid and fleshy changes in the heart lobe, middle lobe, and apex lobe of both lungs . Its main harm is that it causes the growth of pigs to be hindered and the feed conversion rate to decrease. Mycoplasma hyopneumoniae spreads through the respiratory tract. After infecting the respiratory epithelium, the cilia of the respiratory tract will be shed and damaged, and the epithelial cells will b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A61K39/02A61K39/12A61K39/102A61P31/04A61P11/00C12R1/35
CPCA61K39/0241A61K39/102A61K39/12A61K2039/521A61K2039/522A61K2039/552A61K2039/70C12N2750/10034C12N1/205C12R2001/35A61K2300/00
Inventor 田克恭金云云孙进忠张许科
Owner PULIKE BIOLOGICAL ENG INC
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