Vaccine composition and application thereof

A technology of vaccine composition and protein, which is applied in the direction of drug combination, medical preparations containing active ingredients, bacterial antigen components, etc., and can solve problems such as poor immune effect

Inactive Publication Date: 2016-11-23
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current subunit vaccine against Mycoplasma pneumoniae in swine has poor immune effect and cannot provide complete protection to pigs (Fagan PK, Djordievic SP. Chin J, et al. Oral immunization of mice with attenuated Salmonellatyphimurium aroA expressing a recombinant Mycoplasma hyopeumoniaeantigen (NrdF). Infection and immunity, 1997, 65:2502-2507)

Method used

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  • Vaccine composition and application thereof
  • Vaccine composition and application thereof
  • Vaccine composition and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The preparation of embodiment 1 mycoplasma hyopneumoniae whole bacterium antigen

[0049] Dilute the freeze-dried strain of Mycoplasma hyopneumoniae HN0613 strain with liquid medium, inoculate it on a solid medium plate by streaking, culture at 37°C for 7 days, select well-growing colonies, inoculate on the slope of solid medium, and culture at 37°C 7 days, as a primary seed.

[0050] Take a small amount of liquid medium to wash the slant culture of the primary seed, inoculate it in a large tube of liquid medium, culture it at 37°C for 7 days, and use it as the secondary seed after being tested to be pure.

[0051] The secondary seed liquid cultured in the liquid medium is inoculated in the liquid medium at a volume ratio of 10% (V / V). Cultivate at 37°C for 3-6 days, the color will turn yellow, and the bacterial liquid will be harvested when the pH value drops to 6.8-7.0. After passing the pure inspection, expand the culture in the same way (the number of subcultures s...

Embodiment 2

[0054] Example 2 Expression, Identification and Purification of Mycoplasma Hyopneumoniae Immunogenic Protein

[0055] According to the XylF protein nucleoside in the Mycoplasma hyopneumoniae J strain (accession number: AE017243.1) and 7448 strain (accession number: AE017244.1) reported in NCBI (http: / / www.ncbi.nlm.nih.gov) acid sequence, and the P78 protein nucleotide sequence in strain J (accession number: AE017243.1) and strain 232 (accession number: AE017332.1), which were synthesized by Shanghai Bioengineering Co., Ltd. by artificial synthesis. The full lengths of the gene fragments are XylF-I 1341bp, XylF-II 1341bp, P78-I 2076bp, P78-II 2076bp. On the basis of artificially synthesizing XylF-I, XylF-II, P78-I, and P78-II gene fragments, templates for XylF-I, XylF-II, P78-I, and P78-II proteins were prepared respectively.

[0056] The gene fragments synthesized in the previous step were used as templates for PCR reactions respectively, and primers for XylF-I, XylF-II, P78-...

Embodiment 3

[0059] The preparation of embodiment 3 swine mycoplasma pneumonia vaccine composition

[0060] The mycoplasma hyopneumoniae whole bacterium antigen prepared in embodiment 1, the mycoplasma hyopneumoniae immunogenic protein prepared in embodiment 2 are diluted with pH7.2 PBS solution, and each protein solution after dilution is mixed with Montanide TM Gel 01 adjuvant is mixed together by the components and ratio contained in the Mycoplasma pneumoniae vaccine composition in Table 1, stirred at a speed of 500-800r / min for 10-15min, and added 1% (volume ratio) thimerosal solution before terminating the stirring , so that the final concentration does not exceed 1 / 10,000, fully shake and mix, and store at 2-8°C after aliquoting.

[0061] Table 1 Components and ratios contained in the swine mycoplasma pneumonia vaccine composition

[0062]

[0063]

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Abstract

The invention provides a swine mycoplasma pneumonia vaccine composition, the vaccine composition includes swine pneumonia mycoplasma whole cell antigen and an immunogenic protein, and also comprises a veterinary-medicine-acceptable carrier, wherein the immunogenic protein comprises at least one of XylF and P78 proteins. The present invention also discloses the application of the vaccine composition in preparing of medicines for preventing and / or treating swine mycoplasma pneumonia. The content of the whole cell antigen and the immunogenic protein of the vaccine composition is decreased significantly compared with that of single use of a whole cell vaccine or a subunit vaccine for immunization, and immune effect is enhanced; protein part can be recombinationally expressed by means of genetic engineering in quantity, short time is taken, and the swine mycoplasma pneumonia vaccine composition can also be convenient for mass production. In addition, when an animal is immunized with the vaccine composition, the immune effects of the vaccine composition on the swine mycoplasma pneumonia antigen contained in the vaccine composition and other antigens are not affected.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a swine mycoplasma pneumonia vaccine composition and application thereof. Background technique [0002] Mycoplasma pneumonia of swine (MPS) is also known as swine panting disease or porcine enzootic pneumonia (Li Yanming, Zhang Ying. Advances in biological research of Mycoplasma hyopneumoniae, Advances in Veterinary Medicine, 2003,24(3):25- 27), is a kind of pig contact chronic respiratory disease caused by Mycoplasma hyopneumoniae (Mycoplasma hyopneumoniae, Mhp), which is widely distributed and characterized by chronicity, contact, high infectivity, high morbidity and low mortality, etc. The main manifestations are cough and dyspnea, and the dissection shows fleshy or marble-like lesions in the lung tissue. The disease can also lead to growth retardation of pigs and low feed conversion rate, seriously endangering the development of pig industr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/02A61K39/12A61K39/102A61K39/23A61K39/205A61K39/145A61P11/00A61P31/04
Inventor 张许科孙进忠廖永洪王莹田克恭
Owner PU LIKE BIO ENG
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