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Bovine-derived A-type pasteurella multocida inactivated vaccine as well as preparation method and application thereof

A Pasteurella and inactivated vaccine technology, applied in the field of biomedicine, can solve the problems of animal husbandry economic losses, low cross-immunity efficacy, lack of cross-protective vaccines, etc., and achieve the effect of less harm

Pending Publication Date: 2021-01-08
XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, most of the vaccines used in China are traditional inactivated vaccines against type B HS. There are few studies on type A vaccines, and the cross-immunity efficacy of Pm vaccines with different serotypes is low. Resisting the attack caused by the same type of strains, the abuse of antibiotics and the increase of drug-resistant bacteria make the treatment of Pm disease more difficult and lack of ideal cross-protective vaccines. cause major economic losses

Method used

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  • Bovine-derived A-type pasteurella multocida inactivated vaccine as well as preparation method and application thereof
  • Bovine-derived A-type pasteurella multocida inactivated vaccine as well as preparation method and application thereof
  • Bovine-derived A-type pasteurella multocida inactivated vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation of embodiment 1 bovine type A Pm inactivated vaccine and the establishment of serum antibody ELISA method

[0042] In this example, one Pm (Pm12) strain of type A was selected as the research object according to the research of the previous experiment, and Pm12 (type A) and M6 (type B) were prepared into inactivated vaccines according to the preparation procedure of bacterial inactivated vaccines. The ELISA conditions were screened according to the checkerboard method, and an indirect ELISA method for detecting serum antibodies of different animals was established. The results show that: the prepared inactivated vaccine is qualified through the safety test, and the bacterial content of the Pm12 (type A) inactivated vaccine is 8×10 9 CFU / mL, the bacterial content of M6 (type B) inactivated vaccine is 1.4×10 10 CFU / mL.

[0043] 1. Preparation of seedling oil phase

[0044] Medicinal white oil 94mL, aluminum stearate 2g, first take a small amount of whit...

Embodiment 2

[0096] Example 2 Analysis of changes in antibody levels and immune protection effects of Pm12 inactivated vaccines after immunization of mice and rabbits

[0097] 1. The median lethal dose (LD) 50 ) determination

[0098] Take 30 mice, 10 in each group, and challenge the virus by intraperitoneal injection, each group is challenged with 1×10 6 , 1×10 7 , 1×10 8 CFU, LD was calculated according to the modified Cole's method 50 . The groups of mice are shown in Table 7, and the preparation of the four inactivated vaccines refers to Example 1.

[0099] Table 7 Grouping situation of mouse median lethal dose

[0100]

[0101] Experimental results: According to the modified Cole's method, the median lethal dose of Pm12 is about 3.8×10 1 CFU, the median lethal dose of Pm8 is about 4.2×10 1 CFU, the median lethal dose of Pm122 is about 3.1×10 1 CFU, the median lethal dose of M6 is about 5.6×10 1 CFU.

[0102] 2. Mouse immune antibody assay and challenge protection test

...

Embodiment 3

[0148] Example 3 Determination and Analysis of Serum Antibody Levels of Pm12 Inactivated Vaccine Immunized Different Day-Old Cattle

[0149] 1. Determination of the immune effect and immune antibody of cows in the dry period

[0150] Select 30 cows in the dry period (the expected delivery date is about 60 days) and divide them into 3 groups, 10 cows in each group, and inoculate Pm12 inactivated vaccine, M6 inactivated vaccine and Pm12-M6 bivalent inactivated vaccine respectively. Each head and neck muscle Inoculate with 2 mL, and perform the second immunization 21 days after the first immunization, and observe the appetite and mental status of the cows during the dry period. Blood was collected before vaccination as a negative control. After the first vaccination, blood was collected every 7 days to separate serum for antibody monitoring. Indirect ELISA was used to detect the regularity of antibody levels produced by inactivated vaccines in cows during the dry period.

[0151...

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Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a bovine-derived A-type pasteurella multocida inactivated vaccine as well as a preparation method andapplication thereof. The inactivated vaccine prepared by the invention meets the safety requirements of inactivated vaccines. And a sensitive and specific indirect ELISA method for determining a serumantibody is established. When the prepared inactivated vaccine is used for immunizing mice and rabbits, a serum antibody with relatively high titer and longer duration can be generated, the generatedantibody can resist attack of A-type Pm, and meanwhile, a part of protective effect on attack of B-type Pm is shown. The inactivated vaccine immunizes cows and calves to obtain antibodies with high titer and relatively long duration, and the antibodies are maintained for at least 6 months. Meanwhile, the maternal antibody level of the born calves is high and is maintained for at least two monthsor more.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to an inactivated vaccine of bovine-derived Pasteurella multocida type A, a preparation method and an application thereof. Background technique [0002] Pasteurellosis (pasteurellosis) is caused by Pasteurella multocida (Pasteurella multocida, Pm), an acute, febrile infectious disease that can infect a variety of animals and humans. Pasteurellosis in cattle manifests as bovine respiratory diseases (BRDs) caused by Pm type A and bovine hemorrhagic septicemia (HS) caused by Pm type B, both of which spread rapidly and are widespread. , High case fatality rate, the disease has become a major infectious disease that endangers the cattle industry all over the world, causing huge economic losses. Due to the large number of Pm serotypes, they can infect each other between the same species and different species of animals. [0003] The capsular structure and lipopolysacchar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/102A61P31/04
CPCA61K39/102A61P31/04A61K2039/521A61K2039/552
Inventor 韩猛立吴桐忠张星星黄新钟发钢张倩何延华
Owner XINJIANG ACADEMY OF AGRI & RECLAMATION SCI
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