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A dual inactivated vaccine for Mycoplasma ovine pneumonia and Chlamydia psittacosis and preparation method thereof

A dual inactivated vaccine, the technology of Chlamydia psittaci, applied in the direction of Chlamydia antigenic components, microorganism-based methods, biochemical equipment and methods, etc. Safety, the effect of important application value

Active Publication Date: 2022-04-19
INNER MONGOLIA HUAXI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, it has also been reported that some sheep reproductive disorders caused by Chlamydia psittaci have been reported. Some township sheep farms and large family sheep in Helen City have been affected by Chlamydia to varying degrees. The main clinical manifestations are fever, abortion, death and delivery of deformed lambs. , Weak lambs are characterized by causing serious economic losses to the sheep industry

Method used

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  • A dual inactivated vaccine for Mycoplasma ovine pneumonia and Chlamydia psittacosis and preparation method thereof
  • A dual inactivated vaccine for Mycoplasma ovine pneumonia and Chlamydia psittacosis and preparation method thereof
  • A dual inactivated vaccine for Mycoplasma ovine pneumonia and Chlamydia psittacosis and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Embodiment 1, the seed batch preparation of ovine mycoplasma pneumoniae

[0076] The inventor of the present invention is located in my country's Chongqing City, Anhui Province, Qinghai Province, Yunnan Province, Xinjiang Uygur Autonomous Region, Guizhou Province, Guangxi Zhuang Autonomous Region, and Hohhot, Keshigten Banner, Ordos, Siziwang Banner, Ewenki Banner, and Lanzhou in the Inner Mongolia Autonomous Region. The sheep farms in Qi and Wuyuan counties collected 2684 samples of lung tissue and throat swabs from sheep suspected of suffering from mycoplasma pneumonia. After separation and testing, the dominant and prevalent strains of Mycoplasma ovis pneumoniae were screened and determined. Specific steps are as follows:

[0077] 1. Primary screening

[0078] (1) Put the sample into a culture bottle filled with mycoplasma liquid medium (throat swabs are put in directly, sheep lung tissue is put in after grinding), and cultured in a constant temperature incubator at ...

Embodiment 2

[0090] Embodiment 2, the screening determination of inactivated bacterium vaccine strain and bacterial strain for inspection

[0091] 1. Morphological observation of Mycoplasma pneumoniae in sheep

[0092] 1. Select 5 strains from the Mycoplasma ovis pneumoniae screened in step 3 in Example 1, and name them successively as SH-01 strain, SH-02 strain, SH-03 strain, SH-04 strain and SH-05 strain, respectively source In Chongqing, Hohhot Inner Mongolia, Xinjiang Uygur Autonomous Region, Wuyuan County Inner Mongolia and Ordos Inner Mongolia.

[0093] 2. Carry out Dienes staining to Mycoplasma ovis pneumoniae (SH-01 strain, SH-02 strain, SH-03 strain, SH-04 strain or SH-05 strain), and then observe the colony morphology under a microscope (200×).

[0094] See the experimental results figure 1 (A-E are SH-01 strain, SH-02 strain, SH-03 strain, SH-04 strain and SH-05 strain). The results showed that the colonies of the 5 strains of Mycoplasma ovis pneumoniae were all blue, without a...

Embodiment 3

[0132] Embodiment 3, the expression of MOMP protein (the main outer membrane protein of Chlamydia psittaci)

[0133] 1. Construction of recombinant plasmid pET-22b-MOMP

[0134] 1. Because the MOMP gene of Chlamydia psittaci is highly conserved among the same species, the MOMP gene (GenBank: L25436.1), a typical representative of Cps, was selected for codon optimization. The nucleotide sequence shown in Sequence 1 in the sequence listing after codon optimization (ie MOMP gene) was artificially synthesized.

[0135] 2. Use the nucleotide sequence of the MOMP gene synthesized in step 1 as a template, and use 5'-TATT CATATG TTGCCTGTAGGGAACCCAGCT-3' (the underline is the recognition site of restriction endonuclease NdeI) and 5'-GCGC CTCGAG A primer pair consisting of CTAGAATCTGAATTGAGCATT-3' (the underline is the recognition site of the restriction endonuclease XhoI) was used for PCR amplification to obtain a PCR amplification product of about 1130 bp.

[0136] 3. Take the PC...

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Abstract

The invention discloses a dual inactivated vaccine for ovine mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. The active components of the dual inactivated vaccine include inactivated Ovis Mycoplasma pneumoniae and inactivated MOMP protein of Chlamydia psittaci. The amino acid sequence of the MOMP protein of Chlamydia psittaci is shown as sequence 2 in the sequence listing. Mycoplasma ovis pneumoniae is Mycoplasma ovis pneumoniae (Mycoplasma ovipneumoniae) SH‑01CGMCC No.16503. Experiments have proved that the dual inactivated vaccine immunization experimental animals (such as guinea pigs, rabbits, Beagle dogs) and target animals (such as sheep) have good safety, and can effectively prevent Mycoplasma ovis pneumoniae and Chlamydia psittaci infection. animals and target animals. It is of great practical significance to prevent and control the prevalence and spread of mycoplasma pneumonia and chlamydia psittaci. The invention has important application value.

Description

technical field [0001] The invention belongs to the field of veterinary biological products, and in particular relates to a dual inactivated vaccine of sheep mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. Background technique [0002] Sheep pneumonia and abortion are important factors that seriously endanger the healthy development of animal husbandry. Especially in recent years, with climate change, ecological environment damage and antibiotic abuse, the incidence and mortality of sheep pneumonia and abortion have increased year by year. Therefore, the prevention and control of sheep pneumonia and abortion has become a major practical problem that needs to be solved urgently for the healthy development of animal husbandry, food safety and human health. [0003] Pneumonia and abortion in sheep are caused by a variety of reasons, the most important of which are pathogenic microorganism infection, especially Mycoplasma ovis and Chlamydia infectio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/118A61K39/02A61P31/04C07K14/295C12N1/20C12R1/01
CPCA61K39/0241A61K39/118A61P31/04C07K14/295A61K2039/70A61K2039/552A61K2039/521A61K2039/55566A61K2039/55505C12R2001/01C12N1/205
Inventor 李敏刘少蓉高强
Owner INNER MONGOLIA HUAXI BIOTECH
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