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Bivalent inactivated vaccine for sheep mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof

A technology for Chlamydia psittacosis and double inactivated vaccine, which can be applied in the directions of Chlamydia antigen components, microorganism-based methods, biochemical equipment and methods, etc., and can solve problems such as weak lambs, miscarriage, fever, etc.

Active Publication Date: 2020-10-20
INNER MONGOLIA HUAXI BIOTECH
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  • 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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  • Bivalent inactivated vaccine for sheep mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof
  • Bivalent inactivated vaccine for sheep mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof
  • Bivalent inactivated vaccine for sheep mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof

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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 bivalent inactivated vaccine for sheep mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. Active ingredients of the bivalent inactivated vaccine comprise inactivated mycoplasma ovipneumoniae and inactivated MOMP protein of chlamydia psittaci. The amino acid sequence of the MOMP protein of the chlamydia psittaci is shown as a sequence 2 in a sequence table. The mycoplasma ovipneumoniae is (Mycoplasma ovipneumoniae) SH-01 CGMCC No.16503. Experiments prove that the bivalent inactivated vaccine has good safety for immune experimental animals (such as guinea pigs, rabbits and Beagle dogs) and target animals (such as sheep), and can effectively prevent mycoplasma ovipneumoniae and chlamydia psittaci from infecting the experimental animals andthe target animals. The vaccine has important practical significance for preventing and controlling prevalence and propagation of mycoplasma pneumonia and chlamydia psittaci of sheep. The vaccine hasimportant 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 Applications(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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