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

A dual inactivated vaccine, Chlamydia psittaci technology, applied in Chlamydia antigen components, microbial-based methods, biochemical equipment and methods, etc., can solve problems such as abortion, weak lambs, and economic losses in the sheep industry

Active Publication Date: 2020-10-20
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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  • Bivalent inactivated vaccine for goat mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof
  • Bivalent inactivated vaccine for goat mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof
  • Bivalent inactivated vaccine for goat mycoplasma pneumonia and chlamydia psittaci disease and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Embodiment 1, the seed batch preparation of mycoplasma mycoplasma goat subspecies and mycoplasma ovis pneumoniae

[0070] 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 2,684 samples of lung tissue and throat swabs from sheep with suspected mycoplasma pneumonia. After separation and testing, the dominant prevalence of Mycoplasma goat subspecies and the prevalence of Mycoplasma ovis pneumoniae were determined. strain. Specific steps are as follows:

[0071] 1. Primary screening

[0072] (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 g...

Embodiment 2

[0089] Embodiment 2, the screening determination of inactivated bacterium vaccine strain and bacterial strain for inspection

[0090] 1. Morphological observation

[0091] A. Morphological observation of Mycoplasma mycoplasma goat subspecies

[0092] 1. Select 5 strains from the mycoplasma goat subspecies screened in step 3 in Example 1, named successively as GT-01 strain, GT-02 strain, GT-03 strain, GT-04 strain and GT-05 strain , respectively from Wuyuan County, Inner Mongolia, Chongqing, Guizhou, Xinjiang Uygur Autonomous Region and Ordos, Inner Mongolia.

[0093] 2. Carry out Dienes staining to mycoplasma goat subspecies (GT-01 strain, GT-02 strain, GT-03 strain, GT-04 strain or GT-05 strain), and then place it under a microscope (200×) to observe the colony form.

[0094] See the experimental results figure 1 (A-E are GT-01 strain, GT-02 strain, GT-03 strain, GT-04 strain and GT-05 strain). The results showed that the colonies of the five strains of Mycoplasma goat s...

Embodiment 3

[0170] Embodiment 3, the expression of MOMP protein (the main outer membrane protein of Chlamydia psittaci)

[0171] 1. Construction of recombinant plasmid pET-22b-MOMP

[0172] 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.

[0173] 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 The primer pair consisting of CTAGAATCTGAATTGAGCATT-3' (the underline is the recognition site of restriction endonuclease XhoI) was subjected to PCR amplification to obtain a PCR amplification product of about 1130bp.

[0174] 3. Take the P...

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Abstract

The invention discloses a bivalent inactivated vaccine for goat mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. The active ingredients of the bivalent inactivated vaccinecomprise inactivated Mycoplasma capricolum subsp. Capripneumoniae, inactivated mycoplasma ovipneumoniae and inactivated MOMP protein of chlamydia psittaci. The amino acid sequence of the MOMP proteinof the chlamydia psittaci is shown as a sequence 2 in a sequence table. The mycoplasma ovipneumoniae is mycoplasma ovipneumoniae SH-01 CGMCC No.16503. The Mycoplasma capricolum subsp. Capripneumoniaeis Mycoplasma capricolum subsp. Capripneumoniae GT-01 CGMCC No.16504. Experiments prove that the bivalent inactivated vaccine has good safety in immunizing experimental animals and target animals, and can effectively prevent goat mycoplasma pneumonia and chlamydia psittaci from infecting the experimental animals and the target animals. The vaccine has important significance for preventing and controlling prevalence and propagation of mycoplasma pneumonia and chlamydia psittaci of goats. The vaccine 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 mycoplasma pneumonia and chlamydia psittaci and a preparation method thereof. Background technique [0002] Mycoplasma pneumonia is caused by Mycoplasma mycoides subsp. capri (Mmc) and / or Mycoplasma ovipneumoniae (Movi) and / or Mycoplasma capricolum subsp. capricolum (Mcc) , a common contagious disease in goats and sheep. Mycoplasma pneumonia in goats is most common in late summer and autumn. It is characterized by high fever, cough, panting, progressive emaciation, pulmonary interstitial hyperplasia, sputum and fibrinous inflammation in the chest and pleura. , high infection rate. Mycoplasma pneumonia in goats currently occurs widely in many countries and regions, including Central Asia, North Africa, the Middle East and other sheep-raising areas. In my country, mycoplasma pneumonia in goats caused by Mmc and Movi is wides...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61K39/118A61K39/39A61P11/00A61P31/04C12N1/20C12R1/35
CPCA61K39/0241A61K39/118A61K39/39A61P11/00A61P31/04A61K2039/552A61K2039/521A61K2039/70A61K2039/55566A61K2039/55505C12R2001/35C12N1/205
Inventor 李敏刘少蓉高强段跃强
Owner INNER MONGOLIA HUAXI BIOTECH
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