Attenuated live vaccine strain for preventing pig-pig infection breeding and respiratory syndrome

A technology for respiratory syndrome and live attenuated vaccines, applied in the field of live attenuated vaccines, can solve the problems of recurrent PRRS in pig farms, prone to mutation, unsatisfactory preventive effect of attenuated vaccines, etc. Good protection and prevention of porcine reproductive and respiratory syndrome

Active Publication Date: 2010-01-27
华威特(江苏)生物制药有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

However, PRRSV is very prone to mutation, which leads to the unsatisfactory clinical preventive...

Method used

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  • Attenuated live vaccine strain for preventing pig-pig infection breeding and respiratory syndrome
  • Attenuated live vaccine strain for preventing pig-pig infection breeding and respiratory syndrome

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Experimental program
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Effect test

Embodiment 1

[0031] Passage weakening of embodiment 1PRRSV TJ strain

[0032] A strain of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) was isolated from diseased materials collected from dead pigs in a high fever pig farm in Tianjin, and named as PRRSV TJ strain. The GenBank accession number of this strain is EU860248. The strain was submitted for deposit, and its microorganism deposit number is: CGMCC NO.2129. The full length of the PRRSV TJ strain genome is 15324bp (including the PolyA tail). Compared with the American standard strain VR-2332, the Nsp2 sequence of the TJ strain Nsp2 sequence lacks the 481st and 532-560th amino acids, and the two nucleotides are identical. The homology is 83.7%, and the homology of the encoded amino acid is 77.9%. The results of this test showed that the isolate was a mutated PRRSV, which was highly pathogenic to pigs. The PRRSV TJ strain was continuously passaged through Marc-145 cells, and the plaque clone was purified every 10 passages...

Embodiment 2

[0033] Embodiment 2 PRRSV TJM strain virulence returns strong test

[0034] The PRRSV TJM strain was attenuated by the prevailing high fever virus (PRRSV TJ strain) and was identified and preserved. This study further tested whether the vaccine virus has virulence reversion. For the first virulence recovery, PRRSV TJM strain virus was used to inoculate 4-week-old healthy PRRS-negative piglets, intramuscularly injected into the neck, 2ml / pig, 10 5.7 TCID 50 / ml, and a negative control pig without inoculation was set at the same time. From the 2nd to the 5th virulence reactivation test, the PRRSV-positive sera from the previous animals were mixed and used as the next animal inoculum to inoculate pigs. All were vaccinated by intramuscular injection in the neck, and pigs not vaccinated each time were set as negative controls. After each virus inoculation, the rectal temperature of the experimental animals was measured at a fixed point for 14 consecutive days, and the clinical ob...

Embodiment 3

[0035] Embodiment 3PRRSV TJM strain genetic variation analysis

[0036] The PRRSV TJ strain was attenuated by continuous passage of Marc-145 cells to obtain the PRRSV TJM strain. The Nsp2 gene of the PRRSV TJM strain was sequenced and compared with the PRRSV TJ strain and the classic American VR-2332 strain. The results showed that : The full-length Nsp2 sequence of PRRSV TJM strain is 2490bp, with 98.6% and 82.9% nucleotide sequence homology and 97.6% and 75.8% amino acid sequence homology with PRRSV TJ strain and American type standard strain VR-2332 respectively , Compared with the PRRSV TJ strain, amino acids 598-717 were continuously deleted, and a total of 120 amino acids were deleted; compared with the VR-2332 strain, amino acids 481, 537-565, and 628-747 were deleted, and a total of 120 amino acids were deleted. 150 amino acids. The nucleotide sequence of Nsp2 of PRRSV TJM strain is shown in SEQ ID NO:1.

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Abstract

The invention provides an attenuated live vaccine strain for preventing pig-pig infection breeding and respiratory syndrome, a formulation thereof and a preparation method of the attenuated live vaccine strain and the formulation thereof. The attenuated live vaccine strain has obvious immunity protection effect on the pig-pig infection breeding and respiratory syndrome. The vaccine formulation also has long protection period and stable storage.

Description

technical field [0001] The invention relates to an attenuated live vaccine for preventing porcine reproductive and respiratory syndrome from being infected. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine Reproductive and Respiratory Syndrome, PRRS) is a pig infectious disease caused by PRRSV (Porcine Reproductive and Respiratory SyndromeVirus, PRRSV). Clinical signs are characterized by reproductive failure in sows and dyspnea in piglets. In the early stage of the onset, all symptoms were fever, lethargy, loss of appetite, fatigue, dyspnea, and cough. The disease was first discovered in North Carolina (1987) in the United States, and has since been found in Canada (1998), Germany (1990), and the United Kingdom (1991). In 1992, the International Office of Epizootics (OIE) named the disease Porcine Reproductive and Respiratory Syndrome (PRRS). my country also reported the prevalence of this disease in 1996. PRRSV is divided into two ge...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N7/08C12N5/06A61K39/12A61P31/14C12N15/40C12Q1/70C12Q1/68C12R1/93
Inventor 武华冷雪李真光王凤雪
Owner 华威特(江苏)生物制药有限公司
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