Duck tembusu virus low virulent strain and application thereof

A technology for duck tambusu virus and attenuated strains, which is applied in the field of viruses, can solve the problems of no vaccine, no effective serological detection method, difficulty in preventing and controlling duck tambusu virus disease, etc., and achieves a good specific effect.

Active Publication Date: 2014-02-12
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Duck Tembusu virus disease is a new infectious disease, highly contagious, and can be popular all year round, which brings great difficulties to the prevention and control of duck Tembusu virus disease. At present, there is no effective prevention There are no control measures, no vaccine available, and no effective, fast and easy serological detection method

Method used

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  • Duck tembusu virus low virulent strain and application thereof
  • Duck tembusu virus low virulent strain and application thereof
  • Duck tembusu virus low virulent strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Subculture and Sequence Analysis of Duck Tembusu Virus

[0036] Use the isolated Duck Tambusu virus virulent strain FX2010 that causes ducks to become ill in nature, inoculate chicken embryo fibroblasts (CEF) into T75 cell culture flasks that grow into a single layer, inoculate 1mL of virus liquid in each bottle, and absorb 1- After 2 hours, discard the virus liquid, add 12mL of DMEM culture medium containing 10% fetal bovine serum, 36-72 hours later, absorb the culture supernatant, and then inoculate the T75 cell culture flask in which CEF grows into a single layer, and continuously subculture 173 passages, and then purified 5 times by limiting dilution method, then passaged to 180 passages. The 60th generation, 80th generation, 100th generation, 130th generation, 145th generation, 168th generation and 180th generation viruses were sequenced. The results are as follows figure 1 as shown ( figure 1 In , the gray shade indicates the newly added mutation point...

Embodiment 2

[0037] Example 2 The pathogenicity analysis of different generations of viruses to ducks

[0038] Infecting ducklings and laying ducks with different generations of viruses in the above-mentioned embodiment 1, by observing the clinical symptoms of ducks, the pathological changes after autopsy and the proliferation and distribution of viruses in ducks, the effects of different generations of viruses on ducks were analyzed. pathogenicity. The specific method is as follows:

[0039] (1) Nasal infection test in ducklings

[0040] Sixty 21-day-old healthy ducklings were randomly divided into 6 groups, 10 in each group. Groups 1-5 were inoculated nasally with FX2010 (dose 10 3.5 TCID 50 ), FX2010-60P, FX2010-80P, FX2010-100P and FX2010-130P (dose of 10 5.5 TCID 50 ), and the sixth group was the blank control group. On the 4th day after inoculation, autopsy, observe the pathological changes of organs, and take spleen, lung, kidney, brain and other tissues, add 1 mL of LPBS per...

Embodiment 3

[0147] The 180th generation poison (FX2010-180P) of embodiment 3 is to the immune effect of duck

[0148] The ducks inoculated with the 180th generation virus (FX2010-180P) were subjected to an immunization test, and challenged with a strong virus 14 days after immunization, to study the immune efficacy of FX2010-180P as a live vaccine on the inoculated ducks.

[0149] (1) The immune effect test of FX2010-180P on ducklings

[0150] Twenty 21-day-old healthy ducklings were randomly divided into 2 groups, 10 in each group. Group 1 was intramuscularly inoculated with FX2010-180P at a dose of 10 3.0 TCID 50 . Group 2 was the blank control group. Blood was collected 7 days and 14 days after inoculation, and the antibody titer was detected by blocking ELISA, and 10 3.5 TCID 50 The FX2010 was challenged, and the clinical symptoms were observed daily. Four days after the challenge, autopsies were performed to observe the pathological changes, and lung, kidney and brain tissues w...

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Abstract

The invention discloses a duck tembusu virus low virulent strain. The low virulent strain is obtained through mutation after continuous subculture of duck tembusu virus virulent strain FX2010 on chicken embryo fibroblast (CEF). The invention further discloses a vaccine for preventing or treating duck tembusu virus disease. The vaccine is duck tembusu virus disease live attenuated vaccine or inactivated vaccine using the duck tembusu virus low virulent strain for vaccination. The invention further discloses a detection antigen for diagnosing the duck tembusu virus disease. The detection antigen is virus particle of the duck tembusu virus low virulent strain. The protection rate of the duck tembusu virus low virulent strain to virulent attack is 100% after the duckling and egg-laying duck are vaccinated whether the duck tembusu virus low virulent strain is used as live attenuated vaccine or the inactivated vaccine; the duck tembusu virus low virulent strain has good specificity as the detection antigen; therefore, the duck tembusu virus low virulent strain has excellent application prospect in the aspect of preventing and treating and diagnosing the duck tembusu virus disease.

Description

technical field [0001] The invention relates to the field of virus technology, in particular to an attenuated strain of duck Tembusu virus which is subcultured from a strong strain of duck Tembusu virus to weaken its pathogenicity, and the attenuated strain is used for duck Tembusu virus disease Application of live vaccines, inactivated vaccines and diagnostic antigens. Background technique [0002] Since April 2010, an infectious disease characterized by slow growth of meat ducks and a sharp drop in egg production of laying ducks has occurred in Shanghai, Zhejiang, Jiangsu and other places in my country. In just a few months, almost all eggs in southern provinces Duck farms have been attacked by the disease, causing huge economic losses to the duck industry. Domestic researchers have proved that the infectious disease is caused by a new flavivirus—Duck Tembusu virus (DTMUV) through virus isolation and identification, virus genome sequence analysis, and animal regression exp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/12A61P31/14G01N33/554C12R1/93
Inventor 李泽君李国新肖亚莉李雪松滕巧泱
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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