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Recombinant koi herpesvirus (KHV) or cyprinid herpesvirus 3 (CyHV-3) and vaccine for the prevention of disease caused by KHV/CyHV-3 in cyprinus carpio carpio or cyprinus carpio koi

A kind of koi herpes virus, the technology of koi herpes virus, be used in recombinant koi herpes virus (KHV) or koi herpes virus 3 (CyHV-3) and be used for preventing common carp or koi from being caused by KHV/CyHV-3 In the field of vaccines for different diseases, it can solve the problems that cannot be ruled out, and the decision mechanism of attenuation is unknown.

Inactive Publication Date: 2010-09-01
UNIV LIEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the vaccine candidate has two major drawbacks. First, the attenuation is due to random mutations of the virus during in vitro replication. Therefore, the mechanism responsible for the attenuation is unknown and reversion to fully pathogenic

Method used

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  • Recombinant koi herpesvirus (KHV) or cyprinid herpesvirus 3 (CyHV-3) and vaccine for the prevention of disease caused by KHV/CyHV-3 in cyprinus carpio carpio or cyprinus carpio koi
  • Recombinant koi herpesvirus (KHV) or cyprinid herpesvirus 3 (CyHV-3) and vaccine for the prevention of disease caused by KHV/CyHV-3 in cyprinus carpio carpio or cyprinus carpio koi
  • Recombinant koi herpesvirus (KHV) or cyprinid herpesvirus 3 (CyHV-3) and vaccine for the prevention of disease caused by KHV/CyHV-3 in cyprinus carpio carpio or cyprinus carpio koi

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0311] Example 1: Production of KHV FL Strains with Parental Strains Derived Recombinant Attenuated Vaccines inherent characteristics

[0312] The growth characteristics of the wild strain in vitro and the virulence in vivo were tested. At the end of this screening process, the KHV FL strain is selected as the parental strain from which the derived attenuated recombinant strain was derived. figure 1 Showing the growth characteristics of the KHV FL strain compared to two KHV reference strains (IS strain and KHV-U strain), the FL strain replicates more efficiently in vitro. figure 2 It is shown that the KHV FL strain is less pathogenic than the KHVIS strain in vivo. Indeed, the IS strain and the FL strain caused 90% and 80% mortality, respectively, when tested on 7 g of koi ( figure 2 ). Interestingly, when assayed on 2 g of common carp, the FL strain induced disease delayed by two weeks compared to the IS strain. With a view to the 2-week delay observed compared with ...

Embodiment 2

[0313] Example 2: Cloning of the KHV genome in Escherichia coli

[0314] The genome of the KHV FL strain was cloned as a BAC to produce a KHV recombinant. use as image 3 The method described to back clone KHV. The first step in the method requires insertion of the BAC cassette into the KHV genome by homologous recombination in eukaryotic cells. Since the genome of KHV was only partially sequenced when the inventors started their research, the insertion site of the BAC cassette was chosen at the end of the TK gene ( image 3 A). KHV FL BAC strain ( image 3 B). The ability of the latter virus to grow in the presence of G418 allowed it to be isolated from the parental strain. In addition, EGFP expression induced by the KHV FL BAC strain can be used to monitor the screening process. The virus was passaged three times in the presence of G418 to obtain a pure KHV FL BAC population. The molecular structure of described KHV FL BAC strain is confirmed by the combination of ...

Embodiment 3

[0315] Example 3: Stability of the KHV genome in E. coli

[0316] BAC plasmids are usually propagated in bacteria with a recA mutation that minimizes recombination. But the large size and complex structure of the KHV genome can lead to the relative instability of the BAC plasmid (Ilouze et al., Microbiol. Mol. Biol. Rev., 70(1), 147-156, (2006)). To assess the stability of the KHV genome as a BAC plasmid, bacteria harboring the KHV FLBAC plasmid were continuously cultured for 20 consecutive days, representing approximately 36 passages per day. After various periods of cultivation, BAC plasmids were extracted and digested with SacI for characterization ( Figure 6 ). No differences among plasmids were observed at various stages of growth, demonstrating the high stability of the KHV genome in E. coli.

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Abstract

The present invention refers to a recombinant koi herpesvirus (KHV) or Cyprinid herpesvirus 3 (CyHV-3), which is immunogenic in fish, preferably in carps, more preferably in Cyprinus carpio, and to a vaccine for preventive and / or therapeutic treatment of a disease caused by koi herpesvirus (KHV) or CyHV-3. The 5 recombinant herpesvirus is used to confer immunity on fish, preferably on carps, more preferably on Cyprinus carpio, against a disease caused by koi herpesvirus (KHV) or Cyprinid herpesvirus 3 (CyHV-3).

Description

technical field [0001] The present invention relates to recombinant koi herpesvirus (koi herpesvirus, KHV) or cyprinid herpesvirus 3 (Cyprinid herpesvirus-3, CyHV-3) and for preventing common carp or koi from being caused by koi herpesvirus / carp herpesvirus 3 vaccines against diseases. Background technique [0002] The common carp (Cyprinus carpio carpio) is the most widely farmed fish for human consumption mainly in Asia, Europe and the Middle East; while worldwide, especially in Japan, the koi (Cyprinus carpio koi) subspecies is cultivated according to personal preference or A precious, beautiful and colorful pet fish bred for competitive exhibitions. A virus was discovered in the United Kingdom in 1996 that caused common carp and koi to suffer from a fatal disease originally known as Koi Herpes Virus Disease (KHVD). Mass die-offs of common carp and koi in Israel, the United States, and Germany were quickly identified as being attributable to the virus. Unfortunately, i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/245C12N7/04C12N15/38C12N15/85C12N15/86A61K39/205
CPCC12N2760/20022A61K39/245C12N7/00C12N2810/6081A61K2039/5254C12N2710/16034C12N2710/16045C12N2800/30C12N2800/204C12N2710/16062C12N2710/16043A61K2039/552A61K39/12A61P31/22A61P37/04
Inventor B·考斯特斯F·莱弗里格A·梵德普拉斯奇恩
Owner UNIV LIEGE
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