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A Fish Rhabdovirus Attenuated Vaccine Strain

A rhabdovirus and attenuated vaccine technology, applied in the field of virus vaccines, can solve rare and other problems

Active Publication Date: 2021-06-15
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Immunization is the most effective way to prevent and control the disease, but currently there are few vaccine strains that can exert effective immunity against Rhabdovirus

Method used

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  • A Fish Rhabdovirus Attenuated Vaccine Strain
  • A Fish Rhabdovirus Attenuated Vaccine Strain
  • A Fish Rhabdovirus Attenuated Vaccine Strain

Examples

Experimental program
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Effect test

Embodiment 1

[0027] 1. Materials and methods

[0028] 1.1 Cell lines and virus strains

[0029] The mandarin fish brain tissue cell line (CPB) was proliferated in L-15 medium (GIBCO, USA) containing 10% fetal bovine serum (FBS), and cultured at 28°C. Six strains of fish rhabdoviruses were preliminarily isolated in our laboratory, namely, the Sanshui strain of mandarin fish rhabdovirus (SCRV-SS), the Qingyuan strain of mandarin fish rhabdovirus (SCRV-QY), the Gaoming strain of mandarin fish rhabdovirus (SCRV- GM), California perch rhabdovirus Sanshui strain (MSRV-SS), snakehead rhabdovirus Foshan strain (CMRV-FS) and bamboo shoot shell rhabdovirus Jiangmen strain (OMBRV-JM), the six strains of viruses can be tested at 28°C Infected under CPB. Viruses were stored at -80°C.

[0030] 1.2 Total RNA extraction, reverse transcription and fluorescent quantitative PCR

[0031] The total RNA in the viral supernatant of the infected cells was extracted using the TRIzol method (Invitrogen, USA), a...

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Abstract

The invention discloses fish rhabdovirus attenuated vaccine strain and application thereof. The present invention screens out an attenuated strain through the fish body challenge experiment, and names it California perch rhabdovirus Sanshui strain (MSRV‑SS). The attenuated strain has immune protection to mandarin fish and bamboo shoot shell fish through fish immune experiment. The attenuated strain was purified by plaque cloning to obtain a pure line clone MSRV-SS-7, and the growth curves of the clone and the wild virus MSRV-SS on the mandarin fish brain tissue (CPB) cell line were determined, and it was found that It has similar in vitro growth characteristics to the wild virus MSRV‑SS. It has been verified that the clone MSRV‑SS‑7 also has good immunogenicity. The screening of the attenuated strain of the present invention and the sequencing of the whole genome provide technical support for the construction of the MSRV-SS-7 reverse genetic operating system and the development of vector live vaccines.

Description

technical field [0001] The invention belongs to the field of virus vaccines, in particular to fish rhabdovirus attenuated vaccine strains and applications thereof. Background technique [0002] Rhabdoviruses are enveloped negative-strand RNA viruses with a wide range of hosts, including mammals, birds, reptiles, fish, insects, and plants. Among them, a class of rhabdoviruses that can cause fatal and epidemic diseases in fish are called fish rhabdoviruses, and more than 20 species have been reported so far. In the latest virus classification system released by ICTV in 2016, Rhabdoviridae is divided into 18 genera. Fish rhabdoviruses are mainly distributed in three genera of Perhabdovirus, Novirhadovirus and Sprivivirus, but some have not yet been classified. The genome of fish rhabdovirus is a non-segmented single-stranded negative-strand RNA with a size of 11-12kb, encoding five structural proteins, namely nucleoprotein (N), phosphorylated protein (P), matrix protein (M), ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/205A61P31/14
CPCA61K39/12A61K2039/552C12N7/00C12N2760/20021C12N2760/20034
Inventor 付小哲李宁求张莉娟林强刘礼辉梁红茹
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI