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Novel rabies virus fake virus system as well as preparation and application thereof

A rabies virus and pseudovirus technology, applied in the biological field, can solve the problems of lowering the biosafety level and the limited number of wild strains

Inactive Publication Date: 2013-06-26
中国疾病预防控制中心病毒病预防控制所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Considering that when the attacking virus is a wild strain, a BSL-3 laboratory is required and the number of wild strains is limited, if a simulated virus of a wild strain of rabies virus (i.e. a pseudovirus of rabies virus) can be used as the attacking virus, on the one hand it can be expanded. On the other hand, it is possible to reduce the biosafety level required by the laboratory

Method used

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  • Novel rabies virus fake virus system as well as preparation and application thereof
  • Novel rabies virus fake virus system as well as preparation and application thereof
  • Novel rabies virus fake virus system as well as preparation and application thereof

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Embodiment 1

[0067] 1. Construction of a novel pseudovirus system for rabies virus

[0068] 1. Construction of the shuttle plasmid

[0069](1) Amplification of target gene fragments: BHK-21 cells were infected with rabies virus CVS-11, PV, CTN1, and aG respectively, and nucleic acid was extracted by conventional Tizol method 72 hours after infection; Superscript II reverse transcription kit of Invitrogen Company was used to construct the reverse transcription kit. Transcribe cDNA library; use reverse transcription cDNA as template, use CVSG-P1 / CVSG-P2, PVG-P1 / PVG-P2, CTNG-P1 / CTNG-P2, aGG-P1 / aGG-P2 as primers (primer sequence See SEQ ID NO: 1-8 in the sequence listing) to amplify CVS-11, PV, CTN1, and aG glycoprotein genes respectively. The PCR reaction conditions were unified at 94°C for 3min; 35 cycles, 72°C extension for 10 min.

[0070] (2) Construction of the pVRC8301 vector carrying the target gene: Dilute the gel-recovered product of the PCR product and the pVRC8301 vector plasmid ...

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Abstract

The invention relates to a novel rabies virus fake virus system as well as preparation and application thereof, belonging to the field of biotechnology. The novel rabies virus fake virus system comprises a carrier system and host cells, wherein the carrier system consists of pVRC-RVG series shuttle plasmids and auxiliary plasmids of a rabies virus glucoprotein gene; and when the shuttle plasmids and auxiliary plasmids in the carrier system are transfected to the host cells together, rabies virus fake viruses can be obtained from culture supernatant. The novel rabies virus fake virus system can be used for evaluating neutralizing antibodies of rabies viruses, screening neutral monoclonal antibody epitopes of the rabies viruses, carrying out high throughput screening on rabies virus medicaments, and preparing a novel rabies vaccine, so that the novel rabies virus fake virus system is a brand-new technical system for construction, preparation and application of rabies virus fake viruses.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to a novel rabies virus pseudovirus system and its preparation and application. Background technique: [0002] Rabies is a zoonotic infectious disease caused by rabies virus (rabies virus, RV). Once onset, the fatality rate is almost 100%. When rabies exposure occurs, post-exposure treatment of rabies needs to be carried out in time, mainly including rabies vaccination and injection of rabies immunoglobulin preparations, so that the body can produce or acquire rabies virus neutralizing antibodies, thereby preventing the virus from invading the body. [0003] The rabies virus vaccine strains commonly used by domestic human rabies vaccine manufacturers are PV, CTN1, and aG. However, domestic research data on the antigenicity and protective properties of these vaccine strains are not sufficient. The reason is that the isolation and cultivation of wild rabies virus strains It ...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N7/04G01N21/64
Inventor 谭文杰邓瑶吕新军唐青
Owner 中国疾病预防控制中心病毒病预防控制所
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