US 12,600,988 B2 · Coronavirus pseudovirus packaging system, packaging method therefor, and application of coronavirus pseudovirus in evaluating disinfection efficacy

Abstract

A packaging system for a coronavirus pseudovirus, including a vesicular stomatitis virus (VSV) vector in which Fluc and EGFP dual-reporter genes replace a GP gene, and packaging cell that expresses a coronavirus spike protein. The packaging system may quickly package pseudoviruses by using a one-step packaging method, and may be used in the research of coronaviruses such as COVID-19 (SARS-CoV-2), SARS (SARS-CoV) and MERS, and other viruses. The packaging system and thereby pseudovirus method may also be used to evaluate the efficacy of disinfectants by means of virus contamination distribution models, setting up scenarios, and sampling and testing steps.

Claims10

Dependent claims

A coronavirus pseudovirus packaging system, comprising a modified vesicular stomatitis virus (VSV) and a packaging cell that expresses a coronavirus spike protein;

wherein the coronavirus is SARS, MERS, or COVID-19 virus, and the amino acid sequence of the spike protein is set forth in SEQ ID NO:
4 or SEQ ID NO:
5 or SEQ ID NO:
6 or SEQ ID NO:
7;
the modified vesicular stomatitis virus VSV is defined as a replication-defective virus with structural gene replaced by Fluc and EGFP dual-reporter genes, the modified vesicular stomatitis virus VSV is named as dVSVΔG-Fluc-EGFP, a gene coding GP in the genetic material of the VSV is replaced by Fluc reporter gene, the EGFP reporter gene is integrated between Fluc and VSV polymerase L gene, and the gene sequence of the dVSVΔG-Fluc-EGFP is set forth in SEQ ID NO:
3.

4 more elements

A method of evaluating efficacy of a virucidal disinfectant using a coronavirus pseudovirus, comprising the following steps:

(1) construction of virus-contaminated environment simulating distribution of a target virus under a virucidal disinfectant evaluation scene, including the existence of medium, temperature, humidity and gas disturbance, through analysis of virus contamination distribution models;
diluting the packaged coronavirus pseudovirus, uniformly smearing the diluted coronavirus pseudovirus on a medium, and setting environmental parameters of evaluation scenarios;
(2) determination of concentration of coronavirus pseudovirus before virucidal disinfection treatment based on the evaluation requirements, performing standard virus characteristic detection before treatment with sampling the coronavirus pseudovirus at different positions and different points;
and
(3) sampling and determination during and after virucidal disinfection treatment uniformly spraying or smearing the virucidal disinfectant on the medium;
based on the evaluation requirements, selecting the positions and points selected in the step (2), sampling the coronavirus pseudovirus at different times, and detecting titer activity of the coronavirus pseudovirus.

3 more elements

Dependencies are inferred from the claim text.

Coronavirus pseudovirus packaging system, packaging method therefor, and application of coronavirus pseudovirus in evaluating disinfection efficacy

Life of this patent

Coronavirus pseudovirus packaging system, packaging method therefor, and application of coronavirus pseudovirus in evaluating disinfection efficacy

Life of this patent

Abstract

Claims10