The invention relates to the technical field of
virus detection, in particular to an SARS-CoV-2 pseudovirus vector construction and
verification method which comprises the following steps: selecting target genes based on novel
coronavirus, connecting
nucleotide sequences of the target genes in series, and
cloning the
nucleotide sequences to a
lentivirus packaging
expression vector to obtain plasmids; transforming the
plasmid and a second-generation
lentivirus packaging
plasmid into an
escherichia coli cloned strain, carrying out enlarged culture, and extracting an endotoxin-free
plasmid; co-transfecting cells with the endotoxin-free plasmids, and collecting pseudovirus particles containing
target gene RNA (Ribonucleic Acid) from supernate of the transfected cells; precipitating pseudovirus particles, discarding supernate, and resuspending and concentrating by using a
virus dissolving solution; according to the method, a
standard curve is drawn for a
target gene,
RNA extraction, reverse transcription and fluorescent quantitative PCR are performed on pseudovirus particles, the
titer of the pseudovirus is calculated through the
standard curve, the pseudovirus is obtained, the pseudovirus is not infectious, but the
genome of the pseudovirus is similar to that of a real
virus, so that the method can be safely used in a laboratory, and the
biological safety risk in operation is reduced.