|
Basic Characteristics of Mutations
|
|
Mutation Site
|
G215C |
|
Mutation Site Sentence
|
By constructing VLP pseudotyped viruses with natural mutations in the N protein, we discovered that point mutations P13L, P80R, P199L, S202R, R203M, G204R, G215C, and S235F enhanced the titers of the VLP pseudotyped viruses (Figure 1D). |
|
Mutation Level
|
Amino acid level |
|
Mutation Type
|
Nonsynonymous substitution |
|
Gene/Protein/Region
|
N |
|
Standardized Encoding Gene
|
N
|
|
Genotype/Subtype
|
- |
|
Viral Reference
|
-
|
|
Functional Impact and Mechanisms
|
|
Disease
|
-
|
|
Immune
|
- |
|
Target Gene
|
-
|
|
Clinical and Epidemiological Correlations
|
|
Clinical Information
|
- |
|
Treatment
|
- |
|
Location
|
China |
|
Literature Information
|
|
PMID
|
38881676
|
|
Title
|
Optimization and validation of a virus-like particle pseudotyped virus neutralization assay for SARS-CoV-2
|
|
Author
|
Liu S,Zhang L,Fu W,Liang Z,Yu Y,Li T,Tong J,Liu F,Nie J,Lu Q,Lu S,Huang W,Wang Y
|
|
Journal
|
MedComm
|
|
Journal Info
|
2024 Jun 14;5(6):e615
|
|
Abstract
|
Spike-protein-based pseudotyped viruses were used to evaluate vaccines during the COVID-19 pandemic. However, they cannot be used to evaluate the envelope (E), membrane (M), and nucleocapsid (N) proteins. The first generation of virus-like particle (VLP) pseudotyped viruses contains these four structural proteins, but their titers for wild-type severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are relatively low, even lower for the omicron variant, rendering them unsuitable for neutralizing antibody detection. By optimizing the spike glycoprotein signal peptide, substituting the complexed M and E proteins with SARS-COV-1, optimizing the N protein with specific mutations (P199L, S202R, and R203M), and truncating the packaging signal, PS9, we increased the titer of the wild-type VLP pseudotyped virus over 100-fold, and successfully packaged the omicron VLP pseudotyped virus. The SARS-CoV-2 VLP pseudotyped viruses maintained stable titers, even through 10 freeze-thaw cycles. The key neutralization assay parameters were optimized, including cell type, cell number, and viral inoculum. The assay demonstrated minimal variation in both intra- and interassay results, at 11.5% and 11.1%, respectively. The correlation between the VLP pseudotyped virus and the authentic virus was strong (r = 0.9). Suitable for high-throughput detection of various mutant strains in clinical serum. In summary, we have developed a reliable neutralization assay for SARS-CoV-2 based on VLP pseudotyped virus.
|
|
Sequence Data
|
-
|
|
|