Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein / 病毒学报
Chinese Journal of Virology
;
(6): 440-446, 2007.
Artículo
en Chino
| WPRIM
| ID: wpr-334868
ABSTRACT
The severe acute respiratory syndrome-associated coronavirus (SARS-CoV) spike protein (S) is a major target for neutralizing antibody. To develop and apply a safe neutralization assay for SARS-CoV, lentiviral SARS-CoV S pseudotypes had been constructed based on a three plasmid system, which contained pVRC8304 (harboring codon optimized full-length SARS-CoV S protein), pCMV delta 8. 2 (HIV-1 gag/pol construct) and pHR'CMV EGFP (the green fluorescent protein reporter construct). The pseudo-typed lentiviral particles were used to develop an in vitro microneutralization assay that was both sensitive and specific for SARS-CoV neutralizing antibody. We used this assay to determine the titers of the neutralizing antibodies (Nabs) in serum samples from mice immunized with various rVVs expressing different S fragments of SARS-CoV. The serum antibodies derived from S and various segments of S1 region neutralized SARS-CoV in vitro. No cross-neutralization occurred with the goat antiserum prepared with inactivated HCoV-OC43 or HCoV-229E. Neutralization titers measured by this assay were highly parallel with those measured by the assay using live SARS-CoV. Because the pseudotype assay does not require handling live SARS virus, it is a useful tool to determine serum neutralizing titers during natural infection and the preclinical evaluation of candidate vaccines.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
Proyectos de Investigación
/
Sangre
/
Proteínas Recombinantes
/
Pruebas de Neutralización
/
Glicoproteínas de Membrana
/
Proteínas del Envoltorio Viral
/
Western Blotting
/
Lentivirus
/
Coronavirus Relacionado al Síndrome Respiratorio Agudo Severo
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Virology
Año:
2007
Tipo del documento:
Artículo
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