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[Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein].
Bing Du Xue Bao. 2007 Nov; 23(6):440-6.BD

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.

Authors+Show Affiliations

National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

chi

PubMed ID

18092680

Citation

Yan, Ke-Xia, et al. "[Development and Application of a Safe SARS-CoV Neutralization Assay Based On Lentiviral Vectors Pseudotyped With SARS-CoV Spike Protein]." Bing Du Xue Bao = Chinese Journal of Virology, vol. 23, no. 6, 2007, pp. 440-6.
Yan KX, Tan WJ, Zhang XM, et al. [Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein]. Bing Du Xue Bao. 2007;23(6):440-6.
Yan, K. X., Tan, W. J., Zhang, X. M., Wang, H. J., Li, Y., & Ruan, L. (2007). [Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein]. Bing Du Xue Bao = Chinese Journal of Virology, 23(6), 440-6.
Yan KX, et al. [Development and Application of a Safe SARS-CoV Neutralization Assay Based On Lentiviral Vectors Pseudotyped With SARS-CoV Spike Protein]. Bing Du Xue Bao. 2007;23(6):440-6. PubMed PMID: 18092680.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Development and application of a safe SARS-CoV neutralization assay based on lentiviral vectors pseudotyped with SARS-CoV spike protein]. AU - Yan,Ke-Xia, AU - Tan,Wen-Jie, AU - Zhang,Xiang-Min, AU - Wang,Hui-Juan, AU - Li,Yan, AU - Ruan,Li, PY - 2007/12/21/pubmed PY - 2008/1/11/medline PY - 2007/12/21/entrez SP - 440 EP - 6 JF - Bing du xue bao = Chinese journal of virology JO - Bing Du Xue Bao VL - 23 IS - 6 N2 - 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. SN - 1000-8721 UR - https://www.unboundmedicine.com/medline/citation/18092680/[Development_and_application_of_a_safe_SARS_CoV_neutralization_assay_based_on_lentiviral_vectors_pseudotyped_with_SARS_CoV_spike_protein]_ DB - PRIME DP - Unbound Medicine ER -