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Differentiation between human coronaviruses NL63 and 229E using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies.
Clin Vaccine Immunol. 2011 Jan; 18(1):113-8.CV

Abstract

Human coronaviruses (HCoVs) are responsible for respiratory tract infections ranging from common colds to severe acute respiratory syndrome. HCoV-NL63 and HCoV-229E are two of the four HCoVs that circulate worldwide and are close phylogenetic relatives. HCoV infections can lead to hospitalization of children, elderly individuals, and immunocompromised patients. Globally, approximately 5% of all upper and lower respiratory tract infections in hospitalized children are caused by HCoV-229E and HCoV-NL63. The latter virus has recently been associated with the childhood disease croup. Thus, differentiation between the two viruses is relevant for epidemiology studies. The aim of this study was to develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) as a potential tool for identification and differentiation between HCoV-NL63 and HCoV-229E. The nucleocapsid (N) proteins of HCoV-NL63 and HCoV-229E were expressed in an Escherichia coli system and used to immunize mice in order to obtain monoclonal antibodies (MAbs) specific for each virus. Three specific MAbs to HCoV-NL63, one MAb specific to HCoV-229E, and four MAbs that recognized both viruses were obtained. After their characterization, three MAbs were selected in order to develop a differential DAS-ELISA. The described assay could detect up to 3 ng/ml of N protein and 50 50% tissue culture infective doses/ml of virus stock. No cross-reactivity with other human coronaviruses or closely related animal coronaviruses was found. The newly developed DAS-ELISA was species specific, and therefore, it could be considered a potential tool for detection and differentiation of HCoV-NL63 and HCoV-229E infections.

Authors+Show Affiliations

Inmunología y Genética Aplicada S.A. (INGENASA), Hnos. García Noblejas 41, 28037 Madrid, Spain. psastre@ingenasa.esNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21084464

Citation

Sastre, Patricia, et al. "Differentiation Between Human Coronaviruses NL63 and 229E Using a Novel Double-antibody Sandwich Enzyme-linked Immunosorbent Assay Based On Specific Monoclonal Antibodies." Clinical and Vaccine Immunology : CVI, vol. 18, no. 1, 2011, pp. 113-8.
Sastre P, Dijkman R, Camuñas A, et al. Differentiation between human coronaviruses NL63 and 229E using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies. Clin Vaccine Immunol. 2011;18(1):113-8.
Sastre, P., Dijkman, R., Camuñas, A., Ruiz, T., Jebbink, M. F., van der Hoek, L., Vela, C., & Rueda, P. (2011). Differentiation between human coronaviruses NL63 and 229E using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies. Clinical and Vaccine Immunology : CVI, 18(1), 113-8. https://doi.org/10.1128/CVI.00355-10
Sastre P, et al. Differentiation Between Human Coronaviruses NL63 and 229E Using a Novel Double-antibody Sandwich Enzyme-linked Immunosorbent Assay Based On Specific Monoclonal Antibodies. Clin Vaccine Immunol. 2011;18(1):113-8. PubMed PMID: 21084464.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Differentiation between human coronaviruses NL63 and 229E using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies. AU - Sastre,Patricia, AU - Dijkman,Ronald, AU - Camuñas,Ana, AU - Ruiz,Tamara, AU - Jebbink,Maarten F, AU - van der Hoek,Lia, AU - Vela,Carmen, AU - Rueda,Paloma, Y1 - 2010/11/17/ PY - 2010/11/19/entrez PY - 2010/11/19/pubmed PY - 2011/4/22/medline SP - 113 EP - 8 JF - Clinical and vaccine immunology : CVI JO - Clin Vaccine Immunol VL - 18 IS - 1 N2 - Human coronaviruses (HCoVs) are responsible for respiratory tract infections ranging from common colds to severe acute respiratory syndrome. HCoV-NL63 and HCoV-229E are two of the four HCoVs that circulate worldwide and are close phylogenetic relatives. HCoV infections can lead to hospitalization of children, elderly individuals, and immunocompromised patients. Globally, approximately 5% of all upper and lower respiratory tract infections in hospitalized children are caused by HCoV-229E and HCoV-NL63. The latter virus has recently been associated with the childhood disease croup. Thus, differentiation between the two viruses is relevant for epidemiology studies. The aim of this study was to develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) as a potential tool for identification and differentiation between HCoV-NL63 and HCoV-229E. The nucleocapsid (N) proteins of HCoV-NL63 and HCoV-229E were expressed in an Escherichia coli system and used to immunize mice in order to obtain monoclonal antibodies (MAbs) specific for each virus. Three specific MAbs to HCoV-NL63, one MAb specific to HCoV-229E, and four MAbs that recognized both viruses were obtained. After their characterization, three MAbs were selected in order to develop a differential DAS-ELISA. The described assay could detect up to 3 ng/ml of N protein and 50 50% tissue culture infective doses/ml of virus stock. No cross-reactivity with other human coronaviruses or closely related animal coronaviruses was found. The newly developed DAS-ELISA was species specific, and therefore, it could be considered a potential tool for detection and differentiation of HCoV-NL63 and HCoV-229E infections. SN - 1556-679X UR - https://www.unboundmedicine.com/medline/citation/21084464/Differentiation_between_human_coronaviruses_NL63_and_229E_using_a_novel_double_antibody_sandwich_enzyme_linked_immunosorbent_assay_based_on_specific_monoclonal_antibodies_ L2 - https://journals.asm.org/doi/10.1128/CVI.00355-10?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -