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Optimized enzyme-linked immunosorbent assay for detecting cytomegalovirus infections during clinical trials of recombinant vaccines.
Pagnon, Anke; Piras, Fabienne; Gimenez-Fourage, Sophie; Dubayle, Joseline; Arnaud-Barbe, Nadège; Hessler, Catherine; Caillet, Catherine.
Afiliação
  • Pagnon A; Sanofi Pasteur, Marcy l'Etoile, France. Electronic address: anke.pagnon@sanofi.com.
  • Piras F; Sanofi Pasteur, Marcy l'Etoile, France.
  • Gimenez-Fourage S; Sanofi Pasteur, Marcy l'Etoile, France.
  • Dubayle J; Sanofi Pasteur, Marcy l'Etoile, France.
  • Arnaud-Barbe N; Sanofi Pasteur, Marcy l'Etoile, France.
  • Hessler C; Sanofi Pasteur, Marcy l'Etoile, France.
  • Caillet C; Sanofi Pasteur, Marcy l'Etoile, France.
J Clin Virol ; 96: 99-104, 2017 11.
Article em En | MEDLINE | ID: mdl-29035763
BACKGROUND: In clinical trials of cytomegalovirus (CMV) glycoprotein B (gB) vaccines, CMV infection is detected by first depleting serum of anti-gB antibodies and then measuring anti-CMV antibodies with a commercially available enzyme-linked immunosorbent assay (ELISA) kit, with confirmation of positive findings by immunoblot. OBJECTIVES: Identification of CMV immunoantigens for the development of an ELISA that detects specifically CMV infection in clinical samples from individuals immunized with gB vaccines. STUDY DESIGN: Sensitivity and specificity of ELISAs using antigenic regions of CMV proteins UL83/pp65, UL99/pp28, UL44/pp52, UL80a/pp38, UL57, and UL32/pp150 were measured. RESULTS: An IgG ELISA using a UL32/pp150 [862-1048] capture peptide was the most specific (93.7%) and sensitive (96.4%) for detecting CMV-specific antibodies in sera. The ELISA successfully detected CMV-specific antibodies in 22 of 22 sera of subjects who had been vaccinated with a gB vaccine but who had later been infected with CMV. The ELISA was linear over a wide range of CMV concentrations (57-16,814 ELISA units/mL) and was reproducible as indicated by a 5% intra-day and 7% inter-day coefficients of variation. The signal was specifically competed by UL32/pp150 [862-1048] peptide but not by CMV-gB or herpes simplex virus 2 glycoprotein D. Lipid and hemoglobin matrix did not interfere with the assay. CONCLUSION: The UL32/pp150 [862-1048] IgG ELISA can be used for the sensitive and specific detection of CMV infection in gB-vaccinated individuals.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ensaio de Imunoadsorção Enzimática / Ensaios Clínicos como Assunto / Infecções por Citomegalovirus / Vacinas contra Citomegalovirus / Testes Diagnósticos de Rotina / Anticorpos Antivirais Tipo de estudo: Diagnostic_studies / Evaluation_studies / Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Ensaio de Imunoadsorção Enzimática / Ensaios Clínicos como Assunto / Infecções por Citomegalovirus / Vacinas contra Citomegalovirus / Testes Diagnósticos de Rotina / Anticorpos Antivirais Tipo de estudo: Diagnostic_studies / Evaluation_studies / Prognostic_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article