title: Recombinant Envelope-Proteins with Mutations in the Conserved Fusion Loop Allow Specific Serological Diagnosis of Dengue-Infections.
abstract: Dengue virus (DENV) is a mosquito-borne flavivirus and a major international public health concern in many tropical and sub-tropical areas worldwide. DENV is divided into four major serotypes, and infection with one serotype leads to immunity against the same, but not the other serotypes. The specific diagnosis of DENV-infections via antibody-detection is problematic due to the high degree of cross-reactivity displayed by antibodies against related flaviviruses, such as West Nile virus (WNV), Yellow Fever virus (YFV) or Tick-borne encephalitis virus (TBEV). Especially in areas where several flaviviruses co-circulate or in the context of vaccination e.g. against YFV or TBEV, this severely complicates diagnosis and surveillance. Most flavivirus cross-reactive antibodies are produced against the highly conserved fusion loop (FL) domain in the viral envelope (E) protein. We generated insect-cell derived recombinant E-proteins of the four DENV-serotypes which contain point mutations in the FL domain. By using specific mixtures of these mutant antigens, cross-reactivity against heterologous flaviviruses was strongly reduced, enabling sensitive and specific diagnosis of the DENV-infected serum samples in IgG and IgM-measurements. These results have indications for the development of serological DENV-tests with improved specificity.
authors.affiliation: Department of Immunology, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Rockstroh
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Barzon
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Monia
authors.initials: M
authors.lastname: Pacenti
authors.affiliation: Department of Molecular Medicine, University of Padova, Padova, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palù
authors.affiliation: Robert Koch Institute, Berlin, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Niedrig
authors.affiliation: Department of Immunology, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Ulbert
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D002460:Cell Line; D003429:Cross Reactions; D003715:Dengue / Q000175:diagnosis*; D003716:Dengue Virus / Q000276:immunology*; D004330:Drosophila; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D050505:Mutant Proteins / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D050505:Mutant Proteins; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins