Antigen-capture ELISA for the detection of Rift Valley fever virus nucleoprotein using new monoclonal antibodies

J Virol Methods. 2012 Mar;180(1-2):68-74. doi: 10.1016/j.jviromet.2011.12.013. Epub 2012 Jan 5.

Abstract

Monoclonal antibodies (MAbs) raised against the nucleoprotein (NP) of Rift Valley fever virus (RVFV) were developed, and an antigen-capture enzyme-linked immunosorbent assay (Ag-capture ELISA) system was developed for the detection of RVFV NP. The assay detected RVFV antigen from culture supernatants containing as little as 7.8-31.3 pfu per 100 μl. Reactivity with various truncated NPs indicated that MAb C10-54 bound only to the full-length NP, probably due to recognition of a conformational epitope, whereas MAbs G2-36 and D5-59 bound to a linear epitope ranging from amino acid residues 195-201 in the C-terminal region. Based on the alignments of the amino acid sequence of RVFV NP, the epitope regions of MAbs G2-36 and D5-59 were completely conserved among all RVFV strains. These results suggest that the MAbs are applicable to the Ag-capture ELISA for the diagnosis of RVFV infections.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal* / chemistry
  • Antibodies, Monoclonal* / immunology
  • Antibodies, Viral* / chemistry
  • Antibodies, Viral* / immunology
  • Antigens, Viral / analysis*
  • Antigens, Viral / chemistry
  • Antigens, Viral / immunology
  • Cells, Cultured
  • Chlorocebus aethiops
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitope Mapping
  • Epitopes / chemistry
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Nucleocapsid Proteins / analysis*
  • Nucleocapsid Proteins / chemistry
  • Nucleocapsid Proteins / immunology
  • Rift Valley Fever / diagnosis*
  • Rift Valley Fever / immunology
  • Rift Valley fever virus / immunology*
  • Sequence Alignment
  • Spodoptera
  • Vero Cells

Substances

  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Epitopes
  • Nucleocapsid Proteins