Defining the levels of secreted non-structural protein NS1 after West Nile virus infection in cell culture and mice.
Infection with West Nile virus (WNV) causes a febrile illness that can progress to meningitis or encephalitis, primarily in humans that are immunocompromised or elderly. For successful treatment of WNV infection, accurate and timely diagnosis is essential. Previous studies have suggested that the flavivirus non-structural protein NS1, a highly conserved and secreted glycoprotein, is a candidate protein for rapid diagnosis. Herein, we developed a capture enzyme-linked immunosorbent assay (ELISA) to detect WNV NS1 using two anti-NS1 monoclonal antibodies (mAbs) that map to distinct sites on the protein. The capture ELISA efficiently detected as little as 0.5 ng/ml of soluble NS1 and exhibited no cross-reactivity for yellow fever, Dengue, and St. Louis encephalitis virus NS1. The capture ELISA reliably detected NS1 in plasma at day 3 after WNV infection, prior to the development of clinical signs of disease. As the time course of infection continued, the levels of detectable NS1 diminished, presumably because of interference by newly generated anti-NS1 antibodies. Indeed, treatment of plasma with a solution that dissociated NS1 immune complexes extended the window of detection. Overall, the NS1-based capture ELISA is a sensitive readout of infection and could be an important tool for diagnosis or screening small molecule inhibitors of WNV infection.
Chung
Kyung Min
KM
Department of Medicine, Washington University School of Medicine, St Louis, Missouri 63110, USA.
Diamond
Michael S
MS
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D012367:RNA, Viral / Q000097:blood / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D014766:Viremia / Q000276:immunology / Q000821:virology; D014901:West Nile Fever / Q000175:diagnosis* / Q000821:virology; D014902:West Nile virus / Q000276:immunology*
D000914:Antibodies, Viral; C100315:NS1 protein, Flavivirus; D012367:RNA, Viral; D011994:Recombinant Proteins; D017361:Viral Nonstructural Proteins