Rapid and specific detection of Sin Nombre virus antibodies in patients with hantavirus pulmonary syndrome by a strip immunoblot assay suitable for field diagnosis

Abstract

To develop a rapid antibody test for Sin Nombre hantavirus (SNV) infection for diagnosis of hantavirus pulmonary syndrome (HPS) in field settings where advanced instrumentation is not available, a strip immunoblot assay bearing four immobilized antigens of SNV and a recombinant nucleocapsid protein antigen of Seoul hantavirus (SEOV) was prepared. The SNV antigens included a full-length recombinant-expressed nucleocapsid (N) protein (rN), a recombinant-expressed G1 protein (residues 35 to 117), and synthetic peptides derived from N (residues 17 to 59) and G1 (residues 58 to 88). On the basis of the observed reactivities of hantavirus-infected patient and control sera, we determined that a positive assay requires reactivity with SNV or SEOV rN antigen and at least one other antigen. Isolated reactivity to either viral rN antigen is indeterminate, and any pattern of reactivity that does not include reactivity to an rN antigen is considered indeterminate but is unlikely to represent hantavirus infection. Fifty-eight of 59 samples from patients with acute SNV-associated HPS were positive according to these criteria, and one was initially indeterminate. Four of four samples from patients with HPS due to other hantaviruses were positive, as were most samples from patients with SEOV and Puumala virus infections. Of 192 control serum samples, 2 (1%) were positive and 2 were indeterminate. Acute SNV infection was distinguishable from remote SNV infection or infection with hantaviruses other than SNV by the presence of G1 peptide antigen reactivities in the former. The strip immunoblot assay shows promise for the detection of SNV antibodies early in the course of HPS.

Human immune response to Puumala virus glycoproteins and nucleocapsid protein expressed in mammalian cells

“…IgG antibodies against N develop early after onset of symptoms and persist in serum for years, whereas a slower IgG response against the glycoproteins has been described earlier using EIA [Lundkvist et al, 1993;Vapalahti et al, 1995]. In other studies, the G1-antibody responses have been shown to become detectable in the acute phase of hantavirus pulmonary syndrome, but not of PUUV or Hantaan virus infections when recombinant G1 antigens expressed in bacterial or yeast systems were used [Jenison et al, 1994;Hjelle et al, 1997]. In some reports, an early antibody response to PUUV-G1 has been shown, G2 antibodies appearing later according to inhibition EIA or slot-blot assay/ EIA [Go Ètt et al, 1997].…”

High Seroprevalence of Hantavirus Infection on the Azuero Peninsula of Panama

“…Antibody to all known hantaviruses in the Americas cross-react to the N protein of Sin Nombre virus in binding assays. 15 A strip immunoblot assay (SIA) for IgG antibody containing recombinant N protein of the 3H226 genotype of Sin Nombre virus was used as described. 15 An enzyme immunoassay (EIA) used recombinant nucleocapsid protein from Sin Nombre virus.…”

A Global Perspective on Hantavirus Ecology, Epidemiology, and Disease

“…The N protein is suitable for use as an antigen in immunoenzymatic assays (EIAs) for the diagnosis of hantavirus infection (106,147) as well as strip immunoblot tests (147). However, the most common serological tests for hantaviruses are indirect IgG and IgM enzyme-linked immunosorbent assays (ELISAs) as well as IgM capture ELISAs.…”