A newly identified hantavirus: The development of immunologic diagnostic assays and phylogenetic analysis for detection and characterization

We developed an enzyme linked immunosorbent assay (ELISA) utilizing a 59 amino acid synthetic peptide as an antigen for the serodiagnosis of hantavirus. We also developed a one-step real-time detection-PCR (RTD-PCR) assay for hantavirus detection in rodents and insectivores captured at various time points and trapsites between September 2000 and May 2004 in the Great Smoky Mountains National Park.;Blood samples from 305 rodents and insectivores were tested for anti-hantavirus IgG antibodies with our synthetic peptide ELISA and immunofluorescent assay (IFA). The sensitivity and specificity of this ELISA were comparable to those of the IFA using virus infected cells. The only seropositive animals detected were deer mice (Peromyscus maniculatus) and white-footed mice (Peromyscus leucopus). The overall estimated seroprevalence in GSMNP was 8.9% (27/305).;A total of 345 rodents and insectivores were tested by RTD-PCR and the overall estimated viral prevalence was 5.2% (18/345). We detected virus in 16 deer mice and white-footed mice, one smoky shrew (Sorex cinerus ) and one Southern red-backed vole (Clethrionomys gapperi ). This is the first reported case of a New World hantavirus being detected in a shrew and the first evidence of a hantavirus detected in a Clethrionomys spp.;We performed phylogenetic analysis on partial G1 and G 2 segments of the M gene and partial N segment of the S gene. We have designated this hantavirus strain Newfound Gap virus (NGV). The incongruous homologies of the NGV S and M segments to other closely related hantaviruses suggest that genetic reassortment resulting in a hybrid virus may have occurred. NGV possesses unique characteristics and is closely related to pathogenic strains that have resulted in HCPS case fatalities in this region.