Detection by Enzyme-Linked Immunosorbent Assay of Antibodies to<i>West Nile virus</i>in Birds

Ebel, Gregory D.; Dupuis, Alan P.; Nicholas, David; Young, Donna M.; Maffei, Joseph G.; Kramer, Laura D.

doi:10.3201/eid0809.020152

Cited by 113 publications

(79 citation statements)

References 13 publications

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“…Blood was tested for flavivirus antibodies using an enzyme-linked immunosorbent assay (ELISA; Ebel et al 2002). We tested 346 flavivirus positive samples from birds trapped in 2003 by plaquereduction neutralization test (PRNT; Calisher et al 1989) to distinguish between WNV and St Louis encephalitis virus (SLEV), and found no evidence of SLEV exposure.…”

Section: Methodsmentioning

confidence: 99%

Host heterogeneity dominates West Nile virus transmission

Kilpatrick¹,

Daszak²,

et al. 2006

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Heterogeneity in host populations and communities can have large effects on the transmission and control of a pathogen. In extreme cases, a few individuals give rise to the majority of secondary infections, which have been termed super spreading events. Here, we show that transmission of West Nile virus (WNV) is dominated by extreme heterogeneity in the host community, resulting in highly inflated reproductive ratios. A single relatively uncommon avian species, American robin (Turdus migratorius), appeared to be responsible for the majority of WNV-infectious mosquitoes and acted as the species equivalent of a super spreader for this multi-host pathogen. Crows were also highly preferred by mosquitoes at some sites, while house sparrows were significantly avoided. Nonetheless, due to their relative rarity, corvids (crows and jays) were relatively unimportant in WNV amplification. These results challenge current beliefs about the role of certain avian species in WNV amplification and demonstrate the importance of determining contact rates between vectors and host species to understand pathogen transmission dynamics.

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Section: Methodsmentioning

confidence: 99%

Host heterogeneity dominates West Nile virus transmission

Kilpatrick¹,

Daszak²,

et al. 2006

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“…Mice that exhibited severe disease were euthanized. Serum was harvested from surviving mice at the termination of the study, and infection was confirmed by an enzyme-linked immunosorbent assay (ELISA) detecting antibodies against WNV, essentially as described (12). For tissue tropism and viral load studies, mice were sacrificed at various times postinoculation (p.i.)…”

Section: Cellsmentioning

confidence: 99%

Nonconsensus West Nile Virus Genomes Arising during Mosquito Infection Suppress Pathogenesis and Modulate Virus Fitness In Vivo

Ebel

Fitzpatrick

Lim

et al. 2011

J Virol

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West Nile virus (WNV) is similar to other RNA viruses in that it forms genetically complex populations within hosts. The virus is maintained in nature in mosquitoes and birds, with each host type exerting distinct influences on virus populations. We previously observed that prolonged replication in mosquitoes led to increases in WNV genetic diversity and diminished pathogenesis in mice without remarkable changes to the consensus genome sequence. We therefore sought to evaluate the relationships between individual and group phenotypes in WNV and to discover novel viral determinants of pathogenesis in mice and fitness in mosquitoes and birds. Individual plaque size variants were isolated from a genetically complex population, and mutations conferring a small-plaque and mouse-attenuated phenotype were localized to the RNA helicase domain of the NS3 protein by reverse genetics. The mutation, an Asp deletion, did not alter type I interferon production in the host but rendered mutant viruses more susceptible to interferon compared to wild type (WT) WNV. Finally, we used an in vivo fitness assay in Culex quinquefasciatus mosquitoes and chickens to determine whether the mutation in NS3 influenced fitness. The fitness of the NS3 mutant was dramatically lower in chickens and moderately lower in mosquitoes, indicating that RNA helicase is a major fitness determinant of WNV and that the effect on fitness is host specific. Overall, this work highlights the complex relationships that exist between individual and group phenotypes in RNA viruses and identifies RNA helicase as an attenuation and fitness determinant in WNV.West Nile virus (WNV; Flaviviridae: Flavivirus) is a positivesense, single-stranded RNA (ssRNA) virus belonging to the Japanese encephalitis virus (JEV) serological complex of the flaviviruses. WNV perpetuates in nature in enzootic transmission cycles through alternating replication in (mainly avian) vertebrates and mosquitoes. The specific mosquito and avian hosts that are most important in a particular locality differ but tend to include Culex species mosquitoes and passerine birds (2, 10). WNV infection of mammals, including humans and horses, occurs via spillover from this enzootic cycle. Since its introduction into North America in 1999 (20), molecular epidemiologic studies have clearly demonstrated that the virus has evolved to maximize its transmission potential within local transmission cycles (8,11,23). This finding stimulated subsequent efforts to understand the details of the underlying evolutionary mechanisms that lead to population-level genetic and phenotypic changes in the virus. In brief, these studies demonstrated that WNV populations are genetically diverse within hosts (15), that genetic diversity may be shared between hosts (15), and that mosquito infection drives genetic diversification of the virus population both through relaxation of purifying selection and through selection of rare genotypes resulting from RNA interference (RNAi) (4,16,17). Thus, in the WNV transmission cycle, different...

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“…Assays were developed for both antibody and virus detection. WNV-specific antibody was detected by standard indirect enzyme-linked immunosorbent assay (6) and plaque reduction neutralization assays. Detection of virus was accomplished through molecular procedures that detected viral nucleic acid, cell culture procedures that detected live virus, and immunologic procedures that detected viral antigen.…”

Section: West Nile Virus (Wnv) Belongs To the Family Flaviviridaementioning

confidence: 99%

Virus Detection Protocols for West Nile Virus in Vertebrate and Mosquito Specimens

et al. 2003

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The recent outbreaks of West Nile virus (WNV) infection in the northeastern West Nile virus (WNV) belongs to the family Flaviviridae.The virion contains single-stranded, positive-sense RNA approximately 11 kb in length. In the summer and fall of 1999, an outbreak of WNV infection in the northeastern United States was responsible for 62 human cases, including seven deaths. Extensive mortality in crows (Corvus spp.) and deaths of several exotic birds at a zoological park in the same geographic area were concurrently observed (13). This was the first evidence

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Detection by Enzyme-Linked Immunosorbent Assay of Antibodies toWest Nile virusin Birds

Cited by 113 publications

References 13 publications

Host heterogeneity dominates West Nile virus transmission

Host heterogeneity dominates West Nile virus transmission

Nonconsensus West Nile Virus Genomes Arising during Mosquito Infection Suppress Pathogenesis and Modulate Virus Fitness In Vivo

Virus Detection Protocols for West Nile Virus in Vertebrate and Mosquito Specimens

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