High-Throughput Detection of West Nile Virus RNA

Shi, Pei–Yong; Kauffman, Elizabeth B.; Ren, Ping; Felton, Andy; Tai, J Y; Dupuis, Alan P.; Jones, Susan; Ngo, Kiet; Nicholas, David; Maffei, Joseph G.; Ebel, Gregory D.; Bernard, Kristen A.; Kramer, Laura D.

doi:10.1128/jcm.39.4.1264-1271.2001

Cited by 215 publications

(170 citation statements)

References 22 publications

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“…Extracted RNA was tested for WNV RNA by RT-PCR using a ViiA 7 Real-Time PCR system (Life Technologies) and SensiFAST One Step reagents (Bioline, Taunton, MA). Primers and probe that were specific for the envelope (Lanciotti et al 2001) or NS1 (Shi et al 2001) genes were used for screening and confirmation, respectively. Threshold cycle (C t ) scores were recorded and analyzed (see below) for kidney sample and cloacal swab tests.…”

Section: Methodsmentioning

confidence: 99%

West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

et al. 2015

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Since its emergence in North America, West Nile virus (WNV) has had a large impact on equines, humans, and wild bird communities, yet gaps remain in our understanding of how the virus persists at temperate latitudes when winter temperatures preclude virus replication and host-seeking activity by mosquito vectors. Bird-to-bird transmission at large communal American Crow roosts could provide one mechanism for WNV persistence. Herein, we describe seasonal patterns of crow and Culex mosquito abundance, WNV infection rates, and the prevalence of WNV-positive fecal samples at a winter crow roost to test the hypothesis that bird-to-bird transmission allows WNV to persist at winter crow roosts. Samples were collected from large winter crow roosts in the Sacramento Valley of California from January 2013 until August 2014, encompassing two overwintering roost periods. West Nile virus RNA was detected in local crow carcasses in both summer [13/18 (72% WNV positive)] and winter [18/44 (41% WNV positive)] 2013-2014. Winter infections were unlikely to have arisen by recent bites from infected mosquitoes because Culex host-seeking activity was very low in winter and all Culex mosquitoes collected during winter months tested negative for WNV. Opportunities existed for fecal-oral transfer at the overwintering roost: most carcasses that tested positive for WNV had detectable viral RNA in both kidney and cloacal swabs, suggesting that infected crows were shedding virus in their feces, and >50% of crows at the roost were stained with feces by mid-winter. Moreover, 2.3% of fecal samples collected in late summer, when mosquitoes were active, tested positive for WNV RNA. Nevertheless, none of the 1,119 feces collected from three roosts over two winters contained detectable WNV RNA. This study provided evidence of WNV infection in overwintering American crows without mosquito vector activity, but did not elucidate a mechanism of WNV transmission during winter.

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Section: Methodsmentioning

confidence: 99%

West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

et al. 2015

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“…A sample was confirmed positive when at least two different tests were positive. Details on virus testing are described elsewhere (4,5).…”

Section: Laboratory Testing Of Mosquitoes and Birdsmentioning

confidence: 99%

West Nile Virus Infection in Mosquitoes, Birds, Horses, and Humans, Staten Island, New York, 2000

Kulasekera

Kramer

Nasci

et al. 2001

Emerg. Infect. Dis.

124

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“…To determine viremia, sera were tested by standard Vero cell plaque assay. 27 Dead birds reported by the public were necropsied at the California Animal Health and Food Safety Laboratory at the University of California, Davis, and kidney samples sent to the Center for Vectorborne Diseases laboratory for testing for WNV RNA by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) 28 using previously published primers. …”

Section: Introductionmentioning

confidence: 99%

Migratory Birds and the Dispersal of Arboviruses in California

Reisen

Wheeler

Garcia

et al. 2010

The American Society of Tropical Medicine and Hygiene

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MATERIALS AND METHODS Bird collections.Migratory and resident birds were captured while transiting sites on the northeastern (35.51, −115.92) and northwestern (33.46, −116.06) shores of the Salton Sea, Coachella Valley, Riverside County, CA, from 1996 through 2005. In addition, migratory transients were targeted during collections made during the springs of 2006 and 2007. Migrants and other birds also were collected during 1996 through 2008 in Kern County, primarily from wetlands at the Kern National Wildlife Refuge (35.73,) and the Kern River (35.32,. The ecology of these areas, descriptions of arbovirus activity, and bird sampling methods have been described previously. 20,23 Briefly, birds were collected using multiple mist nets and grain-baited traps. Mist nets were 10 m long × 2.5 m tall and had 28-, 32-, or 60-mm mesh depending on the target species. Captured birds were banded with U.S. Geological Survey (USGS) bands, aged, sexed, and a 0.1 mL sample of blood collected from the jugular vein with 28-g needles. For antibody detection, blood samples were expelled into 0.9 mL of 0.9% saline solution, clarified through centrifugation, and stored at −70°C. During 2004During , 2006During , and 2007 blood samples from migratory species collected in Coachella Valley were expelled into 0.4 mL of virus diluent (phosphate buffered saline, 15% fetal bovine sera, antibiotics) and tested for infectious virus and antibody. Data from the California WNV Dead Bird Surveillance Program 24 on neotropical migrants that were found dead by the public, sent for necropsy, and tested positive for WNV RNA were included for comparison.Experimental infection. Abstract. Each spring large numbers of neotropical migrants traversing the Pacific flyway pass through the Coachella Valley enroute to northern destinations, providing an opportunity to test the hypothesis that mosquito-borne encephalitis viruses are introduced annually into California by migratory birds. A total of 5,632 sera were collected from 43 species of migrants during spring (April-June), of which 34 (0.61%) comprised of 14 species tested positive by enzyme immunoassay; only 10 were confirmed by plaque reduction neutralization tests (PRNT). In addition, of 1,109 migrants comprised of 76 species that were reported dead by the public and necropsied, 126 (11%) were positive for West Nile virus (WNV) RNA; however, only three (0.7%) of 428 birds tested during the spring were positive. Limited experimental infection studies with WNV showed that Orange-crowned Warblers were highly susceptible and frequently died, whereas most Yellow Warblers survived. Our results indicated that birds entering California rarely exhibited a history of infection and that most birds probably became infected after entering California.

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High-Throughput Detection of West Nile Virus RNA

Abstract: The recent outbreaks of West Nile virus (WNV) in the northeastern

Cited by 215 publications

References 22 publications

West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

West Nile Virus Activity in a Winter Roost of American Crows (Corvus brachyrhynchos): Is Bird-To-Bird Transmission Important in Persistence and Amplification?

West Nile Virus Infection in Mosquitoes, Birds, Horses, and Humans, Staten Island, New York, 2000

Migratory Birds and the Dispersal of Arboviruses in California

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