scholarly journals Rapid assessment of West Nile virus circulation in a German zoo based on honey-baited FTA cards in combination with box gravid traps

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Noelle Fynmore ◽  
Renke Lühken ◽  
Heike Maisch ◽  
Tina Risch ◽  
Sabine Merz ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Rapid Assessment ◽  
Gravid Female ◽  
Virus Detection ◽  
Turnaround Time ◽  
Viral Rna ◽  
Mass Trapping ◽  
Vector Surveillance ◽  
West Nile ◽  
Fta Cards

Abstract Background For over a decade, monitoring of West Nile virus (WNV) in Germany has consisted of a bird monitoring programme as well as a mosquito-based surveillance programme employing CO2-baited encephalitis vector surveillance (EVS) traps for mass trapping and screening of mosquitoes. In contrast to the EVS traps, the Reiter/Cummings type box gravid trap collects gravid female mosquitoes, which have already taken a blood meal, increasing the likelihood of being infected with pathogens. The traps can be equipped with a honey-baited Flinders Technology Associates® (FTA) card to encourage sugar feeding by the trapped mosquitoes. FTA cards contain nucleic acid preserving substances, which prevent the degradation of viral RNA in the expectorated mosquito saliva and allows for testing the card for flavivirus RNA. This study aimed to assess the suitability of the method for WNV surveillance in Germany as an alternative to previous methods, which are expensive, time-consuming, and predominantly target host-seeking populations less likely to be infected with WNV. Methods In the Thüringer Zoopark Erfurt, snowy owls (Nyctea scandiaca) and greater flamingos (Phoenicopterus roseus) died of WNV infections in July and August 2020. In response, five Reiter/Cummings type box gravid traps were positioned during the daytime on the 10th, 13th, and 16th of September in five different locations. The FTA cards and mosquitoes in the chamber were collected, kept in a cool chain, and further processed for virus detection using a modified generic flavivirus reverse transcription PCR. Results A total of 15 trappings during September collected a total of 259 female mosquitoes, 97% of which were Culex pipiens sensu lato, as well as 14 honey-baited FTA cards. Eight mosquitoes tested PCR-positive for WNV. Four FTA cards tested PCR-positive for mosquito-borne flaviviruses, two of which were confirmed as WNV, and the remaining two confirmed as Usutu virus. Conclusion The suitability of the FTA cards in preserving viral RNA in the field and rapid turnaround time from collection to result is combined with a simple, cost-effective, and highly specific trapping method to create an arbovirus surveillance system, which circumvents many of the difficulties of previous surveillance programmes that required the analysis of mosquitoes in the laboratory. Graphical Abstract

Stability of West Nile Virus (Flaviviridae: Flavivirus) RNA in Mosquito Excreta

2019 ◽  
Vol 56 (4) ◽  
pp. 1135-1138 ◽  
Author(s):  
Ana L Ramírez ◽  
Sonja Hall-Mendelin ◽  
Glen R Hewitson ◽  
Jamie L McMahon ◽  
Kyran M Staunton ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Control Measures ◽  
Viral Rna ◽  
Limited Area ◽  
Sample Type ◽  
West Nile ◽  
Fta Cards ◽  
Tropical Conditions ◽  
Significant Difference ◽  
Arbovirus Surveillance

Abstract Arbovirus surveillance is crucial for the implementation of vector-borne disease control measures. Recently, it has been demonstrated that mosquitoes with a disseminated arbovirus infection excrete viral RNA, which can be detected by molecular methods. Thereby, mosquito excreta has been proposed as a sample type that could be utilized for arbovirus surveillance. In this study, we evaluated if West Nile virus (Kunjin strain, WNVKUN) RNA in Culex annulirostris Skuse (Diptera: Culicidae) excreta deposited on different substrates could be detected after storage for up to 2 wk at tropical conditions of high heat and humidity. No significant drop in relative quantity of WNVKUN RNA (determined by comparison of Ct values) in excreta deposited on Flinders Associate Technologies (FTA) cards was observed over 14 d, suggesting that RNA was stable for that time. There was no significant difference in relative quantity of WNVKUN RNA in excreta deposited on FTA cards or polycarbonate substrates after 24 h. However, after 7 and 14 d, there was a significant decline in the relative quantity of viral RNA in the excreta stored on polycarbonate substrates. For incorporation in arbovirus surveillance programs, we recommend the use of polycarbonate substrates for excreta collection in mosquito traps deployed overnight, and the integration of FTA cards in traps serviced weekly or fortnightly. Polycarbonate substrates facilitate the collection of the majority of excreta from a trap, and while FTA cards offer limited area coverage, they enable preservation of viral RNA in tropical conditions for extended periods of time.

scholarly journals First record of West Nile Virus detection inside wild mosquitoes in Khartoum capital of Sudan using PCR

2020 ◽  
Vol 27 (12) ◽  
pp. 3359-3364
Author(s):  
Rania Ali El Hadi Mohamed ◽  
Deena M. Abdelgadir ◽  
Hind M. Bashab ◽  
Laila A. Al-Shuraym ◽  
Fadilah Sfouq Aleanizy ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Virus Detection ◽  
First Record ◽  
West Nile

Establishment and evaluation of real-time PCR for West Nile virus detection

2009 ◽  
Vol 6 (1) ◽  
pp. 55-59 ◽  
Author(s):  
Shi Li-Jun ◽  
Lu Mao-Min ◽  
Li Gang ◽  
Li Cheng-Yao ◽  
Zhang Jin-Gang
Keyword(s):  
West Nile Virus ◽  
Real Time ◽  
Virus Detection ◽  
Rt Pcr ◽  
Pcr Assay ◽  
West Nile ◽  
Capsid Protein Gene ◽  
Specific Test ◽  
Intercalating Dye ◽  
Polymerase Chain

AbstractA rapid real-time polymerase chain reaction (RT-PCR) for detecting West Nile virus (WNV) was established. Primers were designed according to the sequence of the capsid protein gene of WNV by Primer Premier 5.0. In this way, an inexpensive assay using the intercalating dye SYBR Green I was developed and validated. The amplifying curve showed that this method could successfully amplify 102 copies/μl of the WNV gene, while reference to Japanese encephalitis virus (JEV) and blank control were all negative. Tenfold successive dilutions of positive WNV DNA were used to measure the sensitivity of RT-PCR. The assay system showed high reproducibility with coefficient of variation (CV) <2%. Thus the newly established RT-PCR assay was shown to be a rapid, sensitive and specific test for detecting WNV.

scholarly journals West Nile virus strain Kunjin NS5 polymerase is a phosphoprotein localized at the cytoplasmic site of viral RNA synthesis

2007 ◽  
Vol 88 (4) ◽  
pp. 1163-1168 ◽  
Author(s):  
Jason M. Mackenzie ◽  
Mark T. Kenney ◽  
Edwin G. Westaway
Keyword(s):  
West Nile Virus ◽  
Rna Polymerase ◽  
Virus Strain ◽  
Viral Rna ◽  
West Nile ◽  
Replication Complex ◽  
Virus Rna ◽  
West Nile Virus Strain ◽  
First Time ◽  
Cytoplasmic Location

Using West Nile virus strain Kunjin virus (WNVKUN) as a model system for flavivirus replication, we showed that the virus replication complex (RC) is associated with the dsRNA template located in induced membranes only in the cytoplasm. In this report we established for the first time that the RNA-dependent RNA polymerase NS5 is located in flavivirus-induced membranes, including the site of viral RNA replication. We found no evidence for nuclear localization of the essential RC components NS5 and its dsRNA template for WNVKUN or the closely related WNV strain Sarafend, by immuno-electron microscopy or by immunofluorescence. Metabolic radiolabelling with [32P]orthophosphate revealed that WNVKUN NS5 was phosphorylated and this was confirmed by Western blotting with antibodies specific for phosphorylated serine and threonine only. These observations of a cytoplasmic location for the WNV polymerase and its phosphorylation state correspond to the characteristics of the hepatitis C virus RNA polymerase NS5B.

scholarly journals West Nile Virus Detection and Commercial Assays

2005 ◽  
Vol 11 (7) ◽  
pp. 1154-1155 ◽  
Author(s):  
Peter A.G. Tilley ◽  
Gail A. Zachary ◽  
Roberta Walle ◽  
Paul F. Schnee
Keyword(s):  
West Nile Virus ◽  
Virus Detection ◽  
West Nile

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

2003 ◽  
Vol 41 (8) ◽  
pp. 3661-3667 ◽  
Author(s):  
E. B. Kauffman ◽  
S. A. Jones ◽  
A. P. Dupuis ◽  
K. A. Ngo ◽  
K. A. Bernard ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Virus Detection ◽  
West Nile

scholarly journals Development of a Double Antibody Sandwich ELISA for West Nile Virus Detection Using Monoclonal Antibodies against Non-Structural Protein 1

PLoS ONE ◽  
2014 ◽  
Vol 9 (10) ◽  
pp. e108623 ◽  
Author(s):  
Xi-Xia Ding ◽  
Xiao-Feng Li ◽  
Yong-Qiang Deng ◽  
Yong-Hui Guo ◽  
Wei Hao ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
West Nile Virus ◽  
Sandwich Elisa ◽  
Virus Detection ◽  
Structural Protein ◽  
West Nile ◽  
Double Antibody

scholarly journals Cell Proteins TIA-1 and TIAR Interact with the 3′ Stem-Loop of the West Nile Virus Complementary Minus-Strand RNA and Facilitate Virus Replication

2002 ◽  
Vol 76 (23) ◽  
pp. 11989-12000 ◽  
Author(s):  
W. Li ◽  
Y. Li ◽  
N. Kedersha ◽  
P. Anderson ◽  
M. Emara ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Rna Binding ◽  
Peptide Sequencing ◽  
Viral Rna ◽  
Cell Protein ◽  
Affinity Column ◽  
Rna Recognition Motif ◽  
The West ◽  
West Nile ◽  
Stem Loop

ABSTRACT It was reported previously that four baby hamster kidney (BHK) proteins with molecular masses of 108, 60, 50, and 42 kDa bind specifically to the 3′-terminal stem-loop of the West Nile virus minus-stand RNA [WNV 3′(−) SL RNA] (P. Y. Shi, W. Li, and M. A. Brinton, J. Virol. 70:6278-6287, 1996). In this study, p42 was purified using an RNA affinity column and identified as TIAR by peptide sequencing. A 42-kDa UV-cross-linked viral RNA-cell protein complex formed in BHK cytoplasmic extracts incubated with the WNV 3′(−) SL RNA was immunoprecipitated by anti-TIAR antibody. Both TIAR and the closely related protein TIA-1 are members of the RNA recognition motif (RRM) family of RNA binding proteins. TIA-1 also binds to the WNV 3′(−) SL RNA. The specificity of these viral RNA-cell protein interactions was demonstrated using recombinant proteins in competition gel mobility shift assays. The binding site for the WNV 3′(−) SL RNA was mapped to RRM2 on both TIAR and TIA-1. However, the dissociation constant (Kd ) for the interaction between TIAR RRM2 and the WNV 3′(−) SL RNA was 1.5 × 10−8, while that for TIA-1 RRM2 was 1.12 × 10−7. WNV growth was less efficient in murine TIAR knockout cell lines than in control cells. This effect was not observed for two other types of RNA viruses or two types of DNA viruses. Reconstitution of the TIAR knockout cells with TIAR increased the efficiency of WNV growth, but neither the level of TIAR nor WNV replication was as high as in control cells. These data suggest a functional role for TIAR and possibly also for TIA-1 during WNV replication.

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