scholarly journals Evaluation of West Nile Virus Diagnostic Capacities in Veterinary Laboratories of the Mediterranean and Black Sea Regions

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1038
Author(s):  
Elisa Pérez-Ramírez ◽  
Cristina Cano-Gómez ◽  
Francisco Llorente ◽  
Ani Vodica ◽  
Ljubiša Veljović ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Black Sea ◽  
Simultaneous Detection ◽  
Mediterranean Area ◽  
Rt Pcr ◽  
West Nile ◽  
Detection Rates ◽  
Animal Populations ◽  
Surveillance Programs ◽  
L1 And L2

The increasing incidence of West Nile virus (WNV) in the Euro-Mediterranean area warrants the implementation of effective surveillance programs in animals. A crucial step in the fight against the disease is the evaluation of the capacity of the veterinary labs to accurately detect the infection in animal populations. In this context, the animal virology network of the MediLabSecure project organized an external quality assessment (EQA) to evaluate the WNV molecular and serological diagnostic capacities of beneficiary veterinary labs. Laboratories from 17 Mediterranean and Black Sea countries participated. The results of the triplex real time RT-PCR for simultaneous detection and differentiation of WNV lineage 1 (L1), lineage 2 (L2) and Usutu virus (USUV) were highly satisfactory, especially for L1 and L2, with detection rates of 97.9% and 100%, respectively. For USUV, 75% of the labs reported correct results. More limitations were observed for the generic detection of flaviviruses using conventional reverse-transcription polymerase chain reaction (RT-PCR), since only 46.1% reported correct results in the whole panel. As regards the serological panel, the results were excellent for the generic detection of WNV antibodies. More variability was observed for the specific detection of IgM antibodies with a higher percentage of incorrect results mainly in samples with low titers. This EQA provides a good overview of the WNV (and USUV) diagnostic performance of the involved veterinary labs and demonstrates that the implemented training program was successful in upgrading their diagnostic capacities.

A novel quantitative multiplex real-time RT-PCR for the simultaneous detection and differentiation of West Nile virus lineages 1 and 2, and of Usutu virus

2013 ◽  
Vol 189 (2) ◽  
pp. 321-327 ◽  
Author(s):  
Javier Del Amo ◽  
Elena Sotelo ◽  
Jovita Fernández-Pinero ◽  
Carmina Gallardo ◽  
Francisco Llorente ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Real Time ◽  
Simultaneous Detection ◽  
Rt Pcr ◽  
West Nile ◽  
Usutu Virus

scholarly journals Comparison of immunohistochemistry and polymerase chain reaction for detection of West Nile virus in naturally infected dead birds

2010 ◽  
Vol 4 (09) ◽  
pp. 587-589 ◽  
Author(s):  
Tejbir S Sandhu ◽  
Dalbinder S Sidhu ◽  
Major S Dhillon ◽  
Ying Fang
Keyword(s):  
West Nile Virus ◽  
Disease Surveillance ◽  
Polyclonal Antibodies ◽  
Kidney Tissue ◽  
Rt Pcr ◽  
West Nile ◽  
Fixed Tissue ◽  
Riverside County ◽  
Surveillance Programs ◽  
Sensitivity Specificity

Introduction: Credible vector-borne disease surveillance programs, especially in developing countries with limited resources, must include diagnostic tests that are efficient, inexpensive and simple and safe to administer while maintaining high levels of sensitivity and specificity. Since immunohistochemistry (IHC) includes most of these features, its sensitivity, specificity, predictive positive value (PPV) and predictive negative value (PNV) for West Nile virus (WNv) screening were compared to those of the gold standard, RT-PCR testing of kidney tissue in dead birds. Methodology: IHC and RT-PCR were performed for WNv antigen on 41 dead birds (belonging to five orders) collected from the northwest region of the Riverside County of California. Fixed tissue sections were screened by IHC using polyclonal antibodies, and frozen kidney tissues were tested with RT-PCR. Results: Kidney screening with IHC showed sensitivity, specificity, PPV and NPV of 95.45%, 73.68%, 80.77% and 93.33%, respectively. Based on WNv screening of kidney tissue, IHC and RT-PCR were in agreement with 95.45% (21/22) for positive dead birds and were in 100% (22/22) agreement when multi-organ screening by IHC was performed. Conclusions: The present study showed that IHC is as equally effective as RT-PCR in screening for WNv in dead birds. Therefore, IHC can effectively serve as a competent screening technique for those disease surveillance agencies that lack expensive RT-PCR technology while promoting safer biohazardous conditions, except at the initial stage of tissue collection.

scholarly journals Laboratory and Field Evaluations of a Commercially Available Real-Time Loop-Mediated Isothermal Amplification Assay for the Detection of West Nile Virus in Mosquito Pools

2021 ◽  
Vol 37 (4) ◽  
pp. 256-262
Author(s):  
Kristen L. Burkhalter ◽  
Michael O'Keefe ◽  
Zachary Holbert-Watson ◽  
Theodore Green ◽  
Harry M. Savage ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Real Time ◽  
Lamp Assay ◽  
Isothermal Amplification ◽  
Rt Pcr ◽  
Vector Surveillance ◽  
West Nile ◽  
Loop Mediated Isothermal Amplification ◽  
Surveillance Programs ◽  
Kappa Agreement

ABSTRACT Although the specific cDNA amplification mechanisms of reverse-transcriptase polymerase chain reaction (RT-PCR) and RT loop-mediated isothermal amplification (RT-LAMP) are very different, both molecular assays serve as options to detect arboviral RNA in mosquito pools. Like RT-PCR, RT-LAMP uses a reverse transcription step to synthesize complementary DNA (cDNA) from an RNA template and then uses target-specific primers to amplify cDNA to detectable levels in a single-tube reaction. Using laboratory-generated West Nile virus (WNV) samples and field-collected mosquito pools, we evaluated the sensitivity and specificity of a commercially available WNV real-time RT-LAMP assay (Pro-AmpRT™ WNV; Pro-Lab Diagnostics, Inc., Round Rock, Texas) and compared the results to a validated real-time RT-PCR assay. Laboratory generated virus stock samples containing ≥ 2.3 log10 plaque-forming units (PFU)/ml and intrathoracically inoculated mosquitoes containing ≥ 2.4 log10 PFU/ml produced positive results in the Pro-AmpRT WNV assay. Of field-collected pools that were WNV positive by real-time RT-PCR, 74.5% (70 of 94) were also positive by the Pro-AmpRT WNV assay, resulting in an overall Cohen's kappa agreement of 79.4% between the 2 tests. The Pro-AmpRT WNV assay shows promise as a suitable virus screening tool for vector surveillance programs provided agencies are aware of its characteristics and limitations.

scholarly journals Screening of Mosquitoes for West Nile Virus and Usutu Virus in Croatia, 2015–2020

2021 ◽  
Vol 6 (2) ◽  
pp. 45
Author(s):  
Ana Klobucar ◽  
Vladimir Savic ◽  
Marcela Curman Posavec ◽  
Suncica Petrinic ◽  
Urska Kuhar ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Culex Pipiens ◽  
Rt Pcr ◽  
West Nile ◽  
Usutu Virus ◽  
Culex Pipiens Complex ◽  
Mammalian Origin ◽  
Mosquito Pool ◽  
The City ◽  
Generation Sequencing

In the period from 2015 to 2020, an entomological survey for the presence of West Nile virus (WNV) and Usutu virus (USUV) in mosquitoes was performed in northwestern Croatia. A total of 20,363 mosquitoes were sampled in the City of Zagreb and Međimurje county, grouped in 899 pools and tested by real-time RT-PCR for WNV and USUV RNA. All pools were negative for WNV while one pool each from 2016 (Aedes albopictus), 2017 (Culex pipiens complex), 2018 (Cx. pipiens complex), and 2019 (Cx. pipiens complex), respectively, was positive for USUV. The 2018 and 2019 positive pools shared 99.31% nucleotide homology within the USUV NS5 gene and both clustered within USUV Europe 2 lineage. The next-generation sequencing of one mosquito pool (Cx. pipiens complex) collected in 2018 in Zagreb confirmed the presence of USUV and revealed several dsDNA and ssRNA viruses of insect, bacterial and mammalian origin.

The survey of wild birds for West Nile virus in Poland

2011 ◽  
Vol 14 (4) ◽  
pp. 573-577 ◽  
Author(s):  
J. Niczyporuk ◽  
E. Samorek-Salamonowicz ◽  
W. Kozdruń ◽  
Z. Mizak
Keyword(s):  
West Nile Virus ◽  
Genetic Material ◽  
Early Spring ◽  
Wild Birds ◽  
Rt Pcr ◽  
West Nile ◽  
Late Autumn ◽  
The Brain

The survey of wild birds for West Nile virus in PolandTwo thousand one hundred and forty birds belonging to 39 different species from different locations in Poland were examined. The study has taken place from the early spring till late autumn 2007-2010 when the activity of the mosquitoes was the highest. The brain samples were taken from the birds and whole cellular RNA was isolated, then the RT-PCR and NRT-PCR were performed to detect the presence of West Nile virus (WNV). The obtained results were confirmed by the commercial WNV Kit. No genetic material of WNV was found in the examined samples.

Detection of West Nile virus in formalin-fixed, paraffin-embedded human tissues by RT-PCR: A useful adjunct to conventional tissue-based diagnostic methods

2007 ◽  
Vol 38 (2) ◽  
pp. 106-111 ◽  
Author(s):  
Julu Bhatnagar ◽  
Jeannette Guarner ◽  
Christopher D. Paddock ◽  
Wun-Ju Shieh ◽  
Robert S. Lanciotti ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Diagnostic Methods ◽  
Human Tissues ◽  
Rt Pcr ◽  
West Nile ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

scholarly journals West Nile Virus and Related Flavivirus in European Wild Boar (Sus scrofa), Latium Region, Italy: A Retrospective Study

Animals ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 494
Author(s):  
Angela Petruccelli ◽  
Tiziana Zottola ◽  
Gianmarco Ferrara ◽  
Valentina Iovane ◽  
Cristina Di Russo ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Wild Boar ◽  
Specific Antibody ◽  
Central Italy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
West Nile ◽  
Wild Boars ◽  
European Wild Boar

Background: A retrospective sero-survey for evidence of West Nile virus (WNV) infection in European wild boar (Sus scorfa) was conducted in the Latium region, Italy, on stored serum samples of the period November 2011 to January 2012. Methods: Sera were collected from 168 European wild boars and screened for antibodies to WNV and other Flaviviruses by competitive enzyme linked immunosorbent assay (cELISA). All sera positive for Flavivirus antibodies by cELISA were further examined by virus neutralization test (VNT). To test the presence of Flavivirus RNA in samples, an RT-PCR was performed using a pan-Flavivirus primers pair. Results: Thirteen wild boars (7.73%) were seropositive for Flaviviruses. The hemolysis of serum samples limited the interpretation of the VNT for 7 samples, confirming the presence of specific antibody against WNV in a single European wild boar serum sample. The presence of ELISA positive/VNT negative samples suggests the occurrence of non-neutralizing antibodies against WNV or other antigen-related Flaviviruses. No samples resulted positive for Flavivirus by RT-PCR assay. Conclusion: Although a moderately high percentage of animals with specific antibody for WNV has been detected in wild boar in other surveillance studies in Europe, this has not been reported previously in Italy. Together, these data indicate that European wild boar are exposed to WNV and/or other related-Flavivirus in central Italy and confirm the usefulness of wild ungulates, as suitable Flavivirus sentinels.

Simultaneous detection of Dengue virus, Chikungunya virus, Zika virus, Yellow fever virus and West Nile virus

2019 ◽  
Vol 268 ◽  
pp. 53-55 ◽  
Author(s):  
José A. Boga ◽  
Marta E. Alvarez-Arguelles ◽  
Susana Rojo-Alba ◽  
Mercedes Rodríguez ◽  
María de Oña ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Dengue Virus ◽  
Yellow Fever ◽  
Zika Virus ◽  
Chikungunya Virus ◽  
Yellow Fever Virus ◽  
Simultaneous Detection ◽  
Fever Virus ◽  
West Nile ◽  
Virus Zika

Attempts to detect West Nile virus in wild birds in Poland

2011 ◽  
Vol 59 (3) ◽  
pp. 405-408 ◽  
Author(s):  
Jowita Niczyporuk ◽  
Elżbieta Samorek-Salamonowicz ◽  
Wojciech Kozdruń ◽  
Zbigniew Mizak
Keyword(s):  
West Nile Virus ◽  
Early Spring ◽  
Wild Birds ◽  
Rt Pcr ◽  
West Nile ◽  
Late Autumn ◽  
The Brain

The aim of the study was to attempt the detection of West Nile virus (WNV) in wild birds in Poland. Forty-eight species of 1912 wild birds were used for the investigations. The birds were derived from various locations in Poland from early spring till late autumn of the years 2009–2011. The brain samples were homogenised and cellular RNA was isolated. Two methods (RT-PCR and nested RT-PCR) were used. The presence of WNV RNA was not detected in the samples examined. Additionally, a short analysis of the epizootiological situation regarding the presence of WNV in Poland is presented.

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