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Transboundary and emerging diseases2022; 69(5); e2506-e2515; doi: 10.1111/tbed.14593

West Nile virus in the Republic of Serbia-Diagnostic performance of five serological tests in dog and horse sera.

Abstract: West Nile virus (WNV) is a zoonotic mosquito-borne virus classified as family Flaviviridae and genus Flavivirus. The first WNV outbreak in humans in the Republic of Serbia was recorded in 2012. Equids and dogs can show clinical symptoms after WNV infection and are often used as sentinels. This study aimed to (i) give insight into seropositivity for WNV in clinically healthy dog and horse sera in different regions of Serbia and (ii) compare diagnostic value of 'in-house' and commercially available indirect immunofluorescence (IFA) and enzyme-linked immunoassay (ELISA) tests to 'gold standard' virus neutralization test (VNT). Due to cross-reactivity, sera were tested for Usutu virus and tick-borne encephalitis virus in VNT based on the epidemiological data of field presence. Blood sera of dogs (n = 184) and horses (n = 232) were collected from 2011 to 2013. The seropositivity was confirmed by VNT in 36.9 % tested dog sera and 34.9% tested horse sera with highest positivity in regions near two big rivers, while in four dog and seven horse sera, positivity resulted from Usutu virus infection. Comparative results of diagnostic tests in dogs ranged from 18.7 % seropositivity by 'in-house' ELISA to 31.9% by commercially available ELISA. In horses, seropositivity ranged from 36.2% by 'in-house' IFA to 32.5% by commercially available IFA and from 26.3% by 'in-house' IgG ELISA to 20.9% by commercially available ELISA. There were no statistically significant differences according to the McNemar test between 'in-house' and commercially available IFA and ELISA test in horse sera, while the same was not true for two ELISAs used in dog sera (χ  = 8.647, p = .003). Established seropositivity in dogs and horses was in accordance with the epidemiological situation and WNV spread in the Republic of Serbia and proven Usutu virus co-circulation. 'In-house' tests remain a valuable tool in early diagnostics of WNV.
© 2022 The Authors. Transboundary and Emerging Diseases published by Wiley-VCH GmbH.
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Publication Date: 2022-05-24 PubMed ID: 35538046DOI: 10.1111/tbed.14593Google Scholar: Lookup
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Summary

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The research paper’s abstract discusses a study on the West Nile virus (WNV) in Serbia and the performance of five different tests to detect its presence in dogs and horses. The study also compared the efficacy of ‘in-house’ and commercially available diagnostic tools.

Study Objective

  • The study has two main objectives. First, to explore the rate of seropositivity for the West Nile Virus (WNV) in clinically healthy dogs and horses in different regions of Serbia. Seropositivity refers to the presence of antibodies in the blood serum of an organism, indicating exposure to a particular pathogen, in this case, the WNV.
  • The second objective was to compare the diagnostic value of both ‘in-house’ and commercially available testing methods. The key methods of testing mentioned are Indirect Immunofluorescence (IFA) and Enzyme-Linked Immunoassay (ELISA) tests. These were compared against the ‘gold standard’ Virus Neutralization Test (VNT).

Methodology and Findings

  • Between 2011 and 2013, blood samples were collected from dogs (n = 184) and horses (n = 232). These samples were tested for Usutu virus and tick-borne encephalitis virus in addition to WNV due to the intertwined epidemiological presence of these viruses.
  • The seropositivity was identified by VNT, revealing a presence of 36.9% in tested dog sera and 34.9% in horse sera. The occurrence was particularly high in regions closer to two large rivers.
  • The diagnostic outcomes in dogs ranged from 18.7% seropositivity using ‘in-house’ ELISA to 31.9% using commercially available ELISA. For horses, seropositivity ranged from 36.2% with ‘in-house’ IFA to 32.5% with commercially available IFA and from 26.3% by ‘in-house’ IgG ELISA to 20.9% by commercially available ELISA.
  • Interestingly, certain positivity emerged not from a WNV infection, but was owing to the Usutu virus infection in four dog sera and seven horse sera.

Comparison of Diagnostic Tests and Conclusion

  • There was no statistically significant difference between ‘in-house’ and commercially available IFA and ELISA tests in horse sera based upon the McNemar test. However, this was not the case for the two ELISA tests applied on dog sera, where differences were statistically significant.
  • The results point out that the rate of seropositivity among horses and dogs aligns with the epidemiological status and WNV proliferation in Serbia, along with confirmed Usutu virus co-circulation.
  • Even though commercially available tests are useful, ‘in-house’ tests still have significant value for early diagnosis of WNV.

Cite This Article

APA
Vasić A, Răileanu C, Körsten C, Vojinović D, Manić M, Urošević A, Nikolić N, Dulović O, Tews BA, Petrović T, Silaghi C, Valčić M, Gligić A. (2022). West Nile virus in the Republic of Serbia-Diagnostic performance of five serological tests in dog and horse sera. Transbound Emerg Dis, 69(5), e2506-e2515. https://doi.org/10.1111/tbed.14593

Publication

Transboundary and emerging diseases
ISSN: 1865-1682
NlmUniqueID: 101319538
Country: Germany
Language: English
Volume: 69
Issue: 5
Pages: e2506-e2515

Researcher Affiliations

Vasić, Ana
  • Faculty of Veterinary Medicine, Department of Infectious Diseases of Animals and Bee Diseases, University of Belgrade, Belgrade, Serbia.
  • Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald Isle of Riems, Germany.
  • Institute for Medical Research, National Institute of Serbia, University of Belgrade, Belgrade, Serbia.
Răileanu, Cristian
  • Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald Isle of Riems, Germany.
Körsten, Christin
  • Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald Isle of Riems, Germany.
Vojinović, Dragica
  • Serbian Institute of Veterinary Science, Department of Immunology, Belgrade, Serbia.
Manić, Marija
  • Specialized Veterinary Institute Niš, Department of Microbiology, Niš, Serbia.
Urošević, Aleksandar
  • Clinic for Infectious and Tropical Diseases, Clinical Centre Serbia, Belgrade, Serbia.
  • Parexel International Serbia, Belgrade, Serbia.
Nikolić, Nataša
  • Clinic for Infectious and Tropical Diseases, Clinical Centre Serbia, Belgrade, Serbia.
  • Faculty of Medicine, University of Belgrade, Belgrade, Serbia.
Dulović, Olga
  • Clinic for Infectious and Tropical Diseases, Clinical Centre Serbia, Belgrade, Serbia.
  • Faculty of Medicine, University of Belgrade, Belgrade, Serbia.
Tews, Birke Andrea
  • Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald Isle of Riems, Germany.
Petrović, Tamaš
  • Scientific Veterinary Institute 'Novi Sad', Department of Virology, Novi Sad, Serbia.
Silaghi, Cornelia
  • Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Greifswald Isle of Riems, Germany.
  • Faculty of Mathematics and Natural Sciences, University of Greifswald, Greifswald, Germany.
Valčić, Miroslav
  • Faculty of Veterinary Medicine, Department of Infectious Diseases of Animals and Bee Diseases, University of Belgrade, Belgrade, Serbia.
Gligić, Ana
  • Institute of Virology, Vaccines and Sera 'Torlak', Belgrade, Serbia.

MeSH Terms

  • Animals
  • Antibodies, Viral
  • Dog Diseases / diagnosis
  • Dog Diseases / epidemiology
  • Dogs
  • Encephalitis Viruses, Tick-Borne
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Flavivirus
  • Horse Diseases / diagnosis
  • Horse Diseases / epidemiology
  • Horses
  • Humans
  • Immunoglobulin G
  • Serbia / epidemiology
  • Serologic Tests / veterinary
  • West Nile Fever / diagnosis
  • West Nile Fever / epidemiology
  • West Nile Fever / veterinary
  • West Nile virus

Grant Funding

  • TR37015 / Ministry of Education, Science and Technological Development of the Republic of Serbia-Grant
  • Institutional Funding of IMR

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Citations

This article has been cited 5 times.
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