Abstract
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.
Highlights
West Nile virus (WNV) is an enveloped spherical, single-stranded positive-sense RNA virus belonging to the Flaviviridae family [1]
Four labs did not carry out this technique due to lack of specific equipment to perform conventional reverse-transcription polymerase chain reaction (RT-PCR) assays and were unable to detect the flavivirus positive sample of the panel corresponding to Japanese encephalitis virus (JEV)
Despite the increasing incidence of WNV in Europe and the Mediterranean area, few external quality assessments have been organized to evaluate the diagnostic capacities of labs, especially in the veterinary sector
Summary
West Nile virus (WNV) is an enveloped spherical, single-stranded positive-sense RNA virus belonging to the Flaviviridae family [1]. A training curriculum was implemented to improve diagnostic performance of the veterinary labs for this pathogen, including two diagnostic workshops (molecular and serological diagnosis) that were organized in 2015 and 2016 at Centro de Investigación en Sanidad Animal (INIA-CISA) (Madrid, Spain) After these training sessions, an external quality assessment (EQA) was organized between October. In this study we report the results of this inter-laboratory trial and provide relevant information about the current WNV (and USUV) diagnostic capacities of veterinary labs in the Mediterranean and Black Sea regions. This exercise enabled an extensive reproducibility assessment of the recommended tests for WNV and USUV diagnostics
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