Abstract
High impact, mosquito-borne flaviviruses such as West Nile virus (WNV), Usutu virus (USUV), Japanese encephalitis virus (JEV), Tembusu virus (TMUV), and Bagaza/Israel turkey meningoencephalomyelitis virus (BAGV/ITV) are emerging in different areas of the world. These viruses belong to the Japanese encephalitis (JE) serocomplex (JEV, WNV, and USUV) and the Ntaya serocomplex (TMUV and BAGV/ITV). Notably, they share transmission route (mosquito bite) and reservoir host type (wild birds), and some of them co-circulate in the same areas, infecting overlapping mosquito and avian population. This may simplify epidemiological surveillance, since it allows the detection of different infections targeting the same population, but also represents a challenge, as the diagnostic tools applied need to detect the whole range of flaviviruses surveyed, and correctly differentiate between these closely related pathogens. To this aim, a duplex real-time RT-PCR (dRRT-PCR) method has been developed for the simultaneous and differential detection of JE and Ntaya flavivirus serocomplexes. The method has been standardized and evaluated by analyzing a panel of 49 flaviviral and non-flaviviral isolates, and clinical samples of different bird species obtained from experimental infections or from the field, proving its value for virus detection in apparently healthy or suspicious animals. This new dRRT-PCR technique is a reliable, specific and highly sensitive tool for rapid detection and differentiation of JE and Ntaya flavivirus groups in either domestic or wild animals. This novel method can be implemented in animal virology diagnostic laboratories as screening tool in routine surveillance and in the event of bird encephalitis emergence.
Highlights
The genus Flavivirus, within the family Flaviviridae, comprises more than 70 different viruses, many of which represent relevant pathogens for humans and animals [1, 2]
This is significant in the current epidemiological situation, where the recent and potential emergence of bird-pathogenic flaviviruses in different territories poses a complex challenge for the diagnostic laboratories and for the veterinary authorities
The combination of the relevant bird-pathogenic flaviviruses species in a single assay makes very difficult to develop a PCR test with the high sensitivity level demanded for a screening tool
Summary
The genus Flavivirus, within the family Flaviviridae, comprises more than 70 different viruses, many of which represent relevant pathogens for humans and animals [1, 2]. Relevant viruses within the JE group are, for instance, West Nile virus (WNV), Saint Louis encephalitis virus (SLEV), Japanese encephalitis virus (JEV), Murray Valley encephalitis virus (MVEV), and Usutu virus (USUV), while the Ntaya serocomplex includes Bagaza virus (BAGV), its synonymous Turkey meningoencephalomyelitis virus (ITV) [8], and Tembusu virus (TMUV). All these viruses are maintained in nature in a cycle involving avian reservoir hosts and Culex spp. mosquitoes. USUV and BAGV were able to reach Europe likely from Sub-Saharan Africa [12, 20, 26]
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