Establishment and Application of Rapid RT-PCR Assays for Wesselsbron Virus
In order to rapidly detect Wesselsbron virus(WSLV),specific primers and probe were designed based on conserved sequence of NS5 gene of WSLV to establish TaqMan-based fluorescent RT-PCR assay and RT-PCR assay,and their sensitivity,specificity and clinical feasibility were verified.The results showed that the established methods could detect nucleic acids at the lowest limits of 10 and 100 copies/μL,respectively,and failed to crossly react with the nucleic acids of Japanese encephalitis virus,West Nile virus and Tambusu virus;100 samples collected from entry horses were tested by the two methods,respectively,and no WSLV-positive samples were detected,which was consistent with the results by dual fluorescence RT-PCR assay provided by the reference laboratory in South Africa.In conclusion,the established methods were with high sensitivity,strong specificity and good stability,and could be used to detect clinical samples,supporting entry-exit inspection and quarantine,epidemiological investigation,prevention and control of Wesselsbron disease.
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