Establishment of RT-PCR method and application for detecting bovine ephemeral fever virus in cattle in Viet Nam

Abstract

Bovine ephemeral fever (three-day sickness) is an arthropod-borne disease that affects cattle. The etiology is bovine ephemeral fever virus (BEFV). Up to date, BEFV has been known circulating in Africa, Asia, Middle East, Australia; and continues to threaten cattle populations in the previously BEFV-free areas. Facing the risk that a new disease can occur at any time, it is essential to prepare the diagnostic testing methods. Therefore, in this study, bioinformatic and experiment-based analyses in order to choose the most suitable primers used in RT-PCR for BEFV detection were carried out.
As a result, 2/6 primer sets for BEFV were determined to be the most suitable in terms of nucleotide sequence match (differ 0% - 0.7%, compared to BEFV strains in GenBank). The detection limit of the two primer sets was from 4.07 x 102 to 8.11 x 102 copies. By using suitable primers, 2/152 pooled
samples collected in Viet Nam were detected to be positive with BEFV.