In September 2012, several cows and a calf showed decreased activity, anorexia and fever
on Ishigaki Island, Okinawa Prefecture, Japan, and the cases were diagnosed as bovine
ephemeral fever (BEF). We isolated BEF virus (BEFV) from one of the affected cows and then
determined the complete genome sequence of the G gene, which encodes a class I
transmembrane glycoprotein of BEFV. The BEFV isolate in this case, ON-3/E/12, was sorted
into the same cluster as other BEFV isolates in Japan, Taiwan and China obtained in
1996−2004 and was most closely related to a 2002 Chinese isolate, JT02L, according to the
phylogenetic analysis of the complete G gene. Since inactivated vaccines for BEF available
in Japan are considered effective against the ON-3/E/12 isolate as well as other isolates
in East Asia from 1996−2004, annual vaccination should be conducted to prevent BEF in
Okinawa. Additionally, in this study, we developed an RT-PCR assay to detect the BEFV gene
in Japan and neighboring countries. Our assay was able to amplify target sequences in all
of the tested BEFV isolates, including 18 isolates in Japan and another isolate in
Australia. The assay was found to be useful also for testing RNA samples extracted from
bovine peripheral blood mononuclear cells, and the detection limit of the assay was 10
copies per tube. We believe that our assay would be an important tool for the screening of
BEFV infection and the diagnosis of BEF.