Broad-range detection of arboviruses belonging to Simbu serogroup lineage 1 and specific detection of Akabane, Aino and Peaton viruses by newly developed multiple TaqMan assays.
TaqMan assays were developed for the broad-range detection of arboviruses belonging to Simbu serogroup lineage 1 in the genus Orthobunyavirus and also for the specific detection of three viruses in the lineage, Akabane, Aino and Peaton viruses (AKAV, AINOV and PEAV, respectively). A primer and probe set was designed for the broad-range detection of Simbu serogroup lineage 1 (Pan-Simbu1 set) mainly targeting AKAV, AINOV, PEAV, Sathuperi and Shamonda viruses (SATV and SHAV), and the forward and reverse primers of the Pan-Simbu1 set were also used for the specific detection of AKAV with another probe (AKAV-specific set). In addition, two more primer and probe sets were designed for AINOV- and PEAV-specific detection, respectively (AINOV- and PEAV-specific sets). All of the four primer and probe sets successfully detected targeted viruses, and thus broad-range and specific detection of all the targeted viruses can be achieved by using two multiplex assays and a single assay in a dual (two-color) assay format when another primer and probe set for a bovine β-actin control is also used. The assays had an analytical sensitivity of 10 copies/tube for AKAV, at least 100 copies/tube for AINOV, 100 copies/tube for PEAV, one copy/tube for SATV and at least 10 copies/tube for SHAV, respectively. Diagnostic sensitivity of the assays was tested with field-collected bovine samples, and the results suggested that the sensitivity was higher than that of a conventional RT-PCR. These data indicate that the newly developed TaqMan assays will be useful tools for the diagnosis and screening of field-collected samples for infections of AKAV and several other arboviruses belonging to the Simbu serogroup lineage 1.
Kyushu Research Station, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Japan. Electronic address: shirah@affrc.go.jp.
Hiroaki
H
Shirafuji
Kusu Livestock Hygiene Service Center, Oita Prefectural Government, Japan.
Ryu
R
Yazaki
Oita Livestock Hygiene Service Center, Oita Prefectural Government, Japan.
Yozo
Y
Shuto
Kyushu Research Station, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Japan.
Tohru
T
Yanase
Kyushu Research Station, National Institute of Animal Health (NIAH), National Agriculture and Food Research Organization (NARO), Japan.
Tomoko
T
Kato
Domestic Livestock Disease Identification Office, Food Safety Promotion Division, Shimane Prefectural Government, Japan.
Youji
Y
Ishikura
Kagoshima Central Livestock Hygiene Service Center, Kagoshima Prefectural Government, Japan.
Zenjiro
Z
Sakaguchi
Okinawa Prefectural Institute of Animal Health, Okinawa Prefectural Government, Japan.
Moemi
M
Suzuki
Viral Disease and Epidemiology Research Division, NIAH, NARO, Japan.
Makoto
M
Yamakawa
D000818:Animals; D001103:Arboviruses / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002044:Bunyaviridae Infections / Q000175:diagnosis* / Q000662:veterinary / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000821:virology; D017931:DNA Primers / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D015345:Oligonucleotide Probes / Q000235:genetics; D016851:Orthobunyavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D014773:Virology / Q000379:methods*
D017931:DNA Primers; D015345:Oligonucleotide Probes
Akabane virus; Arbovirus; Diagnosis; Orthobunyavirus; TaqMan assay