Novel virus discovery in field-collected mosquito larvae using an improved system for rapid determination of viral RNA sequences (RDV ver4.0).
In this study, we improved a method for rapid determination of viral RNA sequences (RDV) to overcome the limitations of previous versions. The RDV ver4.0 method can detect RNA sequences with at least 1,000 copies as starting material. A novel virus, which was isolated from field-collected Aedes aegypti larvae in the Phasi Charoen district of Thailand using C6/36 cells, was identified using the RDV ver4.0 protocol. The virus was named Phasi Charoen virus (PhaV). We used a high-throughput pyrosequencing approach to obtain more information about the genome sequence of PhaV. Analysis of a phylogenic tree based on amino acid sequences strongly suggested that PhaV belongs to the family Bunyaviridae.
Yamao
Takuya
T
Faculty of Pharmaceutical Sciences, Fukuoka University, Nanakuma, Fukuoka, Japan.
Eshita
Yuki
Y
Kihara
Yuki
Y
Satho
Tomomitsu
T
Kuroda
Makoto
M
Sekizuka
Tsuyoshi
T
Nishimura
Miho
M
Sakai
Kouji
K
Watanabe
Shumpei
S
Akashi
Hiroomi
H
Rongsriyam
Yupha
Y
Komalamisra
Narumon
N
Srisawat
Raweewan
R
Miyata
Takeshi
T
Sakata
Akira
A
Hosokawa
Masato
M
Nakashima
Manabu
M
Kashige
Nobuhiro
N
Miake
Fumio
F
Fukushi
Shuetsu
S
Nakauchi
Mina
M
Saijo
Masayuki
M
Kurane
Ichiro
I
Morikawa
Shigeru
S
Mizutani
Tetsuya
T
D000330:Aedes / Q000821:virology*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002043:Bunyaviridae / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*; D002522:Chlorocebus aethiops; D018076:DNA, Complementary / Q000737:chemistry; D007814:Larva / Q000821:virology; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D010802:Phylogeny; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D014709:Vero Cells; D014773:Virology / Q000379:methods*
D018076:DNA, Complementary; D012367:RNA, Viral