| RRC ID |
45087
|
| 著者 |
Hanaki K, Ike F, Kajita A, Yasuno W, Yanagiba M, Goto M, Sakai K, Ami Y, Kyuwa S.
|
| タイトル |
A broadly reactive one-step SYBR Green I real-time RT-PCR assay for rapid detection of murine norovirus.
|
| ジャーナル |
PLoS One
|
| Abstract |
A one-step SYBR Green I real-time RT-PCR assay was developed for the detection and quantification of a broad range of murine noroviruses (MNVs). The primer design was based on the multiple sequence alignments of 101 sequences of the open reading frame (ORF)1-ORF2 junction of MNV. The broad reactivity and quantitative capacity of the assay were validated using 7 MNV plasmids. The assay was completed within 1 h, and the reliable detection limit was 10 copies of MNV plasmid or 0.063 median tissue culture infective doses per milliliter of RAW264 cell culture-propagated viruses. The diagnostic performance of the assay was evaluated using 158 mouse fecal samples, 91 of which were confirmed to be positive. The melting curve analysis demonstrated the diversity of MNV in the samples. This is the first report of a broadly reactive one-step SYBR Green I real-time RT-PCR assay for detecting of MNVs. The rapid and sensitive performance of this assay makes it a powerful tool for diagnostic applications.
|
| 巻・号 |
9(5)
|
| ページ |
e98108
|
| 公開日 |
2014-1-1
|
| DOI |
10.1371/journal.pone.0098108
|
| PII |
PONE-D-13-44231
|
| PMID |
24847970
|
| PMC |
PMC4029972
|
| MeSH |
Animals
Base Sequence
Benzothiazoles
Cell Line
DNA Primers
Diamines
Feces / virology
Mice
Mice, Inbred C57BL
Molecular Sequence Data
Norovirus / genetics
Norovirus / isolation & purification*
Open Reading Frames
Organic Chemicals / chemistry*
Plasmids
Quinolines
RNA, Viral / genetics
Real-Time Polymerase Chain Reaction / methods*
Reproducibility of Results
Temperature
Time Factors
|
| IF |
2.74
|
| 引用数 |
7
|
|
WOS 分野
|
VIROLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
RAW 264(RCB0535) |
