RRC ID 45014
著者 Hanaki K, Ike F, Kajita A, Yasuno W, Yanagiba M, Goto M, Sakai K, Ami Y, Kyuwa S.
タイトル Detection of murine norovirus by reverse transcription loop-mediated isothermal amplification.
ジャーナル J Virol Methods
Abstract Murine norovirus (MNV) has considerable genetical and biological diversity and is recognized worldwide as the most common contaminant in laboratory mouse colonies. This study developed a reverse transcription loop-mediated isothermal amplification (RT-LAMP) method with the potential to detect a broad range of MNV. RT-LAMP, using a set of five primers containing mixed bases, obtained results under isothermal conditions at 62°C for 90min. Sensitivity of RT-LAMP was 50-fold less than that of two-step TaqMan real-time reverse transcription-polymerase chain reaction (TaqMan RT-PCR). Diagnostic performance of RT-LAMP on RNA extracted from mouse fecal specimens was compared with TaqMan RT-PCR and nested RT-PCR. MNV was detected in 54 of 120 mouse fecal specimens by RT-LAMP, and RT-LAMP had an estimated sensitivity and specificity of 96.4% and 100% compared with TaqMan RT-PCR, and 94.7% and 100% compared with nested RT-PCR. RT-LAMP, which does not require expensive instruments, might be useful for the screening of mice actively or persistently infected with MNV.
巻・号 204
ページ 17-24
公開日 2014-8-1
DOI 10.1016/j.jviromet.2014.03.025
PII S0166-0934(14)00131-1
PMID 24717164
PMC PMC7172776
MeSH Animals Caliciviridae Infections / veterinary* Caliciviridae Infections / virology DNA Primers / genetics Feces / virology Mice Norovirus / genetics Norovirus / isolation & purification* Nucleic Acid Amplification Techniques / methods* Reverse Transcription* Rodent Diseases / diagnosis* Rodent Diseases / virology* Sensitivity and Specificity Temperature Time Factors
IF 1.786
引用数 5
WOS 分野 BIOTECHNOLOGY & APPLIED MICROBIOLOGY BIOCHEMICAL RESEARCH METHODS VIROLOGY
リソース情報
ヒト・動物細胞 RAW 264(RCB0535)