| RRC ID |
45014
|
| Author |
Hanaki K, Ike F, Kajita A, Yasuno W, Yanagiba M, Goto M, Sakai K, Ami Y, Kyuwa S.
|
| Title |
Detection of murine norovirus by reverse transcription loop-mediated isothermal amplification.
|
| Journal |
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.
|
| Volume |
204
|
| Pages |
17-24
|
| Published |
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
|
| Times Cited |
5
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
BIOCHEMICAL RESEARCH METHODS
VIROLOGY
|
| Resource |
| Human and Animal Cells |
RAW 264(RCB0535) |
