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Reverse transcription-loop-mediated isothermal amplification for the detection of rodent coronaviruses

Hanaki, Ken-Ichi, Ike, Fumio, Hatakeyama, Rika, Hirano, Norio
Journal of virological methods 2013 v.187 no.2 pp. 222-227
Murine hepatitis virus, cattle, feces, humans, mice, monitoring, rats, reverse transcriptase polymerase chain reaction, reverse transcription loop-mediated isothermal amplification, swine, viruses
Mouse hepatitis virus (MHV) is one of the most prevalent viruses detected in laboratory mouse colonies. Enterotropic strains predominate in natural infections, and molecular techniques for the detection of MHV shedding in feces are powerful enough to diagnose active infections. A reverse transcription-loop-mediated isothermal amplification (RT-LAMP) technique was developed for the detection of rodent coronaviruses within 90min. The specificity of this technique was confirmed by its ability to detect all 17 different strains of MHV and 6 strains of rat coronaviruses as well as its failure to detect human, bovine, and porcine coronaviruses nonspecifically. The sensitivity of RT-LAMP was 3.2-fold higher than that of reverse transcription-polymerase chain reaction (RT-PCR) and 31.6-fold lower than that of nested RT-PCR. An evaluation of the diagnostic performance of RT-LAMP performed in duplicate using mouse fecal specimens showed that the sensitivity and specificity with respect to nested RT-PCR were 85.7% and 100%, respectively. RT-LAMP assays would be suitable for monitoring active MHV infection in mouse colonies.