Peer-reviewed veterinary case report
Reverse transcription-loop-mediated isothermal amplification for the detection of rodent coronaviruses.
- Journal:
- Journal of virological methods
- Year:
- 2013
- Authors:
- Hanaki, Ken-Ichi et al.
- Affiliation:
- Department of Laboratory Animal Medicine · Japan
- Species:
- rodent
Plain-English summary
Researchers have developed a new test called reverse transcription-loop-mediated isothermal amplification (RT-LAMP) to quickly detect mouse hepatitis virus (MHV), which is a common virus found in laboratory mice. This test can identify all known strains of MHV and certain strains of rat coronaviruses, while not mistakenly detecting coronaviruses that affect humans or other animals like cows and pigs. The RT-LAMP test is more sensitive than another common test called RT-PCR, making it a reliable option for checking if mice are actively shedding the virus in their feces. Overall, this new method shows great promise for monitoring MHV infections in mouse colonies.
Abstract
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 90 min. 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.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/23123121/