Highlights ► A RT-LAMP assay detecting rodent coronaviruses was developed. ► The detection sensitivity was 3.2-fold higher than that of RT-PCR. ► The detection sensitivity was 31.6-fold lower than that of nested RT-PCR. ► In a practical diagnostic test, the detection sensitivity of duplicate RT-LAMP with regard to the nested RT-PCR was 87.5%. ► In a practical diagnostic test, the detection specificity of duplicate RT-LAMP with regard to the nested RT-PCR was 100%. 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.
【저자키워드】 RT-LAMP, Loop-mediated isothermal amplification, mouse hepatitis virus, Rat coronavirus, Rodent coronavirus, 【초록키워드】 coronavirus, Infection, diagnostic, RT-PCR, virus, Laboratory, RT-LAMP, sensitivity, specificity, infections, Sensitivity and specificity, Hepatitis, bovine, isothermal amplification, molecular, assay, mouse, MHV, diagnose, Detection sensitivity, strain, fecal, specimen, colonies, active infections, detected, different, performed, detect, prevalent virus,