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.

译文

小鼠肝炎病毒 (MHV) 是在实验室小鼠菌落中检测到的最普遍的病毒之一。肠溶性菌株在自然感染中占主导地位,用于检测粪便中MHV脱落的分子技术足以诊断活动性感染。开发了一种逆转录环介导的等温扩增 (rt-lamp) 技术,用于在90分钟内检测啮齿动物冠状病毒。该技术的特异性通过其检测所有17种不同的MHV菌株和6种大鼠冠状病毒菌株的能力以及未能非特异性检测人,牛和猪冠状病毒的能力得到证实。Rt-lamp的敏感性比逆转录聚合酶链反应 (rt-pcr) 高3.2倍,比巢式rt-pcr低31.6倍。对使用小鼠粪便标本进行的rt-lamp的诊断性能的评估显示,相对于巢式rt-pcr的敏感性和特异性分别为85.7% 和100%。RT-LAMP检测将适用于监测小鼠菌落中活跃的MHV感染。

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