Guangdong Key Laboratory of Laboratory Animals, Guangdong Laboratory Animals, Monitoring Institute, Guangzhou, China.
School of Science, RMIT University, Melbourne, Australia.
Mol Cell Probes. 2018 Oct;41:27-31. doi: 10.1016/j.mcp.2018.08.006. Epub 2018 Aug 26.
Theiler's murine encephalomyelitis virus (TMEV) is one of the most common viral pathogens that circulate widely in captive mouse colonies. A molecular biology detection method would be a useful tool to use in an integrated program to monitor and prevent TMEV infection and transmission. Thus, a reverse transcription recombinase polymerase amplification (RT-RPA) assay was developed to detect TMEV infection. The sensitivity of the RT-RPA assay approached 8 copies per reaction, which is equivalent to the sensitivity of RT-qPCR reactions. This assay did not detect RNA extracts from other murine pathogens included in this study or TMEV negative samples. Brain tissues and contaminated biological materials were used to assess the clinical performance of the RT-RPA. The detection results of RT-RPA and RT-qPCR were very similar, except that a contaminated biological material sample which was positive by RT-qPCR, with a CT value of 38, was negative by RT-RPA. In summary, the developed RT-RPA assay offers a rapid, sensitive and specific alternative method for monitoring of TMEV, especially in resource-limited conditions.
泰勒鼠脑脊髓炎病毒（TMEV）是广泛存在于封闭鼠群中的最常见的病毒病原体之一。分子生物学检测方法将是监测和预防 TMEV 感染和传播的综合计划中有用的工具。因此，开发了逆转录重组酶聚合酶扩增（RT-RPA）检测方法来检测 TMEV 感染。RT-RPA 检测方法的灵敏度接近每个反应 8 个拷贝，相当于 RT-qPCR 反应的灵敏度。该检测方法未检测到本研究中包括的其他鼠病原体或 TMEV 阴性样本的 RNA 提取物。脑组织和污染的生物材料用于评估 RT-RPA 的临床性能。RT-RPA 和 RT-qPCR 的检测结果非常相似，除了一个 RT-qPCR 检测为阳性的污染生物材料样本，CT 值为 38，而 RT-RPA 为阴性。总之，开发的 RT-RPA 检测方法为监测 TMEV 提供了一种快速、敏感和特异的替代方法，特别是在资源有限的情况下。
