Key Laboratory of Testing and Evaluation for Aquatic Product Safety and Quality, Ministry of Agriculture and Rural Affairs, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, People's Republic of China.
Food Environ Virol. 2019 Dec;11(4):383-392. doi: 10.1007/s12560-019-09400-5. Epub 2019 Aug 21.
Foodborne viruses are a global threat to food safety. Real-time reverse transcription polymerase chain reaction (RT-PCR) is the most commonly used method to detect viral RNA in food. Armored RNA (AR) prepared using the MS2 phage system is a successful positive control for detecting foodborne viruses and is an important quality control process when using real-time RT-PCR. In this study, we report a novel technology for preparing AR using bacteriophage Qβ and compare its stability with AR prepared using the MS2 phage system for packaging norovirus detection target RNA. AR could be successfully and efficiently produced using the developed bacteriophage Qβ system. Two types of AR-AR-QNoV prepared using the Qβ system and AR-MNoV prepared using the MS2 system-were stored at different temperatures for different durations. After incubating at - 20 °C for 360 days, the copy numbers of AR-QNoV and AR-MNoV decreased by 8.9% and 35.9%, respectively. After incubating at 4 °C for 60 days, the copy numbers of AR-QNoV and AR-MNoV decreased by 12.0% and 38.9%, respectively. After incubating at 45 °C, the copy numbers of AR-QNoV decreased by 71.8% after 5 days, whereas those of AR-MNoV decreased by 92.9% after only 4 days. After 5 days, AR-MNoV could not be detected using real-time RT-PCR. There was a significant difference in copy numbers decrease rate between AR-QNoV and AR-MNoV at three different temperatures (P < 0.05 ). Therefore, AR prepared using the new bacteriophage Qβ system is more stable than the traditional AR, making the developed strategy a good candidate for AR preparation and quality control.
食源性病原体是食品安全的全球性威胁。实时逆转录聚合酶链反应（RT-PCR）是检测食品中病毒 RNA 最常用的方法。使用 MS2 噬菌体系统制备的装甲 RNA（AR）是检测食源性病原体的成功阳性对照，也是使用实时 RT-PCR 时重要的质量控制过程。在本研究中，我们报告了一种使用噬菌体 Qβ制备 AR 的新技术，并比较了其用于包装诺如病毒检测靶标 RNA 的稳定性与使用 MS2 噬菌体系统制备的 AR。可以使用开发的噬菌体 Qβ系统成功且有效地生产 AR。使用 Qβ系统制备的两种类型的 AR-AR-QNoV 和使用 MS2 系统制备的 AR-MNoV 在不同温度下储存不同的时间。在-20°C 孵育 360 天后，AR-QNoV 和 AR-MNoV 的拷贝数分别下降了 8.9%和 35.9%。在 4°C 孵育 60 天后，AR-QNoV 和 AR-MNoV 的拷贝数分别下降了 12.0%和 38.9%。在 45°C 孵育时，AR-QNoV 的拷贝数在 5 天后下降了 71.8%，而 AR-MNoV 的拷贝数在仅 4 天后下降了 92.9%。5 天后，实时 RT-PCR 无法检测到 AR-MNoV。在三种不同温度下，AR-QNoV 和 AR-MNoV 的拷贝数下降率有显著差异（P<0.05）。因此，与传统 AR 相比，使用新噬菌体 Qβ系统制备的 AR 更稳定，这使得所开发的策略成为 AR 制备和质量控制的良好候选方案。
