Institute of Biophysics, Johannes Kepler University, 4020 Linz, Austria.
Institute of Laboratory Medicine, Johannes Kepler Universitätsklinikum, 4020 Linz, Austria.
Viruses. 2021 Aug 28;13(9):1712. doi: 10.3390/v13091712.
The SARS-CoV-2 pandemic has required the development of multiple testing systems to monitor and control the viral infection. Here, we developed a PCR test to screen COVID-19 infections that can process up to ~180 samples per day without the requirement of robotics. For this purpose, we implemented the use of multichannel pipettes and plate magnetics for the RNA extraction step and combined the reverse transcription with the qPCR within one step. We tested the performance of two RT-qPCR kits as well as different sampling buffers and showed that samples taken in NaCl or PBS are stable and compatible with different COVID-19 testing systems. Finally, we designed a new internal control based on the human gene that does not require a DNA digestion step. Our protocol is easy to handle and reaches the sensitivity and accuracy of the standardized diagnostic protocols used in the clinic to detect COVID-19 infections.
SARS-CoV-2 大流行要求开发多种检测系统来监测和控制病毒感染。在这里，我们开发了一种用于筛查 COVID-19 感染的 PCR 检测方法，该方法每天可处理多达约 180 个样本，而无需使用机器人。为此，我们实施了多通道移液器和板磁体用于 RNA 提取步骤，并将逆转录与一步内的 qPCR 相结合。我们测试了两种 RT-qPCR 试剂盒以及不同的采样缓冲液的性能，并表明在 NaCl 或 PBS 中采集的样本是稳定的，并且与不同的 COVID-19 检测系统兼容。最后，我们设计了一种基于人类基因的新内参，该内参不需要 DNA 消化步骤。我们的方案易于操作，达到了临床使用的标准化诊断方案检测 COVID-19 感染的灵敏度和准确性。
