Servicio de Microbiología, Hospital Universitario La Paz, IdiPAZ, Madrid, Spain.
PLoS One. 2021 Feb 16;16(2):e0246302. doi: 10.1371/journal.pone.0246302. eCollection 2021.
Two automatable in-house protocols for high-troughput RNA extraction from nasopharyngeal swabs for SARS-CoV-2 detection have been evaluated.
One hundred forty one SARS-CoV-2 positive samples were collected during a period of 10-days. In-house protocols were based on extraction with magnetic beads and designed to be used with either the Opentrons OT-2 (OT-2in-house) liquid handling robot or the MagMAXTM Express-96 system (MMin-house). Both protocols were tested in parallel with a commercial kit that uses the MagMAXTM system (MMkit). Nucleic acid extraction efficiencies were calculated from a SARS-CoV-2 DNA positive control.
No significant differences were found between both in-house protocols and the commercial kit in their performance to detect positive samples. The MMkit was the most efficient although the MMin-house presented, in average, lower Cts than the other two. In-house protocols allowed to save between 350€ and 400€ for every 96 extracted samples compared to the commercial kit.
The protocols described harness the use of easily available reagents and an open-source liquid handling system and are suitable for SARS-CoV-2 detection in high throughput facilities.
我们评估了两种自动化的、高通量的鼻咽拭子 RNA 提取方法，用于 SARS-CoV-2 的检测。
在 10 天的时间里，我们收集了 141 份 SARS-CoV-2 阳性样本。我们的提取方案基于磁珠提取法，并设计用于 Opentrons OT-2（OT-2 内建方案）液体处理机器人或 MagMAXTM Express-96 系统（MM 内建方案）。这两种方案都与使用 MagMAXTM 系统的商业试剂盒（MMkit）进行了平行测试。从 SARS-CoV-2 DNA 阳性对照中计算核酸提取效率。
在检测阳性样本的性能方面，两种内建方案与商业试剂盒之间没有显著差异。尽管 MM 内建方案的平均 Ct 值比其他两种方案低，但 MMkit 的效率最高。与商业试剂盒相比，每个 96 个样本的提取，内建方案可以节省 350 到 400 欧元。
所描述的方案利用了易于获得的试剂和开源的液体处理系统，适用于高通量设施中的 SARS-CoV-2 检测。
