ICAR-International Centre for Foot-and-Mouth Disease, DFMD, Jatni, Khordha, Odisha, India.
Mol Biol Rep. 2021 Oct;48(10):6871-6877. doi: 10.1007/s11033-021-06688-0. Epub 2021 Sep 12.
RT-qPCR technique is the current world-wide method used for the early detection of SARS-CoV2 RNA in the suspected clinical samples. Viral RNA extraction is the key pre-analytical step for SARS-CoV2 detection which often achieved using commercial RNA-extraction kits. However, due to the COVID-19 pandemic, bulk production and the supply chains for the commercial RNA-extraction kit have been seriously compromised. The shortage of commercial RNA-extraction kit is even more acute in developing country. Furthermore, use of one-off design RNA-columns can generate plastic wastes that have an environmental pollution effect.
To address these issues, in this study, we used warm alkaline solution containing Triton X-100 for the complete removal of the residual SARS-CoV2 RNA from the used RNA-binding silica column. Columns regenerated using the alkaline solution have the viral RNA purification capability that is comparable to the fresh silica columns. We also demonstrated that RNA-binding silica columns can be regenerated and reused for a minimum of five-times.
Therefore, the use of the RNA-column regeneration method may benefits several SARS-CoV2 diagnostic laboratories throughout the world by cutting down the requirement of commercial RNA-purification column.
RT-qPCR 技术是目前全球用于检测疑似临床样本中 SARS-CoV2 RNA 的方法。病毒 RNA 提取是 SARS-CoV2 检测的关键分析前步骤，通常使用商用 RNA 提取试剂盒来实现。然而，由于 COVID-19 大流行，商用 RNA 提取试剂盒的批量生产和供应链受到了严重影响。在发展中国家，商用 RNA 提取试剂盒的短缺更为严重。此外，一次性设计的 RNA 柱会产生塑料废物，对环境造成污染。
为了解决这些问题，在本研究中，我们使用含有 Triton X-100 的碱性溶液从已使用的 RNA 结合硅胶柱上完全去除残留的 SARS-CoV2 RNA。用碱性溶液再生的柱子具有与新鲜硅胶柱相当的病毒 RNA 纯化能力。我们还证明，RNA 结合硅胶柱可以至少再生和重复使用五次。
因此，RNA 柱再生方法的使用可以通过减少对商用 RNA 纯化柱的需求，使全球更多的 SARS-CoV2 诊断实验室受益。
