Silva Lívia Mara, Riani Lorena Rodrigues, Silvério Marcelo Silva, Pereira-Júnior Olavo Dos Santos, Pittella Frederico
Faculdade de Farmácia, Universidade Federal de Juiz de Fora, Juiz de Fora 36036-900, Brazil.
Diagnostics (Basel). 2022 Feb 27;12(3):601. doi: 10.3390/diagnostics12030601.
Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world's response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to collect and process the samples, and to perform reliable mass diagnosis by RT-qPCR. In particular, time-consuming and high cost of nucleic acid extraction procedures have hampered the diagnosis; moreover, several steps in the routine for the preparation of the material makes the extracted sample susceptible to contamination. Here two rapid nucleic acid extraction reagents were compared as extraction procedures for SARS-CoV-2 detection in clinical samples by singleplex and multiplex RT-qPCR analysis, using different transport media, samples with high and low viral load, and different PCR machines. As observed, rapid nucleic acid extraction procedures can be applied for reliable diagnosis using a TaqMan-based assay, over multiple platforms. Ultimately, prompt RNA extraction may reduce costs with reagents and plastics, the chances of contamination, and the overall time to diagnosis by RT-qPCR.
自2020年以来，人类一直面临由严重急性呼吸综合征冠状病毒2（SARS-CoV-2）引起的呼吸道疾病——2019冠状病毒病（COVID-19）大流行。全球对该大流行的应对经历了诊断方法、疫苗和药物的发展。在诊断方面，由于收集和处理样本以及通过逆转录定量聚合酶链反应（RT-qPCR）进行可靠的大规模诊断所需材料短缺，面临着巨大挑战。特别是，核酸提取程序耗时且成本高昂，阻碍了诊断；此外，样本制备常规流程中的几个步骤使提取的样本容易受到污染。本文通过单重和多重RT-qPCR分析，使用不同的运输介质、高低病毒载量的样本以及不同的PCR机器，比较了两种快速核酸提取试剂作为临床样本中SARS-CoV-2检测的提取程序。结果表明，快速核酸提取程序可通过基于TaqMan的检测方法在多个平台上用于可靠诊断。最终，快速的RNA提取可能会降低试剂和塑料制品成本、污染几率以及RT-qPCR诊断的总时间。
