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蛋白酶 K 处理法在无需 RNA 提取经典程序的情况下,是一种快速且更经济的 SARS-CoV-2 分子检测替代方法。

Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection.

机构信息

Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.

Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.

出版信息

J Virol Methods. 2021 Jul;293:114131. doi: 10.1016/j.jviromet.2021.114131. Epub 2021 Mar 30.

Abstract

The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures for monitoring virus circulation and reduce the spreading of SARS-CoV-2. Here, we described quickness and cheaper strategies of direct RT-qPCR (in the absence of RNA isolation) and compared the results to those obtained using standard RNA isolation procedure. The tests varied using pure, diluted samples, combined with Proteinase K (PK) or Lysis Buffer. Our findings showed consistently that PK pre-treated samples in the absence of RNA extraction procedures presents similar results to those obtained by standard RNA isolation procedures. On average, 16 samples extracted with the MagMAX™ CORE Kit, take around 2 h, costing an average of USD 5, the pre-treatment of samples using PK, on the other hand, would cut the value to less than USD 0.30 and reduce the time of procedure in more than 1 ½ hours. The present study suggests the use of PK treatment instead of RNA isolation in order to reduce costs and time in processing samples for molecular diagnosis of SARS-CoV-2.

摘要

世界卫生组织(WHO)已宣布 COVID-19 大流行,该病由最近描述的 SARS-CoV-2 引起。广泛认为,大规模诊断对于发现无症状个体具有重要意义,是加强病毒传播监测和减少 SARS-CoV-2 传播的强制性手段。在这里,我们描述了直接 RT-qPCR(无需 RNA 分离)的快速且更廉价的策略,并将结果与使用标准 RNA 分离程序获得的结果进行了比较。该测试使用纯样、稀释样,结合蛋白酶 K(PK)或裂解缓冲液进行。我们的研究结果一致表明,在没有 RNA 提取程序的情况下,经 PK 预处理的样品与标准 RNA 分离程序获得的结果相似。平均而言,使用 MagMAX™ CORE 试剂盒提取 16 个样本大约需要 2 小时,花费约 5 美元,而使用 PK 预处理样品,则可将成本降低至 0.30 美元以下,且程序时间可缩短一个半小时以上。本研究建议使用 PK 处理代替 RNA 分离,以降低 SARS-CoV-2 分子诊断样本处理的成本和时间。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/66bf/8008826/2e0becf8d7b3/gr1_lrg.jpg

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