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使用含有 IGEPAL-630 的浓缩病毒裂解-扩增缓冲液直接从临床样本中扩增 SARS-CoV-2。

Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630.

机构信息

Infectious Diseases Division, Internal Medicine Department, University of Texas Medical Branch, Galveston, TX, USA.

Infectious Diseases Division, Pathology Department, University of Texas Medical Branch, Galveston, TX, USA.

出版信息

Sci Rep. 2021 Jul 9;11(1):14204. doi: 10.1038/s41598-021-93333-2.

Abstract

The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shortage of kits and supplies to purify viral RNA. Therefore, it is urgent to validate alternative nucleic acid isolation methods for SARS-CoV-2. Our results demonstrate that a concentrated viral lysis amplification buffer (vLAB) prepared with the nonionic detergent IGEPAL enables qualitative detection of SARS-CoV-2 by direct Reverse Transcriptase-Polymerase Chain Reaction (dRT-PCR). Furthermore, vLAB was effective in inactivating SARS-CoV-2. Since this method is inexpensive and no RNA purification equipment or additional cDNA synthesis is required, this dRT-PCR with vLAB should be considered as an alternative method for qualitative detection of SARS-CoV-2.

摘要

由新型冠状病毒(SARS-CoV-2)引起的 2019 年大流行仍在全球范围内迅速蔓延。核酸扩增是确认 COVID-19 感染的金标准方法。然而,来自欠发达国家的诊断实验室面临的挑战包括试剂盒和病毒 RNA 纯化用品的短缺。因此,迫切需要验证 SARS-CoV-2 的替代核酸分离方法。我们的结果表明,用非离子型去污剂 IGEPAL 制备的浓缩病毒裂解扩增缓冲液(vLAB)可通过直接逆转录-聚合酶链反应(dRT-PCR)定性检测 SARS-CoV-2。此外,vLAB 可有效灭活 SARS-CoV-2。由于该方法成本低廉,且不需要 RNA 纯化设备或额外的 cDNA 合成,因此这种使用 vLAB 的 dRT-PCR 可被视为定性检测 SARS-CoV-2 的替代方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b7d9/8270935/09fc7aecbf0f/41598_2021_93333_Fig1_HTML.jpg

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