Detection of SARS-CoV-2 RNA by multiplex RT-qPCR.
Detection of SARS-CoV-2 RNA by multiplex RT-qPCR.
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DOI：
10.1371/journal.pbio.3000867
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发表时间：
2020-10
期刊：
影响因子：
9.8
通讯作者：
Iwasaki A
中科院分区：
文献类型：
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作者：
Kudo E;Israelow B;Vogels CBF;Lu P;Wyllie AL;Tokuyama M;Venkataraman A;Brackney DE;Ott IM;Petrone ME;Earnest R;Lapidus S;Muenker MC;Moore AJ;Casanovas-Massana A;Yale IMPACT Research Team;Omer SB;Dela Cruz CS;Farhadian SF;Ko AI;Grubaugh ND;Iwasaki A
在美国，目前推荐用于严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）检测的定量逆转录PCR（RT-qPCR）检测需要分析每份样本的3个基因组靶标：2个病毒和1个宿主。为了简化检测并减少所需试剂的体积，我们设计了一种多重RT-qPCR检测方法，在单一反应中检测SARS-CoV-2。我们使用疾病控制和预防中心现有的N1、N2和RP引物和探针组，但替换荧光团以允许检测的多路复用。我们的多重RT-qPCR的循环阈值（Ct）值与适用于研究目的的单一测定法获得的值相当。通过多重RT-qPCR一致地检测到低拷贝数（≥500拷贝/反应）的SARS-CoV-2 RNA。我们的新型多重RT-qPCR通过节省试剂、成本、时间和劳动力来改进当前的单一诊断。目前在美国推荐用于SARS-CoV-2检测的RT-qPCR检测需要分析每个样本的三个基因组靶标：两个病毒和一个宿主。本研究描述了一种多重PCR，节省试剂和劳动力，同时准确检测SARS-CoV-2 RNA。
The current quantitative reverse transcription PCR (RT-qPCR) assay recommended for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) testing in the United States requires analysis of 3 genomic targets per sample: 2 viral and 1 host. To simplify testing and reduce the volume of required reagents, we devised a multiplex RT-qPCR assay to detect SARS-CoV-2 in a single reaction. We used existing N1, N2, and RP primer and probe sets by the Centers for Disease Control and Prevention, but substituted fluorophores to allow multiplexing of the assay. The cycle threshold (Ct) values of our multiplex RT-qPCR were comparable to those obtained by the single assay adapted for research purposes. Low copy numbers (≥500 copies/reaction) of SARS-CoV-2 RNA were consistently detected by the multiplex RT-qPCR. Our novel multiplex RT-qPCR improves upon current single diagnostics by saving reagents, costs, time, and labor. The current RT-qPCR assay recommended for SARS-CoV-2 testing in the United States requires analysis of three genomic targets per sample: two viral and one host. This study describes a multiplex PCR that saves both reagents and labor while accurately detecting SARS-CoV-2 RNA.
影响因子：
19
作者：
Singanayagam, Anika;Patel, Monika;Gopal, Robin
通讯作者：
Gopal, Robin
影响因子：
9.3
作者：
Chu, Daniel K. W.;Pan, Yang;Poon, Leo L. M.
通讯作者：
Poon, Leo L. M.
影响因子：
28.3
作者：
通讯作者：
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