US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2
US CDC Real-Time Reverse Transcription PCR Panel for Detection of Severe Acute Respiratory Syndrome Coronavirus 2
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DOI：
10.3201/eid2608.201246
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发表时间：
2020-08-01
影响因子：
11.8
通讯作者：
Lindstrom, Stephen
中科院分区：
文献类型：
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作者：
Lu, Xiaoyan;Wang, Lijuan;Lindstrom, Stephen
严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）被确定为与2019年底出现的冠状病毒病相关的病原。为此，我们开发了一个由3种针对核衣壳基因的实时反转录PCR检测组成的诊断小组，并评估了这些检测在检测SARS-CoV-2感染中的应用。所有试验均显示出8个数量级的线性动态范围和定量RNA转录本的5拷贝/反应和细胞培养的SARS-CoV-2的1 × 10(-1.5) 50%组织培养感染剂量/mL的分析限。所有检测方法均可与含有培养病毒的鼻咽和口咽分泌物、血清和粪便标本进行比较。我们没有获得与其他人类冠状病毒或常见呼吸道病原体的假阳性扩增。在临床标本检测中，所有3种检测结果高度相关。2020年2月4日，美国食品和药物管理局发布了紧急使用授权，允许紧急使用该面板。
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was identified as the etiologic agent associated with coronavirus disease, which emerged in late 2019. In response, we developed a diagnostic panel consisting of 3 real-time reverse transcription PCR assays targeting the nucleocapsid gene and evaluated use of these assays for detecting SARS-CoV-2 infection. All assays demonstrated a linear dynamic range of 8 orders of magnitude and an analytical limit of detection of 5 copies/reaction of quantified RNA transcripts and 1 x 10(-1.5) 50% tissue culture infectious dose/mL of cell-cultured SARS-CoV-2. All assays performed comparably with nasopharyngeal and oropharyngeal secretions, serum, and fecal specimens spiked with cultured virus. We obtained no false-positive amplifications with other human coronaviruses or common respiratory pathogens. Results from all 3 assays were highly correlated during clinical specimen testing. On February 4, 2020, the Food and Drug Administration issued an Emergency Use Authorization to enable emergency use of this panel.