Highly sensitive detection of SARS-CoV-2 RNA by multiplex rRT-PCR for molecular diagnosis of COVID-19 by clinical laboratories
Highly sensitive detection of SARS-CoV-2 RNA by multiplex rRT-PCR for molecular diagnosis of COVID-19 by clinical laboratories
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DOI：
10.1016/j.cca.2020.04.023
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发表时间：
2020-08-01
影响因子：
5
通讯作者：
Matsushita, Kazuyuki
中科院分区：
文献类型：
--
作者：
Ishige, Takayuki;Murata, Shota;Matsushita, Kazuyuki
背景：实时定量逆转录聚合酶链式反应检测SARS-CoV-2RNA用于分子诊断实验室对新冠肺炎的临床诊断。建立了检测SARS-CoV-2RNA的多重RRT-PCR方法。方法：以SARS-CoV-2特异性E基因、SARS-CoV-2特异性N基因和人ABL1基因为内对照，建立多重RRT-PCR方法。结论：所建立的多重RRT-PCR方法可以实现对SARS-CoV-2RNA的高灵敏度检测，减少试剂的使用，减少成本，减少临床检验人员所需的时间。
Background: The detection of SARS-CoV-2 RNA by real-time reverse transcription-polymerase chain reaction (rRT-PCR) is used to confirm the clinical diagnosis of COVID-19 by molecular diagnostic laboratories. We developed a multiplex rRT-PCR methodology for the detection of SARS-CoV-2 RNA.Methods: Three genes were used for multiplex rRT-PCR: the Sarbecovirus specific E gene, the SARS-CoV-2 specific N gene, and the human ABL1 gene as an internal control.Results: Good correlation of C-q values was observed between the simplex and multiplex rRT-PCR methodologies. Low copies (< 25 copies/reaction) of SARS-CoV-2 RNA were detected by the novel multiplex rRT-PCR method.Conclusion: The proposed multiplex rRT-PCR methodology will enable highly sensitive detection of SARS-CoV-2 RNA, reducing reagent use and cost, and time required by clinical laboratory technicians.