使用联合逆转录聚合酶链反应熔解曲线筛选和SIRPH技术对严重急性呼吸综合征冠状病毒2（SARS-CoV-2）变异株进行快速碱基特异性检测
Rapid Base-Specific Calling of SARS-CoV-2 Variants of Concern Using Combined RT-PCR Melting Curve Screening and SIRPH Technology.
作者信息
Tierling Sascha, Kattler Kathrin, Vogelgesang Markus, Pfuhl Thorsten, Lohse Stefan, Lo Porto Christina, Schmitt Beate, Nastasja Seiwert, Salhab Abdulrahman, Smola Sigrun, Walter Jörn
机构信息
Department of Genetics, Saarland University, Saarbrücken, Saarland, Germany.
Institute of Virology, Saarland University Medical Center, Homburg, Saarland, Germany.
出版信息
Open Forum Infect Dis. 2021 Jul 14;8(8):ofab364. doi: 10.1093/ofid/ofab364. eCollection 2021 Aug.
BACKGROUND
The emergence of novel variants of concern of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) demands fast and reliable detection of such variants in local populations.
METHODS
Here we present a cost-efficient and fast workflow combining a prescreening of SARS-CoV-2-positive samples using reverse transcription polymerase chain reaction melting curve analysis with multiplexed IP-RP-HPLC-based single nucleotide primer extensions.
RESULTS
The entire workflow from positive SARS-CoV-2 testing to base-specific identification of variants requires about 24 hours.
CONCLUSIONS
We applied the sensitive method to monitor local variant of concern outbreaks in SARS-CoV-2-positive samples collected in a confined region of Germany.
背景
严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的新型关注变体的出现需要在当地人群中快速可靠地检测此类变体。
方法
在此，我们展示了一种经济高效且快速的工作流程，该流程将使用逆转录聚合酶链反应熔解曲线分析对SARS-CoV-2阳性样本进行预筛选与基于多重IP-RP-HPLC的单核苷酸引物延伸相结合。
结果
从SARS-CoV-2阳性检测到变体的碱基特异性鉴定的整个工作流程大约需要24小时。
结论
我们应用这种灵敏的方法来监测在德国一个受限区域收集的SARS-CoV-2阳性样本中当地关注变体的爆发情况。
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