采用数字液滴PCR或实时PCR定量血浆中SARS-CoV-2 RNA的方法——有差异吗?
Droplet Digital PCR or Real-Time PCR as a Method for Quantifying SARS-CoV-2 RNA in Plasma-Is There a Difference?
作者信息
Granerud Beathe Kiland, Kaarbø Mari, Al-Baldawi Huda, Otterdal Kari, Halvorsen Bente, Lind Andreas, Rayner Simon, Holter Jan Cato, Dudman Susanne
机构信息
Institute of Clinical Medicine, University of Oslo, 0318 Oslo, Norway.
Department of Microbiology, Oslo University Hospital, 0450 Oslo, Norway.
出版信息
Viruses. 2025 May 28;17(6):772. doi: 10.3390/v17060772.
The aim of this study is to ascertain whether qRT-PCR (reverse transcriptase real-time PCR) or RT-ddPCR (reverse transcriptase digital droplet PCR) is more effective for detecting SARS-CoV-2 RNA (severe acute respiratory syndrome coronavirus 2 RNA) in blood plasma from COVID-19 (coronavirus infectious disease-19) patients. The E-gene of SARS-CoV-2 RNA was quantified using both methods in 128 plasma samples from 70 hospitalized patients, followed by a statistical analysis to compare the sensitivity and concordance between the methods. Out of the 128 samples, 89 yielded consistent results irrespective of the method used, whereas 39 samples showed discrepancies between the two different methods. RT-ddPCR frequently registered higher viral quantities compared to qRT-PCR; however, the results did not demonstrate a clear superiority in sensitivity for RT-ddPCR. Although RT-ddPCR registered higher viral quantities, this study concludes that both methods provide comparable results for detecting SARS-CoV-2 E-gene RNA in plasma.
本研究的目的是确定定量逆转录聚合酶链反应(qRT-PCR)或逆转录数字液滴聚合酶链反应(RT-ddPCR)在检测新型冠状病毒肺炎(COVID-19,冠状病毒感染疾病-19)患者血浆中的严重急性呼吸综合征冠状病毒2(SARS-CoV-2)RNA时哪种方法更有效。使用这两种方法对70名住院患者的128份血浆样本中的SARS-CoV-2 RNA的E基因进行定量,随后进行统计分析以比较两种方法之间的敏感性和一致性。在128份样本中,89份无论使用哪种方法都产生了一致的结果,而39份样本在两种不同方法之间显示出差异。与qRT-PCR相比,RT-ddPCR经常记录到更高的病毒量;然而,结果并未显示RT-ddPCR在敏感性方面有明显优势。尽管RT-ddPCR记录到更高的病毒量,但本研究得出结论,两种方法在检测血浆中的SARS-CoV-2 E基因RNA时提供了可比的结果。