Inference of Active Viral Replication in Cases with Sustained Positive Reverse Transcription-PCR Results for SARS-CoV-2.
Inference of Active Viral Replication in Cases with Sustained Positive Reverse Transcription-PCR Results for SARS-CoV-2.
复制标题
SARS-COV-2的持续呈阳性逆转录PCR结果的病毒复制的推断。
DOI：
10.1128/jcm.02277-20
复制
发表时间：
2021-01-21
影响因子：
9.4
通讯作者：
García de Viedma D
中科院分区：
文献类型：
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作者：
Rodríguez-Grande C;Adán-Jiménez J;Catalán P;Alcalá L;Estévez A;Muñoz P;Pérez-Lago L;García de Viedma D
本研究的目的是检测2019年冠状病毒病（COVID-19）病例，其严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的逆转录-PCR（RT-PCR）结果持续阳性，由于存在仅在复制病毒中表达的亚基因组（SG）病毒RNA，因此可以推断出活病毒。从鼻咽部诊断标本中提取残留的RNA作为模板，进行特异性RT-PCR检测。本研究的目的是检测2019年冠状病毒病（COVID-19）病例，其严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的逆转录-PCR（RT-PCR）结果持续阳性，由于存在仅在复制病毒中表达的亚基因组（SG）病毒RNA，因此可以推断出活病毒。从鼻咽部诊断标本中提取残留的RNA作为模板，进行特异性RT-PCR检测。作为对照，我们还检测了E基因和/或人类管家基因（RNase P）的病毒基因组RNA。我们评估了60例RT-PCR阳性病例的样本，这些病例自首次诊断性RT-PCR以来具有延长的病毒SARS-CoV-2脱落（24至101天）。在症状发作后28至79天，在12/60（20%）的持续病例中检测到SG病毒RNA。具有延长的病毒脱落和存在SG RNA的病例的年龄范围相当宽（40至100岁），并且病例在男性（42%）和女性（58%）之间均匀分布。没有艾滋病毒阳性病例，但有7例免疫抑制。根据COVID-19疫情的严重程度，其为轻度（40%）、中度（20%）及重度（40%）。在一定比例的持续SARS-CoV-2 PCR阳性病例中，可以推断存在活跃复制的病毒，远远超出诊断范围。我们不应假设COVID-19病例普遍缺乏传染性，病毒脱落时间较长。
The purpose of this study was to detect coronavirus disease 2019 (COVID-19) cases with persistent positive reverse transcription-PCR (RT-PCR) results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which viable virus can be inferred due to the presence of subgenomic (SG) viral RNA, which is expressed only in replicating viruses. RNA remnants purified from diagnostic nasopharyngeal specimens were used as the templates for RT-PCR-specific detection of SG E gene RNA. The purpose of this study was to detect coronavirus disease 2019 (COVID-19) cases with persistent positive reverse transcription-PCR (RT-PCR) results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), for which viable virus can be inferred due to the presence of subgenomic (SG) viral RNA, which is expressed only in replicating viruses. RNA remnants purified from diagnostic nasopharyngeal specimens were used as the templates for RT-PCR-specific detection of SG E gene RNA. As controls, we also detected viral genomic RNA for the E gene and/or a human housekeeping gene (RNase P). We assessed the samples of 60 RT-PCR-positive cases with prolonged viral SARS-CoV-2 shedding (24 to 101 days) since the first diagnostic RT-PCR. SG viral RNA was detected in 12/60 (20%) of the persistent cases, 28 to 79 days after the onset of symptoms. The age range of the cases with prolonged viral shedding and the presence of SG RNA was quite wide (40 to 100 years), and the cases were equally distributed between males (42%) and females (58%). No case was HIV positive, although seven were immunosuppressed. According to the severities of the COVID-19 episodes, they were mild (40%), intermediate (20%), and severe (40%). In a percentage of persistent SARS-CoV-2 PCR-positive cases, the presence of actively replicating virus may be inferred, far beyond diagnosis. We should not assume a universal lack of infectiousness for COVID-19 cases with prolonged viral shedding.