Prolonged SARS-CoV-2 cell culture replication in respiratory samples from patients with severe COVID-19.
Prolonged SARS-CoV-2 cell culture replication in respiratory samples from patients with severe COVID-19.
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DOI：
10.1016/j.cmi.2021.02.014
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发表时间：
2021-06
期刊：
影响因子：
--
通讯作者：
Delgado R
中科院分区：
文献类型：
--
作者：
Folgueira MD;Luczkowiak J;Lasala F;Pérez-Rivilla A;Delgado R
本研究比较了轻度COVID-19患者和严重双侧肺炎住院患者呼吸道样本中SARS-CoV-2的传染性。在严重的COVID-19中，我们还分析了配对血清中中和活性的存在。我们对193份实时逆转录聚合酶链反应呼吸道样本进行了细胞培养，这些样本是从189名患者在从临床诊断到随访的不同时间获得的，对SARS-CoV-2呈阳性。其中11份样本来自无症状感染者，91份样本来自91名轻度COVID-19门诊患者，91份样本来自87名重症肺炎住院患者。在这些患者中，在症状发作>10天后收集的30对血清中分析中和活性。我们在91/193（47%）样本中检测到细胞病变效应（CPE）。在轻度COVID-19患者中，病毒活力可维持长达10天。在严重的COVID-19患者中，病毒在症状出现后长达32天仍然存活。重度COVID-19患者在中低病毒载量（循环阈值≥ 26）样本中呈现感染性病毒的比率显著更高：32/75（43%）与轻度病例的14/63（22%）（p < 0.01）。我们观察到阳性CPE，尽管存在明显的中和活性（在10%（3/30）的样品中NT 50> 1：1024）。严重COVID-19患者可能在症状发作后长达4周的长时间内排出活病毒，即使在其呼吸道样本中出现高循环阈值时，尽管已产生高中和抗体滴度。
This study compares the infectivity of SARS-CoV-2 in respiratory samples from patients with mild COVID-19 with those from hospitalized patients with severe bilateral pneumonia. In severe COVID-19, we also analysed the presence of neutralizing activity in paired sera. We performed cell cultures on 193 real-time reverse transcription polymerase chain reaction respiratory samples, positive for SARS-CoV-2, obtained from 189 patients at various times, from clinical diagnosis to follow-up. Eleven samples were obtained from asymptomatic individuals, 91 samples from 91 outpatients with mild forms of COVID-19 and 91 samples from 87 inpatients with severe pneumonia. In these patients, neutralizing activity was analysed in 30 paired sera collected after symptom onset >10 days. We detected a cytopathic effect (CPE) in 91/193 (47%) samples. Viral viability was maintained for up to 10 days in patients with mild COVID-19. In patients with severe COVID-19, the virus remained viable for up to 32 days after the onset of symptoms. Patients with severe COVID-19 presented infectious virus at a significantly higher rate in the samples with moderate to low viral load (cycle threshold value ≥ 26): 32/75 (43%) versus 14/63 (22%) for mild cases (p < 0.01). We observed a positive CPE despite the presence of clear neutralizing activity (NT50 > 1:1024 in 10% (3/30) of samples. Patients with severe COVID-19 might shed viable virus during prolonged periods of up to 4 weeks after symptom onset, even when presenting high cycle threshold values in their respiratory samples and despite having developed high neutralizing antibody titres.
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