Department of Respiratory Sciences, University of Leicester, United Kingdom; Department of Clinical Microbiology, University Hospitals of Leicester NHS Trust, United Kingdom; Department of Infectious Diseases and HIV Medicine, University Hospitals of Leicester NHS Trust, Leicester, United Kingdom.
Department of Respiratory Sciences, University of Leicester, United Kingdom; Department of Infectious Diseases and HIV Medicine, University Hospitals of Leicester NHS Trust, Leicester, United Kingdom.
J Infect. 2021 Jun;82(6):253-259. doi: 10.1016/j.jinf.2021.03.018. Epub 2021 Mar 24.
Human to human transmission of SARS-CoV-2 is driven by the respiratory route but little is known about the pattern and quantity of virus output from exhaled breath. We have previously shown that face-mask sampling (FMS) can detect exhaled tubercle bacilli and have adapted its use to quantify exhaled SARS-CoV-2 RNA in patients admitted to hospital with Coronavirus Disease-2019 (COVID-19).
Between May and December 2020, we took two concomitant FMS and nasopharyngeal samples (NPS) over two days, starting within 24 h of a routine virus positive NPS in patients hospitalised with COVID-19, at University Hospitals of Leicester NHS Trust, UK. Participants were asked to wear a modified duckbilled facemask for 30 min, followed by a nasopharyngeal swab. Demographic, clinical, and radiological data, as well as International Severe Acute Respiratory and emerging Infections Consortium (ISARIC) mortality and deterioration scores were obtained. Exposed masks were processed by removal, dissolution and analysis of sampling matrix strips fixed within the mask by RT-qPCR. Viral genome copy numbers were determined and results classified as Negative; Low: ≤999 copies; Medium: 1000-99,999 copies and High ≥ 100,000 copies per strip for FMS or per 100 µl for NPS.
102 FMS and NPS were collected from 66 routinely positive patients; median age: 61 (IQR 49 - 77), of which FMS was positive in 38% of individuals and concomitant NPS was positive in 50%. Positive FMS viral loads varied over five orders of magnitude (<10-3.3 x 10 genome copies/strip); 21 (32%) patients were asymptomatic at the time of sampling. High FMS viral load was associated with respiratory symptoms at time of sampling and shorter interval between sampling and symptom onset (FMS High: median (IQR) 2 days (2-3) vs FMS Negative: 7 days (7-10), p = 0.002). On multivariable linear regression analysis, higher FMS viral loads were associated with higher ISARIC mortality (Medium FMS vs Negative FMS gave an adjusted coefficient of 15.7, 95% CI 3.7-27.7, p = 0.01) and deterioration scores (High FMS vs Negative FMS gave an adjusted coefficient of 37.6, 95% CI 14.0 to 61.3, p = 0.002), while NPS viral loads showed no significant association.
We demonstrate a simple and effective method for detecting and quantifying exhaled SARS-CoV-2 in hospitalised patients with COVID-19. Higher FMS viral loads were more likely to be associated with developing severe disease compared to NPS viral loads. Similar to NPS, FMS viral load was highest in early disease and in those with active respiratory symptoms, highlighting the potential role of FMS in understanding infectivity.
人类 2019 冠状病毒病(COVID-19)的人际传播是由呼吸道驱动的,但人们对呼出的呼吸飞沫中病毒的排放模式和数量知之甚少。我们之前已经证明,面罩采样(FMS)可以检测呼出的结核分枝杆菌,并已将其用于量化 COVID-19 住院患者呼出的 SARS-CoV-2 RNA。
2020 年 5 月至 12 月,我们在英国莱斯特大学医院对 COVID-19 住院患者在常规病毒阳性 NPS 后 24 小时内的两天内同时采集了两次 FMS 和鼻咽样本(NPS)。要求参与者佩戴改良的鸭嘴面罩 30 分钟,然后进行鼻咽拭子采样。收集参与者的人口统计学、临床和影像学数据,以及国际严重急性呼吸和新兴感染联合会(ISARIC)的死亡率和恶化评分。暴露的面罩通过去除、溶解和分析固定在面罩内的采样基质条进行处理,通过 RT-qPCR 进行分析。确定病毒基因组拷贝数,并将结果分类为阴性;低:≤999 拷贝;中:1000-99999 拷贝;高:FMS 或 NPS 每个条带≥100000 拷贝。
从 66 例常规阳性患者中采集了 102 例 FMS 和 NPS;中位年龄:61(IQR 49-77),其中 38%的个体 FMS 阳性,50%的同时采集的 NPS 阳性。FMS 的病毒载量变化超过五个数量级(<10-3.3 x 10 基因组拷贝/条);21(32%)名患者在采样时无症状。FMS 高病毒载量与采样时的呼吸道症状和采样与症状出现之间的时间间隔较短有关(FMS 高:中位数(IQR)2 天(2-3)与 FMS 阴性:7 天(7-10),p=0.002)。多变量线性回归分析显示,FMS 病毒载量较高与 ISARIC 死亡率较高(FMS 中值与 FMS 阴性相比,调整系数为 15.7,95%CI 3.7-27.7,p=0.01)和恶化评分较高(FMS 高与 FMS 阴性相比,调整系数为 37.6,95%CI 14.0-61.3,p=0.002)相关,而 NPS 病毒载量则没有显著相关性。
我们证明了一种简单有效的方法,可以检测和量化 COVID-19 住院患者呼出的 SARS-CoV-2。与 NPS 病毒载量相比,FMS 高病毒载量更有可能与发生严重疾病相关。与 NPS 相似,FMS 病毒载量在疾病早期和有活跃呼吸道症状的患者中最高,这突出了 FMS 在了解传染性方面的潜在作用。