CSIRO Land and Water, Ecosciences Precinct, 41 Boggo Road, Qld 4102, Australia.
Australian Centre for Ecogenomics, School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, QLD 4072, Australia.
J Travel Med. 2020 Aug 20;27(5). doi: 10.1093/jtm/taaa116.
Wastewater-based epidemiology (WBE) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can be an important source of information for coronavirus disease 2019 (COVID-19) management during and after the pandemic. Currently, governments and transportation industries around the world are developing strategies to minimize SARS-CoV-2 transmission associated with resuming activity. This study investigated the possible use of SARS-CoV-2 RNA wastewater surveillance from airline and cruise ship sanitation systems and its potential use as a COVID-19 public health management tool.
Aircraft and cruise ship wastewater samples (n = 21) were tested for SARS-CoV-2 using two virus concentration methods, adsorption-extraction by electronegative membrane (n = 13) and ultrafiltration by Amicon (n = 8), and five assays using reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and RT-droplet digital PCR (RT-ddPCR). Representative qPCR amplicons from positive samples were sequenced to confirm assay specificity.
SARS-CoV-2 RNA was detected in samples from both aircraft and cruise ship wastewater; however concentrations were near the assay limit of detection. The analysis of multiple replicate samples and use of multiple RT-qPCR and/or RT-ddPCR assays increased detection sensitivity and minimized false-negative results. Representative qPCR amplicons were confirmed for the correct PCR product by sequencing. However, differences in sensitivity were observed among molecular assays and concentration methods.
The study indicates that surveillance of wastewater from large transport vessels with their own sanitation systems has potential as a complementary data source to prioritize clinical testing and contact tracing among disembarking passengers. Importantly, sampling methods and molecular assays must be further optimized to maximize detection sensitivity. The potential for false negatives by both wastewater testing and clinical swab testing suggests that the two strategies could be employed together to maximize the probability of detecting SARS-CoV-2 infections amongst passengers.
基于污水的流行病学（WBE）可用于严重急性呼吸综合征冠状病毒 2（SARS-CoV-2），这对于大流行期间和之后的 2019 年冠状病毒病（COVID-19）管理而言是重要的信息来源。目前，世界各地的政府和运输行业正在制定策略，以最大程度地减少与恢复活动相关的 SARS-CoV-2 传播。本研究调查了从航空公司和游轮卫生系统中监测 SARS-CoV-2 污水的可能性，及其作为 COVID-19 公共卫生管理工具的潜在用途。
使用两种病毒浓缩方法（吸附-电负性膜提取，n=13；Amicon 超滤，n=8）和五种使用逆转录定量聚合酶链反应（RT-qPCR）和 RT-微滴数字 PCR（RT-ddPCR）的检测方法，对 21 个飞机和游轮污水样本进行 SARS-CoV-2 检测。从阳性样本中提取代表性 qPCR 扩增子进行测序，以确认检测的特异性。
在飞机和游轮污水样本中均检测到 SARS-CoV-2 RNA，但浓度接近检测限。对多个重复样本的分析以及使用多个 RT-qPCR 和/或 RT-ddPCR 检测方法提高了检测的灵敏度，减少了假阴性结果。代表性 qPCR 扩增子经测序证实为正确的 PCR 产物。然而，分子检测方法和浓缩方法之间存在灵敏度差异。
该研究表明，对具有自身卫生系统的大型运输船舶的污水进行监测，可能成为临床检测和下船乘客接触者追踪的补充数据源。重要的是，必须进一步优化采样方法和分子检测方法，以最大程度地提高检测灵敏度。污水检测和临床拭子检测均存在假阴性的可能性，这表明这两种策略可以一起使用，以最大程度地提高在乘客中检测到 SARS-CoV-2 感染的概率。
