葡萄牙废水中登革热病毒和基孔肯雅热病毒的检测：一项探索性监测研究。

Detection of dengue virus and chikungunya virus in wastewater in Portugal: an exploratory surveillance study.

机构信息

Laboratório de Análises, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal; Civil Engineering Research and Innovation for Sustainability, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal; Department of Nuclear Sciences and Engineering, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.

Laboratório de Análises, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.

出版信息

Lancet Microbe. 2024 Nov;5(11):100911. doi: 10.1016/S2666-5247(24)00150-2. Epub 2024 Sep 12.

DOI:10.1016/S2666-5247(24)00150-2

PMID:39278232

Abstract

BACKGROUND

The global distribution and prevalence of arboviral diseases have increased in recent years, driven by factors such as climate change, biodiversity loss, globalisation, and urbanisation. These diseases are often underestimated due to uneven surveillance and unreported asymptomatic cases. Current surveillance relies on vector and clinical surveillance. In this study, we aimed to explore wastewater-based surveillance (WBS) as an additional tool for dengue virus (DENV) and chikungunya virus (CHIKV) tracking.

METHODS

In this exploratory surveillance study, WBS was done at eleven wastewater treatment plants in three regions in Portugal (North, Lisboa and Vale do Tejo, and south). Using quantitative RT-PCR, we quantified in raw wastewater the RNA concentrations of DENV and CHIKV (non-structural viral protein 1 [nsP1] and envelope protein [E1] genes) once every 2 weeks for a period of 11 months, between May 16, 2022, and April 19, 2023. Results were normalised with crAssphage (concentration of target viral RNA divided by the concentration of crAssphage DNA) and provided as median normalised viral load. Prevalence (proportion of positive samples) and viral quantities were summarised for the total sampling period, by calendar month, and by seasons.

FINDINGS

273 samples were collected from 11 wastewater treatment plants situated across the North (n=75 samples), Lisboa and Vale do Tejo (n=98), and south (n=100) regions of Portugal. DENV was detected in 68 (25%) of 273 samples, with a median viral load of 1·1 × 10 (IQR 3·2 × 10 to 8·0 × 10). CHIKV was detected in 30 (11%) of 273 samples, with median viral loads of 3·1 × 10 (1·6 × 10 to 6·4 × 10; nsP1 gene) and 7·8 × 10 (4·2 × 10 to 2·0 × 10³; E1 gene). The pattern of occurrence of CHIKV was similar between regions whereas slight differences were found for DENV. When combining results for the three studied regions, DENV prevalence and viral load had two seasonal peaks (summer and winter) and CHIKV prevalence and viral load had a single peak during March and April of 2023.

INTERPRETATION

This study highlights the potential of WBS as a potent tool for gauging the epidemiological landscape of DENV and CHIKV in Portugal, where autochthonous cases have not yet been detected. WBS could serve as an additional element to conventional surveillance approaches, especially in areas where real-time clinical surveillance data are scarce or delayed.

FUNDING

EU Emergency Support Instrument and Fundação para a Ciencia e Tecnologia.

摘要

背景

近年来，由于气候变化、生物多样性丧失、全球化和城市化等因素，虫媒病毒病的全球分布和流行率有所增加。由于监测不均衡和无症状病例未报告，这些疾病往往被低估。目前的监测依赖于媒介和临床监测。在这项研究中，我们旨在探索基于废水的监测（WBS）作为追踪登革热病毒（DENV）和基孔肯雅热病毒（CHIKV）的额外工具。

方法

在这项探索性监测研究中，我们在葡萄牙的三个地区（北部、里斯本和特茹河谷，以及南部）的 11 个污水处理厂进行了 WBS。使用定量 RT-PCR，我们在 11 个月的时间内（2022 年 5 月 16 日至 2023 年 4 月 19 日）每隔两周对原始废水中 DENV 和 CHIKV（非结构病毒蛋白 1 [nsP1]和包膜蛋白 [E1]基因）的 RNA 浓度进行定量。结果用 crAssphage（目标病毒 RNA 浓度除以 crAssphage DNA 浓度）归一化，并以中位数归一化病毒载量表示。对整个采样期、逐月和按季节总结了阳性样本的比例（阳性样本的比例）和病毒载量。

结果

从葡萄牙北部（n=75 份样本）、里斯本和特茹河谷（n=98 份样本）和南部（n=100 份样本）的 11 个污水处理厂采集了 273 份样本。在 273 份样本中，DENV 的检出率为 68%（25%），中位病毒载量为 1.1×10（IQR 3.2×10 至 8.0×10）。在 273 份样本中，CHIKV 的检出率为 30%（11%），中位病毒载量分别为 3.1×10（nsP1 基因，1.6×10 至 6.4×10）和 7.8×10（E1 基因，4.2×10 至 2.0×10³）。CHIKV 的发生模式在各地区相似，而 DENV 则存在细微差异。当结合三个研究地区的结果时，DENV 的患病率和病毒载量有两个季节性高峰（夏季和冬季），而 CHIKV 的患病率和病毒载量在 2023 年 3 月和 4 月有一个单一高峰。