Boogaerts Tim, Van den Bogaert Siel, Van Poelvoorde Laura A E, El Masri Diala, De Roeck Naomi, Roosens Nancy H C, Lesenfants Marie, Lahousse Lies, Van Hoorde Koenraad, van Nuijs Alexander L N, Delputte Peter
Toxicological Centre, University of Antwerp, Universiteitsplein 1, 2610 Antwerp, Belgium.
Laboratory for Microbiology, Parasitology and Hygiene, University of Antwerp, Universiteitsplein 1, 2610 Antwerp, Belgium.
Viruses. 2022 Mar 15;14(3):610. doi: 10.3390/v14030610.
Since the beginning of the COVID-19 pandemic, the wastewater-based epidemiology (WBE) of SARS-CoV-2 has been used as a complementary indicator to follow up on the trends in the COVID-19 spread in Belgium and in many other countries. To further develop the use of WBE, a multiplex digital polymerase chain reaction (dPCR) assay was optimized, validated and applied for the measurement of emerging SARS-CoV-2 variants of concern (VOC) in influent wastewater (IWW) samples. Key mutations were targeted in the different VOC strains, including SΔ69/70 deletion, N501Y, SΔ241 and SΔ157. The presented bioanalytical method was able to distinguish between SARS-CoV-2 RNA originating from the wild-type and B.1.1.7, B.1.351 and B.1.617.2 variants. The dPCR assay proved to be sensitive enough to detect low concentrations of SARS-CoV-2 RNA in IWW since the limit of detection of the different targets ranged between 0.3 and 2.9 copies/µL. This developed WBE approach was applied to IWW samples originating from different Belgian locations and was able to monitor spatio-temporal changes in the presence of targeted VOC strains in the investigated communities. The present dPCR assay developments were realized to bring added-value to the current national WBE of COVID-19 by also having the spatio-temporal proportions of the VoC in presence in the wastewaters.
自新冠疫情开始以来，基于污水的新冠病毒流行病学（WBE）已被用作一项补充指标，以跟踪比利时和许多其他国家新冠病毒传播的趋势。为了进一步拓展WBE的应用，一种多重数字聚合酶链反应（dPCR）检测方法得到了优化、验证，并应用于测量流入污水（IWW）样本中出现的新冠病毒变异株（VOC）。针对不同的VOC毒株，对关键突变进行了靶向检测，包括SΔ69/70缺失、N501Y、SΔ241和SΔ157。所提出的生物分析方法能够区分源自野生型以及B.1.1.7、B.1.351和B.1.617.2变异株的新冠病毒RNA。dPCR检测方法被证明具有足够的灵敏度，能够检测IWW中低浓度的新冠病毒RNA，因为不同靶点的检测限在0.3至2.9拷贝/微升之间。这种开发的WBE方法应用于来自比利时不同地点的IWW样本，并能够监测所调查社区中目标VOC毒株存在情况的时空变化。当前dPCR检测方法的发展通过获取废水中存在的VOC的时空比例，为当前国家层面的新冠病毒WBE带来了附加值。
