Cheshomi Naeema, Alum Absar, Smith Matthew F, Lim Efrem S, Conroy-Ben Otakuye, Abbaszadegan Morteza
School of Sustainable Engineering and the Built Environment, Arizona State University, Tempe, Arizona, USA.
Water and Environmental Technology Center, Arizona State University, Tempe, Arizona, USA.
Appl Environ Microbiol. 2025 Jan 31;91(1):e0133924. doi: 10.1128/aem.01339-24. Epub 2024 Dec 6.
Viral detection methodologies used for wastewater-based epidemiology (WBE) studies have a broad range of efficacies. The complex matrix and low viral particle load in wastewater emphasize the importance of the concentration method. This study focused on comparing three commonly used virus concentration methods: polyethylene glycol precipitation (PEG), immuno-magnetic nanoparticles (IMNP), and electronegative membrane filtration (EMF). Influent and effluent wastewater samples were processed by the methods and analyzed by DNA/RNA quantification and sequencing for the detection of human viruses. SARS-COV-2, Astrovirus, and Hepatitis C virus were detected by all the methods in both sample types. PEG precipitation resulted in the detection of 20 types of viruses in influent and 16 types in effluent samples. The corresponding number of virus types detected was 21 and 11 for IMNP, and 16 and 8 for EMF. Certain viruses were unique to only one concentration method. For example, PEG detected three types of viruses in influent and six types in effluent compared to IMNP, which detected seven types in influent and one type in effluent samples. However, the EMF method appeared to be the least effective, detecting three types in influent and none in effluent samples. Rotavirus was detected in influent sample using IMNP method, whereas EMF and PEG methods failed to yield a similar outcome. Consequently, the potential false negative results pose a risk to the credibility of WBE applications. Therefore, implementation of a proper concentration technique is critical to minimize method biases and ensure accurate viral profiling in WBE studies.IMPORTANCEIn recent years, significant research efforts have been focused on the development of viral detection methodology for wastewater-based epidemiology studies, showing a range of variability in detection efficacies. A proper methodology is essential for an appropriate evaluation of disease prevalence and community health in such studies and necessitates designing a concentration method based on the target pathogenic virus. There remains a need for comparative performance evaluations of methods in the context of detection efficiencies. This study highlights the significant impact of sample matrix, viral structure, and nucleic acid composition on the efficacy of viral concentration methods. Assessing WBE techniques to ensure accurate detection and understanding of viral presence within wastewater samples is critical for revealing viral profiles in municipality wastewater samples.
用于基于废水的流行病学（WBE）研究的病毒检测方法具有广泛的效能范围。废水中复杂的基质和低病毒颗粒载量凸显了浓缩方法的重要性。本研究着重比较三种常用的病毒浓缩方法：聚乙二醇沉淀法（PEG）、免疫磁性纳米颗粒法（IMNP）和电负性膜过滤法（EMF）。采用这些方法对进水和出水废水样本进行处理，并通过DNA/RNA定量和测序分析来检测人类病毒。在两种样本类型中，所有方法均检测到了严重急性呼吸综合征冠状病毒2（SARS-CoV-2）、星状病毒和丙型肝炎病毒。PEG沉淀法在进水样本中检测到20种病毒，在出水样本中检测到16种病毒。IMNP检测到的病毒类型数量在进水和出水中分别为21种和11种，EMF则分别为16种和8种。某些病毒仅在一种浓缩方法中被检测到。例如，与IMNP相比，PEG在进水样本中检测到3种病毒，在出水样本中检测到6种病毒，而IMNP在进水样本中检测到7种病毒，在出水样本中检测到1种病毒。然而，EMF方法似乎效果最差，在进水样本中检测到3种病毒，在出水样本中未检测到任何病毒。使用IMNP方法在进水样本中检测到了轮状病毒，而EMF和PEG方法未能得到类似结果。因此，潜在的假阴性结果对WBE应用的可信度构成风险。所以，实施适当的浓缩技术对于最小化方法偏差并确保WBE研究中准确的病毒谱分析至关重要。
重要性
近年来，大量研究工作集中在开发用于基于废水的流行病学研究的病毒检测方法上，这些方法在检测效能上呈现出一定的变异性。在这类研究中，恰当的方法对于准确评估疾病流行情况和社区健康至关重要，并且需要根据目标致病病毒设计浓缩方法。在检测效率方面，仍然需要对各种方法进行比较性能评估。本研究强调了样本基质、病毒结构和核酸组成对病毒浓缩方法效能的重大影响。评估WBE技术以确保准确检测并了解废水样本中的病毒存在情况，对于揭示城市废水样本中的病毒谱至关重要。
