Department of Public Health, The Welfare Epidemiology and Monitoring Unit, Finnish Institute for Health and Welfare, Mannerheimintie 166, Helsinki, 00271, Finland.
Research Unit of Population Health, Faculty of Medicine, University of Oulu, Oulu, Finland.
Sci Rep. 2024 Oct 26;14(1):25582. doi: 10.1038/s41598-024-77155-6.
Reverse transcription-quantitative PCR (RT-qPCR) is widely used for monitoring viruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in wastewater. Various materials, including plasmid DNA, synthetic nucleic acids, PCR amplicons, genomic DNA, and cDNA, are currently used for SARS-CoV-2 quantification by generating standard curves. We assessed three common standards on quantifying SARS-CoV-2 RNA across nine wastewater treatment plants in Finland, as part of the national wastewater surveillance effort. We pairwise compared RT-qPCR results from 148 wastewater samples, using both IDT (#10006625, IDT, USA) and CODEX standards (#SC2-RNAC-1100, CODEX DNA), and 179 samples using both IDT and EURM019 standards (#EURM-019, European Commission, Joint Research Centre) in our assessment. Amongst the tested standards, the CODEX standard consistently yielded more stable results than either the IDT or EURM019 standards. We found that SARS-CoV-2 levels were higher with the IDT standard (4.36 Log GC/100 mL) compared to the CODEX standard (4.05 Log GC/100 mL). Similarly, quantification using the IDT standard was higher (5.27 Log GC/100 mL) than values obtained with the EURM019 (4.81 Log GC/100 mL). SARS-CoV-2 RNA quantified with IDT and CODEX standards exhibited stronger concordance (Spearman's correlation rho median of 0.79) compared to those quantified with IDT and EURM019 standards (rho median of 0.59). This study highlights the significant impact of standard material selection on SARS-CoV-2 RNA quantification, emphasizing the need for harmonization in standard material.
逆转录定量聚合酶链反应 (RT-qPCR) 广泛用于监测病毒，包括严重急性呼吸系统综合征冠状病毒 2 (SARS-CoV-2)，在废水中。目前，各种材料，包括质粒 DNA、合成核酸、PCR 扩增子、基因组 DNA 和 cDNA，用于通过生成标准曲线来定量 SARS-CoV-2。作为国家废水监测工作的一部分，我们评估了芬兰 9 个污水处理厂中三种常见标准对 SARS-CoV-2 RNA 的定量。我们使用 IDT (#10006625，IDT，美国) 和 CODEX 标准 (#SC2-RNAC-1100，CODEX DNA) 比较了 148 个废水样本的 RT-qPCR 结果，使用 IDT 和 EURM019 标准 (#EURM-019，欧盟委员会，联合研究中心) 比较了 179 个废水样本，在我们的评估中。在所测试的标准中，与 IDT 或 EURM019 标准相比，CODEX 标准始终产生更稳定的结果。我们发现，与 CODEX 标准 (4.05 Log GC/100 mL) 相比，使用 IDT 标准时 SARS-CoV-2 水平更高 (4.36 Log GC/100 mL)。同样，使用 IDT 标准进行定量时的水平也高于使用 EURM019 标准 (4.81 Log GC/100 mL) 的水平 (5.27 Log GC/100 mL)。与使用 IDT 和 EURM019 标准定量的 SARS-CoV-2 RNA 相比，使用 IDT 和 CODEX 标准定量的 SARS-CoV-2 RNA 表现出更强的一致性 (Spearman 相关 rho 中位数为 0.79)。本研究强调了标准物质选择对 SARS-CoV-2 RNA 定量的重大影响，强调了标准物质协调的必要性。
