Laboratory of Bioresources and Food Safety, Faculty of Sciences and Technology, Cadi Ayyad University, Marrakech, Morocco.
Laboratory of Bioresources and Food Safety, Faculty of Sciences and Technology, Cadi Ayyad University, Marrakech, Morocco; Higher School of Technology of El Kelâa des Sraghna, Cadi Ayyad University, El Kelâa des Sraghna, Morocco.
J Virol Methods. 2024 Dec;330:115013. doi: 10.1016/j.jviromet.2024.115013. Epub 2024 Aug 28.
Human enteric viruses, as adenovirus (HAdV), norovirus (HuNoV) and rotavirus (RVA) are significant causes of gastroenteritis associated with consumption of contaminated water worldwide. Various methods have been described for their detection and monitoring in water. The aim of this study was to compare the performance of four conditions for concentrating HAdV, HuNoV and RVA from water matrices, in order to develop a single protocol that could simultaneously concentrate all target viruses from tap water. The tested conditions were based on the adsorption-elution using electronegative filters, in which we evaluated cation-coated filtration by MgCl with or without acid rinse by HSO and two elution buffers, namely NaOH and tris-glycine-beef extract. Genomic material was extracted and amplified by real-time PCR and real-time RT-PCR using commercial kits. Based on the statistical analysis of amplification results (cycles of quantification), the condition involving cation-coated filtration by MgCl using electronegative filters with acid rinse by HSO combined with NaOH elution allowed efficient recovery of both HAdV, HuNoV and RVA from tap water compared to the other conditions. These findings confirm the effectiveness of the approach used to monitor three major enteric viruses in tap water.
人类肠道病毒，如腺病毒（HAdV）、诺如病毒（HuNoV）和轮状病毒（RVA），是全球因饮用受污染水而引起的胃肠炎的重要病原体。已经有多种方法被用于这些病毒在水中的检测和监测。本研究旨在比较四种从水样中浓缩 HAdV、HuNoV 和 RVA 的条件，以开发一种能够同时从自来水中浓缩所有目标病毒的单一方案。测试的条件基于使用带负电荷的过滤器的吸附-洗脱，其中我们评估了 MgCl 阳离子涂层过滤，以及是否用 HSO 进行酸冲洗，还有两种洗脱缓冲液，即 NaOH 和 tris-甘氨酸-牛肉提取物。使用商业试剂盒通过实时 PCR 和实时 RT-PCR 提取和扩增基因组材料。基于对扩增结果（定量循环数）的统计分析，与其他条件相比，使用带负电荷的过滤器进行 MgCl 阳离子涂层过滤，并用 HSO 进行酸冲洗，然后用 NaOH 洗脱的条件，能从自来水中有效回收 HAdV、HuNoV 和 RVA。这些发现证实了用于监测自来水中三种主要肠道病毒的方法的有效性。
