Hmaïed F, Jebri S, Saavedra M E R, Yahya M, Amri I, Lucena F, Hamdi M
Unité de Microbiologie et de Biologie Moléculaire, CNSTN, Technopôle de Sidi Thabet, 2020, Sidi Thabet, Tunisia.
Department of Microbiology, Barcelona University, Diagonal 645, 08028, Barcelona, Spain.
Curr Microbiol. 2016 Jan;72(1):12-8. doi: 10.1007/s00284-015-0909-4. Epub 2015 Sep 12.
Human enteric viruses are a major causative agent of emerging waterborne diseases and constitute a serious public health concern. Environmental contamination occurs through discharge of waste materials from infected persons. Methods for viral detection should be developed to detect low infective dose of enteric viruses in environment. In this study, we aimed at comparing two concentration methods for the detection of naturally occurring enteroviruses in raw and treated sewage. In the first method, polyethylene glycol is used to concentrate viral particles from the collected samples. The second method is based on ultracentrifugation of viral particles at high speed (110,000×g). Genomes of enteroviruses were quantified by the quantitative real-time PCR method in raw and treated sewage samples. PEG-based method yielded higher genomic copies of enteric viruses (with an average of 5.9 log10 genomic copies/100 mL) when applied to raw sewage samples. While the ultracentrifugation assay in the second method decreases genomic copies number (with an average of 5.4 log10 genomic copies/100 mL). The recovery differences between the two methods were not significant when applied to clean samples (treated sewage). This could be explained by the presence of inhibitors, which interfere with qRT-PCR, in less quantity comparatively to raw sewage. PEG-based method would be more accurate for samples with high-organic matter load. This report emphasizes the importance of matrices nature on the recovery of enteroviruses from sewage samples. This should be taken into consideration for establishing standardized virological assays to ensure the virological quality control of discharged water in environment.
人类肠道病毒是新出现的水传播疾病的主要病原体，严重威胁公众健康。环境污染是由感染者排放的废物造成的。应开发病毒检测方法，以检测环境中低感染剂量的肠道病毒。在本研究中，我们旨在比较两种浓缩方法，用于检测原污水和处理后污水中天然存在的肠道病毒。第一种方法是使用聚乙二醇从采集的样本中浓缩病毒颗粒。第二种方法是基于对病毒颗粒进行高速（110,000×g）超速离心。通过定量实时PCR方法对原污水和处理后污水样本中的肠道病毒基因组进行定量。基于聚乙二醇的方法应用于原污水样本时，产生的肠道病毒基因组拷贝数更高（平均为5.9 log10基因组拷贝/100 mL）。而第二种方法中的超速离心检测降低了基因组拷贝数（平均为5.4 log10基因组拷贝/100 mL）。当应用于清洁样本（处理后污水）时，两种方法的回收率差异不显著。这可以解释为，与原污水相比，抑制剂的含量较少，而抑制剂会干扰定量逆转录PCR。基于聚乙二醇的方法对于高有机物负荷的样本更准确。本报告强调了基质性质对从污水样本中回收肠道病毒的重要性。在建立标准化病毒学检测方法以确保环境中排放水的病毒学质量控制时，应考虑这一点。
