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离子强度对噬菌体 MS2 行为的影响及其对超滤膜评估病毒截留的意义。

Effects of ionic strength on bacteriophage MS2 behavior and their implications for the assessment of virus retention by ultrafiltration membranes.

机构信息

Laboratoire de Microbiologie Industrielle (LU49), Université de Toulouse, Université Paul Sabatier, Toulouse, France.

出版信息

Appl Environ Microbiol. 2011 Jan;77(1):229-36. doi: 10.1128/AEM.01075-10. Epub 2010 Nov 12.

Abstract

Bacteriophage MS2 is widely used as a surrogate to estimate pathogenic virus elimination by membrane filtration processes used in water treatment. Given that this water technology may be conducted with different types of waters, we focused on investigating the effects of ionic strength on MS2 behavior. For this, MS2 was analyzed while suspended in solutions of various ionic strengths, first in a batch experiment and second during membrane ultrafiltration, and quantified using (i) quantitative reverse transcriptase PCR (qRT-PCR), which detects the total number of viral genomes, (ii) qRT-PCR without the RNA extraction step, which reflects only particles with a broken capsid (free RNA), and (iii) the PFU method, which detects only infectious viruses. At the beginning of the batch experiments using solutions containing small amounts of salts, losses of MS2 infectivity (90%) and broken particles (20%) were observed; these proportions did not change during filtration. In contrast, in high-ionic-strength solutions, bacteriophage kept its biological activity under static conditions, but it quickly lost its infectivity during the filtration process. Increasing the ionic strength decreased both the inactivation and the capsid breakup in the feed suspension and increased the loss of infectivity in the filtration retentate, while the numbers of MS2 genomes were identical in both experiments. In conclusion, the effects of ionic strength on MS2 behavior may significantly distort the results of membrane filtration processes, and therefore, the combination of classical and molecular methods used here is useful for an effective validation of the retention efficiency of ultrafiltration membranes.

摘要

噬菌 MS2 被广泛用作替代物,用于估计水处理中膜过滤工艺对致病性病毒的去除效果。鉴于该水技术可能在不同类型的水中进行,我们专注于研究离子强度对 MS2 行为的影响。为此,我们在不同离子强度的溶液中分析了 MS2 的悬浮液,首先在批处理实验中,其次在膜超滤过程中进行,并使用 (i) 定量逆转录 PCR(qRT-PCR)进行定量,该方法检测病毒基因组的总数,(ii) 不进行 RNA 提取步骤的 qRT-PCR,该方法仅反映带有破裂衣壳的颗粒(游离 RNA),以及 (iii) PFU 方法,该方法仅检测传染性病毒。在使用含有少量盐的溶液进行批处理实验的开始时,观察到 MS2 感染性(90%)和破裂颗粒(20%)的损失;在过滤过程中,这些比例没有变化。相比之下,在高离子强度溶液中,噬菌体在静态条件下保持其生物活性,但在过滤过程中很快失去其感染性。增加离子强度会降低进料悬浮液中的失活和衣壳破裂,同时增加过滤截留物中感染性的损失,而在两种实验中 MS2 基因组的数量都相同。总之,离子强度对 MS2 行为的影响可能会严重扭曲膜过滤过程的结果,因此,这里使用的经典和分子方法的组合对于超滤膜的保留效率的有效验证是有用的。

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