Investigating norovirus removal by microfiltration, ultrafiltration, and precoagulation-microfiltration processes using recombinant norovirus virus-like particles and real-time immuno-PCR
Investigating norovirus removal by microfiltration, ultrafiltration, and precoagulation-microfiltration processes using recombinant norovirus virus-like particles and real-time immuno-PCR
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使用重组诺如病毒病毒样颗粒和实时免疫 PCR 研究通过微滤、超滤和预凝微滤工艺去除诺如病毒
DOI：
10.1016/j.watres.2013.07.004
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发表时间：
2013
期刊：
影响因子：
12.8
通讯作者：
Y.
中科院分区：
文献类型：
--
作者：
Matsushita;T.;Shirasaki;N.;Tatsuki;Y. and Matsui;Y.
通过饮用水处理工艺去除微生物已在使用人工繁殖微生物的实验室规模实验中得到了广泛研究。然而，这种方法不能应用于去除诺如病毒，因为这种病毒不会在细胞或器官培养物中生长，这一事实阻碍了我们研究其在饮用水处理过程中行为的能力。为了克服这一困难，我们的研究小组之前在实验室规模的饮用水处理实验中使用了重组诺如病毒病毒样颗粒（rNV-VLPs），该颗粒由人工表达的诺如病毒衣壳蛋白组成。然而，通常用于检测rNV-VLP的酶联免疫吸附测定（ELISA）方法不够灵敏，无法评估超滤（UF）获得的高去除率。因此，我们开发并应用了实时免疫聚合酶链反应 (iPCR) 测定法进行 rNV-VLP 定量，以研究通过微滤 (MF)、UF 和混合预凝 - MF 过程去除诺如病毒。与 ELISA 相比，开发的 iPCR 检测的 rNV-VLP 检测限提高了至少 1000 倍。虽然标称孔径为 0.1 μm 的 MF 无法消除 NV-VLP，但截留分子量为 1 kDa 的 UF 却实现了 4 个对数的减少。当 MF 与预凝相结合时（氯化铁≥10 μmol-Fe/L；聚合氯化铝≥20 μmol-Al/L；明矾≥40 μmol-Al/L），混合工艺消除 rNV-VLP 的性能优于 1 kDa UF 所实现的性能。对于所有过程，噬菌体 MS2 和 Qβ 的去除率比 rNV-VLP 去除率大 1-2 个对数，因此这两种噬菌体都不能被推荐作为可能的保守替代品来预测这些过程中天然 NV 的行为。
The removal of microorganisms by drinking water treatment processes has been widely investigated in laboratory-scale experiments using artificially propagated microorganisms. However, this approach cannot be applied to norovirus removal, because this virus does not grow in cell or organ culture, and this fact has hampered our ability to investigate its behavior during drinking water treatment. To overcome this difficulty, our research group previously used recombinant norovirus virus-like particles (rNV-VLPs), which consist of an artificially expressed norovirus capsid protein, in laboratory-scale drinking water treatment experiments. However, the enzyme-linked immunosorbent assay (ELISA) method generally used to detect rNV-VLPs is not sensitive enough to evaluate high removal ratios such as those obtained by ultrafiltration (UF). We therefore developed and applied a real-time immuno-polymerase chain reaction (iPCR) assay for rNV-VLP quantification to investigate norovirus removal by microfiltration (MF), UF, and hybrid precoagulation–MF processes. The rNV-VLP detection limit with the developed iPCR assay was improved at least 1000-fold compared with ELISA. Whereas MF with a nominal pore size of 0.1 μm could not eliminate NV-VLPs, a 4-log reduction was achieved by UF with a molecular weight cutoff of 1 kDa. When MF was combined with precoagulation (≥10 μmol-Fe/L for ferric chloride; ≥20 μmol-Al/L for polyaluminum chloride; ≥40 μmol-Al/L for alum), the performance of the hybrid process in eliminating rNV-VLPs was greater than that achieved by the 1 kDa UF. For all processes, the removal ratios of the bacteriophages MS2 and Qβ were greater than the rNV-VLP removal ratios by 1–2 logs, so neither bacteriophage can be recommended as a possible conservative surrogate for predicting the behavior of native NV during these processes.