Investigating norovirus removal by microfiltration, ultrafiltration, and precoagulation-microfiltration processes using recombinant norovirus virus-like particles and real-time immuno-PCR.

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.