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Review
. 2011 Nov;214(6):424-36.
doi: 10.1016/j.ijheh.2011.07.014. Epub 2011 Sep 15.

Methods to detect infectious human enteric viruses in environmental water samples

Ibrahim Ahmed Hamza  1 , Lars Jurzik, Klaus Überla, Michael Wilhelm
Affiliations
Review

Methods to detect infectious human enteric viruses in environmental water samples

Ibrahim Ahmed Hamza et al. Int J Hyg Environ Health. 2011 Nov.

Abstract

Currently, a wide range of analytical methods is available for virus detection in environmental water samples. Molecular methods such as polymerase chain reaction (PCR) and quantitative real time PCR (qPCR) have the highest sensitivity and specificity to investigate virus contamination in water, so they are the most commonly used in environmental virology. Despite great sensitivity of PCR, the main limitation is the lack of the correlation between the detected viral genome and viral infectivity, which limits conclusions regarding the significance for public health. To provide information about the infectivity of the detected viruses, cultivation on animal cell culture is the gold standard. However, cell culture infectivity assays are laborious, time consuming and costly. Also, not all viruses are able to produce cytopathic effect and viruses such as human noroviruses have no available cell line for propagation. In this brief review, we present a summary and critical evaluation of different approaches that have been recently proposed to overcome limitations of the traditional cell culture assay and PCR assay such as integrated cell culture-PCR, detection of genome integrity, detection of capsid integrity, and measurement of oxidative damages on viral capsid protein. Techniques for rapid detection of infectious viruses such as fluorescence microscopy and automated flow cytometry have also been suggested to assess virus infectivity in water samples.

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Figures

Fig. 1
Fig. 1
Common approaches to estimate viral infectivity in environmental water samples.
Fig. 2
Fig. 2
Schematic representation of the use of antibody coated paramagnetic beads for separation of viral particles from water.
Fig. 3
Fig. 3
Effect of (A) heat treatment (85 °C) and (B) protease treatment on bacteriophage T4 quantitation over time as assessed by plaque assay, qPCR, and PMA-PCR. As shown, PMA-qPCR is not sufficiently effective to discriminate between infectious and non-infectious viruses. Error bars show standard deviations from three independent assays.
Fig. 4
Fig. 4
Schematic representation of FRET for monitoring viral proteolytic activity in the infected cells. The cyan fluorescent protein (CFP)–yellow fluorescent protein (YFP) substrate is linked by a cleavage recognition site for poliovirus 2A protease. A fluorescence protein pair undergoing FRET, in which fluorescence energy is transferred from an excited fluorophore (reporter) to a light-absorbing molecule (acceptor) when, located in close proximity. A disruption of FRET, as shown by a change in CFP and YFP emission signals, indicates active viral replication within the infected cells.
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