Abstract
Quantitative viral risk assessments for wastewaters are notoriously difficult. The often considered quantitative reverse transcriptase PCR reflects poorly on virus infectivity rates leading to inaccurate risk interpretations. Various techniques focused on the degradation of the nucleic acids of non-infective viruses were previously employed. We comparatively assessed the effectiveness of such enzymatic treatments for MS2 bacteriophage in treated wastewaters. The single use of RNase A at an appropriate concentration may be as effective as the combination of RNase followed by Proteinase K and more rapid. While all tested enzymatic treatments minimized recovery of RNA (>95 %) in the absence of infective MS2, none completely eliminated the signal recovery. Selection of any enzymatic protocol for minimizing recovery of RNA from degraded, non-infective viruses should balance the methods efficacy with its expediency.
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Acknowledgments
Funding was provided by the New Mexico State University, Agricultural Experiment Station, and by the U.S. Department of Energy under contract DE-EE0003046 awarded to the National Alliance for Advanced Biofuels and Bioproducts. A. Unc has received support from the University of Leeds through a water@leeds Senior Cheney Fellowship during data analysis and manuscript preparation.
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Unnithan, V.V., Unc, A. & Smith, G.B. Enzymatic Pre-treatment of Wastewater to Minimize Recovery by Reverse Transcriptase PCR of RNA from Inactive Bacteriophages. Curr Microbiol 71, 49–53 (2015). https://doi.org/10.1007/s00284-015-0830-x
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DOI: https://doi.org/10.1007/s00284-015-0830-x