Enzymatic Pre-treatment of Wastewater to Minimize Recovery by Reverse Transcriptase PCR of RNA from Inactive Bacteriophages.
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.
Department of Biology, New Mexico State University, Las Cruces, NM, 88003, USA, vvunnithan85@gmail.com.
Veena V
VV
Unnithan
Adrian
A
Unc
Geoffrey B
GB
Smith
D019286:Endopeptidase K / Q000378:metabolism; D017909:Levivirus / Q000302:isolation & purification*; D012367:RNA, Viral / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism*; D062065:Wastewater / Q000737:chemistry* / Q000821:virology*
D019286:Endopeptidase K; D012367:RNA, Viral; D012259:Ribonuclease, Pancreatic; D062065:Wastewater