Using propidium monoazide to distinguish between viable and nonviable bacteria, MS2 and murine norovirus.
OBJECTIVE
The ability to distinguish between viable and/or infectious micro-organisms and inactivated cells is extremely important for correctly performing microbial risk assessments. In this study, we evaluated whether propidium monoazide (PMA)-qPCR could distinguish between viable and nonviable bacteria and viruses.


RESULTS
A PMA-qPCR combined assay was applied to viable and inactivated bacteria (Escherichia coli and Bacillus subtilis) and viruses (MS2 and murine norovirus [MNV]). PMA, a DNA-intercalating agent, in combination with PCR was better able to distinguish between viable and nonviable bacteria and viruses than conventional PCR.


CONCLUSIONS
These results suggest that a combined PMA-qPCR assay can be used to measure the viability of bacterial cells and bacteriophage MS2, but not MNV.


CONCLUSIONS
PMA-qPCR could potentially be used to measure the viability of some micro-organisms, including virus. However, a thorough evaluation should be performed prior to measuring the viability of micro-organisms by PMA-qPCR in a quantitative way.
Department of Environmental Health, School of Public Health, Institute of Health and Environment, Seoul National University, Seoul, Korea.
S Y
SY
Kim
G
G
Ko
D001386:Azides / Q000737:chemistry*; D001412:Bacillus subtilis / Q000254:growth & development; D004926:Escherichia coli / Q000254:growth & development; D007364:Intercalating Agents / Q000737:chemistry*; D017909:Levivirus / Q000254:growth & development; D050296:Microbial Viability; D029322:Norovirus / Q000254:growth & development; D016133:Polymerase Chain Reaction / Q000379:methods*; D011419:Propidium / Q000031:analogs & derivatives* / Q000737:chemistry
D001386:Azides; D007364:Intercalating Agents; C533957:propidium monoazide; D011419:Propidium