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dc.contributor.authorNocker, Andreas
dc.contributor.authorSossa, Katherine E.
dc.contributor.authorCamper, Anne K.
dc.identifier.citationNocker A, Sossa KE, Camper AK, "Molecular monitoring of disinfection efficacy using propidium monoazide in combination with quantitative PCR," Journal of Microbiological Methods 2007 70(2):252-260en_US
dc.description.abstractOne of the major drawbacks of DNA-based microbial diagnostics is its inability to discriminate between live and dead bacteria. Due to the persistence of DNA in the environment after cells have lost their viability, DNA-based assays cannot assess pathogenic risk since signals can originate from both live and dead cells. Presented here is a potential application of the novel chemical propidium monoazide (PMA), which results in the selective suppression of DNA detection from dead cells. PMA can only penetrate dead cells with permeabilized cell membranes. Upon intercalation into the DNA, covalent crosslinkage of PMA to DNA is achieved through light exposure. This modification prevents the DNA from being amplified by PCR. The method, in combination with quantitative PCR as a diagnostic tool, successfully monitored the disinfection efficacy of hypochlorite, benzalkonium and heat on several model pathogens. Threshold cycle numbers increased with increasing disinfection strength after PMA treatment of samples compared to non-PMA treated samples. With some disinfectant-specific differences, monitoring viability loss with membrane integrity as an indicator seemed to be more conservative than monitoring viability loss with plate counts. Loss of viability after short UV-exposure could not be monitored with PMA as UV light affects viability by inducing DNA damage without directly affecting membrane permeability.en_US
dc.titleMolecular monitoring of disinfection efficacy using propidium monoazide in combination with quantitative PCRen_US
mus.citation.journaltitleJournal of Microbiological Methodsen_US
mus.identifier.categoryEngineering & Computer Scienceen_US
mus.relation.collegeCollege of Engineeringen_US
mus.relation.departmentCenter for Biofilm Engineering.en_US
mus.relation.departmentChemical & Biological Engineering.en_US
mus.relation.departmentChemical Engineering.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.relation.researchgroupCenter for Biofilm Engineering.en_US
mus.contributor.orcidNocker, Andreas|0000-0002-5343-9418en_US

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