

@article {9980,
	title = {{Development of a real-time RT-PCR assay combined with ethidium monoazide treatment for RNA viruses and its application to detect viral RNA after heat exposure.}},
	journal = {Water science and technology : a journal of the International Association on Water Pollution Research},
	volume = {63},
	number = {3},
	year = {2011},
	month = {jan},
	pages = {502{\textendash}7},
	abstract = {A method was developed for discriminating damaged viruses or naked viral RNA from intact viruses by ethidium monoazide (EMA) treatment before RT-PCR. The applied EMA treatment consisted of three steps: (1) EMA dose, (2) exposure to light, and (3) additional purification by spin-column gel filtration. Approximately 4-log reduction in viral RNA concentration was observed by adding a dose of 10 $μ$g/mL-EMA with 300 s of light irradiation. Although residual EMA can be an inhibitor of RT-PCR, its effect was reduced by spin-column gel filtration or a QIAamp{\textregistered} Viral RNA Mini Kit. EMA-RT-PCR was applied to the thermally treated PV1. Results of EMA-RT-PCR were similar to the plaque assay when PV1 was thermally inactivated. Although this is a preliminary study investigating applicability of the EMA-RT-PCR method for RNA viruses, the results suggest that the method is potentially applicable for the selective detection of epidemiologically important enteric viruses in water such as enteroviruses and noroviruses.},
	keywords = {Animals, Azides, Azides: pharmacology, Hot Temperature, Light, Mice, Norovirus, Norovirus: drug effects, Norovirus: genetics, Norovirus: isolation {\&} purification, Norovirus: radiation effects, Reverse Transcriptase Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction: m, RNA, RNA Viruses, RNA Viruses: drug effects, RNA Viruses: genetics, RNA Viruses: isolation {\&} purification, RNA Viruses: radiation effects, Viral, Viral: genetics, Viral: isolation {\&} purification},
	issn = {0273-1223},
	doi = {10.2166/wst.2011.249},
	url = {http://www.ncbi.nlm.nih.gov/pubmed/21278473},
	author = {Kim, K. and Katayama, H. and Kitajima, M. and Tohya, Y. and Ohgaki, S.}
}
