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dc.contributor.authorMcKinney, Michelle D.
dc.contributor.authorMoon, Steven J.
dc.contributor.authorKulesh, David A.
dc.contributor.authorLarsen, Thomas
dc.contributor.authorSchoepp, Randal J.
dc.date.accessioned2012-02-06T11:40:36Z
dc.date.available2012-02-06T11:40:36Z
dc.date.issued2009
dc.identifier.citationMcKinney, M. D., Moon, S. J., Kulesh, D. A., Larsen, T., Schoepp, R. J. (2009). Detection of viral RNA from paraffin-embedded tissues after prolonged formalin fixation. Journal of Clinical Virology, 44:39-42en_US
dc.identifier.issn1386-6532
dc.identifier.issn1873-5967
dc.identifier.urihttp://dx.doi.org/10.1016/j.jcv.2008.09.003
dc.identifier.urihttp://hdl.handle.net/10570/393
dc.description.abstractBackground: Isolating amplifiable RNA from formalin-fixed, paraffin-embedded (FFPE) tissues is more difficult than isolating DNA because of RNases, chemical modification of the RNA, and cross-linking of nucleic acids and proteins. Tissues containing infectious disease agents that require bio safety level (BSL)-3 and -4 necessitate fixation times of 21 and 30 days, respectively. Objective: To improve procedures for extracting RNA fromthese FFPE tissues and detect the RNA with the more sensitive TaqMan®-based reverse transcriptase (RT)-PCR. Study design: Through a single modification of a commercially available kit,we were able to extract amplifiable RNA and detect West Nile virus (WNV), Marburg virus (MARV), and Ebola virus (EBOV)-infected tissues using TaqMan® assays. Results: Formalin fixation results in an approximately 2 log10 reduction in detection limit when compared to fresh tissues. Increasing proteinase K digestion (24 h) improved extraction of amplifiable RNA from FFPE tissues. The TaqMan® results were comparable to more traditional detection results such as virus isolation. Conclusion: This improved extraction procedure for obtaining RNA combined with the TaqMan® RT-PCR assays permit retrospective and prospective studies on FFPE tissues infected with BSL-3 and -4 pathogens.en_US
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.subjectVirus Detectionen_US
dc.subjectRNAen_US
dc.subjectFormalin-fixeden_US
dc.subjectParaffin-embeddeden_US
dc.subjectPCRen_US
dc.subjectDetectionen_US
dc.titleDetection of viral RNA from paraffin-embedded tissues after prolonged formalin fixationen_US
dc.typeJournal article, peer revieweden_US


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