Abstract
Reverse transcriptase polymerase chain reaction (RT-PCR)-based techniques allow for highly sensitive and specific detection of RNA viruses. Detection of the amplification products can be achieved using several methods. The following are descriptions of the detection of ebolavirus RNA using end-point RT-PCR (agarose gel visualization of amplification products) and quantitative RT-PCR (Q-RT-PCR), with fluorescent detection using an intercalating dye or detection with the use of 5′ hydrolysis probe assays. All of these techniques can be used to accurately detect the presence of ebolavirus in samples.
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References
Sanchez A, Ksaizek TG, Rollin PE, Miranda MEG, Trappier SG, Khan AS et al (1999) Detection and molecular characterization of ebola viruses causing disease in human and nonhuman primates. J Infect Dis 179(Suppl 1):S164–S169
Towner J, Sealy T, Khristova M, Albariño C, Conlan S, Reeder S et al (2008) Newly discovered ebola virus associated with hemorrhagic fever outbreak in Uganda. PLoS Pathog 4(11):e1000212
Grolla A, Jones SM, Fernando L, Strong JE, Ströher U, Möller P et al (2011) The use of a mobile laboratory unit in support of patient management and epidemiological surveillance during the 2005 Marburg outbreak in Angola. PLoS Negl Trop Dis 5(5):e1183
Towner J, Rollin PE, Bausch DG, Sanchez A, Crary SM, Vincent M et al (2004) Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome. J Virol 78(8):4330–4341
Trombley A, Wachter L, Garrison J, Buckley-Beason V, Jarhling J, Hensley L et al (2010) Comprehensive panel of real-time TaqMan polymerase chain reaction assays for detection and absolute quatification of filoviruses, arenaviruses and New World hantaviruses. Am J Trop Med Hyg 82(5):954–960
Panning M, Laue T, Olschlager S, Eickmann M, Becker S, Raith S et al (2007) Diagnostic reverse-transcription polymerase chain reaction kit for filoviruses based on the strain collections of all European biosafety level 4 laboratories. J Infect Dis 196(Suppl 2):S199–S204
Pinsky BA, Sahoo MK, Sandlund J, Kleman M, Kulkarni M, Grufman P et al (2015) Analytical performance characteristics of the Cepheid GeneXpert Ebola assay for the detection of ebola virus. PLoS One 10(11):e0142216
Dreier J, Störmer M, Kleesiek K (2005) Use of bacteriophage MS2 as an internal control in viral reverse transcription-PCR assays. J Clin Microbiol 43(9):4551–4557
Boddicker JD, Rota PA, Kreman T, Wangeman A, Lowe L, Hummel KB et al (2007) Real-time reverse transcription PCR assay for detection of mumps virus RNA in clinical specimens. J Clin Microbiol 45(9):2902–2908
Smither SJ, Weller SA, Phelps A, Eastaugh L, Ngugi S, O’Brien LM et al (2015) Buffer AVL alone does not inactivate ebola virus in a representative clinical sample type. J Clin Microbiol 53(10):3148–3154
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Grolla, A. (2017). Real-Time and End-Point PCR Diagnostics for Ebola Virus. In: Hoenen, T., Groseth, A. (eds) Ebolaviruses. Methods in Molecular Biology, vol 1628. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7116-9_27
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DOI: https://doi.org/10.1007/978-1-4939-7116-9_27
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