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  • The aim of this study was to optimise a two-tube reverse transcription triplex quantitative real time PCR (qRT-PCR) combining amplification of two loci with an internal amplification control (IAC) for detection and quantitation of hepatitis E virus (HEV) RNA and to validate its performance on a pool of biological samples. Optimisation was performed on serially diluted %22home-made%22 RNA standards. The limit of detection was determined experimentally as 10 copies/mu l of the RNA standard for both amplification targets. The qRT-PCR was validated on a cohort of samples originating from 48 wild boars (Sus scrofa), 17 fallow deer (Dama dama) and 28 mouflons (Ovis musimon), with nested RT-PCR used as a reference method. qRT-PCR was found to be more specific for the detection of HEV RNA in examined samples. HEV RNA was detected in samples of five more animals (one wild boar and four mouflons) in comparison with nested RT-PCR. (C) 2011 Elsevier B.V. All rights reserved.
  • The aim of this study was to optimise a two-tube reverse transcription triplex quantitative real time PCR (qRT-PCR) combining amplification of two loci with an internal amplification control (IAC) for detection and quantitation of hepatitis E virus (HEV) RNA and to validate its performance on a pool of biological samples. Optimisation was performed on serially diluted %22home-made%22 RNA standards. The limit of detection was determined experimentally as 10 copies/mu l of the RNA standard for both amplification targets. The qRT-PCR was validated on a cohort of samples originating from 48 wild boars (Sus scrofa), 17 fallow deer (Dama dama) and 28 mouflons (Ovis musimon), with nested RT-PCR used as a reference method. qRT-PCR was found to be more specific for the detection of HEV RNA in examined samples. HEV RNA was detected in samples of five more animals (one wild boar and four mouflons) in comparison with nested RT-PCR. (C) 2011 Elsevier B.V. All rights reserved. (en)
Title
  • Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples
  • Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples (en)
skos:prefLabel
  • Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples
  • Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples (en)
skos:notation
  • RIV/00027162:_____/12:#0000871!RIV13-MZE-00027162
http://linked.open...avai/predkladatel
http://linked.open...avai/riv/aktivita
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  • P(ED0006/01/01), P(OC08045), Z(MZE0002716202)
http://linked.open...iv/cisloPeriodika
  • 1-2
http://linked.open...vai/riv/dodaniDat
http://linked.open...aciTvurceVysledku
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  • 156910
http://linked.open...ai/riv/idVysledku
  • RIV/00027162:_____/12:#0000871
http://linked.open...riv/jazykVysledku
http://linked.open.../riv/klicovaSlova
  • Internal amplification control; RNA quantitation; Zoonosis (en)
http://linked.open.../riv/klicoveSlovo
http://linked.open...odStatuVydavatele
  • NL - Nizozemsko
http://linked.open...ontrolniKodProRIV
  • [10D32540937C]
http://linked.open...i/riv/nazevZdroje
  • Journal of Virological Methods
http://linked.open...in/vavai/riv/obor
http://linked.open...ichTvurcuVysledku
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http://linked.open...vavai/riv/projekt
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http://linked.open...v/svazekPeriodika
  • 180
http://linked.open...iv/tvurceVysledku
  • Králík, Petr
  • Pavlík, Ivo
  • Slaná, Iva
  • Vašíčková, Petra
http://linked.open...ain/vavai/riv/wos
  • 000301884200007
http://linked.open...n/vavai/riv/zamer
issn
  • 0166-0934
number of pages
http://bibframe.org/vocab/doi
  • 10.1016/j.jviromet.2011.12.007
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