-
Something wrong with this record ?
Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples
P. Vasickova, P. Kralik, I. Slana, I. Pavlik,
Language English Country Netherlands
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Hepatitis E diagnosis veterinary virology MeSH
- Sheep virology MeSH
- Reverse Transcriptase Polymerase Chain Reaction methods standards MeSH
- Reproducibility of Results MeSH
- RNA, Viral analysis genetics standards MeSH
- Sensitivity and Specificity MeSH
- Sus scrofa virology MeSH
- Hepatitis E virus genetics MeSH
- Deer virology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
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 "home-made" RNA standards. The limit of detection was determined experimentally as 10 copies/μl 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.
References provided by Crossref.org
- 000
- 00000naa a2200000 a 4500
- 001
- bmc12024157
- 003
- CZ-PrNML
- 005
- 20170113094338.0
- 007
- ta
- 008
- 120815s2012 ne f 000 0#eng||
- 009
- AR
- 024 7_
- $a 10.1016/j.jviromet.2011.12.007 $2 doi
- 035 __
- $a (PubMed)22207081
- 040 __
- $a ABA008 $b cze $d ABA008 $e AACR2
- 041 0_
- $a eng
- 044 __
- $a ne
- 100 1_
- $a Vašíčková, Petra $u Veterinary Research Institute, Hudcova 70, 621 00 Brno, Czech Republic. vasickova@vri.cz $7 xx0209778
- 245 10
- $a Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples / $c P. Vasickova, P. Kralik, I. Slana, I. Pavlik,
- 520 9_
- $a 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 "home-made" RNA standards. The limit of detection was determined experimentally as 10 copies/μl 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.
- 650 _2
- $a zvířata $7 D000818
- 650 _2
- $a vysoká zvěř $x virologie $7 D003670
- 650 _2
- $a hepatitida E $x diagnóza $x veterinární $x virologie $7 D016751
- 650 _2
- $a virus hepatitidy E $x genetika $7 D016752
- 650 _2
- $a RNA virová $x analýza $x genetika $x normy $7 D012367
- 650 _2
- $a reprodukovatelnost výsledků $7 D015203
- 650 _2
- $a polymerázová řetězová reakce s reverzní transkripcí $x metody $x normy $7 D020133
- 650 _2
- $a senzitivita a specificita $7 D012680
- 650 _2
- $a ovce $x virologie $7 D012756
- 650 _2
- $a Sus scrofa $x virologie $7 D034421
- 655 _2
- $a hodnotící studie $7 D023362
- 655 _2
- $a časopisecké články $7 D016428
- 655 _2
- $a práce podpořená grantem $7 D013485
- 700 1_
- $a Králík, Petr $7 uzp2011673690
- 700 1_
- $a Slana, Iva
- 700 1_
- $a Pavlik, Ivo
- 773 0_
- $w MED00003047 $t Journal of virological methods $x 1879-0984 $g Roč. 180, č. 1-2 (2012), s. 38-42
- 856 41
- $u https://pubmed.ncbi.nlm.nih.gov/22207081 $y Pubmed
- 910 __
- $a ABA008 $b sig $c sign $y m $z 0
- 990 __
- $a 20120815 $b ABA008
- 991 __
- $a 20170113094439 $b ABA008
- 999 __
- $a ok $b bmc $g 946305 $s 781485
- BAS __
- $a 3
- BAS __
- $a PreBMC
- BMC __
- $a 2012 $b 180 $c 1-2 $d 38-42 $i 1879-0984 $m Journal of virological methods $n J Virol Methods $x MED00003047
- LZP __
- $a Pubmed-20120815/12/02
