Optimisation of a triplex real time RT-PCR for detection of hepatitis E virus RNA and validation on biological samples
Language English Country Netherlands Media print-electronic
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
22207081
DOI
10.1016/j.jviromet.2011.12.007
PII: S0166-0934(11)00492-7
Knihovny.cz E-resources
- 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
- Names of Substances
- RNA, Viral 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
Evidence of Hepatitis E Virus in Goat and Sheep Milk
Preliminary Study of Sars-Cov-2 Occurrence in Wastewater in the Czech Republic
Changes in Microbial Composition of Wastewater During Treatment in a Full-Scale Plant
