Tick-borne encephalitis virus-specific RT-PCR--a rapid test for detection of the pathogen without viral RNA purification
Language English Country Switzerland Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
15595210
Knihovny.cz E-resources
- MeSH
- 5' Untranslated Regions genetics MeSH
- DNA Primers MeSH
- Species Specificity MeSH
- Ixodes physiology virology MeSH
- Animals, Suckling MeSH
- Brain virology MeSH
- Mice MeSH
- Nymph virology MeSH
- Reverse Transcriptase Polymerase Chain Reaction * MeSH
- RNA, Viral genetics MeSH
- Life Cycle Stages MeSH
- Encephalitis Viruses, Tick-Borne genetics isolation & purification MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 5' Untranslated Regions MeSH
- DNA Primers MeSH
- RNA, Viral MeSH
Among diseases transmitted by ticks in the Czech Republic, tick-borne encephalitis (TBE) caused by Tick-borne encephalitis virus (TBEV) and Lyme disease caused by Borrelia burgdorferi spirochete are most important. We propose an effective and specific test for detection of TBEV in a single tick or a pool of ticks based on the detection of TBEV RNA using an RT-PCR technique without RNA purification. The method is very sensitive with the detection limit of about 14 fg TBEV RNA in total RNA obtained from brain suspension from suckling mice infected with TBEV per reaction. The primers were derived from the 5'-terminal non-coding region, a highly conserved part of the virus. The method was successfully applied to field-collected ticks in detecting TBEV RNA. This method can be used in studies of several aspects of TBEV: epidemiology, screening of natural foci, circulation and detection of virus genome sequences in clinical materials.
