20933016
not annotated - annotated - LINNAEUS only
Detection and differentiation of tick-borne encephalitis virus subtypes by a reverse transcription quantitative real-time PCR and pyrosequencing.
Tick-borne encephalitis (TBE) virus causes one of the most important flaviviral infections of the human central nervous system in Europe and Asia. In recent years the rate of TBE infection has been raising and the virus has been spreading to new areas. Currently, the diagnosis of TBE is based on detection of specific antibodies in patients' sera which appear as late as about 2 weeks post-infection. For a timely diagnosis of TBE virus infections and epidemiological studies, a TBE virus-specific reverse transcription quantitative real-time PCR (RT-qPCR) followed by pyrosequencing was developed. The assay is based on one degenerated primer pair detecting all three human-pathogenic TBE virus subtypes with a detection limit of 10 copies. Even though primers and probe are highly degenerated, the assay is specific for TBE virus species and detects all subtypes with a comparable sensitivity. Furthermore, TBE virus RT-qPCR could be carried out as one-step or two-step assay. RT-qPCR can be followed by pyrosequencing which allows a rapid subtyping of TBE viruses. For detection purposes an internal control to monitor RNA extraction, cDNA synthesis and amplification is included. In summary, the method is sensitive, highly specific and easy-to-handle tool for the detection and differentiation of TBE virus in the early phase of illness or in TBE host animal species and ticks.
Ann file
T1 Species 33 62 tick-borne encephalitis virus
N1 Reference T1 Taxonomy:11084
T2 Species 148 183 Tick-borne encephalitis (TBE) virus
N2 Reference T2 Taxonomy:11084
T3 Species 246 251 human
N3 Reference T3 Taxonomy:9606
T4 Species 323 326 TBE
N4 Reference T4 Taxonomy:11084
T5 Species 429 432 TBE
N5 Reference T5 Taxonomy:11084
T6 Species 576 585 TBE virus
N6 Reference T6 Taxonomy:11084
T7 Species 628 637 TBE virus
N7 Reference T7 Taxonomy:11084
T8 Species 818 823 human
N8 Reference T8 Taxonomy:9606
T9 Species 835 844 TBE virus
N9 Reference T9 Taxonomy:11084
T10 Species 971 980 TBE virus
N10 Reference T10 Taxonomy:11084
T11 Species 1058 1067 TBE virus
N11 Reference T11 Taxonomy:11084
T12 Species 1204 1215 TBE viruses
N12 Reference T12 Taxonomy:11084
T13 Species 1451 1460 TBE virus
N13 Reference T13 Taxonomy:11084
T14 Species 1497 1500 TBE
N14 Reference T14 Taxonomy:11084