title: [Highly-sensitive nonradioactive detection of the tick-borne encephalitis virus].
abstract: The non-radioactive reverse dot-blot method was used for the detection of tick-borne encephalitis virus (TBEV) in clinical specimens. The method involves reverse transcription (RT) and polymerase chain reaction (PCR) using a pair of biotin-labelled oligonucleotide primers. These primers flank a region in the gene of the envelope protein E, which is more conserved than other regions, and initiate the polymerisation with RNAs of all the investigated strains. The amplified cDNA was captured from solution on a solid support using complementary oligonucleotides covalently bound to a polyamide membrane. The biotin labels of the resulting hybrids were visualized by means of the streptavidin-horseradish peroxidase conjugate. The detection limit of the test was about 10(3)-10(4) molecules of target RNA. The sensitivity was comparable to that obtained by dot-hybridization of PCR-product with 32P-labelled DNA probe. The method was used for the detection of RNA in specimens of tick and blood.
authors.forename: T S
authors.initials: TS
authors.lastname: Godovikova
authors.forename: T N
authors.initials: TN
authors.lastname: Orlova
authors.forename: E Iu
authors.initials: EIu
authors.lastname: Dobrikova
authors.forename: V A
authors.initials: VA
authors.lastname: Shamanin
authors.forename: V F
authors.initials: VF
authors.lastname: Zarytova
authors.forename: N V
authors.initials: NV
authors.lastname: Vorob'eva
authors.forename: N A
authors.initials: NA
authors.lastname: Serdiukova
authors.forename: M Iu
authors.initials: MIu
authors.lastname: Shamanina
authors.forename: I O
authors.initials: IO
authors.lastname: Petruseva
authors.forename: N D
authors.initials: ND
authors.lastname: Pitsenko
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D004669:Encephalitis Viruses, Tick-Borne / Q000235:genetics / Q000302:isolation & purification*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D012367:RNA, Viral
vernacular_title: Vysokochuvstvitel'naia neradioaktivnaia detektsiia virusa kleshchevogo éntsefalita.