nRT-PCRNested Reverse-transcriptase Polymerase Chain Reaction Assay
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Results of nested reverse transcription-PCR (nRT-PCR) analysis of the tickborne encephalitis virus (TBEV)-infected macaque brain TBEV results by TBEV strain Sample no.
The threshold of detection of the nRT-PCR method was determined by preparing 10-fold dilutions of a pretitrated suspension of Strain 8918FRA (4) in TRIzol (GIBCO-BRL).
Various organs and tissues (medulla, liver, kidney, spleen, heart, tongue, esophagus-larynx-pharynx, and lung) were collected and subjected to nRT-PCR. Esophagus-larynx-pharynx and lung of bat No.
Twenty-seven blood pellets of bats collected in 2000 were also analyzed by nRT-PCR. These samples were obtained from 8 R.
The threshold of detection of the nRT-PCR for the amplification of the EBL1a genomic and antigenomic RNAs of the N gene was 5 x [10.sup.-2] fluorescent forming units of EBL1a/mL.
Nucleotide (nt) sequences were determined by using the positive nRT-PCR products obtained from the four brains and from one blood sample.