nRT-PCRNested Reverse-transcriptase Polymerase Chain Reaction Assay
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Specimen nRT-PCR * (sequencing) 06FO927 T653 Cerebellum + Neudoerfl ([dagger]) T654 Cerebellum - -- T655 Neocortex + Neudoerfl ([dagger]) T656 Neocortex - -- T659 Neocortex + Neudoerfl ([dagger]) T660 Neocortex - -- T657 Brain stem + Neudoerfl ([dagger]) T658 Brain stem - -- T661 Brain stem + Neudoerfl T662 Brain stem + Neudoerfl ([dagger]) 06FO926 T663 Cerebrospinal fluid - -- Total 6/11 * +, virus detected; -, no virus detected.
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.
Twenty-seven blood pellets of bats collected in 2000 were also analyzed by nRT-PCR.
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.