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Single-step reverse transcriptase-polymerase chain reaction for detection of Borna disease virus RNA in vitro and in vivo

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/jjvr.46.4.165

Title: Single-step reverse transcriptase-polymerase chain reaction for detection of Borna disease virus RNA in vitro and in vivo
Authors: MIZUTANI, Tetsuya Browse this author
OGINO, Michiko Browse this author
NISHINO, Yoshii Browse this author
KIMURA, Takashi Browse this author
INAGAKI, Hisae Browse this author
HAYASAKA, Daisuke Browse this author
KARIWA, Hiroaki Browse this author
TAKASHIMA, Ikuo Browse this author
Keywords: Borna disease virus
single-step reverse transcriptase-polymerase chain reaction
Issue Date: 26-Feb-1999
Publisher: The Graduate School of Veterinary Medicine
Journal Title: Japanese Journal of Veterinary Research
Volume: 46
Issue: 4
Start Page: 165
End Page: 169
Abstract: There are few copies of Borna disease virus (BDV) genome in peripheral blood mononuclear cells and no reliable standard reverse transcriptase-polymerase chain reaction (RT-PCR) method for the detection of BDV RNA, which is both highly sensitive and free of contamination. Single-step RT-PCR, in which both reverse transcription and amplification by Taq DNA polymerase work efficiently in a single buffer, was applied to detect the p24 region of BDV RNA in vitro and in vivo. Using in vitro synthesized RNA, it was demonstrated that at least 100 copies of BDV RNA could be detected and the sensitivity and specificity were nearly equal to those obtained by RT-nested PCR. We could detect BDV RNA from more than 1 pg of cellular RNA obtained from BDV-persistently infected MDCK cells. Furthermore, this method was successfully performed on brain specimens obtained from a BDV-infected rat at 11 weeks post-inoculation. This single-step RT-PCR method will be convenient for detecting limited amounts of BDV RNA in various cells and tissue samples.
Type: bulletin
URI: http://hdl.handle.net/2115/2716
Appears in Collections:Volume 46, Number 4

 

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