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Volume 46, Number 2-3 >

A single-tube RT-PCR method for the detection of Borna disease viral genomic RNA

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Please use this identifier to cite or link to this item:http://doi.org/10.14943/jjvr.46.2-3.73

Title: A single-tube RT-PCR method for the detection of Borna disease viral genomic RNA
Authors: MIZUTANI, Tetsuya Browse this author
OGINO, Michiko Browse this author
NISHINO, Yoshii Browse this author
KIMURA, Takashi Browse this author
KARIWA, Hiroaki Browse this author
TSUJIMURA, Kouji Browse this author
INAGAKi, Hisae Browse this author
TAKASHIMA, Ikuo Browse this author
Keywords: Borna disease virus
Single-tube RT-PCR method
Issue Date: 30-Nov-1998
Publisher: The Graduate School of Veterinary Medicine
Journal Title: Japanese Journal of Veterinary Research
Volume: 46
Issue: 2-3
Start Page: 73
End Page: 81
Abstract: For detecting Borna disease virus (BDV) genomic stranded RNA, single-tube reverse transcription-polymerase chain reaction (St RT-PCR) was developed to equal the sensitivity of RT-nested PCR but with reduced risk of contamination. BDV-genomic stranded RNA was synthesized in vitro using plasmid cDNA of BDV p24 region as a template and RNA was also extracted from BDV-persistently infected MDCK (MDCK/BDV) cells. Both RNAs were amplified by St RT-PCR in which a single round of RT and a single round of PCR were performed in the same tube. Ten copies of synthesized RNA could be amplified by St RT-PCR, indicating that St RT-PCR method is as sensitive as the ordinary RT-nested PCR method. Furthermore, this method was applied to quantify the exact copy number of genomic RNA in MDCK/BDV cells. Signals were obtained from the samples containing more than 1 pg total cellular RNA. From the results, approximately 100 copies of BDV genomic RNA exist in one MDCK/BDV cell. BDV genomic RNA from the in vivo RNA samples using St RT-PCR, indicating this method is applicable for the epidemiological study of BDV without contamination.
Type: bulletin (article)
URI: http://hdl.handle.net/2115/2644
Appears in Collections:Japanese Journal of Veterinary Research > Volume 46, Number 2-3

Submitter: 獣医学部図書室

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