Reverse Transcription-Nested Polymerase Chain Reaction for Detecting p40 RNA of Borna Disease Virus, without Risk of Plasmid Contamination

Journal of Veterinary Medical Science

Yoshii NISHINO¹  Hiroaki KARIWA²  Ikuo TAKASHIMA²  Tetsuya MIZUTANI² 

[1] Research Institute of Biosciences, School of Veterinary Medicine, Azabu University, Kanagawa 229-8501, Japan;Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818, Japan

关键词: Borna disease virus;    mRNA selactive PCR;    plasmid contamination;   

DOI  :  10.1292/jvms.61.77

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(19)Several methods for the detection of Borna disease virus (BDV) RNA have been reported, one being the reverse transcription-nested polymerase chain reaction (RT-nested PCR) method. However, due to the possibility of contamination of the cloned DNA in a reaction tube, false-positive results might be obtained by RT-nested PCR. To detect only BDV RNA without anxiety of contamination, we developed an RT-nested PCR system using “mRNA selective PCR kit”. Using this system, cDNA of BDV p40 in the plasmid (up to 5 × 107 molecules) was not amplified. BDV specific sequence was amplified from total RNA (more than 50 pg) of MDCK/BDV cells, which were persistently infected with BDV. These results indicate that this mRNA selective RT-nested PCR system can specifically amplify target RNA as distinguished from plasmid contaminated.

【 授权许可】

Unknown