Research results will be presented as a poster at Prion 2008 in Madrid in October (Prion Molecular Biology Lab, Foreign Animal Disease Division) ( 08/06/24 ) | |||||
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Part | Charger | leeyh | date | 08/06/24 | |
In vitro amplification of chronic wasting disease in elk
Hyun-Joo Sohn1 , Yoon-Hee Lee1, Chan-Lan Kim1, Dong-Seob Tark1, Young-Pyo Choi4, In-Soo Cho2, Yi-Seok Joo3
1 Foreign Animal Disease Division; 2 Virology Division, ;3Animal Disease Diagnostic Center, National Veterinary Research and Quarantine Service, Anyang, Gyeonggi-do 430-824, Korea; 4University of Edinburgh, Edinburgh EH8 9YL UK
Chronic wasting disease(CWD) of cervids is a transmissible spongiform encephalopathy(TSE). TSE pathogenesis is associated with refolding of the normal prion protein, PrPc, into a partially protease-resistant isomer termed PrPsc. Soto and colleagues greatly extended the process and power of in vitro PrPres amplification in developing protein misfolding cyclic amplification(PMCA). In PMCA, normal brain homogenates(NBH) supply PrPc, which upon addition of infected brain homogenate is refold into protease-resistant isoform, PrPres. To enhance sensitivity of CWD prion(PrPCWD) detection in elk, we established in vitro serial PMCA modeled after Soto et al. Here we report amplification using CWD-negative brain homogenate cervid PrP transgenic mice[Tg(elkPrP)]. The serial PMCA- diluting amplified material into fresh Tg(elkPrP) NBH for each successive round – resulted in a yield of 109 fold after just 5 rounds. The magnitude of PrPCWD conversion obtained with serial PMCA may make it possible in vitro detection of PrPCWD in retropharyngeal lymph node and brain of infected animals.
Corresponding author: Dr. Hyun-Joo Sohn(Tel: +82 31 467 1867, Fax: +82 31 467 1830, E.mail: shonhj@nvrqs.go.kr)
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