Establishment of a stable PrP(Sc) panel from brain tissues of experimental hamsters with scrapie strain 263K.
OBJECTIVE
To establish a stable PrP(Sc) panel from brain tissues of experimental hamsters infected with scrapie agent 263K for evaluating diagnostic techniques of human and animals' prion diseases.


METHODS
Thirty brain tissue samples from hamsters intracerebrally infected with scrapie strain 263K and another 30 samples from normal hamsters were selected to prepare 10%, 1%, and 0.5% brain homogenates, which were aliquoted into stocks. PrP(Sc) in each brain homogenate was determined by proteinase K digestions followed by Western blot assay and partially by immunohistochemistry. Stability and glycoforms of PrP(Sc) were repeatedly detected by PrP(Sc)-specific Western blots in half a year and 3 years later.


RESULTS
PrP(Sc) signals were observed in all 10% brain homogenates of infected hamsters. Twenty out of 30 stocks and 19 out of 30 stocks were PrP(Sc) positive in 1% and 0.5% brain homogenatesof infected hamsters, respectively. Twenty-seven out of 30 stocks presented three positive bands in 10% brain homogenates, whereas none of 1% and 0.5% homogenates contained 3 bands. The detection of PrP(Sc)-specific signals stored in half a year and 3 years later demonstrated that the ratio of PrP(Sc) positive samples and glycoforms was almost unchanged. All normal hamsters' brain homogenates were PrP(Sc) negative.


CONCLUSIONS
A PrP(Sc) panel of prion disease can be established, which displays reliably stable PrP(Sc)-specific signals and glycoforms.
Zhang
Bao-Yun
BY
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 100052, China.
Tian
Chan
C
Han
Jun
J
Gao
Chen
C
Shi
Qi
Q
Chen
Jian-Ming
JM
Jiang
Hui-Ying
HY
Zhou
Wei
W
Dong
Xiao-Ping
XP
D000818:Animals; D001921:Brain; D006224:Cricetinae; D007150:Immunohistochemistry; D008297:Male; D018620:PrPSc Proteins / Q000145:classification*; D012608:Scrapie
D018620:PrPSc Proteins