朊病毒病实验动物脑组织PrPSc样本库的建立

目的 建立朊病毒病实验动物脑组织检测样本库,检测其在特定保存条件下的稳定性,以用于动物和人朊病毒病诊断技术的评估和考核.方法 选用30只经颅内注射感染羊瘙痒因子263K的仓鼠和30只正常仓鼠,每只分别制备10%、1%和0.5%3种浓度的脑组织匀浆,分装冷冻保存.以Western Blot方法 确定脑组织匀浆中PrPSc的存在情况,并分别在建库半年和3年检测PrPSc的稳定性.结果 30只感染仓鼠10%脑组织匀浆全部可检出PrPSc,1%脑组织匀浆有26份为PrPSc阳性,0.5%脑组织匀浆有19份为PrPSc阳性.半年及3年后复检,PrPSc阳性检出率基本不变.所有正常仓鼠脑匀浆均为PrPSc阴性.结论 建立了稳定性良好的含有90份PrPSc阳性标本和90份PrPSc阴性样本的朊病毒病实验动物脑组织检测样本库.

Establishment of a prion disease PrPSc panel from the brain tissues of experimental hamsters infected with scrapie agent 263K

Objective To establish a prion disease PrPSc panel from the brain tissues of experimental hamsters and to address the stability of the panel conserved under the specific condition, for evaluating the diagnostic techniques of human and animal's prion diseases. Methods 30 brain tissues of hamsters infected with scrapie strain 263K intracerebrally and 30 ones of normal hamsters were enrolled in this panel. Each brain sample was prepared to 10%, 1% and 0.5% homogenates and aliquoted into stocks. The presences of PrPSc in each brain sample were evaluated with PrP-specific Western Blots and partially with immunohistochemistry, and the stabihty of PrPSc signals in each sample were repeatedly assessed half a year later and 3 years later. Results PrPSc signals were detected in all stocks of 10% brain homogenates from infected hamsters, 26 out of 30 stocks of 1% homogenates and 19 out of 30 stocks of 0.5 % homogenates. The assessments of PrPSc signals in all samples half-year and three years later demonstrated almost unchanged. All homogenates of brain tissues of normal hamsters were PrPSc negative. Conclusion A prion disease PrPSc panel of the brain tissues, which includes 90 PrPSc positive stocks and PrPSc negative ones, was successfully estabhshed, with a reliable stability of PrPSc signals.