朊病毒病实验动物脑组织PrP^Sc样本库的建立

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

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PrP^Sc的存在情况，并分别在建库半年和3年检测PrP^Sc的稳定性。结果30只感染仓鼠10％脑组织匀浆全部可检出PrP^Sc，1％脑组织匀浆有26份为PrP^Sc阳性，0．5％脑组织匀浆有19份为PrP^Sc阳性。半年及3年后复检，PrP^Sc阳性检出率基本不变。所有正常仓鼠脑匀浆均为PrP^Sc阴性。结论建立了稳定性良好的含有90份PrP^Sc阳性标本和90份PrP^Sc阴性样本的朊病毒病实验动物脑组织榆测样本库。

Objective To establish a prion disease PrP^Sc 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 PrP^Sc in each brain sample were evaluated with PrP-specific Western Blots and partially with immunohistochemistry, and the stability of PrP^Sc signals in each sample were repeatedly assessed half a year later and 3 years later. Results PrP^Sc 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 PrP^Sc signals in all samples halfyear and three years later demonstrated almost unchanged. All homogenates of brain tissues of normal hamsters were PrP^Sc negative. Conclusion A prion disease PrP^Sc panel of the brain tissues, which includes 90 PrP^Sc positive stocks and PrP^Sc negative ones, was successfully established, with a reliable stability of PrP^Sc signals.