Peretz David, Supattapone Surachai, Giles Kurt, Vergara Julie, Freyman Yevgeniy, Lessard Pierre, Safar Jiri G, Glidden David V, McCulloch Charles, Nguyen Hoang-Oanh B, Scott Michael, Dearmond Stephen J, Prusiner Stanley B
Institute for Neurodegenerative Diseases, 513 Parnassus Ave., HSE-774, San Francisco, CA 94143-0518, USA.
J Virol. 2006 Jan;80(1):322-31. doi: 10.1128/JVI.80.1.322-331.2006.
Prompted by the discovery that prions become protease-sensitive after exposure to branched polyamine dendrimers in acetic acid (AcOH) (S. Supattapone, H. Wille, L. Uyechi, J. Safar, P. Tremblay, F. C. Szoka, F. E. Cohen, S. B. Prusiner, and M. R. Scott, J. Virol. 75:3453-3461, 2001), we investigated the inactivation of prions by sodium dodecyl sulfate (SDS) in weak acid. As judged by sensitivity to proteolytic digestion, the disease-causing prion protein (PrPSc) was denatured at room temperature by SDS at pH values of < or =4.5 or > or =10. Exposure of Sc237 prions in Syrian hamster brain homogenates to 1% SDS and 0.5% AcOH at room temperature resulted in a reduction of prion titer by a factor of ca. 10(7); however, all of the bioassay hamsters eventually developed prion disease. When various concentrations of SDS and AcOH were tested, the duration and temperature of exposure acted synergistically to inactivate both hamster Sc237 prions and human sporadic Creutzfeldt-Jakob disease (sCJD) prions. The inactivation of prions in brain homogenates and those bound to stainless steel wires was evaluated by using bioassays in transgenic mice. sCJD prions were more than 100,000 times more resistant to inactivation than Sc237 prions, demonstrating that inactivation procedures validated on rodent prions cannot be extrapolated to inactivation of human prions. Some procedures that significantly reduced prion titers in brain homogenates had a limited effect on prions bound to the surface of stainless steel wires. Using acidic SDS combined with autoclaving for 15 min, human sCJD prions bound to stainless steel wires were eliminated. Our findings form the basis for a noncorrosive system that is suitable for inactivating prions on surgical instruments, as well as on other medical and dental equipment.
鉴于朊病毒在乙酸(AcOH)中暴露于支化多胺树枝状大分子后对蛋白酶变得敏感这一发现(S. 苏帕塔蓬、H. 维勒、L. 上池、J. 萨法尔、P. 特伦布莱、F. C. 绍卡、F. E. 科恩、S. B. 普鲁辛纳和M. R. 斯科特,《病毒学杂志》75:3453 - 3461, 2001),我们研究了十二烷基硫酸钠(SDS)在弱酸中对朊病毒的灭活作用。根据对蛋白水解消化的敏感性判断,致病朊病毒蛋白(PrPSc)在室温下,当pH值≤4.5或≥10时会被SDS变性。将叙利亚仓鼠脑匀浆中的Sc237朊病毒在室温下暴露于1% SDS和0.5% AcOH中,导致朊病毒滴度降低约10⁷倍;然而,所有生物测定仓鼠最终都患上了朊病毒疾病。当测试不同浓度的SDS和AcOH时,暴露的持续时间和温度协同作用以灭活仓鼠Sc237朊病毒和人类散发性克雅氏病(sCJD)朊病毒。通过在转基因小鼠中进行生物测定来评估脑匀浆中和与不锈钢丝结合的朊病毒的灭活情况。sCJD朊病毒对灭活的抗性比Sc237朊病毒高100,000倍以上,这表明在啮齿动物朊病毒上验证有效的灭活程序不能外推至人类朊病毒的灭活。一些能显著降低脑匀浆中朊病毒滴度的程序对与不锈钢丝表面结合的朊病毒影响有限。使用酸性SDS并结合高压灭菌15分钟,可以消除与不锈钢丝结合的人类sCJD朊病毒。我们的发现为一种非腐蚀性系统奠定了基础,该系统适用于对外科器械以及其他医疗和牙科设备上的朊病毒进行灭活。