Dickinson J, Murdoch H, Dennis M J, Hall G A, Bott R, Crabb W D, Penet C, Sutton J M, Raven N D H
Health Protection Agency, Porton Down, Salisbury, UK.
J Hosp Infect. 2009 May;72(1):65-70. doi: 10.1016/j.jhin.2008.12.007. Epub 2009 Feb 6.
A previous study has demonstrated the potential of alkaline proteases to inactivate bovine spongiform encephalopathy (BSE301V). Here we explored the use of MC3, a genetically engineered variant of Bacillus lentus subtilisin. MC3 was used to digest BSE301V infectious mouse brain homogenate (iMBH). MC3 eliminated all detectable 6H4-immunoreactive material at pH 10 and 12; however, Proteinase K was only partially effective at pH 12. When bioassayed in VM mice, MC3- and Proteinase K-digested iMBH gave respectively 66.6% and 22.7% survival rates. Using a titration series for disease incubation, this equates to a >7log reduction in infectivity for MC3 and >6log reduction for Proteinase K. This study demonstrates the potential for thermostable proteases to be developed as effective inactivation processes for prion agents in healthcare management.
先前的一项研究已证明碱性蛋白酶使牛海绵状脑病(BSE301V)失活的潜力。在此,我们探索了MC3的用途,它是迟缓芽孢杆菌枯草杆菌蛋白酶的一种基因工程变体。MC3被用于消化BSE301V感染性小鼠脑匀浆(iMBH)。MC3在pH 10和12时消除了所有可检测到的6H4免疫反应性物质;然而,蛋白酶K在pH 12时仅部分有效。当在VM小鼠中进行生物测定时,经MC3和蛋白酶K消化的iMBH的存活率分别为66.6%和22.7%。使用疾病潜伏期的滴定系列,这相当于MC3使感染力降低>7个对数,蛋白酶K使感染力降低>6个对数。这项研究证明了热稳定蛋白酶有潜力被开发为医疗管理中朊病毒剂的有效灭活方法。