Langeveld Jan P M, Wang Jeng-Jie, Van de Wiel Dick F M, Shih Giles C, Garssen G Jan, Bossers Alex, Shih Jason C H
Division of Infectious Diseases and Food Chain Quality, Institute for Animal Science and Health, Lelystad, The Netherlands.
J Infect Dis. 2003 Dec 1;188(11):1782-9. doi: 10.1086/379664. Epub 2003 Nov 18.
Prions-infectious agents involved in transmissible spongiform encephalopathies-normally survive proteolytic and mild protein-destructive processes. Using bacterial keratinase produced by Bacillus licheniformis strain PWD-1, we tested conditions to accomplish the full degradation of prion protein (PrP) in brain-stem tissue from animals with bovine spongiform encephalopathy and scrapie. The detection of PrPSc, the disease-associated isoform of PrP, in homogenates was done by Western blotting and various antibodies. The results indicated that only in the presence of detergents did heat pretreatment at >100 degrees C allow the extensive enzymatic breakdown of PrPSc to a state where it is immunochemically undetectable. Proteinase K and 2 other subtilisin proteases, but not trypsin and pepsin, were also effective. This enzymatic process could lead to the development of a method for the decontamination of medical and laboratory equipment. The ultimate effectiveness of this method of prion inactivation has to be tested in mouse bioassays.
朊病毒——与传染性海绵状脑病相关的感染因子——通常能在蛋白水解和轻度蛋白质破坏过程中存活下来。我们使用地衣芽孢杆菌PWD - 1菌株产生的细菌角蛋白酶，测试了使患有牛海绵状脑病和羊瘙痒症的动物脑干组织中的朊病毒蛋白（PrP）完全降解的条件。通过蛋白质印迹法和各种抗体对匀浆中与疾病相关的PrP异构体PrPSc进行检测。结果表明，只有在存在去污剂的情况下，100℃以上的热预处理才能使PrPSc大量酶解至免疫化学检测不到的状态。蛋白酶K和其他两种枯草杆菌蛋白酶有效，但胰蛋白酶和胃蛋白酶无效。这一酶促过程可能会促成一种医疗和实验室设备去污方法的开发。这种朊病毒灭活方法的最终有效性必须在小鼠生物测定中进行测试。
