Lytle C D, Routson L B
Division of Life Sciences, Food and Drug Administration, Rockville, Maryland 20857, USA.
Appl Environ Microbiol. 1995 Feb;61(2):643-9. doi: 10.1128/aem.61.2.643-649.1995.
Viruses are used to test the barrier properties of materials. Binding of virus particles during passage through holes in the material may yield misleading test results. The choices of challenge virus and suspending medium may be important for minimizing confounding effects that might arise from such binding. In this study, different surrogate viruses, as well as different support media, were evaluated to determine optimal test parameters. Two membranes with high-binding properties (nitrocellulose and cationic polysulfone) were used as filters to compare binding activities of different surrogate challenge viruses (MS2, phi X174, T7, PRD1, and phi 6) in different media. The media consisted of buffered saline with surfactants, serum, or culture broth as additives. In addition, elution rates of viruses that bound to the membranes were determined. The results suggest that viruses can bind by hydrophobic and electrostatic interactions, with phi X174 displaying the lowest level of binding by either process. The nonionic detergents Triton X-100 and Tween 80 (0.1%) equally minimized hydrophobic interactions. Neither anionic nor cationic surfactants were as effective at nontoxic levels. Serum was effective at reducing both hydrophobic and electrostatic binding, with 2% being sufficient for eliminating binding under our test conditions. Thus, phi X174 remains the best choice as a surrogate virus to test barrier materials, and Triton X-100 (0.1%) remains a good choice for reducing hydrophobic binding. In addition, binding of viruses by barrier materials is unlikely to prevent passage of blood-borne pathogens.
病毒被用于测试材料的阻隔性能。病毒颗粒在穿过材料上的孔洞时发生结合,可能会产生误导性的测试结果。选择挑战性病毒和悬浮介质对于将此类结合可能产生的混杂效应降至最低可能很重要。在本研究中,对不同的替代病毒以及不同的支持介质进行了评估,以确定最佳测试参数。使用两种具有高结合特性的膜(硝酸纤维素和阳离子聚砜)作为过滤器,比较不同替代挑战性病毒(MS2、phi X174、T7、PRD1和phi 6)在不同介质中的结合活性。介质由添加了表面活性剂、血清或培养液的缓冲盐水组成。此外,还测定了与膜结合的病毒的洗脱率。结果表明,病毒可通过疏水和静电相互作用发生结合,其中phi X174在这两种过程中显示出最低的结合水平。非离子洗涤剂Triton X-100和吐温80(0.1%)同样能最大程度地减少疏水相互作用。在无毒水平下,阴离子和阳离子表面活性剂的效果均不如它们。血清在减少疏水和静电结合方面均有效,在我们的测试条件下,2%的血清足以消除结合。因此,phi X174仍是测试阻隔材料的最佳替代病毒选择,Triton X-100(0.1%)仍是减少疏水结合的良好选择。此外,阻隔材料对病毒的结合不太可能阻止血源性病原体的通过。