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Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment
Published on: March 2, 2016
Minimized virus binding for tests of barrier materials
1Division of Life Sciences, Food and Drug Administration, Rockville, Maryland 20857, USA.
Abstract:
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.
Insights
Choosing the right virus and medium is key for accurate material barrier testing. Phi X174 virus with Triton X-100 is recommended to minimize misleading binding results.
Area of Science:
- Virology
- Materials Science
- Biotechnology
Background:
- Virus binding to materials can skew barrier property test results.
- Optimizing challenge virus and suspending medium selection is crucial for accurate material testing.
Purpose of the Study:
- Evaluate different surrogate viruses and media to minimize virus binding during material barrier testing.
- Determine optimal parameters for reliable assessment of material barrier properties against viral penetration.
Main Methods:
- Tested five surrogate viruses (MS2, phi X174, T7, PRD1, phi 6) on nitrocellulose and cationic polysulfone membranes.
- Assessed virus binding in various media including buffered saline with surfactants, serum, and culture broth.
- Determined virus elution rates from bound membranes.
Main Results:
- Phi X174 showed the lowest binding via hydrophobic and electrostatic interactions.
- Nonionic detergents Triton X-100 and Tween 80 (0.1%) effectively reduced hydrophobic binding.
- 2% serum minimized both hydrophobic and electrostatic binding, eliminating it under test conditions.
Conclusions:
- Phi X174 is the optimal surrogate virus for material barrier testing.
- Triton X-100 (0.1%) is effective for reducing hydrophobic virus binding.
- Virus binding to barrier materials is unlikely to impede blood-borne pathogen passage.
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