Inactivation of West Nile virus in serum with heat, ionic detergent, and reducing agent for proteomic applications

J Virol Methods. 2017 Oct:248:1-6. doi: 10.1016/j.jviromet.2017.05.010. Epub 2017 May 19.

Abstract

Research involving biosafety level 3 pathogens such as West Nile virus (WNV) is often limited by the limited space and technical constraints of these environments. To conduct complex analytical studies outside of high containment, robust and reliable inactivation methods are needed that maintain compatibility with downstream assays. Here we report the inactivation of WNV in spiked serum samples using a commercially available SDS-PAGE sample buffer for proteomic studies. Using this method, we demonstrate its utility by identification proteins differentially expressed in the serum of mice experimentally infected with WNV.

Keywords: BSL-3; Dithiothreitol; Lithium dodecyl sulfate; Sample buffer; Virus inactivation; West Nile virus.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Blood Proteins / metabolism*
  • Buffers
  • Detergents / pharmacology*
  • Dithiothreitol / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Hot Temperature*
  • Mice
  • Proteomics / methods*
  • Reducing Agents / pharmacology*
  • Serum / virology*
  • Sodium Dodecyl Sulfate / pharmacology
  • Surface-Active Agents / pharmacology
  • Viral Plaque Assay
  • Virus Inactivation*
  • West Nile Fever / virology
  • West Nile virus / drug effects
  • West Nile virus / physiology*

Substances

  • Blood Proteins
  • Buffers
  • Detergents
  • Reducing Agents
  • Surface-Active Agents
  • Sodium Dodecyl Sulfate
  • dodecyl sulfate
  • Dithiothreitol