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A simple method for isolation of cell-associated viral particles from cell culture

Katarina Laposova1, Ingrid Oveckova1, Jana Tomaskova1

  • 1Department of Molecular Medicine, Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Dubravska cesta 9, 845 05 Bratislava, Slovak Republic.

Journal of Virological Methods
|September 20, 2017
PubMed

Insights

Deionized water lysis is the most effective method for releasing lymphocytic choriomeningitis virus (LCMV) from infected cells. This simple technique improves virus yield compared to traditional methods like freeze-thaw cycles and sonication.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • Cell-associated viruses, such as lymphocytic choriomeningitis virus (LCMV) strain MX, are challenging to isolate due to limited extracellular virion release.
  • Traditional isolation methods often involve cell disruption techniques like hypotonic lysis, freeze-thaw cycles (F-T), and sonication.

Purpose of the Study:

  • To compare different methods for releasing infectious LCMV MX from persistently infected cells.
  • To identify the most efficient and effective protocol for isolating cell-associated viruses.

Main Methods:

  • Persistently LCMV MX-infected cells were subjected to different lysis methods, including deionized water lysis, F-T cycles, and sonication.
  • Virus yield was quantified to assess the effectiveness of each isolation approach.

Main Results:

  • Deionized water lysis demonstrated the fastest and most effective release of infectious LCMV MX viral particles.
  • Freeze-thaw cycles and sonication did not significantly improve virus yield compared to deionized water lysis alone.

Conclusions:

  • Deionized water lysis is a superior method for isolating LCMV MX, offering higher yields and efficiency.
  • This simplified protocol has potential applications for isolating other cell-associated viruses that are difficult to release from host cells.
Keywords:
Cell-associated virusCell-free extractInfectionLCMVViral particlesVirus isolation

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