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Fully Automated Cell Culture System Increases the Efficiency and Consistency of Arterivirus Plaque Assays

Thursday, November 07, 2013 — Poster Session II

12:00 p.m. – 2:00 p.m.

FAES Academic Center (Upper-Level Terrace)

NIAID

RSCHSUPP-11

Authors

  • N.M. Deiuliis
  • S. Mazur
  • K. Lamberton
  • S.L. Agar
  • D.L. Freeburger
  • J. Kuhn
  • J.M. Michelotti

Abstract

The mechanism of arterivirus cell entry has not been fully elucidated, although these viruses are thought to enter cells through receptor-mediated endocytosis. To study cell entry and infectivity of arteriviruses, high quality virus stocks are necessary. The Integrated Research Facility Core Cell Culture Lab has expanded and banked 5 quality-controlled AGM cell lines for use in viral assays.A CompacT Selectâ„¢ cell culture robot was used to passage and seed cells into plates for plaque assays. In a study evaluating the growth of equine arteritis virus (EAV) and porcine reproductive and respiratory syndrome virus (PRRSV), we compared the robotic plating technique with the standard manual plating technique in the MA-104 cell line and evaluated the consistency of the robot performance over 2 months in the MARC-145 cell line. The growth of these arteriviruses was tested in four cell lines (BS-C-1, CL 2621, MA-104, MARC-145) and the viral titers from supernatants were quantified by plaque assay in MA-104 and MARC-145 cells.

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