Development of a High-Content Orthopoxvirus Infectivity and Neutralization Assays

Details

• Alternative Title:
  PLoS One
• Personal Author:
  Gates, Irina ; Olson, Victoria ; Smith, Scott ; Patel, Nishi ; Damon, Inger ; Karem, Kevin
• Description:
  Currently, a number of assays measure Orthopoxvirus neutralization with serum from individuals, vaccinated against smallpox. In addition to the traditional plaque reduction neutralization test (PRNT), newer higher throughput assays are based on neutralization of recombinant vaccinia virus, expressing reporter genes such as β-galactosidase or green fluorescent protein. These methods could not be used to evaluate neutralization of variola virus, since genetic manipulations of this virus are prohibited by international agreements. Currently, PRNT is the assay of choice to measure neutralization of variola virus. However, PRNT assays are time consuming, labor intensive, and require considerable volume of serum sample for testing. Here, we describe the development of a high-throughput, cell-based imaging assay that can be used to measure neutralization, and characterize replication kinetics of various Orthopoxviruses, including variola, vaccinia, monkeypox, and cowpox.
• Subjects:
  Animals Chlorocebus Aethiops Green Fluorescent Proteins Humans Immunization Neutralization Tests Orthopoxvirus Sciuridae Vero Cells Viral Plaque Assay
• Source:
  PLoS One. 10(10).
• Pubmed ID:
  26426117
• Pubmed Central ID:
  PMC4591290
• Document Type:
  Journal Article
• Volume:
  10
• Issue:
  10
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:7ff7110d90ac9b8cf25e18b2a9ea44870478b6e3d3edb781a29d1ce6d6e5df08
• Download URL:
  https://stacks.cdc.gov/view/cdc/34883/cdc_34883_DS1.pdf
• File Type:
  [PDF - 8.51 MB ]