Development of a High-Content Orthopoxvirus Infectivity and Neutralization Assays |
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| Authors: | Irina Gates Victoria Olson Scott Smith Nishi Patel Inger Damon Kevin Karem |
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| Institution: | 1. Atlanta Research and Education Foundation, Decatur, Georgia, United States of America.; 2. Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology (DHCPP), National Center for Emerging and Zoonotic Infectious Diseases (NCEZID), Centers for Disease Control and Prevention, Atlanta, Georgia, Unites States of America.; University of Texas HSC at San Antonio, UNITED STATES, |
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| Abstract: | 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. |
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