title: A rapid Orthopoxvirus purification protocol suitable for high-containment laboratories.
abstract: Virus purification in a high-containment setting provides unique challenges due to barrier precautions and operational safety approaches that are not necessary in lower biosafety level (BSL) 2 environments. The need for high risk group pathogen diagnostic assay development, anti-viral research, pathogenesis and vaccine efficacy research necessitates work in BSL-3 and BSL-4 labs with infectious agents. When this work is performed in accordance with BSL-4 practices, modifications are often required in standard protocols. Classical virus purification techniques are difficult to execute in a BSL-3 or BSL-4 laboratory because of the work practices used in these environments. Orthopoxviruses are a family of viruses that, in some cases, requires work in a high-containment laboratory and due to size do not lend themselves to simpler purification methods. Current CDC purification techniques of orthopoxviruses uses 1,1,2-trichlorotrifluoroethane, commonly known as Genetron®. Genetron®is a chlorofluorocarbon (CFC) that has been shown to be detrimental to the ozone and has been phased out and the limited amount of product makes it no longer a feasible option for poxvirus purification purposes. Here we demonstrate a new Orthopoxvirus purification method that is suitable for high-containment laboratories and produces virus that is not only comparable to previous purification methods, but improves on purity and yield.
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA. Electronic address: bkz2@cdc.gov.
authors.forename: Laura
authors.initials: L
authors.lastname: Hughes
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Wilkins
authors.affiliation: Infectious Diseases Pathology Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Cynthia S
authors.initials: CS
authors.lastname: Goldsmith
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Smith
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Hudson
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Nishi
authors.initials: N
authors.lastname: Patel
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Karem
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Inger
authors.initials: I
authors.lastname: Damon
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Yu
authors.initials: Y
authors.lastname: Li
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: Victoria A
authors.initials: VA
authors.lastname: Olson
authors.affiliation: Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
authors.forename: P S
authors.initials: PS
authors.lastname: Satheshkumar
mesh_terms: D000818:Animals; D003264:Containment of Biohazards; D006801:Humans; D007753:Laboratories; D018155:Orthopoxvirus / Q000302:isolation & purification*; D013997:Time Factors; D014773:Virology / Q000379:methods*
keywords: High-containment; Monkeypox; Orthopoxvirus; Purification; Variola