Protocol

Vaccinia Virus and Poxvirology

Volume 269 of the series Methods in Molecular Biology pp 119-133

Orthopoxvirus Diagnostics

  • Hermann MeyerAffiliated withInstitut für Mikrobiologie der Bundeswehr
  • , Inger K. DamonAffiliated withPoxvirus Section, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention
  • , Joseph J. EspositoAffiliated withBiotechnology Core Facility Branch, Scientific Resources Program, National Center for Infectious Diseases, Centers for Disease Control and Prevention

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Abstract

Biologic and antigenic properties are often useful for identifying and differentiating orthopoxviruses (OPV). However, polymerase chain reaction (PCR) amplification, with either restriction cleavage or sequencing of amplicons, has been gaining credibility as a more rapid, specific, sensitive, and often cost-saving technique for research and diagnostic laboratories. This chapter is consolidated using prior research papers from our laboratories with three different methods that should be suitable for the preparation of orthopoxvirus DNA from various sources (e.g., clinical specimens or cell cultures) and four different methods for PCR that should be useful for investigating orthopoxvirus species and strains.

Key Words

Orthopoxvirus polymerase chain reaction (PCR) diagnostic DNA preparation hemagglutinin gene A-type inclusion body gene restriction fragment length polymorphism (RFLP) smallpox variola virus monkeypox virus cowpox virus vaccinia virus camelpox virus ectromelia virus mousepox virus