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Variola virus-specific diagnostic assays: characterization, sensitivity, and specificity.

Ashley V Kondas1, Victoria A Olson1, Yu Li1

  • 1National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.

Journal of Clinical Microbiology
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New real-time PCR assays were developed for variola virus detection. These assays overcome cross-reactivity issues with related viruses, ensuring accurate public health diagnostics.

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Area of Science:

  • Virology
  • Molecular Biology
  • Public Health

Background:

  • Reliable diagnostic assays are crucial for effective public health responses to infectious diseases.
  • Previous diagnostic methods for variola virus exhibited cross-reactivity with newly identified cowpox viruses, compromising specificity.
  • Continuous reassessment of assay specificity is necessary due to the emergence of closely related viral strains.

Purpose of the Study:

  • To design and validate novel real-time PCR assays specifically targeting variola virus.
  • To address the limitations of existing assays, particularly their cross-reactivity with other orthopoxviruses.
  • To enhance the accuracy and reliability of variola virus diagnostics for public health surveillance.

Main Methods:

  • Development of two distinct real-time PCR assays.
  • In silico design and primer/probe selection for variola virus.
  • In vitro validation using characterized viral samples, including variola virus and related orthopoxviruses.
  • Assessment of analytical sensitivity and specificity.

Main Results:

  • Successful design and validation of two specific real-time PCR assays for variola virus.
  • Demonstrated absence of cross-reactivity with newly identified cowpox virus strains.
  • Established high sensitivity and specificity for the developed assays.

Conclusions:

  • The newly developed real-time PCR assays provide a reliable and specific tool for variola virus detection.
  • These assays are essential for accurate diagnosis and effective public health management of potential variola virus outbreaks.
  • Ongoing monitoring and validation are recommended as new viral variants emerge.