Species-specific differentiation of variola, monkeypox, and varicella-zoster viruses by multiplex real-time PCR assay

J Virol Methods. 2016 Oct:236:215-220. doi: 10.1016/j.jviromet.2016.07.024. Epub 2016 Jul 28.

Abstract

A method of one-stage rapid detection and differentiation of epidemiologically important variola virus (VARV), monkeypox virus (MPXV), and varicella-zoster virus (VZV) utilizing multiplex real-time TaqMan PCR assay was developed. Four hybridization probes with various fluorescent dyes and the corresponding fluorescence quenchers were simultaneously used for the assay. The hybridization probes specific for the VARV sequence contained FAM/BHQ1 as a dye/quencher pair; MPXV-specific, JOE/BHQ1; VZV-specific, TAMRA/BHQ2; and internal control-specific, Cy5/BHQ3. The specificity and sensitivity of the developed method were assessed by analyzing DNA of 32 strains belonging to orthopoxvirus and herpesvirus species.

Keywords: Monkeypox virus; Real-time PCR; Varicella-zoster virus; Variola virus.

Publication types

  • Evaluation Study

MeSH terms

  • Herpesvirus 3, Human / classification
  • Herpesvirus 3, Human / genetics
  • Herpesvirus 3, Human / isolation & purification*
  • Monkeypox virus / classification
  • Monkeypox virus / genetics
  • Monkeypox virus / isolation & purification*
  • Multiplex Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Variola virus / classification
  • Variola virus / genetics
  • Variola virus / isolation & purification*
  • Virology / methods*