Identification of human pathogenic variola and monkeypox viruses by real-time polymerase chain reaction


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Abstract

A kit of specific oligonucleotide primers and hybridization probes has been proposed to detect orthopoxviruses (OPV) and to discriminate human pathogenic viruses, such as variola virus and monkey virus by real-time polymerase chain reaction (PCR). For real-time PCR, the following pairs of fluorophore and a fluorescence quencher were used: TAMRA-BHQ2 for genus-specific probes and FAM-BHQ1 for species-specific ones (variola virus, monkeypox virus, ectomelia virus). The specificity of this assay was tested on 38 strains of 6 OPV species and it was 100%.

References

  1. Маренникова С. С., Щелкунов С. Н. Патогенные для человека ортопоксвирусы. - М., 1998.
  2. Михеев М. В., Фещенко М. В., Щелкунов С. Н. Филогенетический анализ гена хемокинсвязывающего белка ортопоксвирусов // Молекул. генетика. - 2004. - № 1. - С. 29-36.
  3. Рябинин В. А., Шундрин Л. А., Костина Е. В. и др. // Молекул. генетика. - 2006. - № 4. - С. 43-49.
  4. Breman J. G. Monkeypox: an emerging infection for humans? // Emerging infections 4 / Eds W. M. Scheld et al. - 2000. - P. 45-67.
  5. Breman J. G., Henderson D. A. Current concepts: diagnosis and management of smallpox // N. Engl. J. Med. - 2002. - Vol. 346. - P. 1300-1308.
  6. Carletti F., Di Caro A., Calcaterra S. et al. Rapid, differential diagnosis of orthopox- and herpesviruses based upon real-time PCR product melting temperature and restriction enzyme analysis of amplicons // J. Virol. Meth. - 2005. - Vol. 129, N 1. - P. 97-100.
  7. Douglass N. J., Richardson M., Dumbell K. R. Evidence for recent genetic variation in monkeypox viruses // J. Gen. Virol. - 1994. - Vol. 75. - P. 1303-1309.
  8. Ibrahim M. S., Esposito J. J., Jahrling P. B. et al. The potential of 5' nuclease PCR for detecting a single-base polymorphism in orthopoxvirus // Mol. Cell. Prob. - 1997. - Vol. 11. -P. 143-147.
  9. Ibrahim M. S., Kulesh D. A., Saleh S. S et al. Real-time PCR assay to detect smallpox virus // J. Clin. Microbiol. - 2003. - Vol. 41, N 8. - P. 3835-3839.
  10. Lewis-Jones S. Zoonotic poxvirus infections in humans // Curr. Opin. Infect. Dis. - 2004. - Vol. 17, N 2. - P. 81-89.
  11. Li Y., Olson V. A., Laue T. et al. Detection of monkeypox virus with real-time PCR assays // J. Clin. Virol. - 2006. - Vol. 36. - P. 194-203.
  12. Likos A. M., Sammons S. A., Olson V. A. et al. A tale of two clades: monkeypox viruses // J. Gen. Virol. - 2005. - Vol. 86. - P. 2661-2672.
  13. Nitsche A., Ellerbrok H., Pauli G. Detection of orthopoxvirus DNA by real-time PCR and identification of variola virus DNA by melting analysis // J. Clin. Microbiol. - 2004. - Vol. 42, N 3. - P. 1207-1213.
  14. Olson V. A., Laue Т., Laker M. T. et al. Real-time PCR system for detection of orthopoxviruses and simultaneous identification of smallpox virus // J. Clin. Microbiol. - 2004. - Vol. 42, N 5. - P. 1940-1946.
  15. Panning M., Asper M., Kramme S. et al. Rapid detection and differentiation of human pathogenic orthopox viruses by a fluorescence resonance energy transfer real-time PCR assay // Clin. Chem. - 2004. - Vol. 50, N 4. - P. 702-708.
  16. Ryabinin V. A., Shundrin L. A., Kostina E. V. et al. Microarray assay for detection and discrimination of Orthopoxvirus species // J. Med. Virol. - 2006. - Vol. 78, N 10. - P. 1325-1340.
  17. Shchelkunov S. N., Marennikova S. S., Moyer R. W. Orthopoxviruses pathogenic for humans. - New York, 2005.
  18. Spencer R. C., Lightfoot N. F. Preparedness and response to bioterrorism // J. Infect. 2001. - Vol. 43. - P. 104-110.
  19. Stephenson J. Monkeypox outbreak a reminder of emerging infections vulnerabilities // J. A. M. A. - 2003. - Vol. 290, N 1. - P. 23-24.
  20. Virus Taxonomy: The Classification and Nomenclature of Viruses // The Seventh report of the International committee on taxonomy of viruses. VII Report of the ICTV/ Eds M. H. V. Regenmortel et al. - San Diego, 2000.

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Copyright (c) 2009 Kostina E.V., Gavrilova E.V., Ryabinin V.A., Shchelkunov S.N., Sinyakov A.N.

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