Poxviruses pp 355-373 | Cite as

Poxvirus diagnostics

  • Martin Pfeffer
  • Hermann Meyer
Part of the Birkhäuser Advances in Infectious Diseases book series (BAID)


Members of the family Poxviridae form a large group of viruses that can infect humans as well as animals including the major domestic animal species (cattle, sheep, goat, swine, dog, cat and chicken). Poxviruses can be highly pathogenic for humans (i.e., Variola virus), of zoonotic importance (e.g., Monkeypox virus) or highly contagious among animal populations (e.g., Sheeppox virus). Therefore, laboratory confirmation of the specific poxvirus involved is, indeed, essential. This is especially true for the most notorious member, Variola virus, the smallpox virus, which might reemerge as a weapon, and also for those “exotic” poxviruses which are absent in many countries but still enzootic in other parts of the world. Today, poxvirus diagnostics covers the entire spectrum of either traditional (such as inoculation of embryonated eggs) or more advanced laboratory tests (such as genome sequencing or microarray assays). This chapter presents methods of sample collection and handling, and reviews techniques used in the diagnosis of poxvirus infections by briefly describing the principle and procedure of the method, and critically weighting the pros and cons as well as providing some examples of application for each method.


Restriction Fragment Length Polymorphism Myxoma Virus Variola Virus Plaque Reduction Neutralization Test Lumpy Skin Disease Virus 
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  1. 1.
    Damon IK, Esposito JJ (2003) Poxviruses that infect humans. In: PR Murray, EJ Baron, JH Jorgensen, MA Pfaller, MH Yolker (eds): Manual of clinical microbiology, 8th edn. ASM Press, Washington DC, 1583–1591Google Scholar
  2. 2.
    Henderson DA, Inglesby TV, Bartlett JG, Ascher MS, Eitzen E, Jahrling PB, Hauer J, Layton M, McDade J, Osterholm TM et al (1999) Smallpox as a biological weapon: medical and public health management. Working Group on Civilian Biodefense. JAMA 281: 2127–2137Google Scholar
  3. 3.
    Hazelton PR, Gelderblom HR (2003) Electron microscopy for rapid diagnosis of infectious agents in emergent situations. Emerg Infect Dis 9: 294–303PubMedGoogle Scholar
  4. 4.
    Renner-Müller IC, Meyer H, Munz E (1995) Characterization of camelpoxvirus isolates from Africa and Asia. Vet Microbiol 45: 371–381PubMedCrossRefGoogle Scholar
  5. 5.
    Downie AW (1939) A study of the lesions produced experimentally by cowpox virus. J Pathol 48: 361–378CrossRefGoogle Scholar
  6. 6.
    Meyer H, Ropp SL, Esposito JJ (1997) Gene for A-type inclusion body protein is useful for a polymerase chain reaction assay to differentiate orthopoxviruses. J Virol Methods 64: 217–221PubMedCrossRefGoogle Scholar
  7. 7.
    Tripathy DN, Hanson LE (1976) A smear technique for staining elementary bodies of fowlpox. Avian Dis 20: 609–610PubMedCrossRefGoogle Scholar
  8. 8.
    Lazarus AS, Eddie A, Meyer KF (1937) Propagation of variola virus in the developing egg. Proc Soc Exp Biol Med 36: 7–8Google Scholar
  9. 9.
    Samour JH, Kaaden OR, Wernery U, Bailey TA (1996) An epornitic of avian pox in houbara bustards (Chlamydotis undulata macqueenii). Zentralbl Veterinaermed B 43: 287–292Google Scholar
  10. 10.
    Niikura M, Narita T, Mikami T (1991) Establishment and characterization of a thymidine kinase deficient avian fibroblast cell line derived from a Japanese quail cell line, QT35. J Vet Med Sci 53: 439–446PubMedGoogle Scholar
  11. 11.
    Czerny C-P, Meyer H, Mahnel H (1989) Establishment of an ELISA for the detection of orthopox viruses based on neutralizing monoclonal and polyclonal antibodies. Zentralbl Veterinarmed B 36: 537–546PubMedGoogle Scholar
  12. 12.
    Carn VM (1995) An antigen trapping ELISA for the detection of capripoxvirus in tissue culture supernatant and biopsy samples. J Virol Methods 51: 95–102PubMedCrossRefGoogle Scholar
  13. 13.
    Bults HG, Brandon MR (1982) An immunological method to determine antigenic variation in three strains of myxoma virus. J Virol Methods 5: 21–26PubMedCrossRefGoogle Scholar
  14. 14.
    Hsu SM, Raine L (1981) Protein A, avidin, and biotin in immunohistochemistry. J Histochem Cytochem 29: 1349–1353PubMedGoogle Scholar
  15. 15.
    Mackett M, Archard LC (1979) Conservation and variation in orthopoxvirus genome structure. J Gen Virol 45: 683–701PubMedCrossRefGoogle Scholar
  16. 16.
    Esposito JJ, Knight JC (1985) Orthopoxvirus DNA: a comparison of restriction profiles and maps. Virology 143: 230–251PubMedCrossRefGoogle Scholar
  17. 17.
    Naidoo J, Baxby D, Bennett M, Gaskell RM, Gaskell CJ (1992) Characterization of orthopoxviruses isolated from feline infections in Britain. Arch Virol 125: 261–272PubMedCrossRefGoogle Scholar
  18. 18.
    Meyer H, Schay C, Mahnel H, Pfeffer M (1999) Characterization of orthopoxviruses isolated from man and animals in Germany. Arch Virol 144: 491–501PubMedCrossRefGoogle Scholar
  19. 19.
    Neubauer H, Reischl U, Ropp S, Esposito JJ, Wolf H, Meyer H (1998) Specific detection of monkeypox virus by polymerase chain reaction. J Virol Methods 74: 201–207PubMedCrossRefGoogle Scholar
  20. 20.
    Ropp SL, Jin Q, Knight JC, Massung RF, Esposito JJ (1995) PCR strategy for identification and differentiation of small pox and other orthopoxviruses. J Clin Microbiol 33: 2069–2076PubMedGoogle Scholar
  21. 21.
    Loparev VN, Massung RF, Esposito JJ, Meyer H (2001) Detection and differentiation of old world orthopoxviruses: restriction fragment length polymorphism of the crmB gene region. J Clin Microbiol 39: 94–100PubMedCrossRefGoogle Scholar
  22. 22.
    Heine HG, Stevens MP, Foord AJ, Boyle DB (1999) A capripoxvirus detection PCR and antibody ELISA based on the major antigen P32, the homolog of the vaccinia virus H3L gene. J Immunol Methods 30: 187–196CrossRefGoogle Scholar
  23. 23.
    Luschow D, Hoffmann T, Hafez HM (2004) Differentiation of avian poxvirus strains on the basis of nucleotide sequences of 4b gene fragment. Avian Dis 48: 453–462PubMedCrossRefGoogle Scholar
  24. 24.
    Nunez A., Funes JM, Agromayor M, Moratilla M, Varas AJ, Lopez-Estebaranz JL, Esteban M, Martin-Gallardo A (1996) Detection and typing of molluscum contagiosum virus in skin lesions by using a simple lysis method and polymerase chain reaction. J Med Virol 50: 342–349PubMedCrossRefGoogle Scholar
  25. 25.
    Thompson CH (1997) Identification and typing of molluscum contagiosum virus in clinical specimens by polymerase chain reaction. J Med Virol 53: 205–211PubMedCrossRefGoogle Scholar
  26. 26.
    Torfason EG, Gunadottir S (2002) Polymerase chain reaction for laboratory diagnosis of orf virus infections. J Clin Virol 24: 79–84PubMedCrossRefGoogle Scholar
  27. 27.
    Espy MJ, Cockerill IF, Meyer RF, Bowen MD, Poland GA, Hadfield TL, Smith TF (2002) Detection of smallpox virus DNA by LightCycler PCR. J Clin Microbiol 40: 1985–1988PubMedCrossRefGoogle Scholar
  28. 28.
    Ibrahim MS, Esposito JJ, Jahrling PB, Lofts RS (1997) The potential of 5’ nuclease PCR for detecting a single-base polymorphism in orthopoxvirus. Mol Cell Probes 11: 143–147PubMedCrossRefGoogle Scholar
  29. 29.
    Ibrahim MS, Kulesh DA, Saleh SS, Damon IK, Esposito JJ, Schmaljohn AL, Jahrling PB (2003) Real-time PCR assay to detect smallpox virus. J Clin Microbiol 41: 3835–3839CrossRefGoogle Scholar
  30. 30.
    Panning M, Asper M, Kramme S, Schmitz H, Drosten C (2004) Rapid detection and differentiation of human pathogenic orthopox viruses by a fluorescence resonance energy transfer real-time PCR assay. Clin Chem 50: 702–708PubMedCrossRefGoogle Scholar
  31. 31.
    Nitsche A, Ellerbrok H, Pauli G (2004) Detection of orthopoxvirus DNA by real-time PCR and identification of variola virus DNA by melting analysis. J Clin Microbiol 42: 1207–1213PubMedCrossRefGoogle Scholar
  32. 32.
    Kulesh DA, Baker RO, Loveless BM, Norwood D, Zwiers SH, Mucker E, Hartmann C, Herrera R, Miller D, Christensen D et al (2004) Smallpox and pan-orthopox virus detection by real-time 3’-minor groove binder TaqMan assays on the Roche LightCycler and the Cepheid smart Cycler platforms. J Clin Microbiol 42: 601–609PubMedCrossRefGoogle Scholar
  33. 33.
    Olson VA, Laue T, Laker MT, Babkin IV, Drosten C, Shchelkunov SN, Niedrig M, Damon IK, Meyer H (2004) Real-time PCR system for detection of orthopoxviruses and simultaneous identification of smallpox virus. J Clin Microbiol 42: 1940–1946PubMedCrossRefGoogle Scholar
  34. 34.
    Zimmermann P, Thordsen I, Frangoulidis D, Meyer H (2005) Real-time PCR assay for the detection of tanapox virus and yaba-like disease virus. J Virol Methods 130: 149–153PubMedCrossRefGoogle Scholar
  35. 35.
    Lapa S, Mikheev M, Shchelkunov S, Mikhailovich V, Sobolev A, Blinov V, Babkin L, Guskov A, Sokunova E, Zasedatelev A et al (2002) Species-level identification of orthopoxviruses with an oligonucleotide microchip. J Clin Microbiol 40: 753–757PubMedCrossRefGoogle Scholar
  36. 36.
    Laassri M, Chizhikov V, Mikheev M, Shchelkunov S, Chumakov K (2003) Detection and discrimination of orthopoxviruses using microarrays of immobilized oligonucleotides. J Virol Methods 112: 67–78PubMedCrossRefGoogle Scholar
  37. 37.
    Gubser C, Hue S, Kellam P, Smith GL (2004) Poxvirus genomes: a phylogenetic analysis. J Gen Virol 85: 105–117PubMedCrossRefGoogle Scholar
  38. 38.
    Likos AM, Sammons SA, Olson VA, Frace AM, Li Y, Olsen-Rasmussen M, Davidson W, Galloway R, Khristova ML, Reynolds MG et al (2005) A tale of two clades: monkeypox viruses. J Gen Virol 86: 2661–2672PubMedCrossRefGoogle Scholar
  39. 39.
    Kaerber G (1931) DeBeitrag zur kollektiven Behandlung pharmakologischer Reihenversuche. Arch Exp Pathol Pharmakol 162: 480CrossRefGoogle Scholar
  40. 40.
    Czerny C-P, Wagner K, Gessler K, Mayr A, Kaaden O-R (1996) A monoclonal blocking ELISA for detection of orthopox virus antibodies in feline sera. Vet Microbiol 52: 185–200PubMedCrossRefGoogle Scholar
  41. 41.
    Karem KL, Reynolds M, Braden Z, Lou G, Bernard N, Patton J, Damon IK (2005) Characterization of acute-phase humoral immunity to monkeypox: use of immunoglobulin M enzyme-linked immunosorbent assay for detection of monkeypox infection during the 2003 North American outbreak. Clin Diag Lab Immunol 12: 867–872CrossRefGoogle Scholar

Copyright information

© Birkhäuser Verlag Basel/Switzerland 2007

Authors and Affiliations

  • Martin Pfeffer
    • 1
  • Hermann Meyer
    • 1
  1. 1.Bundeswehr Institute of MicrobiologyMünchenGermany

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