Development of molecular diagnostics for Orthopoxvirus

Johansson Marie (1), Vennema Harry (2), Koopmans Marion (2), Lundkvist Åke (1,3), Nilsson Mikael (1)

1. Kunskapscentrum för mikrobiologisk beredskap, Smittskyddsinstitutet, Stockholm
2. Diagnostic Lab for Infectious Diseases National Institute of Public Health and the Environment, Bilthoven, The Netherlands
3. Mikrobiologiskt och Tumörbiologiskt Centrum, Karolinska Institutet

In view of the potential use of smallpox virus as biological warfare agent, the demand for re-establishment of fast and reliable methods for diagnostics and characterisation of orthopoxviruses is increasing. In addition to electron microscopy (EM). direct detection by PCR is a fast and reliable method to detect orthopoxvirus in clinical samples.

By the use of an orthopox-specific PCR and hybridisation using specific probes for smallpox, monkeypox, camelpox, cowpox and vaccinia viruses we have established a method that makes it possible to rapidly identify and type different orthopoxviruses from clinical samples.

The method has been evaluated using genomic DNA from vaccinia, monkeypox, camelpox and cowpox viruses. The method shows that genotyping of different orthopoxviruses works and that it is possible to rapidly detect and differentiate orthopoxviruses.

Summary: Through the establishment and combination of orthopox specific PCR detection and virus typing using hybridisation with specific probes it is possible to rapidly both detect and identify orthopoxviruses.

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