Un nuovo metodo rapido, basato su Real-time PCR multiplex, per la diagnosi differenziale di infezioni da Orthopoxvirus e Herpesvirus

Translated title of the contribution: Rapid differential diagnosis of Orthopoxviruses and Herpesviruses based upon multiplex Real-time PCR

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: Variola virus, belonging to Orthopoxviridae family, is one of the most dangerous human pathogens that could be used as biological weapon. We have developed a new rapid assay, based upon Real-time PCR and melting temperatures analysis of amplicons, for the contemporary detection of Orthopoxvirus, VZV and HSV1-2, that are the most important infectious agents to be considered for differential diagnosis. Methods: The target for detection of orthopoxvirus DNA has been a region of the crmB gene which is common to Variola virus and to other old world orthopoxviruses pathogenic for humans. The targets for VZV and HSV1-2 have been ORF 29 and DNA polymerase, respectively. Suitability of the amplified fragments to RFLP or sequencing analysis, to recognize the involved viral species, has been also tested. Result: The selected primers have showed high sensitivity, specificity and compatibility with common amplification conditions. A mean melting temperature difference of 8.7°C was observed between the amplicene from the two virus types. Further identification of individual pathogens was made using RFLP analysis. Conclusion: The PCR-based protocol set up in this study for presumptive differential diagnosis of variola and herpesviral infections is rapid and specific and it can be used also to detect other orthopoxviral infections, like monkeypox.

Translated title of the contributionRapid differential diagnosis of Orthopoxviruses and Herpesviruses based upon multiplex Real-time PCR
Original languageItalian
Pages (from-to)47-55
Number of pages9
JournalInfezioni in Medicina
Volume15
Issue number1
Publication statusPublished - Mar 2007

ASJC Scopus subject areas

  • Microbiology (medical)

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