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 journalArticle

5 Citations (Scopus)

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.

Original languageItalian
Pages (from-to)47-55
Number of pages9
JournalInfezioni in Medicina
Volume15
Issue number1
Publication statusPublished - Mar 2007

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Orthopoxvirus
Herpesviridae
Multiplex Polymerase Chain Reaction
Variola virus
Real-Time Polymerase Chain Reaction
Differential Diagnosis
Restriction Fragment Length Polymorphisms
Freezing
Monkeypox
Biological Warfare Agents
Temperature
Smallpox
DNA-Directed DNA Polymerase
Infection
Open Reading Frames
Viruses
Sensitivity and Specificity
Polymerase Chain Reaction
DNA
Genes

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

@article{72245e04c09940499eba23f55604e692,
title = "Un nuovo metodo rapido, basato su Real-time PCR multiplex, per la diagnosi differenziale di infezioni da Orthopoxvirus e Herpesvirus",
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.",
keywords = "Diagnosi differenziale, Diagnosi rapida, Herpesvirus, Othopoxvirus, Vaiolo",
author = "Catia Sias and Fabrizio Carletti and Capobianchi, {Maria R.} and Damiano Travaglini and Roberta Chiappini and Douglas Horejsh and {Di Caro}, Antonino",
year = "2007",
month = "3",
language = "Italian",
volume = "15",
pages = "47--55",
journal = "Infezioni in Medicina",
issn = "1124-9390",
publisher = "EDIMES Edizioni Medico Scientifiche",
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T1 - Un nuovo metodo rapido, basato su Real-time PCR multiplex, per la diagnosi differenziale di infezioni da Orthopoxvirus e Herpesvirus

AU - Sias, Catia

AU - Carletti, Fabrizio

AU - Capobianchi, Maria R.

AU - Travaglini, Damiano

AU - Chiappini, Roberta

AU - Horejsh, Douglas

AU - Di Caro, Antonino

PY - 2007/3

Y1 - 2007/3

N2 - 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.

AB - 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.

KW - Diagnosi differenziale

KW - Diagnosi rapida

KW - Herpesvirus

KW - Othopoxvirus

KW - Vaiolo

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