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 language | Italian |
|---|---|
| Pages (from-to) | 47-55 |
| Number of pages | 9 |
| Journal | Infezioni in Medicina |
| Volume | 15 |
| Issue number | 1 |
| Publication status | Published - Mar 2007 |
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ASJC Scopus subject areas
- Microbiology (medical)
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Un nuovo metodo rapido, basato su Real-time PCR multiplex, per la diagnosi differenziale di infezioni da Orthopoxvirus e Herpesvirus. / Sias, Catia; Carletti, Fabrizio; Capobianchi, Maria R.; Travaglini, Damiano; Chiappini, Roberta; Horejsh, Douglas; Di Caro, Antonino.
In: Infezioni in Medicina, Vol. 15, No. 1, 03.2007, p. 47-55.Research output: Contribution to journal › Article
}
TY - JOUR
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
UR - http://www.scopus.com/inward/record.url?scp=34248562253&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34248562253&partnerID=8YFLogxK
M3 - Articolo
C2 - 17515675
AN - SCOPUS:34248562253
VL - 15
SP - 47
EP - 55
JO - Infezioni in Medicina
JF - Infezioni in Medicina
SN - 1124-9390
IS - 1
ER -