Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples

Jnatas S. Abrahão, Larissa S. Lima, Felipe L. Assis, Pedro A. Alves, André T. Silva-Fernandes, Marcela Mg Cota, Vanessa M. Ferreira, Rafael Kroon Campos, Carlos Mazur, Zélia Ip Lobato, Giliane S. Trindade, Erna G. Kroon

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Background. Orthopoxvirus (OPV) and Parapoxvirus (PPV) have been associated with worldwide exanthematic outbreaks. Some species of these genera are able to infect humans and domestic animals, causing serious economic losses and public health impact. Rapid, useful and highly specific methods are required to detect and epidemiologically monitor such poxviruses. In the present paper, we describe the development of a nested-multiplex PCR method for the simultaneous detection of OPV and PPV species directly from exanthematic lesions, with no previous viral isolation or DNA extraction. Methods and Results. The OPV/PPV nested-multiplex PCR was developed based on the evaluation and combination of published primer sets, and was applied to the detection of the target pathogens. The method showed high sensitivity, and the specificity was confirmed by amplicon sequencing. Exanthematic lesion samples collected during bovine vaccinia or contagious ecthyma outbreaks were submitted to OPV/PPV nested-multiplex PCR and confirmed its applicability. Conclusion. These results suggest that the presented multiplex PCR provides a highly robust and sensitive method to detect OPV and PPV directly from clinical samples. The method can be used for viral identification and monitoring, especially in areas where OPV and PPV co-circulate.

Original languageEnglish (US)
Article number140
JournalVirology journal
Volume6
DOIs
StatePublished - Oct 12 2009
Externally publishedYes

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Parapoxvirus
Orthopoxvirus
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Disease Outbreaks
Contagious Ecthyma
Poxviridae
Vaccinia
Domestic Animals
Public Health
Economics
Sensitivity and Specificity
DNA

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Abrahão, J. S., Lima, L. S., Assis, F. L., Alves, P. A., Silva-Fernandes, A. T., Cota, M. M., ... Kroon, E. G. (2009). Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples. Virology journal, 6, [140]. https://doi.org/10.1186/1743-422X-6-140

Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples. / Abrahão, Jnatas S.; Lima, Larissa S.; Assis, Felipe L.; Alves, Pedro A.; Silva-Fernandes, André T.; Cota, Marcela Mg; Ferreira, Vanessa M.; Kroon Campos, Rafael; Mazur, Carlos; Lobato, Zélia Ip; Trindade, Giliane S.; Kroon, Erna G.

In: Virology journal, Vol. 6, 140, 12.10.2009.

Research output: Contribution to journalArticle

Abrahão, JS, Lima, LS, Assis, FL, Alves, PA, Silva-Fernandes, AT, Cota, MM, Ferreira, VM, Kroon Campos, R, Mazur, C, Lobato, ZI, Trindade, GS & Kroon, EG 2009, 'Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples', Virology journal, vol. 6, 140. https://doi.org/10.1186/1743-422X-6-140
Abrahão, Jnatas S. ; Lima, Larissa S. ; Assis, Felipe L. ; Alves, Pedro A. ; Silva-Fernandes, André T. ; Cota, Marcela Mg ; Ferreira, Vanessa M. ; Kroon Campos, Rafael ; Mazur, Carlos ; Lobato, Zélia Ip ; Trindade, Giliane S. ; Kroon, Erna G. / Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples. In: Virology journal. 2009 ; Vol. 6.
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