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André T Silva-fern

Carlos Mazur

Erna G Kroon

Felipe L Assis

Giliane S Trindade

Jônatas S Abrahão

Larissa S Lima

Pedro A Alves

Rafael K Campos

Vanessa M Ferreira

Zélia Ip Lobato

Virology Journal

English

PubMed

André T Silva-Fernandes

Marcela MG Cota

Zélia IP Lobato

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

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

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.</p

Trindade Giliane S

Lobato Zélia IP

Mazur Carlos

Campos Rafael K

Ferreira Vanessa M

Cota Marcela MG

Silva-Fernandes André T

Alves Pedro A

Assis Felipe L

Lima Larissa S

Abrahão Jônatas S

Kroon Erna G

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

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