Nested-multiplex PCR detection of Orthopoxvirus and Parapoxvirus directly from exanthematic clinical samples
| Autor: | Vanessa Ferreira, Pedro Augusto Alves, Zélia Inês Portela Lobato, Carlos Mazur, Rafael K. Campos, Giliane de Souza Trindade, Felipe L. Assis, Jônatas Santos Abrahão, Erna Geessien Kroon, Marcela M.G. Cota, André T. Silva-Fernandes, Larissa S. Lima |
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| Rok vydání: | 2009 |
| Předmět: |
Molecular Sequence Data
Cattle Diseases Sheep Diseases Orthopoxvirus Poxviridae Infections Polymerase Chain Reaction lcsh:Infectious and parasitic diseases law.invention Monkeypox chemistry.chemical_compound law Virology Multiplex polymerase chain reaction medicine Animals Humans lcsh:RC109-216 Polymerase chain reaction DNA Primers Parapoxvirus Goat Diseases Sheep biology Goats Research medicine.disease biology.organism_classification Viral Identification DNA extraction Infectious Diseases chemistry Cattle Vaccinia |
| Zdroj: | Virology Journal Virology Journal, Vol 6, Iss 1, p 140 (2009) |
| ISSN: | 1743-422X |
| DOI: | 10.1186/1743-422x-6-140 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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