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Validation of a real-time PCR assay for detection of swinepox virus

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Abstract

Swine are the only known hosts of swinepox virus (SWPV), the sole member of the genus Suipoxvirus, family Poxviridae. Rapid diagnosis is recommended for appropriate interventions because of the high morbidity associated with this virus. This study describes a real-time quantitative PCR (qPCR) assay for rapid detection and quantification of SWPV. The detection limit, repeatability, reproducibility, and specificity of this assay were determined. The efficiency was 96%, and the R2 value was 0.996. The detection limit was 1 fg or 10−0.5 TCID50/50 μL. Tests showed that the greatest source of error in the SWPV qPCR assay was variation between analysts rather than different qPCR kits or equipment. All nucleic acids from other viruses or samples collected from swine were negative in the specificity test. qPCR for SWPV is a new method with tested variables that allows main sources of error in laboratory diagnosis and viral quantification to be identified.

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Authors and Affiliations

  1. Centro Universitário Newton Paiva, Belo Horizonte, Minas Gerais, Brazil

    Felipe Augusto Souza

  2. Centro Universitário de Belo Horizonte, Belo Horizonte, Minas Gerais, Brazil

    Erlânio Marcelo dos Santos Júnior

  3. Laboratório Federal de Defesa Agropecuária de Minas Gerais, Avenida Rômulo Joviano, 26, Centro, Pedro Leopoldo, Minas Gerais, Brazil

    Mateus Laguardia-Nascimento, Tânia Rosária Pereira Freitas, Anselmo V. Rivetti Júnior, Marcelo Fernandes Camargos & Antônio Augusto Fonseca Júnior

  4. Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Clarissa R. Damaso

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  1. Felipe Augusto Souza
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  2. Erlânio Marcelo dos Santos Júnior
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  3. Mateus Laguardia-Nascimento
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  4. Tânia Rosária Pereira Freitas
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  5. Clarissa R. Damaso
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  6. Anselmo V. Rivetti Júnior
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  7. Marcelo Fernandes Camargos
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  8. Antônio Augusto Fonseca Júnior
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Correspondence to Antônio Augusto Fonseca Júnior.

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Souza, F.A., dos Santos Júnior, E.M., Laguardia-Nascimento, M. et al. Validation of a real-time PCR assay for detection of swinepox virus. Arch Virol 164, 3059–3063 (2019). https://doi.org/10.1007/s00705-019-04403-w

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  • DOI: https://doi.org/10.1007/s00705-019-04403-w

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