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Genetic differentiation of Megalocytivirus by real time PCR and sequencing

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Abstract

Background

Megalocytiviruses (MCV) are double-stranded DNA viruses that infect fish. Two species within the genus are epidemiologically important for fish farming: red sea bream iridovirus (RSIV) and infectious spleen and kidney necrosis virus (ISKNV). The objective of this work was to study regions that allow the differentiation and correct diagnosis of RSIV and ISKNV.

Methods

The regions ORF450L, ORF342L, ORF077, and the intergenic region between ORF37 and ORF42R were sequenced and compared with samples from the database.

Results

The tree constructed using the sequencing of the PCR product Megalocytivirus. ORF077 separated the three major clades of MCV. RISV genotypes were well divided, but not ISKNV. All qPCRs tests showed acceptable repeatability values, that is, less than 5%.

Conclusion

Two qPCRs for ISKNV detection and two for RSIV were considered suitable for use in the diagnosis and typing of MCV. The results of this study demonstrate the importance of an accurate evaluation of methodologies for the differentiation of MCV.

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Data availability

Data is available upon request.

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Funding

Brazilian Ministry of Agriculture.

Author information

Authors and Affiliations

  1. Laboratório Federal de Defesa Agropecuária de Minas Gerais, Pedro Leopoldo, Brazil

    Antônio Augusto Fonseca Jr., Mateus Laguardia-Nascimento, Amanda Petrina Scotá Ferreira, Carla do Amaral Pinto, Valdênia Lopes da Silva Gonçalves, Aline Aparecida Silva Barbosa, Anselmo Vasconcelos Rivetti Junior & Marcelo Fernandes Camargos

  2. UNIFEMM - Centro Universitário, Sete Lagoas, Brazil

    Antônio Augusto Fonseca Jr.

Authors
  1. Antônio Augusto Fonseca Jr.
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  2. Mateus Laguardia-Nascimento
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  3. Amanda Petrina Scotá Ferreira
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  4. Carla do Amaral Pinto
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  8. Marcelo Fernandes Camargos
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Contributions

Manuscript preparation: Antônio Augusto Fonseca Júnior.

Manuscript revision: Anselmo Vasconcelos Rivetti Júnior, Marcelo Fernandes Camargos.

Sample preparation: Aline Aparecida Silva Barbosa, Valdenia Lopes da Silva Gonçalves, Anselmo Vasconcelos Rivetti Júnior, Marcelo Fernandes Camargos.

DNA extraction and sequencing: Mateus Laguardia-Nascimento, Anapolino Macedo de Oliveira Bioinformatics, PCR, qPCR and validation tests: Antônio Augusto Fonseca Júnior.

Corresponding author

Correspondence to Antônio Augusto Fonseca Jr..

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The experiment was done on samples sent to routine diagnosis.

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Fonseca Jr., A.A., Laguardia-Nascimento, M., Ferreira, A.P.S. et al. Genetic differentiation of Megalocytivirus by real time PCR and sequencing. Mol Biol Rep 50, 3439–3450 (2023). https://doi.org/10.1007/s11033-023-08282-y

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  • DOI: https://doi.org/10.1007/s11033-023-08282-y

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