Skip to main content
SpringerLink
Log in

Evaluation of three different genomic regions for detection of bovine leukemia virus by real-time PCR

  • Veterinary Microbiology - Research Paper
  • Published:
Brazilian Journal of Microbiology Aims and scope Submit manuscript

Abstract

Bovine leukemia virus (BLV) is an oncogenic member of the genus Deltaretrovirus. BLV infects cattle worldwide and is responsible for significant economic losses. The objective of this study was to validate real-time quantitative PCR (qPCR) for the detection of BLV. After identification of the most efficient qPCR, the limits of detection, repeatability, and reproducibility were determined. The results indicate that qPCR can be easily reproduced between laboratories with high sensitivity. The test variation was low in samples from lesions suggestive of bovine leukosis or whole blood.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Data availability

Data are available upon request.

References

  1. Kettmann R, Portetelle D, Mammerickx M, Cleuter Y, Dekegel D, Galoux M, Ghysdael J, Burny A, Chantrenne H (1976) Bovine leukemia virus: an exogenous RNA oncogenic virus. Proc Natl Acad Sci USA 73(4):1014–1018

    Article  CAS  Google Scholar 

  2. Nekouei O, VanLeeuwen J, Stryhn H, Kelton D, Keefe G (2016) Lifetime effects of infection with bovine leukemia virus on longevity and milk production of dairy cows. Prev Vet Med 133:1–9

    Article  Google Scholar 

  3. Delarmelina E, Buzelin MA, Souza BS, Souto FM, Bicalho JM, Câmara RJF, Resende CF, Bueno BL, Victor RM, Galinari GCF, Nunes CB, Leite RC, Costa ÉA, Reis JKPD (2020) High positivity values for bovine leukemia virus in human breast cancer cases from Minas Gerais, Brazil. PLOS One 15(10):e0239745

    Article  CAS  Google Scholar 

  4. Murakami H, Uchiyama J, Suzuki C, Nikaido S, Shibuya K, Sato R, Maeda Y, Tomioka M, Takeshima SN, Kato H, Sakaguchi M, Sentsui H, Aida Y, Tsukamoto K (2018) Variations in the viral genome and biological properties of bovine leukemia virus wild-type strains. Virus Res 253:103–111. https://doi.org/10.1016/j.virusres.2018.06.005

    Article  CAS  PubMed  Google Scholar 

  5. Camargos MF, Stancek D, Rocha MA, Lessa LM, Reis JK, Leite RC (2002) Partial sequencing of env gene of bovine leukaemia virus from Brazilian samples and phylogenetic analysis. J Vet Med B Infect Dis Vet Public Health 49(7):325–331

    Article  CAS  Google Scholar 

  6. Yu C, Wang X, Zhou Y, Wang Y, Zhang X, Zheng Y (2019) Genotyping bovine leukemia virus in dairy cattle of Heilongjiang, northeastern China. BMC Vet Res 15(1):179

    Article  Google Scholar 

  7. Hirsch C, Camargos MF, Barbosa-Stancioli EF et al (2015) Genetic variability and phylogeny of the 5′ long terminal repeat from Brazilian bovine leukemia virus. Genet Mol Res 14(4):14530–14538

    Article  CAS  Google Scholar 

  8. Jimba M, Takeshima SN, Murakami H et al (2012) BLV-CoCoMo-qPCR: a useful tool for evaluating bovine leukemia virus infection status. BMC Vet Res 8:167

    Article  CAS  Google Scholar 

  9. Naif HM, Brandon RB, Daniel RCW, Lavin MF (1990) Bovine leukemia proviral DNA detection in cattle using the polymerase chain-reaction. Vet Microbiol 25(2–3):117–129

    Article  CAS  Google Scholar 

  10. Burridge MJ, Thurmond MC, Miller JM, Schmerr MJ, Van Der Maaten MJ (1982) Fall in antibody titer to bovine leukemia virus in the periparturient period. Can J Comp Med 46(3):270–271

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Dias NL, Fonseca Júnior AA, Rodrigues DS, Camargos MF (2012) PCR em tempo real para diagnóstico da leucose enzoótica bovina. Cien Rural 42(8):1434–1439

    Article  CAS  Google Scholar 

  12. Miyasaka T, Takeshima SN, Jimba M, Matsumoto Y, Kobayashi N, Matsuhashi T, Sentsui H, Aida Y (2013) Identification of bovine leukocyte antigen class II haplotypes associated with variations in bovine leukemia virus proviral load in Japanese Black cattle. Tissue Antigens 81(2):72–82

    Article  CAS  Google Scholar 

  13. OIE (2019) Manual of diagnostic tests and vaccines for terrestrial animals: Chapter 2.4.11. Enzootic Bovine Leukosis, 7th edn. World organization for animal health, France

    Google Scholar 

  14. Untergasser A, Nijveen H, Rao X, Bisseling T, Geurts R, Leunissen JA (2007) Primer3Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35(Web Server issue):W71–W74

    Article  Google Scholar 

  15. Pinheiro de Oliveira TF, Fonseca AA, Camargos MF et al (2013) Detection of contaminants in cell cultures, sera and trypsin. Biologicals 41(6):407–414

    Article  CAS  Google Scholar 

  16. Klintevall K, Ballagi-Pordány A, Näslund K, Belák S (1994) Bovine leukaemia virus: rapid detection of proviral DNA by nested PCR in blood and organs of experimentally infected calves. Vet Microbiol 42(2–3):191–204

    Article  CAS  Google Scholar 

  17. de Oliveira AM, Fonseca AA, Camargos MF et al (2018) Development and validation of rt-qpcr for vesicular stomatitis virus detection (Alagoas Vesiculovirus). J Virol Methods 257:7–11

    Article  Google Scholar 

  18. Barez PY, de Brogniez A, Carpentier A et al (2015) Recent advances in BLV research. Viruses 7(11):6080–6088

    Article  CAS  Google Scholar 

  19. Bustin S, Huggett J (2017) qPCR primer design revisited. Biomol Detect Quantif 14:19–28

    Article  CAS  Google Scholar 

  20. Jimba M, Takeshima SN, Matoba K et al (2010) BLV-CoCoMo-qPCR: quantitation of bovine leukemia virus proviral load using the CoCoMo algorithm. Retrovirology 7:91

    Article  Google Scholar 

  21. Taylor GM, Worth DR, Palmer S, Jahans K, Hewinson RG (2007) Rapid detection of Mycobacterium bovis DNA in cattle lymph nodes with visible lesions using PCR. BMC Vet Res 3:12

    Article  Google Scholar 

  22. Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT (2009) The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 55(4):611–622

    Article  CAS  Google Scholar 

  23. Stenman J, Orpana A (2001) Accuracy in amplification. Nat Biotechnol 19(11):1011–1012

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the Ministério da Agricultura, Pecuária e Abastecimento, and Universidade Federal de Minas Gerais.

Author information

Authors and Affiliations

  1. Laboratório Federal de Defesa Agropecuária de Minas Gerais, Avenida Rômulo Joviano, Centro, Pedro Leopoldo, Minas Gerais, 33600-000, Brazil

    Antônio Augusto Fonseca Júnior, Luciana Rabello Ferreira, Mateus Laguardia- Nascimento, Anselmo Vasconcelos Rivetti Júnior & Marcelo Fernandes Camargos

  2. Centro Universitário de Sete Lagoas, UNIFEMM, Sete Lagoas, Brazil

    Antônio Augusto Fonseca Júnior

  3. Universidade Federal de Minas Gerais, Belo Horizonte, Brazil

    Bruna Lopes Bueno & Jenner Karlisson Pimenta dos Reis

Authors
  1. Antônio Augusto Fonseca Júnior
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Luciana Rabello Ferreira
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mateus Laguardia- Nascimento
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Anselmo Vasconcelos Rivetti Júnior
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Bruna Lopes Bueno
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jenner Karlisson Pimenta dos Reis
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marcelo Fernandes Camargos
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Antônio Augusto Fonseca Júnior.

Ethics declarations

Competing interests

The authors declare no competing interests.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Responsible Editor: Jorge Luiz Mello Sampaio

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Fonseca Júnior, A.A., Ferreira, L.R., Laguardia- Nascimento, M. et al. Evaluation of three different genomic regions for detection of bovine leukemia virus by real-time PCR. Braz J Microbiol 52, 2483–2488 (2021). https://doi.org/10.1007/s42770-021-00613-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s42770-021-00613-0

Keywords

Search

Navigation