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Indian Journal of Microbiology

, Volume 59, Issue 2, pp 234–236 | Cite as

Importance of the 3′-Terminal Nucleotide of the Forward Primer for Nucleoprotein Gene Detection of Viral Hemorrhagic Septicemia Virus by Conventional Reverse-Transcription PCR

  • Hyoung Jun KimEmail author
Short Communications
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Abstract

Viral hemorrhagic septicemia virus (VHSV), causing severe diseases in farmed fish, is detected and genotyped using conventional reverse-transcription PCR (cRT-PCR) targeting the nucleoprotein gene with corresponding VN F (forward) and VN R (reverse) primers. However, these primers have low sensitivity to VHSV subtype IVa; I investigated the cause for the poor cRT-PCR performance using various primer combinations. The results demonstrated that a 3′-end mismatch in the VN F primer reduced sensitivity and plays a critical role in VHSV detection by cRT-PCR.

Keywords

Viral hemorrhagic septicemia virus Low sensitivity Conventional RT-PCR 3′-Terminal region of forward primer 

Notes

Acknowledgements

I thank Niels Jørgen Olesen and Argelia Cuenca for technical and scientific support.

Funding

This study was funded by the National Fishery Products Quality Management Service, Republic of Korea.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    OIE (2018) Viral haemorrhagic septicaemia. In: Manual of diagnostic tests for aquatic animals. Chapter 2.3.10. http://www.oie.int/fileadmin/Home/eng/Health_standards/aahm/current/chapitre_vhs.pdf. Accessed 12 Dec 2018
  2. 2.
    Snow M, Bain N, Black J et al (2004) Genetic population structure of marine viral haemorrhagic septicaemia virus (VHSV). Dis Aquat Organ 61:11–21.  https://doi.org/10.3354/dao061011 CrossRefGoogle Scholar
  3. 3.
    Kim HJ (2015) Validation of the sensitivities of one-step and two-step reverse-transcription PCR methods for detection of viral hemorrhagic septicemia virus (VHSV) IVa isolates from cultured olive flounder in Korea. Aquaculture 448:359–364.  https://doi.org/10.1016/j.aquaculture.2015.06.034 CrossRefGoogle Scholar
  4. 4.
    Kim MS, Kim KH (2011) Inhibition of viral hemorrhagic septicemia replication using a short hairpin RNA targeting the G gene. Arch Virol 156:457–464.  https://doi.org/10.1007/s00705-010-0882-y CrossRefGoogle Scholar
  5. 5.
    Kim HJ, Park JS, Choi MC, Kwon SR (2016) Comparison of the efficacy of Poly(I:C) immunization with live vaccine and formalin-killed vaccine against viral hemorrhagic septicemia virus (VHSV) in olive flounder (Paralichthys olivaceus). Fish Shellfish Immunol 48:206–211.  https://doi.org/10.1016/j.fsi.2015.11.035 CrossRefGoogle Scholar
  6. 6.
    Kim HJ, Cuenca A, Olesen NJ (2018) Validation of a novel one-step reverse transcription polymerase chain reaction method for detecting viral haemorrhagic septicaemia virus. Aquaculture 492:170–183.  https://doi.org/10.1016/j.aquaculture.2018.03.047 CrossRefGoogle Scholar

Copyright information

© Association of Microbiologists of India 2019

Authors and Affiliations

  1. 1.OIE Reference Laboratory for Viral Hemorrhagic Septicemia (VHS)National Fishery Products Quality Management ServicesBusanKorea

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