Importance of the 3′-Terminal Nucleotide of the Forward Primer for Nucleoprotein Gene Detection of Viral Hemorrhagic Septicemia Virus by Conventional Reverse-Transcription PCR
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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.
KeywordsViral hemorrhagic septicemia virus Low sensitivity Conventional RT-PCR 3′-Terminal region of forward primer
I thank Niels Jørgen Olesen and Argelia Cuenca for technical and scientific support.
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.
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