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Importance of the 3′-Terminal Nucleotide of the Forward Primer for Nucleoprotein Gene Detection of Viral Hemorrhagic Septicemia Virus by Conventional Reverse-Transcription PCR
- Kim, Hyoung Jun
- Indian journal of microbiology 2019 v.59 no.2 pp. 234-236
- Viral hemorrhagic septicemia virus, disease severity, farmed fish, genes, genotyping, nucleoproteins, reverse transcriptase polymerase chain reaction
- 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.