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Updated: Jan 28, 2026

High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
An optimized molecular method for detection of influenza A virus using improved generic primers and concentration of
Ji-Woon Kim1,2,3,4,5, Chung-Young Lee1,2,3,4,5, Thanh Trung Nguyen1,2,3,4,5
1Laboratory of Avian Diseases (J-W Kim, Lee, Nguyen, J-H Kim), College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea.
Abstract:
For reported primer sets used to detect influenza A viruses (IAVs), we verified the nucleotide identities with 9,103 complete sequences of matrix (M) genes. At best, only 93.2% and 85.3% of the sequences had a 100% match with reported forward and reverse primers, respectively. Therefore, we designed new degenerate forward and reverse primers with 100% identity to 94.4% and 96.2% of compared genes, respectively, and the primer set was used with SYBR-based reverse-transcription real-time PCR (SYBR-RT-rtPCR) for lower detection limits. The sensitivity of SYBR-RT-rtPCR with the new primers was 10-fold higher than that with a conventional method in ~2.37% of all M genes in the database used in our study. We successfully increased the sensitivity of SYBR-RT-rtPCR by concentrating the viral ribonucleoprotein (RNP) using immunomagnetic beads and Triton X-100. The improved generic primer set and RNP concentration method may be useful for sensitive detection of IAVs.
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