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Rapid Detection Method of Avian Influenza Subtype H5N1 using Quick Real-Time PCR  

Kim, Eul-Hwan (Department of Life Science, College of Natural Science, Kyonggi University)
Lee, Dong-Woo (Department of Life Science, College of Natural Science, Kyonggi University)
Han, Sang-Hoon (Department of Life Science, College of Natural Science, Kyonggi University)
Kwon, Soon-Hwan (Chronic Inflammatory Disease Research Center, School of Medicine, Ajou University)
Yoon, Byoung-Su (Department of Life Science, College of Natural Science, Kyonggi University)
Publication Information
Korean Journal of Microbiology / v.43, no.1, 2007 , pp. 23-30 More about this Journal
Abstract
The most rapid Real-time PCR based detection method for Avian influenza A virus (AIV) subtype H5N1 was developed. The target DNA sequence in this study was deduced from H5N1 subtype-specific 387 bp partial gene of hemagglutinin, and was synthesized by using PCR-based gene synthesis on the ground of safety. Real-Time PCR was performed by $GenSpector^{TM}$ using microchip-based, total $1{\mu}l$ of reaction mixture with extremely short time in each steps in PCR. The detection including PCR-amplication and analysis of melting temperature was totally completed within 13 min. The H5N1-specific 189 bp PCR product was correctly amplified until 2.4 molecules of hemagglutinin gene as minimum of templates. This kind of PCR was designated as Quick Real-Time PCR in this study and it could be applied to detect not only AIV H5N1, but also other pathogens using PCR-based detection.
Keywords
avian influenza; datection; hemagglutinin; H5N1; Real-time PCR;
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