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Development of Ultra-rapid Multiplex Real-time PCR for the Detection of Genes from Avian Influenza Virus subtype H5N1  

Kim, Eul-Hwan (Department of Biology, Kyonggi University)
Lee, Dong-Woo (Department of Biological engineering, Kyonggi University)
Han, Sang-Hoon (Department of Biology, Kyonggi University)
Lim, Yoon-Kyu (Department of Veterinary Medicine, Cheju National University)
Yoon, Byoung-Su (Department of Biology, Kyonggi University)
Publication Information
Korean Journal of Veterinary Research / v.47, no.4, 2007 , pp. 399-407 More about this Journal
Abstract
Cause of high lethality and dissemination to human being, new development of rapid method for the detection of highly pathogenic Avian Influenza Virus (AIV) is still necessary. For the detection of AIV subtype H5N1, typical pathogenic AIV, new method to confirm sub-typing of this virus is also needed. For the purpose of ultra-rapid detection and sub-typing of hemagglutinin and neuraminidase of AIV, this study was planned. As the results we could demonstrate an ultra-rapid multiplex real-time PCR (URMRT PCR) for the detection of AIV In this study, the URMRT PCR were optimized with synthesized AIV H5- and AIV Nl-specific DNA templates and GenSpector TMC, which is a semiconductor process technology based real-time PCR system with high frequencies of temperature monitoring. Under eight minutes, the amplifications of two AIV subtype-specific PCR products were successfully and independently detected by 30 cycled ultra-rapid PCR, including melting point analysis, from $1{\times}10^3$ copies of mixed template DNA. The URMRT PCR for the detection of AIV H5N 1 developed in this study could be expected to apply not only detections of different AIVs, but also various pathogens. It was also discussed that this kind of the fastest PCR based detection method could be improved by advance of related technology in near future.
Keywords
AIV; detection; H5N1; real-time PCR; ultra-rapid PCR;
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