| Development of Ultra-rapid Multiplex Real-time PCR for the Detection of Genes from Avian Influenza Virus subtype H5N1 |
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Eul-Hwan Kim1, Dong-Woo Lee2, Sang-Hoon Han1, Yoon-Kyu Lim3, Byoung-Su Yoon1 |
1Department of Biology, Kyonggi University
2Department of Biological engineering, Kyonggi University
3Department of Veterinary Medicine, Cheju National University |
| 조류인플루엔자 H5N1 바이러스 유전자의 신속 검출을 위한 초고속 다중 실시간 PCR법의 개발 |
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김을환1, 이동우2, 한상훈1, 임윤규3, 윤병수1 |
1경기대학교 생물학과
2경기대학교 생명공학과
3제주대학교 수의학과 |
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| Abstract |
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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 $1{ imes}10^3$ 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. |
| Key Words:
AIV, detection, H5N1, real-time PCR, ultra-rapid PCR |
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