Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Ultra-Rapid Two-Step Real-Time PCR for the Detection of Human Immunodeficiency Virus (HIV)

Human Immunodeficiency Virus (HIV) 검출물 위한 초고속 이단계 PCR 진단법

  • Lee, Dong-Woo (Department of Life Science, Kyonggi University) ;
  • Kim, Eul-Hwan (Department of Life Science, Kyonggi University) ;
  • Yoo, Mi-Sun (Department of Life Science, Kyonggi University) ;
  • Kim, Il-Uk (Department of Life Science, Kyonggi University) ;
  • Yoon, Byoung-Su (Department of Life Science, Kyonggi University)
  • 이동우 (경기대학교 자연과학대학 생명과학과) ;
  • 김을환 (경기대학교 자연과학대학 생명과학과) ;
  • 유미선 (경기대학교 자연과학대학 생명과학과) ;
  • 김일욱 (경기대학교 자연과학대학 생명과학과) ;
  • 윤병수 (경기대학교 자연과학대학 생명과학과)
  • Published : 2007.12.30
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Abstract

For the detection of human immunodeficiency virus (HIV), ultra-rapid real-time PCR methods were developed. The target DNA sequences were used 495 bp HIV-1-specific env gene (gi_1184090) and 294 bp HIV-2-specific env gene (gi_1332355). Ultra-rapid real-time PCR was peformed by $Genspector^{TM}$ (Samsung, Korea) using microchip-based, $6\;{\mu}l$ of reaction volume with extremely short running time in only 2 steps (denaturation, annealing/extension) in each cycle of PCR. Total reaction for 30 cycled ultra-rapid PCR detection including melting temperature analysis was completed in 7 min and 30 sec. The HIV-1-specific 117 bp-long or HIV-2-spe-cific 119 bp-long PCR products were successfully amplified from the minimum of template, $2.3{\times}10^3$ copies of each euv gene using 30 cycled two-steps ultra-rapid PCR. This kind of ultra-rapid real-time PCR method would be useful not only for the rapid-detection of HIV, but also rapid-detection of other pathogens.

인간면역결핍바이러스(Human Immunodeficiency Virus; HIV) 진단을 위한 초고속 실시간 PCR법을 개발하였다. 검출 대상의 DNA 염기서열은 495염기 HIV-1 특이 env 유전자(gi_l184090)및 294염기 HIV-2 특이 env 유전자(gi_1332355)를 사용하였다. 초고속 실시간 PCR은 microchip에 $6\;{\mu}l$의 PCR 용액을 탑재하는 $Genspector^{TM}$ (Samsung, Korea)을 사용하였으며, PCR의 각 회전 중 단지 두 단계(denaturation, annealing/extension)를 극단적으로 짧은 시간을 주어 수행하게 하였다. 융점분석을 포함한 30회전의 PCR 검색 시간은 총7분30초 이내에 완료되었으며, HIV-1 특이 117염기와 HIV-2 특이 119염기의 PCR산물은 최소 $2.3{\times}10^3$개의 각 env 유전자로부터 30회전의 2단계 초고속 PCR에 의해 성공적으로 증폭시킬 수 있었다. 이러한 초고속 실시간 PCR법은 HIV의 빠른검색뿐 아니라, 다른 병원채의 빠른 검색에도 유용하계 적용될 수 있을 것이다.

Keywords

References

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