The Korean Journal of Nuclear Medicine (대한핵의학회지)
- Volume 28 Issue 2
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- Pages.220-226
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- 1994
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- 1225-6714(pISSN)
Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers
역전사 중합효소 연쇄반응(RT-PCR)에 의한 HCV-RNA의 검출 : Biotin 및 방사성옥소 표지 Primer로 구성된 Kit의 이용
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Ryu, Jin-Sook
(Department of Nuclear Medicine, Asan Medical Center, University of Ulsan) ;
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Moon, Dae-Hyuk
(Department of Nuclear Medicine, Asan Medical Center, University of Ulsan) ;
- Cheon, Jun-Hong (Asan Institute for Life Sciences) ;
- Chung, Yoon-Young (Department of Nuclear Medicine, Asan Medical Center, University of Ulsan) ;
- Park, Hung-Dong (Department of Internal Medicine, Asan Medical Center, University of Ulsan) ;
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Chung, Young-Hwa
(Department of Internal Medicine, Asan Medical Center, University of Ulsan) ;
- Lee, Young-Sang (Department of Internal Medicine, Asan Medical Center, University of Ulsan)
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류진숙
(울산의대 서울중앙병원 핵의학과) ;
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문대혁
(울산의대 서울중앙병원 핵의학과) ;
- 천준홍 (아산생명과학연구소) ;
- 정윤영 (울산의대 서울중앙병원 핵의학과) ;
- 박흥동 (울산의대 서울중앙병원 내과) ;
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정영화
(울산의대 서울중앙병원 내과) ;
- 이영상 (울산의대 서울중앙병원 내과)
- Published : 1994.09.15
Abstract
This study was performed to evaluate the clinical applicability of the reverse transcription polymerase chain reaction (RT-PCR ) kit of HCV-RNA using biotinylated and radioiodinated primers. Study subjects were 118 patients with positive anti-HCV. HCV-RNA in patient's serum was extracted by guanidium thiocyanate method. After first amplification, the product was reamplified by primers labelled with biotin and I-125. The final amplification product was defected by counting the radioactivity after incubation in avidin coated tubes. In 51 samples, the test was repeated for evaluation of reproducibility. This new method was also compared with conventional RT-PCR methods in 34 samples from patients with chronic liver disease. The results were as follows ; 1) HCV-RNA was positive in 85(97%)of 88 patients with chronic liver disease, and in 23 (73%) of 30 patients with normal liver function. 2) In comparison with conventional method, HCV-RNA was detected in 32(94%) of 34 patients with new method, whereas in 27(79%) of the same group with conventional method. 3) Repeated test with new method in 52 samples demonstrated 82% of concordant result. In conclusion, new method with biotinylated and radioiodinated primers was more sensitive than conventional method. However, great care must be taken for quality control because there were considerable interassay variation and possiblity of false positivity and false negativity.
RT-PCR법에 의한 HCV-RNA검출을 임상검사에 적용하는 노력의 일환으로 biotin 및