원저 : 역전사 중합효소 연쇄반응 ( RT-PCR ) 에 의한 HCV-RNA 의 검출 Biotin 및 방사성옥소 표지 Primer 로 구성된 Kit 의 이용 ( Detection of HCV-RNA by Reverse Transcription Polymerase Chain Reaction Using Biotinylated and Radioiodinated Primers ) () |
Author |
류진숙(Jin Sook Ryu),문대혁(Dae Hyuk Moon),천준홍(Jun Hong Cheon),정윤영(Yoon Young Chung),박흥동(Hung Dong Park),정영화(Young Hwa Chung),이영상(Young Sang Lee), |
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Affiliation |
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Abstract |
This study was performed to evaiuate the clinical
applicability of the reverse t.ranscription polymerase
chain reaction (RT-PCR) kit of HCV-RNA using
biotinylated and radioiodinat.ed primers. Study
subjects were 118 patients with positive anti-HCV. HCV-
RNA in patient.s serurn was ext.racted by guanidium
thiocyanate method. After first amplification, the
product. was reamplified by primers labelled with
biotin and I-125. The final amplification product was
de- tected by counting t,he radioactivity after
incubation in avidin coated tubes. In 51 samples, t.he
test was repeated for evaluation of reproducibility.
This new method was also compared with conventional RT-
PCR rnethods 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 pat.ients with
normal liver function. 2) In comparison with
conventional method, HCV-RNA was detected in 32(94%) of
34 patient.s with new method, whereas in 27(79% ) of
the same group with conventional method, 3) Repeated
test with new rnet.hod in 52 samples demonstrat.ed 82%
of concordant result. In conclusion, new rnethod with
biotinylated and radioiodinated primers was rnore
sensitive than conventional method. However, great care
must be taken for quality control because there were
considerable interassay variat,ion and possiblity of
false positivity and false negativity. |
Keyword |
Polymerase chain reaction, HCV-RNA, Viral hepatitis |
Full text Article |
28501371.pdf
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