Title Biotin으로 표지된 cDNA Probe를 이용한 일본 뇌염 바이러스의 검색
Author 황동연 · 신영오 * · 임정빈
Address 서울대학교 미생물학과; * 국립보건원
Bibliography Korean Journal of Microbiology, 26(3),149-154, 1988
DOI
Key Words Japanese Encephalitis Virus, Biotinylated Probe, Aminoallyl cDNA
Abstract Japanese Encephalitis Virus(JEV) can be detected conveniently by the use of biotinylated cDNA probe. To prepare biotinylated probe aminoallyl dUTP was first synthesized chemically to reverse transcribe the virial RNA. The allylamine-labeled cDNA was then converted to the biotin-cDNA by the reaction with an activated biotin ester, NHS-ACA-biotin. The JEV genomic RNA was hybridized to the biotinylated cDNA probe on nitrocellulose filter and visualized colorimetrically by streptavidin complexes with alkaline phosphatase polymer. Sensitivity of the detection system was determined by estimating the amount of the JEV genomic RNA through comparison with signals generated from the biotinylated and ^32P-labeled probes. It was found that the biotin probe was as sensitive as ^32P-cDNA probe which can detect 50pgs of the target RNA.
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