Detection of Japanese encephalitis virus by biotinylated cDNA probe
1988
Hwang, D.Y. | Yim, J.B. (Seoul National Univ., Seoul (Korea R.). Coll. of Natural Sciences) | Shin, Y.O. (National Inst. of Health, Seoul (Korea R.))
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 viral 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 50 pgs of the target RNA
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